Biomarkers for amyotrophic lateral sclerosis and methods using the same

Abstract
The present invention provides various biomarkers of amyotrophic lateral sclerosis (ALS). The present invention also provides various methods of using the biomarkers, including methods for diagnosis of ALS, methods of determining predisposition to ALS, methods of monitoring progression/regression of ALS, methods of assessing efficacy of compositions for treating ALS, methods of screening compositions for activity in modulating biomarkers of ALS, methods of treating ALS, as well as other methods based on biomarkers of ALS.
Description
FIELD

The invention generally relates to biomarkers for amyotrophic lateral sclerosis and methods based on the same biomarkers.


BACKGROUND

Amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig's Disease, is a fatal neurological disease that rapidly attacks and destroys the nerve cells that are responsible for voluntary movement. The destruction of the neurons in the brain and spinal cord that control movement eventually progresses to the point that all voluntary motor control is lost. Death typically occurs from respiratory failure within 3-5 years of disease onset.


Approximately 20,000 people in the United States have ALS, and 5,000 people are diagnosed with ALS each year. ALS is common worldwide, affecting people of all races and ethnic backgrounds. The average age of onset of ALS is between 40 and 60 years of age, but ALS can strike both younger and older men and women. In 90-95% of ALS cases, the disease is apparently random (known as sporadic ALS (SALS)). In such SALS cases, there is no family history of the disease and no clearly associated risk factors. In 5-10% of ALS cases there is an inherited genetic link (known as familial ALS (FALS)).


SUMMARY

In one aspect, a method of diagnosing whether a subject has amyotrophic lateral sclerosis (ALS) is provided. The method comprises:

    • analyzing a biological sample from a subject to determine the level(s) of one or more biomarkers for amyotrophic lateral sclerosis in the sample, wherein the one or more biomarkers are selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, and 8, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; and
    • comparing the level(s) of the one or more biomarkers in the sample to ALS-positive and/or ALS-negative reference levels of the one or more biomarkers in order to diagnose whether the subject has amyotrophic lateral sclerosis.


In another aspect, a method of determining whether a subject is predisposed to developing amyotrophic lateral sclerosis (ALS) is provided, comprising:

    • analyzing a biological sample from a subject to determine the level(s) of one or more biomarkers for amyotrophic lateral sclerosis in the sample, wherein the one or more biomarkers are selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, and 8, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; and
    • comparing the level(s) of the one or more biomarkers in the sample to ALS-positive and/or ALS-negative reference levels of the one or more biomarkers in order to determine whether the subject is predisposed to developing amyotrophic lateral sclerosis.


In yet another aspect, a method of monitoring progression/regression of amyotrophic lateral sclerosis (ALS) in a subject is provided. The method comprises:

    • analyzing a first biological sample from a subject to determine the level(s) of one or more biomarkers for amyotrophic lateral sclerosis in the sample, wherein the first sample is obtained from the subject at a first time point and the one or more biomarkers are selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, 8, 16, 17, and 18, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof;
    • analyzing a second biological sample from a subject to determine the level(s) of the one or more biomarkers, wherein the second sample is obtained from the subject at a second time point; and
    • comparing the level(s) of one or more biomarkers in the first sample to the level(s) of the one or more biomarkers in the second sample in order to monitor the progression/regression of ALS in the subject.


In a further aspect, a method of assessing the efficacy of a composition for treating amyotrophic lateral sclerosis (ALS) is provided. The method comprises:

    • analyzing, from a subject having amyotrophic lateral sclerosis and currently or previously being treated with a composition, a biological sample to determine the level(s) of one or more biomarkers for ALS selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, 8, 16, 17, and 18, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; and
    • comparing the level(s) of the one or more biomarkers in the sample to (a) levels of the one or more biomarkers in a previously-taken biological sample from the subject, wherein the previously-taken biological sample was obtained from the subject before being treated with the composition, (b) ALS-positive reference levels of the one or more biomarkers, (c) ALS-negative reference levels of the one or more biomarkers, (d) ALS-progression-positive reference levels of the one or more biomarkers, and/or (e) ALS-regression-positive reference levels of the one or more biomarkers.


In yet a further aspect, a method for assessing the efficacy of a composition in treating amyotrophic lateral sclerosis (ALS) is provided comprising:

    • analyzing a first biological sample from a subject to determine the level(s) of one or more biomarkers for ALS, the first sample obtained from the subject at a first time point wherein the one or more biomarkers are selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, 8, 16, 17, and 18, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof;
    • administering the composition to the subject;
    • analyzing a second biological sample from the subject to determine the level(s) of the one or more biomarkers, the second sample obtained from the subject at a second time point after administration of the composition;
    • comparing the level(s) of one or more biomarkers in the first sample to the level(s) of the one or more biomarkers in the second sample in order to assess the efficacy of the composition for treating amyotrophic lateral sclerosis.


In another aspect, a method of assessing the relative efficacy of two or more compositions for treating amyotrophic lateral sclerosis (ALS) is provided. The method comprises:

    • analyzing, from a first subject having ALS and currently or previously being treated with a first composition, a first biological sample to determine the level(s) of one or more biomarkers selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, 8, 16, 17, and 18, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof;
    • analyzing, from a second subject having ALS and currently or previously being treated with a second composition, a second biological sample to determine the level(s) of the one or more biomarkers; and
    • comparing the level(s) of one or more biomarkers in the first sample to the level(s) of the one or more biomarkers in the second sample in order to assess the relative efficacy of the first and second compositions for treating amyotrophic lateral sclerosis.


In yet another aspect, a method for screening a composition for activity in modulating one or more biomarkers of amyotrophic lateral sclerosis is provided comprising:

    • contacting one or more cells with a composition;
    • analyzing at least a portion of the one or more cells or a biological sample associated with the cells to determine the level(s) of one or more biomarkers of amyotrophic lateral sclerosis selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, and 8, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; and
    • comparing the level(s) of the one or more biomarkers with predetermined standard levels for the biomarkers to determine whether the composition modulated the level(s) of the one or more biomarkers.


In a further aspect, a method for identifying a potential drug target for amyotrophic lateral sclerosis (ALS) is provided. The method comprises:

    • identifying one or more biochemical pathways associated with one or more biomarkers for ALS, wherein the one or more biomarkers are selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, and 8, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; and
    • identifying a protein affecting at least one of the one or more identified biochemical pathways, the protein being a potential drug target for amyotrophic lateral sclerosis.


In another aspect, a method for treating a subject having amyotrophic lateral sclerosis (ALS) is provided. The method comprises administering to the subject an effective amount of one or more biomarkers selected from Tables 3, 4, 5, 6, 7, and 8 that are decreased in subjects having ALS as compared to subjects not having ALS.


In yet a further aspect, a method of distinguishing whether a subject has amyotrophic lateral sclerosis (ALS) or has peripheral neuropathy or myopathy is provided. The method comprises:

    • analyzing a biological sample from a subject to determine the level(s) of one or more biomarkers for amyotrophic lateral sclerosis in the sample, wherein the one or more biomarkers comprise one or more xenobiotics, metabolites of xenobiotics, and/or biomarkers listed in Tables 14 and 15; and
    • comparing the level(s) of the one or more biomarkers in the sample to ALS-positive and/or ALS-negative reference levels of the one or more biomarkers in order to determine whether a subject has ALS or has peripheral neuropathy or myopathy.




BRIEF DESCRIPTION OF THE DRAWINGS


FIG. 1 is an importance plot for a random forest analysis as explained in Example 1 below.




DETAILED DESCRIPTION

The present invention relates to biomarkers of amyotrophic lateral sclerosis, methods for diagnosis of ALS, methods of determining predisposition to ALS, methods of monitoring progression/regression of ALS, methods of assessing efficacy of compositions for treating ALS, methods of screening compositions for activity in modulating biomarkers of ALS, methods of treating ALS, as well as other methods based on biomarkers of ALS. Prior to describing this invention in further detail, however, the following terms will first be defined.


Definitions:


“Biomarker” means a compound, preferably a xenobiotic or a metabolite, that is differentially present (i.e., increased or decreased) in a biological sample from a subject or a group of subjects having a first phenotype (e.g., having a disease) as compared to a biological sample from a subject or group of subjects having a second phenotype (e.g., not having the disease). A biomarker may be differentially present at any level, but is generally present at a level that is increased by at least 5%, by at least 10%, by at least 15%, by at least 20%, by at least 25%, by at least 30%, by at least 35%, by at least 40%, by at least 45%, by at least 50%, by at least 55%, by at least 60%, by at least 65%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, by at least 95%, by at least 100%, by at least 110%, by at least 120%, by at least 130%, by at least 140%, by at least 150%, or more; or is generally present at a level that is decreased by at least 5%, by at least 10%, by at least 15%, by at least 20%, by at least 25%, by at least 30%, by at least 35%, by at least 40%, by at least 45%, by at least 50%, by at least 55%, by at least 60%, by at least 65%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, by at least 95%, or by 100% (i.e., absent). A biomarker is preferably differentially present at a level that is statistically significant (i.e., a p-value less than 0.05 and/or a q-value of less than 0.10 as determined using either Welch's T-test or Wilcoxon's rank-sum Test).


The “level” of one or more biomarkers means the absolute or relative amount or concentration of the biomarker in the sample.


“Sample” or “biological sample” means biological material isolated from a subject. The biological sample may contain any biological material suitable for detecting the desired biomarkers, and may comprise cellular and/or non-cellular material from the subject. The sample can be isolated from any suitable biological tissue or fluid such as, for example, blood, blood plasma, urine, or cerebral spinal fluid (CSF).


“Subject” means any animal, but is preferably a mammal, such as, for example, a human, monkey, mouse, or rabbit.


A “reference level” of a biomarker means a level of the biomarker that is indicative of a particular disease state, phenotype, or lack thereof, as well as combinations of disease states, phenotypes, or lack thereof. A “positive” reference level of a biomarker means a level that is indicative of a particular disease state or phenotype. A “negative” reference level of a biomarker means a level that is indicative of a lack of a particular disease state or phenotype. For example, an “ALS-positive reference level” of a biomarker means a level of a biomarker that is indicative of a positive diagnosis of ALS in a subject, and an “ALS-negative reference level” of a biomarker means a level of a biomarker that is indicative of a negative diagnosis of ALS in a subject. As another example, an “ALS-progression-positive reference level” of a biomarker means a level of a biomarker that is indicative of progression of ALS in a subject, and an “ALS-regression-positive reference level” of a biomarker means a level of a biomarker that is indicative of regression of ALS in a subject. A “reference level” of a biomarker may be an absolute or relative amount or concentration of the biomarker, a presence or absence of the biomarker, a range of amount or concentration of the biomarker, a minimum and/or maximum amount or concentration of the biomarker, a mean amount or concentration of the biomarker, and/or a median amount or concentration of the biomarker; and, in addition, “reference levels” of combinations of biomarkers may also be ratios of absolute or relative amounts or concentrations of two or more biomarkers with respect to each other. Appropriate positive and negative reference levels of biomarkers for a particular disease state, phenotype, or lack thereof may be determined by measuring levels of desired biomarkers in one or more appropriate subjects, and such reference levels may be tailored to specific populations of subjects (e.g., a reference level may be age-matched so that comparisons may be made between biomarker levels in samples from subjects of a certain age and reference levels for a particular disease state, phenotype, or lack thereof in a certain age group). Such reference levels may also be tailored to specific techniques that are used to measure levels of biomarkers in biological samples (e.g., LC-MS, GC-MS, etc.), where the levels of biomarkers may differ based on the specific technique that is used.


“Metabolite”, or “small molecule”, means organic and inorganic molecules which are present in a cell. The term does not include large macromolecules, such as large proteins (e.g., proteins with molecular weights over 2,000, 3,000, 4,000, 5,000, 6,000, 7,000, 8,000, 9,000, or 10,000), large nucleic acids (e.g., nucleic acids with molecular weights of over 2,000, 3,000, 4,000, 5,000, 6,000, 7,000, 8,000, 9,000, or 10,000), or large polysaccharides (e.g., polysaccharides with a molecular weights of over 2,000, 3,000, 4,000, 5,000, 6,000, 7,000, 8,000, 9,000, or 10,000). The small molecules of the cell are generally found free in solution in the cytoplasm or in other organelles, such as the mitochondria, where they form a pool of intermediates which can be metabolized further or used to generate large molecules, called macromolecules. The term “small molecules” includes signaling molecules and intermediates in the chemical reactions that transform energy derived from food into usable forms. Examples of small molecules include sugars, fatty acids, amino acids, nucleotides, intermediates formed during cellular processes, and other small molecules found within the cell.


“Xenobiotic” means a chemical foreign to a given organism (i.e., not produced in vivo). Xenobiotics include, but are not limited to, drugs, pesticides, and carcinogens. The metabolism of xenobiotics occurs in two phases. Phase I enzymes include Cytochrome P450 enzymes and Phase II enzymes include UDP-glucuronosyltransferases and glutathione S-transferases.


“Phase I metabolite of a xenobiotic” means the product of phase I metabolism of a xenobiotic. For example, paraxanthine (i.e., 1-7-dimethylxanthine), theophylline, and theobromine are Phase I metabolites of caffeine.


“Metabolic profile”, or “small molecule profile”, means a complete or partial inventory of small molecules within a targeted cell, tissue, organ, organism, or fraction thereof (e.g., cellular compartment). The inventory may include the quantity and/or type of small molecules present. The “small molecule profile” may be determined using a single technique or multiple different techniques.


“Non-biomarker compound” means a compound that is not differentially present in a biological sample from a subject or a group of subjects having a first phenotype (e.g., having a first disease) as compared to a biological sample from a subject or group of subjects having a second phenotype (e.g., not having the first disease). Such non-biomarker compounds may, however, be biomarkers in a biological sample from a subject or a group of subjects having a third phenotype (e.g., having a second disease) as compared to the first phenotype (e.g., having the first disease) or the second phenotype (e.g., not having the first disease).


“Metabolome” means all of the small molecules present in a given organism.


A “neurodegenerative disease” includes, but is not limited to, amyotrophic lateral sclerosis, multiple sclerosis, Huntington's Disease, Alzheimer's Disease, and Parkinson's Disease.


I. Biomarkers


The ALS biomarkers described herein were discovered using metabolomic profiling techniques. Such metabolomic profiling techniques are described in more detail in the Examples set forth below as well as in U.S. Pat. No. 7,005,255 and U.S. patent application Ser. Nos. 11/357,732, 10/695,265 (Publication No. 2005/0014132), 11/301,077 (Publication No. 2006/0134676), 11/301,078 (Publication No. 2006/0134677), 11/301,079 (Publication No. 2006/0134678), and 11/405,033, the entire contents of which are hereby incorporated herein by reference.


Generally, metabolic profiles were determined for biological samples from human subjects diagnosed with ALS as well as from one or more other groups of human subjects (e.g., healthy control subjects not diagnosed with ALS). The metabolic profile for ALS was compared to the metabolic profile for biological samples from the one or more other groups of subjects. Those molecules differentially present, including those molecules differentially present at a level that is statistically significant, in the metabolic profile of ALS samples as compared to another group (e.g., healthy control subjects not diagnosed with ALS) were identified as biomarkers to distinguish those groups.


The biomarkers are discussed in more detail herein. The biomarkers that were discovered correspond with the following group(s):

    • Biomarkers for distinguishing ALS vs. control subjects not diagnosed with ALS (see Tables 3, 4, 5, 6, 7, 8, and 9);
    • Biomarkers for distinguishing subjects having ALS vs. subjects having peripheral neuropathy (see Table 14);
    • Biomarkers for distinguishing subjects having ALS vs. subjects having myopathy (see Table 15);
    • Biomarkers for distinguishing early stage ALS vs. later stages of ALS (i.e., biomarkers for distinguishing progression/regression of ALS) (see Tables 16, 17, and 18);


Non-biomarker compounds associated with the compared groups were also identified. The non-biomarker compounds that were discovered correspond with the following group(s):

    • Non-biomarker compounds present at the same levels between ALS and control subjects not diagnosed with ALS (see Tables 3, 4, 5, 6, 7, 8, 16, 17, and 18).


Although the identities of some of the biomarkers and non-biomarker compounds are not known at this time, such identities are not necessary for the identification of the biomarkers or non-biomarker compounds in biological samples from subjects, as the “unnamed” compounds have been sufficiently characterized by analytical techniques to allow such identification. The analytical characterization of all such “unnamed” compounds is listed in the Examples. Such “unnamed” biomarkers and non-biomarker compounds are designated herein using the nomenclature “Metabolite” followed by a specific metabolite number.


In addition, where the identity of a previously “unnamed” metabolite has been determined, the actual metabolite identity is followed by the nomenclature of “previously Metabolite-xxxx” (e.g. Lactate (previously: Metabolite-2694)-35), or the “Metabolite” number is followed by the nomenclature “retired” and the actual metabolite identity (e.g. Metabolite-1713 retired: n-acetyl-L-aspartic acid).


Finally, where the potential identity of a compound is proposed for an “unnamed” metabolite and such identity has not been confirmed, the nomenclature of “possible” (along with the potential compound identity) follows the “Metabolite” number. Such proposed identity should not be considered as limiting the analytical characterization of the otherwise “unnamed” compounds.


II. Xenobiotics and Xenobiotic Metabolites as Biomarkers


In addition to the biomarkers listed above, it has been determined that xenobiotics (including, but not limited to, caffeine) and the metabolites of xenobiotics are biomarkers for ALS.


Without being limited by theory, it is believed that at least one factor in the etiology of ALS (especially sporadic ALS) is an imbalance in the metabolism of xenobiotics in the body due to higher Phase I enzyme activity relative to Phase II enzyme activity (i.e., (1) higher than normal Phase I enzyme activity coupled with normal Phase II enzyme activity, (2) normal Phase I enzyme activity coupled with under active Phase II enzyme activity, or (3) higher than normal Phase I enzyme activity coupled with under active Phase II enzyme activity). Such an imbalance may result in more toxins and free radicals (produced by Phase I enzymes) being present in the body, which may lead to onset and progression of ALS, especially SALS. Thus, xenobiotics and their corresponding metabolites may be used as biomarkers for various methods described herein (including, e.g., distinguishing ALS versus normal subjects as well as distinguishing ALS versus subjects having a neurodegenerative disease similar to ALS).


An example of a xenobiotic and corresponding metabolites that may be used as biomarkers for ALS is caffeine and its corresponding metabolites. As shown in Examples 4-7, caffeine and the Phase I metabolites of caffeine (e.g., paraxanthine (i.e., 1-7-dimethylxanthine), theophylline, and theobromine) may be used as biomarkers to distinguish subjects having ALS from normal subjects not having ALS and to distinguish subjects having ALS from subjects having other neurodegenerative diseases with symptoms similar to ALS (e.g., myopathy, neuropathy). As explained below, the level of caffeine and the caffeine metabolites (i.e., paraxanthine/1,7-dimethylxanthine, theophylline, theobromine) have been shown to be lower in plasma from ALS patients compared to healthy control subjects and the levels of caffeine and paraxanthine can be used to distinguish between ALS and diseases (e.g. myopathy, PN) with symptoms that resemble those of ALS. Such biomarkers may therefore be useful in ALS diagnosis.


Lower levels of caffeine and caffeine metabolites (i.e. paraxanthine/1,7-dimethylxanthine, theobromine, theophylline) were measured in plasma from ALS patients compared to levels in plasma from control subjects (see Table 9 and Example 4). It is believed that the lower levels of caffeine and caffeine metabolites in ALS subjects are due to increased activity of enzymes that metabolize caffeine (e.g. liver Phase I Cytochrome P450 enzymes, which react with toxins, drugs, alcohol, paint fumes and many other substances to form compounds that are capable of being transformed to less toxic, water soluble substances by Phase II reactions in the liver) rather than lower caffeine intake in such subjects.


The activity of CYP1A2, an inducible Cytochrome P450, is reflected in how fast caffeine is removed from the body. A high rate caffeine clearance (i.e. high ratio of caffeine metabolites/caffeine) indicates that the liver is working at a high rate to clear toxic substances. Individuals with very high rates of caffeine clearance are susceptible to environmental toxins, pesticides and even car exhaust because the metabolism of these xenobiotics and toxins by the Phase I Cytochrome P450 activity generates additional toxic substances and free radicals. If the activity of the Phase II system is also high, the systems are in balance. However, if the Phase II system activity is out of balance with Phase I activity, Phase I toxins and free radicals can accumulate.


It is possible that the generation of toxins and free-radicals through rapid hepatic phase I metabolism could contribute to the onset of sporadic ALS. The levels of caffeine and caffeine metabolites are much lower in ALS subjects compared to control subjects (see Table 9 and Example 4). However, the ratios of caffeine metabolites to caffeine are higher in ALS compared to control subjects (see Table 10 and Example 5) indicating that the Phase I hepatic detoxification system in ALS patients may be working at higher than normal levels. This is unlikely to be an age effect because in a demographic study of individuals of various ages, the levels of both caffeine and the caffeine metabolite paraxanthine increase with age (see Table 11 and Example 5). Further, as shown in Table 11, the paraxanthine/caffeine ratio is lowest in the oldest age group, which is the age when ALS typically is diagnosed. Thus, the measurement of the levels of caffeine, caffeine metabolites and/or the ratio of caffeine/caffeine metabolites (i.e., caffeine metabolism/clearance) may provide valuable information useful to diagnose individuals who have ALS as well as to identify individuals who are at risk for ALS, and also implicates an overactive Cytochrome P450 system and/or other mechanisms of xenobiotic metabolism in the etiology of some cases of sporadic ALS. Therapeutics that target these enzyme systems and decrease the rate of formation of toxic substances may be able to prevent or substantially delay ALS onset.


The level of caffeine is significantly lower in plasma from patients with ALS compared to plasma from individuals with myopathy as well as in plasma from peripheral neuropathy (PN) patients (see Table 12 and Example 6). Thus, caffeine and/or caffeine metabolites are also useful biomarker(s) to distinguish ALS from illnesses that have symptoms resembling ALS, such as myopathy and PN. Further, the ratios of paraxanthine to caffeine are higher in ALS than the ratios in myopathy or PN. This supports the idea that caffeine clearance is higher in ALS than in diseases resembling ALS and provides potential useful therapeutic targets for ALS. In addition, the levels of caffeine, caffeine metabolites and/or the ratio of caffeine metabolites/caffeine are useful in diagnostic tests to distinguish ALS from these ALS “mimics”.


In some embodiments of the methods described herein, at least one of the biomarkers used includes a xenobiotic and/or a metabolite of a xenobiotic (preferably a xenobiotic and a Phase I metabolite of the xenobiotic) such as, for example, caffeine and/or a metabolite of caffeine (e.g., paraxanthine, theophylline, and/or theobromine).


III. Diagnosis of ALS


The identification of biomarkers for ALS allows for the diagnosis of (or for aiding in the diagnosis of) ALS in subjects presenting one or more symptoms of ALS. A method of diagnosing (or aiding in diagnosing) whether a subject has ALS comprises (1) analyzing a biological sample from a subject to determine the level(s) of one or more biomarkers of amyotrophic lateral sclerosis in the sample and (2) comparing the level(s) of the one or more biomarkers in the sample to ALS-positive and/or ALS-negative reference levels of the one or more biomarkers in order to diagnose (or aid in the diagnosis of) whether the subject has amyotrophic lateral sclerosis. The one or more biomarkers that are used are selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, and/or 8 and combinations thereof. When such a method is used to aid in the diagnosis of ALS, the results of the method may be used along with other methods (or the results thereof) useful in the clinical determination of whether a subject has ALS.


Any suitable method may be used to analyze the biological sample in order to determine the level(s) of the one or more biomarkers in the sample. Suitable methods include chromatography (e.g., HPLC, gas chromatography, liquid chromatography), mass spectrometry (e.g., MS, MS-MS), enzyme-linked immunosorbent assay (ELISA), antibody linkage, other immunochemical techniques, and combinations thereof. Further, the level(s) of the one or more biomarkers may be measured indirectly, for example, by using an assay that measures the level of a compound (or compounds) that correlates with the level of the biomarker(s) that are desired to be measured.


The levels of one or more of the biomarkers selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, and/or 8 may be determined in the methods of diagnosing and methods of aiding in diagnosing whether a subject has ALS. For example, the level(s) of one biomarker, two or more biomarkers, three or more biomarkers, four or more biomarkers, five or more biomarkers, six or more biomarkers, seven or more biomarkers, eight or more biomarkers, nine or more biomarkers, ten or more biomarkers, etc., including a combination of all of the biomarkers selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, and/or 8 or any fraction thereof, may be determined and used in such methods. Determining levels of combinations of the biomarkers may allow greater sensitivity and specificity in diagnosing ALS and aiding in the diagnosis of ALS, and may allow better differentiation of ALS from other neurodegenerative diseases that may have similar or overlapping biomarkers to ALS (as compared to a subject not having a neurodegenerative disease). For example, ratios of the levels of certain biomarkers (and non-biomarker compounds) in biological samples may allow greater sensitivity and specificity in diagnosing ALS and aiding in the diagnosis of ALS, and may allow better differentiation of ALS from other neurodegenerative diseases that may have similar or overlapping biomarkers to ALS (as compared to a subject not having a neurodegenerative disease).


One or more biomarkers that are specific for diagnosing ALS (or aiding in diagnosing ALS) in a certain type of sample (e.g., CSF sample or blood plasma sample) may also be used. For example, when the biological sample is cerebral spinal fluid, one or more biomarkers listed in Tables 3 and/or 4 may be used to diagnose (or aid in diagnosing) whether a subject has ALS. When the biological sample is blood plasma, one or more biomarkers selected from Tables 5, 6, 7 and/or 8 may be used to diagnose (or aid in diagnosing) whether a subject has ALS.


After the level(s) of the one or more biomarkers in the sample are determined, the level(s) are compared to ALS-positive and/or ALS-negative reference levels to aid in diagnosing or to diagnose whether the subject has ALS. Levels of the one or more biomarkers in a sample corresponding to the ALS-positive reference levels (e.g., levels that are the same as the reference levels, substantially the same as the reference levels, above and/or below the minimum and/or maximum of the reference levels, and/or within the range of the reference levels) are indicative of a diagnosis of ALS in the subject. Levels of the one or more biomarkers in a sample corresponding to the ALS-negative reference levels (e.g., levels that are the same as the reference levels, substantially the same as the reference levels, above and/or below the minimum and/or maximum of the reference levels, and/or within the range of the reference levels) are indicative of a diagnosis of no ALS in the subject. In addition, levels of the one or more biomarkers that are differentially present (especially at a level that is statistically significant) in the sample as compared to ALS-negative reference levels are indicative of a diagnosis of ALS in the subject. Levels of the one or more biomarkers that are differentially present (especially at a level that is statistically significant) in the sample as compared to ALS-positive reference levels are indicative of a diagnosis of no ALS in the subject. In one embodiment, when caffeine and paraxanthine and/or theophylline levels are determined in the sample, the ALS-positive and/or ALS-negative reference levels may comprise the paraxanthine/caffeine and/or the theophylline/caffeine ratios listed in Table 10.


The level(s) of the one or more biomarkers may be compared to ALS-positive and/or ALS-negative reference levels using various techniques, including a simple comparison (e.g., a manual comparison) of the level(s) of the one or more biomarkers in the biological sample to ALS-positive and/or ALS-negative reference levels. The level(s) of the one or more biomarkers in the biological sample may also be compared to ALS-positive and/or ALS-negative reference levels using one or more statistical analyses (e.g., t-test, Welch's T-test, Wilcoxon's rank sum test, random forest).


In addition, the biological samples may be analyzed to determine the level(s) of one or more non-biomarker compounds. The level(s) of such non-biomarker compounds may also allow differentiation of ALS from other neurodegenerative diseases that may have similar or overlapping biomarkers to ALS (as compared to a subject not having a neurodegenerative disease). For example, a known non-biomarker compound present in biological samples of subjects having ALS and subjects not having ALS could be monitored to verify a diagnosis of ALS as compared to a diagnosis of another neurodegenerative disease when biological samples from subjects having the other neurodegenerative disease do not have the non-biomarker compound.


IV. Methods of Determining Predisposition to ALS


The identification of biomarkers for ALS also allows for the determination of whether a subject having no symptoms of ALS is predisposed to developing ALS. A method of determining whether a subject having no symptoms of ALS is predisposed to developing amyotrophic lateral sclerosis comprises (1) analyzing a biological sample from a subject to determine the level(s) of one or more biomarkers selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, and/or 8 in the sample and (2) comparing the level(s) of the one or more biomarkers in the sample to ALS-positive and/or ALS-negative reference levels of the one or more biomarkers in order to determine whether the subject is predisposed to developing amyotrophic lateral sclerosis. The results of the method may be used along with other methods (or the results thereof) useful in the clinical determination of whether a subject is predisposed to developing ALS.


As described above in connection with methods of diagnosing (or aiding in the diagnosis of) ALS, any suitable method may be used to analyze the biological sample in order to determine the level(s) of the one or more biomarkers in the sample.


As with the methods of diagnosing (or aiding in the diagnosis of) ALS described above, the level(s) of one biomarker, two or more biomarkers, three or more biomarkers, four or more biomarkers, five or more biomarkers, six or more biomarkers, seven or more biomarkers, eight or more biomarkers, nine or more biomarkers, ten or more biomarkers, etc., including a combination of all of the biomarkers selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, and/or 8 or any fraction thereof, may be determined and used in methods of determining whether a subject having no symptoms of ALS is predisposed to developing ALS.


After the level(s) of the one or more biomarkers in the sample are determined, the level(s) are compared to ALS-positive and/or ALS-negative reference levels in order to predict whether the subject is predisposed to developing amyotrophic lateral sclerosis. Levels of the one or more biomarkers in a sample corresponding to the ALS-positive reference levels (e.g., levels that are the same as the reference levels, substantially the same as the reference levels, above and/or below the minimum and/or maximum of the reference levels, and/or within the range of the reference levels) are indicative of the subject being predisposed to developing ALS. Levels of the one or more biomarkers in a sample corresponding to the ALS-negative reference levels (e.g., levels that are the same as the reference levels, substantially the same as the reference levels, above and/or below the minimum and/or maximum of the reference levels, and/or within the range of the reference levels) are indicative of the subject not being predisposed to developing ALS. In addition, levels of the one or more biomarkers that are differentially present (especially at a level that is statistically significant) in the sample as compared to ALS-negative reference levels are indicative of the subject being predisposed to developing ALS. Levels of the one or more biomarkers that are differentially present (especially at a level that is statistically significant) in the sample as compared to ALS-positive reference levels are indicative of the subject not being predisposed to developing ALS.


Furthermore, it may also be possible to determine reference levels specific to assessing whether or not a subject that does not have ALS is predisposed to developing ALS. For example, it may be possible to determine reference levels of the biomarkers for assessing different degrees of risk (e.g., low, medium, high) in a subject for developing ALS. Such reference levels could be used for comparison to the levels of the one or more biomarkers in a biological sample from a subject.


As with the methods described above, the level(s) of the one or more biomarkers may be compared to ALS-positive and/or ALS-negative reference levels using various techniques, including a simple comparison, one or more statistical analyses, and combinations thereof.


As with the methods of diagnosing (or aiding in diagnosing) whether a subject has ALS, the methods of determining whether a subject having no symptoms of ALS is predisposed to developing amyotrophic lateral sclerosis may further comprise analyzing the biological sample to determine the level(s) of one or more non-biomarker compounds.


V. Methods of Monitoring Progression/Regression of ALS


The identification of biomarkers for ALS also allows for monitoring progression/regression of ALS in a subject. A method of monitoring the progression/regression of amyotrophic lateral sclerosis in a subject comprises (1) analyzing a first biological sample from a subject to determine the level(s) of one or more biomarkers for ALS selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, 8, 16, 17, and/or 18, the first sample obtained from the subject at a first time point, (2) analyzing a second biological sample from a subject to determine the level(s) of the one or more biomarkers, the second sample obtained from the subject at a second time point, and (3) comparing the level(s) of one or more biomarkers in the first sample to the level(s) of the one or more biomarkers in the second sample in order to monitor the progression/regression of ALS in the subject. The results of the method are indicative of the course of ALS (i.e., progression or regression, if any change) in the subject.


The change (if any) in the level(s) of the one or more biomarkers over time may be indicative of progression or regression of ALS in the subject. In order to characterize the course of ALS in the subject, the level(s) of the one or more biomarkers in the first sample, the level(s) of the one or more biomarkers in the second sample, and/or the results of the comparison of the levels of the biomarkers in the first and second samples may be compared to ALS-positive and/or ALS-negative reference levels of the one or more biomarkers. If the comparisons indicate that the level(s) of the one or more biomarkers are increasing or decreasing over time (e.g., in the second sample as compared to the first sample) to become more similar to the ALS-positive reference levels (or less similar to the ALS-negative reference levels), then the results are indicative of ALS progression. If the comparisons indicate that the level(s) of the one or more biomarkers are increasing or decreasing over time to become more similar to the ALS-negative reference levels (or less similar to the ALS-positive reference levels), then the results are indicative of ALS regression.


The course of ALS in the subject may also be characterized by comparing the level(s) of the one or more biomarkers in the first sample, the level(s) of the one or more biomarkers in the second sample, and/or the results of the comparison of the levels of the biomarkers in the first and second samples to ALS-progression-positive and/or ALS-regression-positive reference levels (e.g., Example 11 below describes biomarkers for distinguishing early stage ALS vs. later stage ALS indicating whether certain biomarkers increase or decrease as ALS progresses; such trends and/or levels of biomarkers at a later stage of ALS versus an earlier stage of ALS are one example of ALS-progression positive reference levels). If the comparisons indicate that the level(s) of the one or more biomarkers are increasing or decreasing over time (e.g., in the second sample as compared to the first sample) to become more similar to the ALS-progression-positive reference levels (or less similar to the ALS-regression-positive reference levels), then the results are indicative of ALS progression. If the comparisons indicate that the level(s) of the one or more biomarkers are increasing or decreasing over time to become more similar to the ALS-regression-positive reference levels (or less similar to the ALS-progression-positive reference levels), then the results are indicative of ALS regression.


As with the other methods described herein, the comparisons made in the methods of monitoring progression/regression of ALS in a subject may be carried out using various techniques, including simple comparisons, one or more statistical analyses, and combinations thereof.


The results of the method may be used along with other methods (or the results thereof) useful in the clinical monitoring of progression/regression of ALS in a subject.


As described above in connection with methods of diagnosing (or aiding in the diagnosis of) ALS, any suitable method may be used to analyze the biological samples in order to determine the level(s) of the one or more biomarkers in the samples. In addition, the level(s) one or more biomarkers, including a combination of all of the biomarkers selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, 8, 16, 17, and/or 18 or any fraction thereof, may be determined and used in methods of monitoring progression/regression of ALS in a subject.


Such methods could be conducted to monitor the course of ALS in subjects having ALS or could be used in subjects not having ALS (e.g., subjects suspected of being predisposed to developing ALS) in order to monitor levels of predisposition to ALS.


VI. Methods of Assessing Efficacy of Compositions for Treating ALS


The identification of biomarkers for ALS also allows for assessment of the efficacy of a composition for treating ALS as well as the assessment of the relative efficacy of two or more compositions for treating ALS. Such assessments may be used, for example, in efficacy studies as well as in lead selection of compositions for treating ALS.


A method of assessing the efficacy of a composition for treating amyotrophic lateral sclerosis comprises (1) analyzing, from a subject (or group of subjects) having amyotrophic lateral sclerosis and currently or previously being treated with a composition, a biological sample (or group of samples) to determine the level(s) of one or more biomarkers selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, 8, 16, 17, and/or 18, and (2) comparing the level(s) of the one or more biomarkers in the sample (or group of samples) to (a) level(s) of the one or more biomarkers in a previously-taken biological sample (or group of samples) from the subject (or group of subjects), wherein the previously-taken biological sample was obtained from the subject (or group of subjects) before being treated with the composition, (b) ALS-positive reference levels of the one or more biomarkers, (c) ALS-negative reference levels of the one or more biomarkers (d) ALS-progression-positive reference levels of the one or more biomarkers, and/or (e) ALS-regression-positive reference levels of the one or more biomarkers. The results of the comparison are indicative of the efficacy of the composition for treating ALS.


Thus, in order to characterize the efficacy of the composition for treating ALS, the level(s) of the one or more biomarkers in the biological sample are compared to (1) ALS-positive reference levels, (2) ALS-negative reference levels, (3) ALS-progression-positive reference levels, (4) ALS-regression-positive reference levels, and/or (5) previous levels of the one or more biomarkers in the subject (or group of subjects) before treatment with the composition.


When comparing the level(s) of the one or more biomarkers in the biological sample (from a subject or group of subjects having amyotrophic lateral sclerosis and currently or previously being treated with a composition) to ALS-positive reference levels, ALS-negative reference levels, ALS-progression-positive reference levels, and/or ALS-regression-positive reference levels, level(s) in the sample(s) corresponding to the ALS-negative reference levels or ALS-regression-positive reference levels (e.g., levels that are the same as the reference levels, substantially the same as the reference levels, above and/or below the minimum and/or maximum of the reference levels, and/or within the range of the reference levels) are indicative of the composition having efficacy for treating ALS. Levels of the one or more biomarkers in the sample(s) corresponding to the ALS-positive reference levels or ALS-progression-positive reference levels (e.g., levels that are the same as the reference levels, substantially the same as the reference levels, above and/or below the minimum and/or maximum of the reference levels, and/or within the range of the reference levels) are indicative of the composition not having efficacy for treating ALS. The comparisons may also indicate degrees of efficacy for treating ALS based on the level(s) of the one or more biomarkers.


When the level(s) of the one or more biomarkers in the biological sample (from a subject or group of subjects having ALS and currently or previously being treated with a composition) are compared to level(s) of the one or more biomarkers in a previously-taken biological sample(s) from the subject (or group of subjects) before treatment with the composition, any changes in the level(s) of the one or more biomarkers are indicative of the efficacy of the composition for treating ALS. That is, if the comparisons indicate that the level(s) of the one or more biomarkers have increased or decreased after treatment with the composition to become more similar to the ALS-negative or ALS-regression-positive reference levels (or less similar to the ALS-positive or ALS-progression positive reference levels), then the results are indicative of the composition having efficacy for treating ALS. If the comparisons indicate that the level(s) of the one or more biomarkers have not increased or decreased after treatment with the composition to become more similar to the ALS-negative or ALS-regression-positive reference levels (or less similar to the ALS-positive or ALS-progression-positive reference levels), then the results are indicative of the composition not having efficacy for treating ALS. The comparisons may also indicate degrees of efficacy for treating ALS based on the amount of changes observed in the level(s) of the one or more biomarkers after treatment. In order to help characterize such a comparison, the changes in the level(s) of the one or more biomarkers, the level(s) of the one or more biomarkers before treatment, and/or the level(s) of the one or more biomarkers in the subject currently or previously being treated with the composition may be compared to ALS-positive, ALS-negative, ALS-progression-positive, and/or ALS-regression-positive reference levels of the one or more biomarkers.


Another method for assessing the efficacy of a composition in treating amyotrophic lateral sclerosis (ALS) comprises (1) analyzing a first biological sample (or group of samples) from a subject (or group of subjects) to determine the level(s) of one or more biomarkers selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, 8, 16, 17, and/or 18, the first sample obtained from the subject at a first time point, (2) administering the composition to the subject, (3) analyzing a second biological sample from a subject to determine the level(s) of the one or more biomarkers, the second sample obtained from the subject at a second time point after administration of the composition, and (4) comparing the level(s) of one or more biomarkers in the first sample to the level(s) of the one or more biomarkers in the second sample in order to assess the efficacy of the composition for treating amyotrophic lateral sclerosis. As indicated above, if the comparison of the samples indicates that the level(s) of the one or more biomarkers have increased or decreased after administration of the composition to become more similar to the ALS-negative or ALS-regression-positive reference levels (or less similar to the ALS-positive or ALS-progression-positive reference levels), then the results are indicative of the composition having efficacy for treating ALS. If the comparison indicates that the level(s) of the one or more biomarkers have not increased or decreased after administration of the composition to become more similar to the ALS-negative or ALS-regression-positive reference levels (or less similar to the ALS-positive or ALS-progression-positive reference levels), then the results are indicative of the composition not having efficacy for treating ALS. The comparison may also indicate a degree of efficacy for treating ALS based on the amount of changes observed in the level(s) of the one or more biomarkers after administration of the composition. In order to help characterize such a comparison, the changes in the level(s) of the one or more biomarkers, the level(s) of the one or more biomarkers before administration of the composition, and/or the level(s) of the one or more biomarkers after administration of the composition may be compared to ALS-positive, ALS-negative, ALS-progression-positive, and/or ALS-regression-positive reference levels of the one or more biomarkers of the two compositions.


A method of assessing the relative efficacy of two or more compositions for treating amyotrophic lateral sclerosis comprises (1) analyzing, from a first subject having ALS and currently or previously being treated with a first composition, a first biological sample to determine the level(s) of one or more biomarkers selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, 8, 16, 17, and/or 18, (2) analyzing, from a second subject having ALS and currently or previously being treated with a second composition, a second biological sample to determine the level(s) of the one or more biomarkers, and (3) comparing the level(s) of one or more biomarkers in the first sample to the level(s) of the one or more biomarkers in the second sample in order to assess the relative efficacy of the first and second compositions for treating amyotrophic lateral sclerosis. The results are indicative of the relative efficacy of the two compositions, and the results (or the levels of the one or more biomarkers in the first sample and/or the level(s) of the one or more biomarkers in the second sample) may be compared to ALS-positive, ALS-negative, ALS-progression-positive, and/or ALS-regression-positive reference levels to aid in characterizing the relative efficacy.


Each of the methods of assessing efficacy may be conducted on one or more subjects or one or more groups of subjects (e.g., a first group being treated with a first composition and a second group being treated with a second composition).


As with the other methods described herein, the comparisons made in the methods of assessing efficacy (or relative efficacy) of compositions for treating ALS may be carried out using various techniques, including simple comparisons, one or more statistical analyses, and combinations thereof. Any suitable method may be used to analyze the biological samples in order to determine the level(s) of the one or more biomarkers in the samples. In addition, the level(s) of one or more biomarkers, including a combination of all of the biomarkers selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, 8, 16, 17, and/or 18 or any fraction thereof, may be determined and used in methods of assessing efficacy (or relative efficacy) of compositions for treating ALS.


Finally, the methods of assessing efficacy (or relative efficacy) of one or more compositions for treating ALS may further comprise analyzing the biological sample to determine the level(s) of one or more non-biomarker compounds. The non-biomarker compounds may then be compared to reference levels of non-biomarker compounds for subjects having (or not having) ALS.


VII. Methods of Screening a Composition for Activity in Modulating Biomarkers Associated with ALS


The identification of biomarkers for ALS also allows for the screening of compositions for activity in modulating biomarkers associated with ALS, which may be useful in treating ALS. Methods of screening compositions useful for treatment of ALS comprise assaying test compositions for activity in modulating the levels of one or more biomarkers selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, and/or 8. Such screening assays may be conducted in vitro and/or in vivo, and may be in any form known in the art useful for assaying modulation of such biomarkers in the presence of a test composition such as, for example, cell culture assays, organ culture assays, and in vivo assays (e.g., assays involving animal models).


In one embodiment, a method for screening a composition for activity in modulating one or more biomarkers of amyotrophic lateral sclerosis comprises (1) contacting one or more cells with a composition, (2) analyzing at least a portion of the one or more cells or a biological sample associated with the cells to determine the level(s) of one or more biomarkers of amyotrophic lateral sclerosis selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, and/or 8; and (3) comparing the level(s) of the one or more biomarkers with predetermined standard levels for the one or more biomarkers to determine whether the composition modulated the level(s) of the one or more biomarkers. As discussed above, the cells may be contacted with the composition in vitro and/or in vivo. The predetermined standard levels for the one or more biomarkers may be the levels of the one or more biomarkers in the one or more cells in the absence of the composition. The predetermined standard levels for the one or more biomarkers may also be the level(s) of the one or more biomarkers in control cells not contacted with the composition.


In addition, the methods may further comprise analyzing at least a portion of the one or more cells or a biological sample associated with the cells to determine the level(s) of one or more non-biomarker compounds of amyotrophic lateral sclerosis. The levels of the non-biomarker compounds may then be compared to predetermined standard levels of the one or more non-biomarker compounds.


Any suitable method may be used to analyze at least a portion of the one or more cells or a biological sample associated with the cells in order to determine the level(s) of the one or more biomarkers (or levels of non-biomarker compounds). Suitable methods include chromatography (e.g., HPLC, gas chromatograph, liquid chromatography), mass spectrometry (e.g., MS, MS-MS), ELISA, antibody linkage, other immunochemical techniques, and combinations thereof. Further, the level(s) of the one or more biomarkers (or levels of non-biomarker compounds) may be measured indirectly, for example, by using an assay that measures the level of a compound (or compounds) that correlates with the level of the biomarker(s) (or non-biomarker compounds) that are desired to be measured.


VIII. Method of Identifying Potential Drug Targets


The identification of biomarkers for ALS also allows for the identification of potential drug targets for ALS. A method for identifying a potential drug target for amyotrophic lateral sclerosis (ALS) comprises (1) identifying one or more biochemical pathways associated with one or more biomarkers for ALS selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, and/or 8 and (2) identifying a protein (e.g., an enzyme) affecting at least one of the one or more identified biochemical pathways, the protein being a potential drug target for amyotrophic lateral sclerosis.


Another method for identifying a potential drug target for amyotrophic lateral sclerosis (ALS) comprises (1) identifying one or more biochemical pathways associated with one or more biomarkers for ALS selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 3, 4, 5, 6, 7, and/or 8 and one or more non-biomarker compounds of ALS selected from Tables 3, 4, 5, 6, 7, and/or 8 and (2) identifying a protein affecting at least one of the one or more identified biochemical pathways, the protein being a potential drug target for amyotrophic lateral sclerosis.


One or more biochemical pathways (e.g., biosynthetic and/or metabolic (catabolic) pathway) are identified that are associated with one or more biomarkers (or non-biomarker compounds). After the biochemical pathways are identified, one or more proteins affecting at least one of the pathways are identified. Preferably, those proteins affecting more than one of the pathways are identified.


A build-up of one metabolite (e.g., a pathway intermediate) may indicate the presence of a ‘block’ downstream of the metabolite and the block may result in a low/absent level of a downstream metabolite (e.g. product of a biosynthetic pathway). In a similar manner, the absence of a metabolite could indicate the presence of a ‘block’ in the pathway upstream of the metabolite resulting from inactive or non-functional enzyme(s) or from unavailability of biochemical intermediates that are required substrates to produce the product. Alternatively, an increase in the level of a metabolite could indicate a genetic mutation that produces an aberrant protein which results in the over-production and/or accumulation of a metabolite which then leads to an alteration of other related biochemical pathways and result in dysregulation of the normal flux through the pathway; further, the build-up of the biochemical intermediate metabolite may be toxic or may compromise the production of a necessary intermediate for a related pathway. It is possible that the relationship between pathways is currently unknown and this data could reveal such a relationship.


For example, it has been proposed that high glutamate levels in ALS lead to hyper-excitability of the glutamate receptors, causing neurotoxicity. The drug Riluzole is thought to work by lowering glutamate levels by pre-synaptically inhibiting glutamate release in the central nervous system. This drug, however, does not lower the overall glutamate levels in the body. The identity of glutamate as a biomarker that is elevated in ALS as compared to a normal subject would suggest that potential drug targets may be in the pathways leading to glutamate production. A composition that would function by inhibiting the synthesis of glutamate may suppress the levels of glutamate. An example of such an enzyme is glutaminase 2, which converts glutamine to glutamate. Pathways leading to the production of any elevated biomarker would provide a number of potential targets for drug discovery.


The proteins identified as potential drug targets may then be used to identify compositions that may be potential candidates for treating ALS, including compositions for gene therapy.


IX. Methods of Treating ALS


The identification of biomarkers for ALS also allows for the treatment of ALS. For example, in order to treat a subject having ALS, an effective amount of one or more ALS biomarkers that are lowered in ALS as compared to a healthy subjects not having ALS may be administered to the subject. The biomarkers that may be administered may comprise one or more of the biomarkers in Tables 3, 4, 5, 6, 7, and/or 8 that are decreased in ALS as compared to subjects not having ALS. Such biomarkers could be isolated based on the identity of the biomarker compound (i.e. compound name). The biomarkers that are currently unnamed metabolites could be isolated based on the analytical characterizations for the biomarkers listed in the Examples below. In some embodiments, the biomarkers that are administered are one or more biomarkers listed in Tables 3, 4, 5, 6, 7, and/or 8 that are decreased in ALS and that have a p-value less than 0.05 and/or a q-value of less than 0.10. In other embodiments, the biomarkers that are administered are one or biomarkers listed in Tables 3, 4, 5, 6, 7, and/or 8 that are decreased in ALS by at least 5%, by at least 10%, by at least 15%, by at least 20%, by at least 25%, by at least 30%, by at least 35%, by at least 40%, by at least 45%, by at least 50%, by at least 55%, by at least 60%, by at least 65%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, by at least 95%, or by 100% (i.e., absent).


X. Methods of Using the ALS Biomarkers for Other Neurodegenerative Diseases


It is believed that some of the biomarkers for ALS described herein may also be biomarkers for neurodegenerative diseases in general. Therefore, it is believed that at least some of the ALS biomarkers may be used in the methods described herein for neurodegenerative diseases in general. That is, the methods described herein with respect to ALS may also be used for diagnosing (or aiding in the diagnosis of) a neurodegenerative disease, methods of monitoring progression/regression of a neurodegenerative disease, methods of assessing efficacy of compositions for treating a neurodegenerative disease, methods of screening a composition for activity in modulating biomarkers associated with a neurodegenerative disease, methods of identifying potential drug targets for neurodegenerative diseases, and methods of treating a neurodegenerative disease. Such methods could be conducted as described herein with respect to ALS.


XI. Methods for Distinguishing ALS from Other Neurodegenerative Diseases


The identification of biomarkers for ALS allows for distinguishing whether a subject has amyotrophic lateral sclerosis or has another neurodegenerative disease with symptoms similar to ALS (e.g., peripheral neuropathy or myopathy). A method of distinguishing whether a subject has ALS or has another neurodegenerative disease with symptoms similar to ALS (e.g., peripheral neuropathy or myopathy) comprises (1) analyzing a biological sample from a subject to determine the level(s) of one or more biomarkers of amyotrophic lateral sclerosis in the sample and (2) comparing the level(s) of the one or more biomarkers in the sample to ALS-positive and/or ALS-negative reference levels of the one or more biomarkers in order to determine whether the subject has amyotrophic lateral sclerosis or the other neurodegenerative disease (e.g., peripheral neuropathy or myopathy). The ALS-positive and/or ALS-negative reference levels may be levels that are specific for comparison with another particular neurodegenerative disease (e.g., reference levels of biomarkers for ALS that distinguish between peripheral neuropathy or myopathy).


The one or more biomarkers that are used are selected from xenobiotics (e.g., caffeine), metabolites of xenobiotics (e.g., paraxanthine, theophylline, and theobromine), and/or Tables 14 and/or 15. For example, in another aspect, a method of distinguishing whether a subject has amyotrophic lateral sclerosis (ALS) or has peripheral neuropathy or myopathy comprises (1) analyzing a biological sample from a subject to determine the level(s) of one or more biomarkers for amyotrophic lateral sclerosis in the sample, wherein the one or more biomarkers comprise one or more xenobiotics, metabolites of xenobiotics, and/or biomarkers listed in Tables 14 and 15; and (2) comparing the level(s) of the one or more biomarkers in the sample to ALS-positive and/or ALS-negative reference levels of the one or more biomarkers in order to determine whether a subject has ALS or has peripheral neuropathy or myopathy.


Any suitable method may be used to analyze the biological sample in order to determine the level(s) of the one or more biomarkers in the sample. Suitable methods include chromatography (e.g., HPLC, gas chromatography, liquid chromatography), mass spectrometry (e.g., MS, MS-MS), enzyme-linked immunosorbent assay (ELISA), antibody linkage, other immunochemical techniques, and combinations thereof. Further, the level(s) of the one or more biomarkers may be measured indirectly, for example, by using an assay that measures the level of a compound (or compounds) that correlates with the level of the biomarker(s) that are desired to be measured.


After the level(s) of the one or more biomarkers in the sample are determined, the level(s) are compared to ALS-positive and/or ALS-negative reference levels to distinguish whether the subject has ALS or has another disease (e.g., PN or myopathy) with symptoms related to ALS. Levels of the one or more biomarkers in a sample corresponding to the ALS-positive reference levels (e.g., levels that are the same as the reference levels, substantially the same as the reference levels, above and/or below the minimum and/or maximum of the reference levels, and/or within the range of the reference levels) are indicative of a diagnosis of ALS in the subject. Levels of the one or more biomarkers in a sample corresponding to the ALS-negative reference levels (e.g., levels that are the same as the reference levels, substantially the same as the reference levels, above and/or below the minimum and/or maximum of the reference levels, and/or within the range of the reference levels) are indicative of a diagnosis of no ALS in the subject. In addition, levels of the one or more biomarkers that are differentially present (especially at a level that is statistically significant) in the sample as compared to ALS-negative reference levels are indicative of a diagnosis of ALS in the subject. Levels of the one or more biomarkers that are differentially present (especially at a level that is statistically significant) in the sample as compared to ALS-positive reference levels are indicative of a diagnosis of no ALS in the subject. As described herein, ratios of biomarkers (e.g., a ratio of paraxanthine to caffeine) may be used as ALS-positive and/or ALS-negative reference levels.


The level(s) of the one or more biomarkers may be compared to ALS-positive and/or ALS-negative reference levels using various techniques, including a simple comparison (e.g., a manual comparison) of the level(s) of the one or more biomarkers in the biological sample to ALS-positive and/or ALS-negative reference levels. The level(s) of the one or more biomarkers in the biological sample may also be compared to ALS-positive and/or ALS-negative reference levels using one or more statistical analyses (e.g., t-test, Welch's T-test, Wilcoxon's rank sum test, random forest).


XII. Other Methods


Other methods of using the biomarkers discussed herein are also contemplated. For example, the methods described in U.S. Pat. No. 7,005,255 and U.S. patent application Ser. No. 10/695,265 may be conducted using a small molecule profile comprising one or more of the biomarkers disclosed herein and/or one or more of the non-biomarker compounds disclosed herein.


In any of the methods listed herein, the biomarkers that are used may be selected from those biomarkers in Tables 3, 4, 5, 6, 7, 8, 14, 15, 16, 17, and/or 18 having p-values of less than 0.05 and/or those biomarkers in Tables 3, 4, 5, 6, 7, 8, 14, 15, 16, 17, and/or 18 having q-values of less than 0.10. The biomarkers that are used in any of the methods described herein may also be selected from those biomarkers in Tables 3, 4, 5, 6, 7, 8, 14, 15, 16, 17, and/or 18 that are decreased as compared to the control group (e.g., subjects not having ALS, subjects having an earlier stage of ALS, etc.) by at least 5%, by at least 10%, by at least 15%, by at least 20%, by at least 25%, by at least 30%, by at least 35%, by at least 40%, by at least 45%, by at least 50%, by at least 55%, by at least 60%, by at least 65%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, by at least 95%, or by 100% (i.e., absent); and/or those biomarkers in Tables 3, 4, 5, 6, 7, 8, 14, 15, 16, 17, and/or 18 that are increased as compared to the control group by at least 5%, by at least 10%, by at least 15%, by at least 20%, by at least 25%, by at least 30%, by at least 35%, by at least 40%, by at least 45%, by at least 50%, by at least 55%, by at least 60%, by at least 65%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, by at least 95%, by at least 100%, by at least 810%, by at least 120%, by at least 130%, by at least 140%, by at least 150%, or more.


In addition, in any of the methods described herein, the biomarkers that are used may be selected from one or more xenobiotics and/or one or more metabolites of xenobiotics (including a xenobiotic and/or one or more Phase I metabolites of the xenobiotic) such as, for example, caffeine and/or one or more metabolites of caffeine (e.g., paraxanthine, theophylline, and/or theobromine).


EXAMPLES

The invention will be further explained by the following illustrative examples that are intended to be non-limiting.


I. General Methods


A. Identification of Metabolic Profiles for ALS


Each sample was analyzed to determine the concentration of several hundred metabolites. Analytical techniques such as GC-MS (gas chromatography-mass spectrometry) and LC-MS (liquid chromatography-mass spectrometry) were used to analyze the metabolites. Multiple aliquots were simultaneously, and in parallel, analyzed, and, after appropriate quality control (QC), the information derived from each analysis was recombined. Every sample was characterized according to several thousand characteristics, which ultimately amount to several hundred chemical species. The techniques used were able to identify novel and chemically unnamed compounds.


B. Statistical Analysis


The data was analyzed using several statistical methods to identify molecules (either known, named metabolites or unnamed metabolites) present at differential levels in a definable population or subpopulation (e.g., biomarkers for ALS biological samples compared to control biological samples; biomarkers for ALS compared to other diseases having symptoms similar to ALS) useful for distinguishing between the definable populations (e.g., ALS and control; ALS and myopathy or peripheral neuropathy). Other molecules (either known, named metabolites or unnamed metabolites) in the definable population or subpopulation were also identified.


C. Biomarker Identification


Various peaks identified in the analyses (e.g. GC-MS, LC-MS, MS-MS), including those identified as statistically significant, were subjected to a mass spectrometry based chemical identification process.


Example 1

In one example, biomarkers were discovered by (1) analyzing CSF samples from different groups of human subjects to determine the levels of metabolites in the samples and then (2) statistically analyzing the results to determine those metabolites that are differentially present in the two groups.


The CSF samples used for the analysis were from 99 ALS subjects (14 FALS subjects and 85 SALS subjects) and 36 control subjects not diagnosed with ALS. After the levels of metabolites were determined, the data was analyzed using univariate T-tests (i.e., Welch's T-test) (Table 3), Wilcoxon's rank-sum tests (Table 4), and random forest analyses.


T-tests were used to determine differences in the unknown means of two populations (e.g., ALS vs. Control). Wilcoxon's rank sum test is a non-parametric test that was used to compare the central values of the two populations. Q-values were used to account for multiple comparisons. Q-values give an estimate of the false discovery rate (for example, if everything with a q-value<0.10 is declared significant, then approximately 10% of those in the list are false discoveries). Random forest analyses were used to classify individuals. The “Out-of-Bag” (OOB) Error rate gives an estimate of how accurately new observations can be predicted using the random forest model (e.g., whether a sample is from an ALS subject or a control subject).


Random Forest Models


The GC-MS data from the CSF samples was collected and the data was partially unblinded. A random forest model was constructed based on the unblinded data. The OOB error from this random forest was approximately 31%, and the model estimated that the identity of control subjects could be predicted correctly 71% of the time and ALS subjects could be predicted 68% of the time. The model was then applied to the blind portion of the GC-MS data. After the predictions were provided, the data was unblinded. The results of the predictions on the blinded portion of the data are shown in Table 1. The OOB error from the initial random forest model was very similar to the actual error (approximately 29%) determined when using the model on the blinded portion of the data.

TABLE 1Results of Metabolomic PredictionsRandom Forest AnalysisOverall71%Control70%ALS71%


Finally, the random forest model was used to classify the complete GC-MS and LC-MS data. The confusion matrix for the random forest is shown in Table 2. In order to see which compounds were more important, an importance plot was constructed (shown in FIG. 1) ranking the compounds based on their importance for the predictions.

TABLE 2Results of Random Forest, CSF: Controls vs. ALS, Full DatasetALSControlErrorALS662932%Control102429%OOB Error30%


Biomarkers


As listed below in Tables 3-4, biomarkers were discovered that were differentially present between samples from ALS subjects and Control subjects not diagnosed with ALS.


Tables 3 and 4 include, for each listed biomarker and non-biomarker compound, the p-value and the q-value determined in the statistical analysis of the data concerning the biomarkers, an indication of whether the mean of a particular compound was higher in the ALS or Control samples (a “+” indicating a higher mean in ALS samples as compared to the control samples and a “−” indicating a lower mean in ALS samples as compared to the control samples), and an indication of the percentage difference in the ALS mean as compared to the control mean. Throughout the tables, names of metabolites ending with the notation “−35” indicate that the levels of those compounds were measured using LC-MS, and names ending with the notation “−9” indicate that the levels of those compounds were measured using GC-MS. The term “Isobar” as used in the tables indicates the compounds that could not be distinguished from each other on the analytical platform used in the analysis (i.e., the compounds in an isobar elute at nearly the same time and have similar (and sometimes exactly) quant ions, and thus cannot be distinguished).


Non-biomarker compounds identified in the analyses are also listed in the Tables 3 and 4 below as those compounds that having a percentage change in ALS of 0%.

TABLE 3ALS Biomarkers from CSF samples - T-tests of ALS vs. ControlsIncrease(+)orDecrease%(−)changeCompoundp-valueq-valuein ALSin ALS2-amino-heptanedioic acid - 92.19E−060.000352124+37%Metabolite - 1113 - 354.55E−060.00036514+63%amino-malinate-mixture - 97.90E−060.000423163+38%trans-4-hydroxyproline - 353.90E−050.001566631+19%Metabolite - 2185 - 357.00E−050.002204109+43%Metabolite - 760 - 98.23E−050.002204109+32%Metabolite - 2389 - 350.0001089960.0024996+16%Metabolite - 3138 - 350.0001244780.0024996+88%alpha-Hydroxyisobutyric acid-tms - 90.0003523890.005660962+27%5-oxoproline - 90.0004289640.005745226+13%Metabolite - 522 - 90.0004291610.005745226+70%Methionine - 350.0005928380.007325901+21%Metabolite - 421 - 90.0008617310.009888077+47%Lactate (previously: Metabolite - 2694) -0.0009599960.010281254+14%35Metabolite - 268 - 90.0010934580.010369188+51%alpha-aminoadipic acid - 90.0011086940.010369188+33%Metabolite - 1086 - 350.0011837490.010369188+61%Pyridoxamine - 350.0012372270.010369188+10%Metabolite - 1116 - 350.0012909420.010369188+29%alpha-2-diamino-gamma-0.0014174410.010843108+76%oxobenzenebutanoic acid - 35Metabolite - 1830 - 350.0018511190.013516981+18%DL-pipecolic acid - 350.0023135390.015578068+108%Metabolite - 2074 - 350.0023273240.015578068+52%Glycerol-2-phosphate (previously0.00244250.015695045+16%Metabolite - 1573) - 35Metabolite - 502 - 90.002611490.016135525+25%Tyrosine - 350.0029017920.016557204+19%Metabolite - 2567 - 350.0029524610.016557204+56%Carnitine (previously: Metabolite - 1336) -0.0029889380.016557204+55%35Urea - 90.003994980.020751305+18%Metabolite - 763 - 90.0040383320.020751305+17%3-amino-isobutyrate - 90.0041335890.020751305+31%Tetradecanoic acid - 90.0047567240.022701843+14%Metabolite - 591 - 90.0048047630.022701843+51%Lactate (previously: Metabolite - 2563) -0.0049896710.022901923+18%35Uric acid - 350.0051883190.023152196+22%Uric acid (previously: Metabolite - 1910) -0.0070142320.029060165+22%35Metabolite - 1068 - 350.0070549610.029060165+8%Metabolite - 2697 - 350.0075066050.030147523+40%Citric acid - 90.0088690790.034433835+12%gamma-L-glutamyl-L-glutamine - 350.0090025730.034433835+16%Metabolite - 547 - 90.0094880080.035446605+18%Alanine - 90.0101871740.037193674+14%Isobar 1: includes-mannose-fructose-0.0108769150.038829452+15%glucose-galactose-alpha-L-sorbopyranose-Inositol- 35Metabolite - 511 - 90.0113903960.039584956+12%Metabolite - 782 - 90.0116293240.039584956+11%Metabolite - 1597 - 350.0120665120.039584956+19%Metabolite - 2526 - 350.0120741950.039584956+9%Tryptophan - 350.0126458490.040199985+15%Metabolite - 609 - 90.0127622720.040199985+6%Phenylalanine - 350.0131098630.040500732+12%Metabolite - 655 - 90.0164390430.049827456+32%Metabolite - 1335 - 350.0173593030.051642419+45%o-phosphoethanolamine - 90.0181007610.052869133+11%Metabolite - 508 - 90.0189771270.054439044+9%Isobar-2-includes-3-amino-isobutyrate-2-0.0201260190.056721941+36%amino-butyrate-4-aminobutanoic acid-dimethylglycine - 351-7-dihydro-6h-purin-6-one - 350.0228681910.063339098−13%Metabolite - 458 - 90.0234836670.063941373+75%Metabolite - 2686 - 350.0253409220.06673607+8%Valine - 90.0278571260.070809473+34%oxalic acid (previously: Metabolite0.0280753440.070809473+8%- 1829) - 35Metabolite - 2141 - 350.0282100130.070809473+32%Citrulline (previously: Metabolite - 2527) -0.0289234770.0714834+267%35Metabolite - 1126 - 350.0298625650.072686077+40%Lactate (previously: Metaboiite-3316) -0.0312286390.073937034+17%35Metabolite - 2390 - 350.0319606380.073937034+55%Serine - 90.0320409230.073937034−11%Metabolite - 2100 - 350.0322349420.073937034+11%Metabolite - 2139 - 350.0326777630.073937034+47%Arginine - 350.0360811990.078737229+7%Cytidine - 350.0362696860.078737229−9%Pantothenic acid - 350.0370212630.079145001+27%Metabolite - 393 - 90.037635870.079145001+16%Lysine - 90.037935530.079145001+17%Glycerate - 90.0401875390.082768458−6%Metabolite - 553 - 90.0409341610.083239001−32%Carnosine - 350.043374190.084597791−8%Metabolite - 1346 - 350.0436610530.084597791+7%Metabolite - 2548-possible-Cl-adduct-of-0.0437530930.084597791+14%uric acid - 35Metabolite - 992 - 90.0441944750.084597791+28%Metabolite - 2105 - 350.044235430.084597791+32%Histamine - 350.0482337850.091159192+19%4-Guanidinobutanoic acid (previously:0.0492045940.091912644−24%Metabolite - 3179) - 35Metabolite - 3441 - 350.0537504380.096784494−44%Metabolite - 753 - 90.0539402010.096784494+35%5-hydroxy-1H-indole-3-acetic acid - 90.0539809790.096784494−15%Metabolite - 988 - 90.0542225870.096784494+16%Metabolite - 1108 - 350.0558866090.09787302+63%Metabolite - 273 - 90.0560509220.09787302+20%Metabolite - 1131 - 350.0621020930.107273226+13%Metabolite - 383 - 90.0641910490.109466474−5%Metabolite - 289 - 90.0647346310.109466474+9%Isobar 4: includes-Gluconic acid-0.0655779410.109737383+12%arabinose-D-ribose- 35Metabolite - 442 - 90.0694023310.114939785−14%Metabolite - 3182 - 350.0719428340.117931419+20%5-S-methyl-5-thioadenosine - 350.0739426220.11917087−7%Metabolite - 783 - 90.0741826010.11917087+5%Metabolite - 568 - 90.076033540.120397213+7%Metabolite - 2687 - 350.0766838140.120397213+9%Metabolite - 3127 - 350.0771943660.120397213+21%Metabolite - 276 - 90.08083860.124144964+4%Metabolite - 780 - 90.0814857570.124144964−7%Metabolite - 3218 - 350.0819156570.124144964+21%Isoleucine - 90.0842750780.126527067+18%N,N-dimethylarginine (previously:0.087169660.129661085+24%Metabolite - 3162) - 35Metabolite - 121 - 90.0886109530.130595726+42%Metabolite - 577 - 90.090630770.132358258+40%Metabolite - 961 - 90.0930029540.133894384+12%Metabolite - 226 - 90.0933495720.133894384−19%Metabolite - 645 - 90.0953132830.135501168+40%Metabolite - 1656 - 350.0976049030.1366745+13%Metabolite - 991 - 90.0981418020.1366745+136%Metabolite - 504 - 90.0986909720.1366745+10%Metabolite - 141 - 90.1034788920.142080332+3%Glutamic acid - 90.1078994890.145713342+8%Metabolite - 863 - 90.1079389590.145713342+18%Praline - 350.1104508120.147366108+156%Metabolite - 1344 retired: Na adduct of0.1109979420.147366108+15%citric acid - 35Metabolite - 283 - 90.1135583570.149529653+12%Metabolite - 549 - 90.116251910.151831905+12%2-ethylhexanoic acid - 90.1173930150.152085786+82%Metabolite - 554 - 90.1250652560.160729176+8%alpha-4-dihydroxybenzenepropanoic acid -0.1264760820.160754983−38%35Metabolite - 2254 - 350.1276763810.160754983−7%Uridine - 350.1280873850.160754983−7%Metabolite - 1289 - 350.1324827660.164982443+28%Metabolite - 706 - 90.1337488060.165277836+13%Metabolite - 3130 - 350.140604030.17043414+36%monoethanolamine- 90.1426257690.17043414−7%Metabolite - 995 - 90.1429048850.17043414+17%Metabolite - 3416 - 350.1448283160.17043414+283%Metabolite - 2056 - 350.1452021340.17043414+14%Metabolite - 998 - 90.1457885490.17043414+7%Creatinine - 350.1464840350.17043414−6%Metabolite - 3334 - 350.1472235190.17043414+16%Metabolite - 386 - 90.1480303110.17043414+40%Metabolite - 595 - 90.1485308180.17043414−13%N-Acetylglutamine - 90.1540403020.175502498+37%Metabolite - 2558 - 350.1590394420.178894231+23%Metabolite - 1979 retired: CL adduct of0.1592444550.178894231+12%Isobar 19 - 35Metabolite - 597 - 90.1607258090.179304498−6%Metabolite - 3180 - 350.1702011340.188565615+19%Metabolite - 761 - 90.1718054750.18884866+4%Glutamine - 350.1764966720.18884866+6%Metabolite - 3056 - 350.177717070.18884866+12%Gamma-L-glutamyl-L-tyrosine - 350.1778959140.18884866+18%Metabolite - 3166 - 350.1779019930.18884866−18%2-amino-butyrate - 90.1783606120.18884866−65%Metabolite - 286 - 90.1795414180.18884866+4%Butanoic acid - 90.180994690.18884866+18%Metabolite - 1127 - 350.181447050.18884866+14%Metabolite - 704 - 90.1822122410.18884866+9%Metabolite - 2052 retired: potassium0.1839579210.189435753+4%adduct of Isobar 1 - 35Metabolite - 441 - 90.1884893850.192170414−10%Metabolite - 996 - 90.1890059860.192170414−17%Metabolite - 593 - 90.2039929150.203543512+5%Metabolite - 413 - 90.2065292070.204157583+9%Glycerol - 90.2071500610.204157583+7%Metabolite - 2366 - 350.2084683640.204204056+9%Caffeine - 350.2137354390.208094522−24%glycerol-2-phosphate (Metabolite - 1820) -0.2170739790.208835572+19%35Metabolite - 1498 - 350.2170965350.208835572+10%Metabolite - 1351 retired: urea adduct of0.2307416260.22064024+153%Isobar 6 - 35Palmitoleic acid - 90.2330038010.221485019−15%Metabolite - 1612 - 350.2366776170.223096352−18%Metabolite - 387 - 90.2429418980.223096352+8%Phosphate - 90.2437300340.223096352−7%Metabolite - 777 - 90.244271330.223096352−12%Metabolite - 1343 retired: p-0.2464745250.223096352−21%hydroxyphenyllactic acid - 35Metabolite - 2181 - 350.247038590.223096352+11%2-aminobutanoic acid - 90.2476632090.223096352−4%Metabolite - 2696 - 350.2486405860.223096352−14%4-hydroxy-2-quinolinecarboxylic acid - 350.2493478170.223096352+12%Isobar 3: includes-inositol-1-phosphate-0.2497999970.223096352+9%mannose-6-phosphate-glucose-6-phosphate- 35Metabolite - 3468 retired: Metabolite - 0.2527219620.223096352−20%1498 - 35Metabolite - 490 - 90.2527526990.223096352−8%Metabolite - 150 - 90.2559773590.224707992+8%Metabolite - 3370 - 350.2579044190.225169215−74%Orotidine-5-phosphate - 350.2650882730.230190206+8%Histidine - 350.2738249310.235233643−10%Metabolite - 1328 - 350.2758597520.235721147+7%Metabolite - 702 - 90.2778274480.236146439+5%Fumaric acid - 90.2832501290.239488448+7%Dulcitol - 90.2853230230.239978042+4%Metabolite - 2806 - 350.293334730.244954506+8%Metabolite - 381 - 90.2942894460.244954506+4%Metabolite - 1132 - 350.2958920180.245018894−6%Metabolite - 984 - 90.2975374730.245117949+14%Metabolite - 571 - 90.2993995690.245393556+2%Malic acid - 350.3082069930.251154317−12%Metabolite - 223 - 90.3095549710.251154317−8%Metabolite - 1114 - 350.3178316170.25541317+8%Lipoate - 90.317983970.25541317−2%Metabolite - 614 - 90.3217138510.256295075+5%Allantoin - 350.3222727430.256295075+7%Metabolite - 1843 - 350.3243465050.256673622+12%2-deoxy-D-ribose- 350.3283668540.25810406+12%Metabolite - 982 - 90.3293674230.25810406+4%Metabolite - 2607 - 350.3324855790.259282764−21%Decanoic acid - 90.341288670.264861955−6%Metabolite - 861 - 90.3450633960.266503927−41%4-hydroxy-3-methoxymandelate - 350.3485267990.26789089−6%N-6-trimethyl-l-lysine - 350.3535595460.270276098+10%Metabolite - 601 - 90.354994840.270276098−8%Metabolite - 285 - 90.3586668510.271223252−5%Metabolite - 990 - 90.3606956440.271223252+9%Metabolite - 1281 - 350.3613038910.271223252+28%Metabolite - 1500 - 350.3645875260.27241524−8%Palmitate - 90.3677157930.273480639−8%Metabolite - 987 - 90.3736596320.27662059−5%Metabolite - 485 - 90.3790636230.279333916−5%pyrophosphate - 350.3828415890.280829709+13%Metabolite - 406 - 90.3907484910.285326877+3%Adenosine - 350.3941766810.286527763+16%Metabolite - 653 - 90.3991370740.288794022−1%Metabolite - 562 - 90.4008898020.288794022+5%Metabolite - 2047 - 350.4142971110.296771543+21%Glutarate - 350.415658550.296771543+8%Metabolite - 1216 - 350.4184753650.297460643−7%Metabolite - 721 - 90.4210481640.297970985+8%Metabolite - 1818 - 350.4355211730.304819961+7%Selenocystine - 350.4366720160.304819961+7%N-acetyl-L-alanine - 350.4377367830.304819961−12%1-Hexadecanol- 90.4384786380.304819961+3%Metabolite - 1252 - 350.4402134760.304819961−12%Metabolit-1142-possible-5-0.4448829660.30673117+12%hydroxypentanoate-or-beta-hydroxyisovaleric acid - 35Adenosine-3-5-cyclic-monophosphate - 350.4509306450.309572196−8%gamma-aminobutyryl-L-histidine - 350.4550699790.309985014−7%Metabolite - 2053 - 350.4589447670.309985014+4%Metabolite - 2174 - 350.4602396240.309985014−7%Metabolite - 1608 - 350.4626097980.309985014+27%Metabolite - 263 - 90.4639045870.309985014+1%Metabolite - 985 - 90.4648534260.309985014−4%Metabolite - 382 - 90.4650393330.309985014−2%Xylitol - 350.4758175670.315858931+8%Metabolite - 2272 - 350.4926502650.322589067+7%Metabolite - 3183 - 350.4944613320.322589067+5%DOPA - 90.4956985650.322589067+3%Metabolite - 688 - 90.4965207860.322589067+7%Metabolite - 642 - 90.4991104440.322589067−4%alpha-L-sorbopyranose - 90.4995569850.322589067+7%Metabolite - 1340 - 350.5000125860.322589067+5%Cholesterol - 90.508662080.324370836+5%Metabolite - 1304 - 350.5094915940.324370836+9%Glucarate - 350.5131966580.324370836+7%Mercaptopyruvate - 350.5136353720.324370836−7%4-acetamidobutyric acid - 350.5145817810.324370836−4%3-nitro-L-tyrosine - 350.514889370.324370836+10%5-6-dihydrouracil - 350.5193182050.325882949+7%Vitamin-B6 - 90.5443646030.337416774−2%N-acetyl-L-valine - 350.546526520.337416774+4%Metabolite - 2051 - 350.5477845750.337416774−7%Metabolite - 138 - 90.5485285740.337416774−9%Hydroorotate - 90.5516139490.337416774+2%Metabolite - 1064 - 350.5534239070.337416774+11%Metabolite - 2753 - 350.5547581410.337416774+8%Metabolite - 147 - 90.5566016480.337416774+4%Metabolite - 2821 - 350.5638672340.3405361 93−7%Metabolite - 505 - 90.5672682760.341307074−9%Metabolite - 1342-possible-0.5738822570.342584549+10%phenylacetylglutamine-or-formyl-N-acetyl-5-methoxykynurenamine - 35N-carbamoyl-L-aspartate - 350.5742864750.342584549+13%Metabolite - 465 - 90.5757891550.342584549+4%Metabolite - 2752 - 350.583774830.346054204−5%Metabolite - 771 - 90.5881856270.346944856+2%Metabolite - 128 - 90.5895967010.346944856+7%Metabolite - 443 - 90.592970.347656388+3%Metabolite - 3249 - 350.5960099460.348168012+5%Metabolite - 394 - 90.6052610430.352291119+1%Metabolite - 2231 - 350.6126498490.355251722−6%Metabolite - 3143 - 350.615013940.355251722+5%Octadecanoic acid - 90.6169817820.355251722−5%Metabolite - 606 - 90.6206073590.35606308−28%Inosine - 350.6232601750.356312544+2%Metabolite - 146 - 90.6284411550.357149158+4%Tetronic acid - 90.6313312580.357149158−3%alpha-keto-glutarate - 350.6359397370.357149158+13%Metabolite - 2313 - 350.6362713580.357149158+6%Sarcosine - 90.6364778080.357149158−4%2-deoxyadenosine - 350.6380628690.357149158+10%Metabolite - 1842 retired: 4-0.6500879120.361421083+11%Guanidinobutanoic acid - 35Metabolite - 2822 - 350.6506406330.361421083−8%alpha-D-ribose-5-phosphate - 350.6524442780.361421083−2%Threonine - 90.6594780250.362815249−3%Metabolite - 651 - 90.6623794080.3631677350%n-dodecanoate - 90.6661435960.363170201−2%Ascorbate - 90.6669052970.363170201+2%Metabolite - 1104 - 350.6738906910.36374061+5%Metabolite - 221 - 90.6761742430.36374061+2%Metabolite - 274 - 90.6763829610.36374061−7%Glyoxylate - 90.6770097520.36374061−4%3-phospho-d-glycerate - 90.6830740.365775452−3%Metabolite - 2593 - 350.6925160590.369599528−8%Metabolite - 501 - 90.6957258740.370083112+1%Metabolite - 2698 - 350.7003199220.371297399−14%2-deoxyguanosine - 350.7047533310.37241881+8%Metabolite - 1713 retired: n-acetyl-L-0.7090126010.373441148−9%aspartic acid - 35Metabolite - 3230 - 350.7207585230.378387181+2%Metabolite - 1286 - 350.7366074810.385448007+2%Guanosine - 350.748930340.390623856−3%Metabolite - 2005 - 350.7519999990.390955579−12%Metabolite - 222 - 90.7600638820.393873223+3%Metabolite - 3313 - 350.7688209990.3971301 89+6%Metaboiite-2038 - 350.7873933150.405420014−6%Pentanedicic acid - 90.7909303250.405940091+1%Metabolite - 2726 - 350.7969085590.407705808+2%Xanthine - 350.8046515420.41036031−1%Metabolite - 691 - 90.8145027550.413496839+1%Saccharopine - 350.8159497260.413496839−1%Metabolite - 2266 retired 4-acetominophen0.8291945150.418887456+5%sulfate - 35Metabolite - 2703 - 350.8338684750.419351988−2%Metabolite - 2026 - 350.8353349060.419351988−3%Metabolite - 594 - 90.8403458430.420553332−1%Isocitrate - 350.8660172080.432054671+2%Metabolite - 3401 - 350.8687978570.432100007−2%Phosphoenolpyruvate - 350.8764784390.434228888−2%Metabolite - 580 - 90.8784843280.4342288880%3-hydroxy-3-methylglutarate - 350.8853114040.436261123+1%Metabolite - 1192 - 350.8983403370.439274785−2%Metabolite - 278 - 90.8993637970.439274785−1%Metabolite - 136 - 90.899630390.439274785−1%Metabolite - 472 - 90.9040694760.440104614−1%Metabolite - 3231 - 350.9160452740.444587246−2%Gulono-1-4-lactone - 90.9205253170.4454158920%Metabolite - 398 - 90.9268062370.445658163−1%Inositol - 90.927301470.4456581630%Tartarate - 90.9293485330.4456581630%Metabolite - 2109 - 350.9450632260.451845168+1%Metabolite - 482 - 90.9539270210.4542978980%12-hydroxydodecanoic acid - 90.9568655940.4542978980%Metabolite - 2867 - 350.9593228760.454297898+2%Metabolite - 388 - 90.9639495350.4542978980%Metabolite - 145 - 90.9643330510.4542978980%Metabolite - 1133 - 350.9744670540.4577297180%3-phospho-d-glycerate - 350.9814025280.4591420630%3-phospho-l-serine - 90.9831900360.4591420630%Metabolite - 770 - 90.9953194040.4622524180%Gluconic acid - 90.9956053810.4622524180%Metabolite - 2854 - 350.9992517870.4626083990%









TABLE 4










ALS Biomarkers from CSF: Wilcoxon's Rank Sum Test ALS v. Controls














Increase






(+) or
%





Decrease
change


Compound
p-value
q-value
(−) in ALS
in ALS














Metabolite - 1113 - 35
4.81E−05
0.00589796
+
63%


Metabolite - 2185 - 35
6.59E−05
0.00589796
+
43%


Metabolite - 1713 retired: n-acetyl-L-
0.000140851
0.008410318

−9%


aspartic acid - 35


alpha-Hydroxyisobutyric acid-tms - 9
0.000278249
0.012019186
+
27%


Metabolite - 1336 retired: carnitine - 35
0.00037254
0.012019186
+
55%


alpha-aminoadipic acid - 9
0.000491208
0.012019186
+
33%


amino-malinate-mixture - 9
0.000519388
0.012019186
+
38%


2-amino-heptanedioic acid - 9
0.000624382
0.012427436
+
37%


Metabolite - 2389 - 35
0.000780424
0.01298768
+
16%


Metabolite - 763 - 9
0.000797537
0.01298768
+
17%


3-amino-isobutyrate - 9
0.000928321
0.013537644
+
31%


Metabolite - 3138 - 35
0.000982455
0.013537644
+
88%


Metabolite - 2139 - 35
0.001196211
0.014291844
+
47%


Metabolite - 1086 - 35
0.001196757
0.014291844
+
61%


5-oxoproline - 9
0.001342087
0.014421736
+
13%


Metabolite - 1116 - 35
0.001368651
0.014421736
+
29%


trans-4-hydroxyproline - 35
0.001455763
0.014487443
+
19%


alpha-2-diamino-gamma-
0.001605672
0.015138292
+
76%


oxobenzenebutanoic acid - 35


Metabolite - 760 - 9
0.002070127
0.018541315
+
32%


DL-pipecolic acid - 35
0.002460098
0.020870333
+
108%


Metabolite - 421 - 9
0.002563176
0.020870333
+
47%


Metabolite - 2567 - 35
0.003042785
0.023653577
+
56%


Metabolite - 753 - 9
0.003171436
0.023653577
+
35%


Urea - 9
0.003301135
0.023653577
+
18%


Metabolite - 1830 - 35
0.003555749
0.024498043
+
18%


Metabolite - 2074 - 35
0.004045965
0.025905285
+
52%


Uric acid - 35
0.004049233
0.025905285
+
22%


Metabolite - 2686 - 35
0.004320203
0.026202897
+
8%


Pyridoxamine - 35
0.004461991
0.026202897
+
10%


Metabolite - 1346 - 35
0.004534583
0.026202897
+
7%


Methionine - 35
0.004754661
0.026401645
+
21%


Metabolite - 2526 - 35
0.004986999
0.026401645
+
9%


Citric acid - 9
0.005011137
0.026401645
+
12%


Valine - 9
0.006160305
0.031397644
+
34%


Glutamine - 35
0.006309951
0.031397644
+
6%


Metabolite - 2694 retired: lactate - 35
0.007031923
0.033150012
+
14%


Metabolite - 268 - 9
0.00703224
0.033150012
+
51%


gamma-L-glutamyl-L-glutamine - 35
0.007475374
0.034335385
+
16%


Metabolite - 1910 retired: uric acid - 35
0.008126877
0.036394628
+
22%


Metabolite - 961 - 9
0.009191951
0.040160347
+
12%


Metabolite - 2527 retired: citrulline - 35
0.010313603
0.043988051
+
267%


Metabolite - 609 - 9
0.010770241
0.044867365
+
6%


Tetradecanoic acid - 9
0.011564259
0.047080248
+
14%


Metabolite - 547 - 9
0.011895473
0.047352487
+
18%


Metabolite - 1573 retired: glycerol-2-
0.013301907
0.051799991
+
16%


phosphate - 35


Serine - 9
0.014264462
0.053295855

−11%


Metabolite - 1829 retired: oxalic acid -
0.01428108
0.053295855
+
8%


35


Metabolite - 121 - 9
0.018224891
0.066182714
+
42%


Metabolite - 782 - 9
0.018473151
0.066182714
+
11%


Metabolite - 2563 retired: lactate - 35
0.019274499
0.067137694
+
18%


Metabolite - 1349 retired: Isobar 7 - 35
0.020162937
0.067137694
+
55%


Metabolite - 591 - 9
0.020199367
0.067137694
+
51%


1-7-dihydro-6h-purin-6-one - 35
0.020491957
0.067137694

−13%


Isobar 2: includes-3-amino-isobutyrate-
0.020613679
0.067137694
+
36%


2-amino-butyrate-4-aminobutanoic acid-


dimethylglycine - 35


Metabolite - 522 - 9
0.02193703
0.069347268
+
70%


Metabolite - 2687 - 35
0.022066355
0.069347268
+
9%


Metabolite - 3316 retired: lactate - 35
0.023746931
0.07334206
+
17%


gamma-L-glutamyl-L-tyrosine - 35
0.026183625
0.079497114
+
18%


Metabolite - 704 - 9
0.026986286
0.080568537
+
9%


Tyrosine - 35
0.027825502
0.080810989
+
19%


Metabolite - 2697 - 35
0.02800104
0.080810989
+
40%


Metabolite - 508 - 9
0.028936572
0.080810989
+
9%


Metabolite - 1335 - 35
0.02932312
0.080810989
+
45%


Metabolite - 1068 - 35
0.03124896
0.083591292
+
8%


Isobar 1: includes-mannose-fructose-
0.031265274
0.083591292
+
15%


glucose-galactose-alpha-L-


sorbopyranose-Inositol - 35


Alanine - 9
0.034246794
0.08935306
+
14%


Histamine - 35
0.034417943
0.08935306
+
19%


Metabolite - 995 - 9
0.036853002
0.094307979
+
17%


Pantothenic acid - 35
0.037556802
0.094347666
+
27%


Metabolite - 393 - 9
0.03869605
0.0949549
+
16%


Caffeine - 35
0.039651027
0.095983442

−24%


Metabolite - 2100 - 35
0.040207393
0.096032506
+
11%


Metabolite - 283 - 9
0.040852014
0.096288292
+
12%


Metabolite - 2005 - 35
0.043644554
0.101534337

−12%


Cytidine - 35
0.044363373
0.10188343

−9%


Metabolite - 1343 retired: p-
0.048835069
0.109317592

−21%


hydroxyphenyllactic acid - 35


Metabolite - 655 - 9
0.049078982
0.109317592
+
32%


Carnosine - 35
0.050049316
0.109317592

−8%


Metabolite - 1126 - 35
0.050906945
0.109317592
+
40%


Metabolite - 863 - 9
0.05109681
0.109317592
+
18%


Metabolite - 996 - 9
0.052123383
0.109317592

−17%


Histidine - 35
0.052173845
0.109317592

−10%


Metabolite - 1597 - 35
0.053116952
0.109367303
+
19%


Metabolite - 3179 retired: 4-
0.055443465
0.111950626

−24%


Guanidinobutanoic acid - 35


Arginine - 35
0.056338742
0.111950626
+
7%


Metabolite - 2390 - 35
0.056389919
0.111950626
+
55%


Metabolite - 458 - 9
0.056871452
0.111950626
+
75%


Metabolite - 3441 - 35
0.057664917
0.112278719

−44%


Creatinine - 35
0.059347516
0.113196789

−6%


Metabolite - 273 - 9
0.059995122
0.113196789
+
20%


Metabolite - 553 - 9
0.060032179
0.113196789

−32%


Metabolite - 549 - 9
0.062403991
0.115890315
+
12%


Glycerate - 9
0.062754558
0.115890315

−6%


Metabolite - 1108 - 35
0.06463188
0.11813928
+
63%


Metabolite - 502 - 9
0.068588529
0.124105178
+
25%


Metabolite - 289 - 9
0.070888141
0.126983471
+
9%


Metabolite - 3056 - 35
0.071674664
0.127121174
+
12%


Lysine - 9
0.074452222
0.130360941
+
17%


Metabolite - 2141 - 35
0.074956812
0.130360941
+
32%


Isobar 4: includes-Gluconic acid-
0.075803406
0.130565665
+
12%


arabinose-D-ribose - 35


Allantoin - 35
0.076640443
0.130688955
+
7%


o-phosphoethanolamine - 9
0.07733412
0.130688955
+
11%


Praline - 35
0.081907325
0.137123714
+
156%


Metabolite - 1351 retired: urea adduct
0.084293113
0.139811193
+
153%


of Isobar 6 - 35


Metabolite - 998 - 9
0.089274784
0.144477301
+
7%


Metabolite - 780 - 9
0.089278687
0.144477301

−7%


Metabolite - 861 - 9
0.089525962
0.144477301

−41%


Glutamic acid - 9
0.092128452
0.146354769
+
8%


Metabolite - 2105 - 35
0.092323385
0.146354769
+
32%


Tryptophan - 35
0.094745188
0.147861757
+
15%


5-S-methyl-5-thioadenosine - 35
0.095257482
0.147861757

−7%


Metabolite - 2548-possible-Cl-adduct-
0.095750323
0.147861757
+
14%


of-uric acid - 35


Metaboiite-1498 - 35
0.098329731
0.149079215
+
10%


Orotidine-5-phosphate - 35
0.098337631
0.149079215
+
8%


Metabolite - 988 - 9
0.099035399
0.149079215
+
16%


5-hydroxy-1H-indole-3-acetic acid - 9
0.103596668
0.153796594

−15%


Uridine - 35
0.104730874
0.153796594

−7%


Metabolite - 1656 - 35
0.104744914
0.153796594
+
13%


Metabolite - 2056 - 35
0.108063805
0.155226181
+
14%


Metabolite - 413 - 9
0.108565144
0.155226181
+
9%


Metabolite - 441 - 9
0.109069778
0.155226181

−10%


Metabolite - 1131 - 35
0.109184732
0.155226181
+
13%


Metabolite - 3182 - 35
0.111442107
0.157187931
+
20%


Metabolite - 761 - 9
0.113020563
0.157474372
+
4%


Metabolite - 1820 retired: glycerol-2-
0.113403378
0.157474372
+
19%


phosphate - 35


Phosphate - 9
0.116453675
0.160466156

−7%


Phenylalanine - 35
0.119736234
0.163729865
+
12%


Metabolite - 141 - 9
0.121159331
0.164420718
+
3%


adenosine - 35
0.123111413
0.16518472
+
16%


Metabolite - 562 - 9
0.123566591
0.16518472
+
5%


Metabolite - 777 - 9
0.126086251
0.166074299

−12%


Metabolite - 286 - 9
0.129743888
0.169644563
+
4%


Metabolite - 595 - 9
0.135916731
0.176381625

−13%


Metabolite - 702 - 9
0.137475446
0.176381625
+
5%


Metabolite - 597 - 9
0.139467115
0.176381625

−6%


Metabolite - 577 - 9
0.140075305
0.176381625
+
40%


Isoleucine - 9
0.140133007
0.176381625
+
18%


Metabolite - 150 - 9
0.140804253
0.176381625
+
8%


Metabolite - 3370 - 35
0.142935013
0.177807361

−74%


Metabolite - 465 - 9
0.148325944
0.182850399
+
4%


Metabolite - 383 - 9
0.149030502
0.182850399

−5%


Metabolite - 2696 - 35
0.153809766
0.186396096

−14%


Monoethanolamine - 9
0.15400149
0.186396096

−7%


Metabolite - 226 - 9
0.158801006
0.190380284

−19%


Metabolite - 2558 - 35
0.159598455
0.190380284
+
23%


Metabolite - 3180 - 35
0.160481625
0.190380284
+
19%


Isobar 3: includes-inositol-1-phosphate-
0.174900751
0.205975696
+
9%


mannose-6-phosphate-glucose-6-


phosphate - 35


Metabolite - 593 - 9
0.17592753
0.205975696
+
5%


Metabolite - 3166 - 35
0.180240282
0.209654767

−18%


Metabolite - 2052 retired: potassium
0.182322211
0.210708223
+
4%


adduct of Isobar 1 - 35


Metabolite - 992 - 9
0.186484988
0.214137576
+
28%


Metabolite - 406 - 9
0.188146172
0.214669002
+
3%


Metabolite - 3127 - 35
0.190857147
0.2163839
+
21%


2-deoxyguanosine - 35
0.193567238
0.218076231
+
8%


Metabolite - 554 - 9
0.194913449
0.21822044
+
8%


Selenocystine - 35
0.201335108
0.220028347
+
7%


Metabolite - 386 - 9
0.201414005
0.220028347
+
40%


Metabolite - 3218 - 35
0.203272805
0.220028347
+
21%


Metabolite - 783 - 9
0.203621448
0.220028347
+
5%


alpha-4-dihydroxybenzenepropanoic
0.203878302
0.220028347

−38%


acid - 35


Metabolite - 442 - 9
0.203898073
0.220028347

−14%


Metabolite - 501 - 9
0.20718613
0.22223774
+
1%


Metabolite - 511 - 9
0.212612601
0.226700941
+
12%


Metabolite - 984 - 9
0.218140976
0.231219337
+
14%


Palmitoleic acid - 9
0.224262761
0.236309862

−15%


Metabolite - 276 - 9
0.227574093
0.238396741
+
4%


Metabolite - 263 - 9
0.229502634
0.239019224
+
1%


Metabolite - 651 - 9
0.231440451
0.239644115

0%


Metabolite - 504 - 9
0.233382993
0.240266688
+
10%


4-hydroxy-2-quinolinecarboxylic acid -
0.239304945
0.244955519
+
12%


35


Octadecanoic acid - 9
0.251419685
0.255235134

−5%


Metabolite - 3162 retired: N,N-
0.252197117
0.255235134
+
24%


dimethylarginine - 35


Butanoic acid - 9
0.257647776
0.25864946
+
18%


Metabolite - 2822 - 35
0.258641334
0.25864946

−8%


alpha-keto-glutarate - 35
0.261385634
0.25864946
+
13%


Metabolite - 982 - 9
0.26185002
0.25864946
+
4%


2-aminobutanoic acid - 9
0.263974958
0.25864946

−4%


Metabolite - 2254 - 35
0.26423422
0.25864946

−7%


Dulcitol - 9
0.27257911
0.26439873
+
4%


Metabolite - 3334 - 35
0.273059625
0.26439873
+
16%


Metabolite - 771 - 9
0.276942999
0.266717217
+
2%


Metabolite - 1281 - 35
0.279811594
0.268038824
+
28%


Metabolite - 706 - 9
0.285832283
0.272349783
+
13%


Metabolite - 1328 - 35
0.289031265
0.27394074
+
7%


Metabolite - 1344 retired: Na adduct of
0.296037148
0.279104099
+
15%


citric acid - 35


Metabolite - 485 - 9
0.312459322
0.29302628

−5%


Metabolite - 2181 - 35
0.314075599
0.29302628
+
11%


Metabolite - 1289 - 35
0.318828456
0.295919355
+
28%


2-deoxy-D-ribose - 35
0.327640183
0.302530409
+
12%


N-Acetylglutamine - 9
0.338148583
0.310632266
+
37%


N-acetyl-L-alanine - 35
0.340262913
0.310979777

−12%


Metabolite - 642 - 9
0.343191395
0.31206407

−4%


Metabolite - 2047 - 35
0.350060981
0.316702958
+
21%


N-carbamoyl-L-aspartate - 35
0.354620559
0.318474197
+
13%


Gulono-1-4-lactone - 9
0.35860987
0.318474197

0%


Metabolite - 1612 - 35
0.359291449
0.318474197

−18%


2-deoxyadenosine - 35
0.359841917
0.318474197
+
10%


Metabolite - 2698 - 35
0.360908147
0.318474197

−14%


Cholesterol - 9
0.366349483
0.31925206
+
5%


Metabolite - 443 - 9
0.366468741
0.31925206
+
3%


Metabolite - 2867 - 35
0.367136298
0.31925206
+
2%


Metabolite - 1818 - 35
0.369855345
0.319462539
+
7%


Metabolite - 2752 - 35
0.370945127
0.319462539

−5%


Glycerol - 9
0.379830663
0.32554973
+
7%


Vitamin-B6 - 9
0.387981091
0.326048061

−2%


Metabolite - 3231 - 35
0.390040532
0.326048061

−2%


Metabolite - 2607 - 35
0.391749243
0.326048061

−21%


Metabolite - 1216 - 35
0.392050899
0.326048061

−7%


N-acetyl-L-valine - 35
0.392051517
0.326048061
+
4%


Metabolite - 990 - 9
0.393153158
0.326048061
+
9%


Metabolite - 645 - 9
0.404507569
0.333918532
+
40%


Decanoic acid - 9
0.407413135
0.334099533

−6%


1-Hexadecanol - 9
0.410232328
0.334099533
+
3%


Metabolite - 991 - 9
0.411063002
0.334099533
+
136%


Metabolite - 381 - 9
0.413060843
0.334099533
+
4%


fumaric acid - 9
0.415906634
0.334099533
+
7%


Metabolite - 147 - 9
0.415917436
0.334099533
+
4%


Metabolite - 614 - 9
0.421638482
0.336484898
+
5%


4-hydroxy-3-methoxymandelate - 35
0.423915743
0.336484898

−6%


Metabolite - 571 - 9
0.424522206
0.336484898
+
2%


Metabolite - 1286 - 35
0.429813623
0.339178194
+
2%


Metabolite - 387 - 9
0.433222915
0.34036914
+
8%


Metabolite - 1252 - 35
0.442432665
0.345077745

−12%


4-acetamidobutyric acid - 35
0.444933181
0.345077745

−4%


Metabolite - 688 - 9
0.444995201
0.345077745
+
7%


Metabolite - 568 - 9
0.450960129
0.34819599
+
7%


Metabolite - 1979 retired: CL adduct of
0.454122164
0.348802529
+
12%


Isobar 19 - 35


Metabolite - 382 - 9
0.458459943
0.348802529

−2%


Metabolite - 2313 - 35
0.45883368
0.348802529
+
6%


Pyrophosphate - 35
0.459568502
0.348802529
+
13%


Metabolite - 222 - 9
0.461481577
0.348802529
+
3%


Metabolite - 1114 - 35
0.46653021
0.351136858
+
8%


Metabolite - 490 - 9
0.46908899
0.35158549

−8%


Metabolite - 1340 - 35
0.477566736
0.354969166
+
5%


Metabolite - 3183 - 35
0.482289383
0.356998129
+
5%


Metabolite - 1127 - 35
0.493684258
0.363928954
+
14%


Metabolite - 2266 retired 4-
0.499552922
0.36673206
+
5%


acetominophen sulfate - 35


Metabolite - 1843 - 35
0.501581325
0.36673206
+
12%


Metabolite - 2174 - 35
0.505912728
0.367626729

−7%


Metabolite - 606 - 9
0.506909499
0.367626729

−28%


Adenosine-3-5-cyclic-monophosphate -
0.510849566
0.368104568

−8%


35


Metabolite - 2593 - 35
0.511678242
0.368104568

−8%


Gluconic acid - 9
0.519338065
0.372120632

0%


Metabolite - 146 - 9
0.525817923
0.375262587
+
4%


Metabolite - 1104 - 35
0.531281422
0.376220557
+
5%


Xylitol - 35
0.53136071
0.376220557
+
8%


gamma-aminobutyryl-L-histidine - 35
0.538066749
0.379468772

−7%


Metabolite - 2051 - 35
0.545176701
0.382975252

−7%


Metabolite - 3468 retired: Metabolite -
0.551648935
0.386008104

−20%


1498 - 35


Metabolite - 388 - 9
0.560462208
0.388789259

0%


5-6-dihydrouracil - 35
0.562006502
0.388789259
+
7%


Lipoate - 9
0.562134731
0.388789259

−2%


Metabolite - 128 - 9
0.565490896
0.389606212
+
7%


12-hydroxydodecanoic acid - 9
0.568868993
0.390431958

0%


Metabolite - 138 - 9
0.578770894
0.394383561

−9%


Metabolite - 601 - 9
0.579029844
0.394383561

−8%


Metabolite - 1342-possible-
0.588057538
0.399015254
+
10%


phenylacetylglutamine-or-formyl-N-


acetyl-5-methoxykynurenamine - 35


Metabolite - 987 - 9
0.594450189
0.401339274

−5%


Metabolite - 2726 - 35
0.596919611
0.401339274
+
2%


Metabolite - 285 - 9
0.599627227
0.401339274

−5%


alpha-D-ribose-5-phosphate - 35
0.600444471
0.401339274

−2%


Metabolite - 3143 - 35
0.604016003
0.401870746
+
5%


Tartarate - 9
0.606556307
0.401870746

0%


3-nitro-L-tyrosine - 35
0.607969903
0.401870746
+
10%


Metabolite - 1132 - 35
0.611167267
0.402171481

−6%


Metabolite - 2053 - 35
0.612959279
0.402171481
+
4%


Metabolite - 721 - 9
0.6151602
0.402171481
+
8%


2-amino-butyrate - 9
0.620541504
0.404214366

−65%


Metabolite - 1500 - 35
0.624875169
0.404544292

−8%


Glutarate - 35
0.625564713
0.404544292
+
8%


Ascorbate - 9
0.631114451
0.406665128
+
2%


N-6-trimethyl-l-lysine - 35
0.643783109
0.411865249
+
10%


Metabolite - 1064 - 35
0.663978387
0.423273644
+
11%


DOPA - 9
0.670522586
0.424427025
+
3%


Metabolite - 770 - 9
0.670526368
0.424427025

0%


3-phospho-d-glycerate - 9
0.677948475
0.427614036

−3%


n-dodecanoate - 9
0.692424536
0.433744571

−2%


Mercaptopyruvate - 35
0.695879606
0.433744571

−7%


3-phospho-l-serine - 9
0.699317446
0.433744571

0%


Phosphoenolpyruvate - 35
0.699652586
0.433744571

−2%


Metabolite - 223 - 9
0.699774788
0.433744571

−8%


Metabolite - 472 - 9
0.710092334
0.43862202

−1%


Metabolite - 3230 - 35
0.714846376
0.440041194
+
2%


Metabolite - 2231 - 35
0.720405761
0.441881344

−6%


Metabolite - 1192 - 35
0.723960835
0.441881344

−2%


Glucarate - 35
0.728249412
0.441881344
+
7%


Metabolite - 2806 - 35
0.730169643
0.441881344
+
8%


3-phospho-d-glycerate - 35
0.734016495
0.442713713

0%


Metabolite - 985 - 9
0.737163153
0.443119599

−4%


Metabolite - 653 - 9
0.740731925
0.44377566

−1%


Metabolite - 594 - 9
0.755810126
0.449140955

−1%


Metabolite - 136 - 9
0.755814739
0.449140955

−1%


Malic acid - 35
0.757209401
0.449140955

−12%


Isocitrate - 35
0.768315843
0.454224718
+
2%


Metabolite - 3416 - 35
0.781343103
0.460406878
+
283%


Metabolite - 3401 - 35
0.785742098
0.461480959

−2%


Xanthine - 35
0.788498648
0.461586534

−1%


Metabolite - 394 - 9
0.793917988
0.461769189
+
1%


Saccharopine - 35
0.796380525
0.461769189

−1%


Guanosine - 35
0.800324952
0.461769189

−3%


Inositol - 9
0.801599262
0.461769189

0%


Metabolite - 505 - 9
0.803385192
0.461769189

−9%


Metabolite - 2366 - 35
0.804277542
0.461769189
+
9%


2-ethylhexanoic acid - 9
0.811185482
0.463898116
+
82%


Glyoxylate - 9
0.813164953
0.463898116

−4%


Metabolite - 3130 - 35
0.821844119
0.467361035
+
36%


Metabolite - 2272 - 35
0.829571496
0.470262494
+
7%


Metabolite - 274 - 9
0.835872048
0.470958082

−7%


3-hydroxy-3-methylglutarate - 35
0.836056775
0.470958082
+
1%


Metabolite - 145 - 9
0.840293993
0.471861104

0%


Hydroorotate - 9
0.844187321
0.472565977
+
2%


Metabolite - 580 - 9
0.867614526
0.48210755

−8%


Palmitate - 9
0.867614526
0.48210755

0%


Metabolite - 398 - 9
0.869575595
0.48210755

−1%


Metabolite - 2753 - 35
0.871997698
0.48210755
+
8%


Metabolite - 1142-possible-5-
0.87816814
0.483474682
+
12%


hydroxypentanoate-or-beta-


hydroxyisovaleric acid - 35


Metabolite - 2854 - 35
0.881164116
0.483474682

0%


Pentanedioic acid - 9
0.883303118
0.483474682
+
1%


Threonine - 9
0.88526639
0.483474682

−3%


Metabolite - 1842 retired: 4-
0.898184283
0.48903862
+
11%


Guanidinobutanoic acid - 35


Metabolite - 1608 - 35
0.901189233
0.489187846
+
27%


Metabolite - 482 - 9
0.91480612
0.49507919

0%


Metabolite - 221 - 9
0.918754567
0.495718391
+
2%


Metabolite - 2026 - 35
0.931706512
0.500287059

−3%


Inosine - 35
0.932807721
0.500287059
+
2%


Metabolite - 3313 - 35
0.94471359
0.50264336
+
6%


Metabolite - 278 - 9
0.9460492
0.50264336

−1%


Tetronic acid - 9
0.946444393
0.50264336

−3%


Metabolite - 691 - 9
0.948425116
0.50264336
+
1%


Metabolite - 3249 - 35
0.953102102
0.503632019
+
5%


Metabolite - 2109 - 35
0.963700048
0.507734375
+
1%


Sarcosine - 9
0.970231636
0.509676555

−4%


Metabolite - 1304 - 35
0.977448006
0.511966051
+
9%


alpha-L-sorbopyranose - 9
0.982136198
0.512921852
+
7%


Metabolite - 1133 - 35
0.989808044
0.51350178

0%


Metabolite - 2703 - 35
0.989808044
0.51350178

−2%


Metabolite - 2821 - 35
0.991846462
0.51350178

−7%


Metabolite - 2038 - 35
0.997806098
0.515098498

−6%









Example 2

In another example, biomarkers were discovered by (1) analyzing plasma samples from different groups of human subjects to determine the levels of metabolites in the samples and then (2) statistically analyzing the results to determine those metabolites that are differentially present in the two groups. As listed below in Tables 5-6, biomarkers were discovered that were differentially present between samples from ALS subjects and Control subjects not diagnosed with ALS.


The plasma samples used for the analysis were from 199 ALS subjects and 94 control subjects not diagnosed with ALS. After the levels of metabolites were determined, the data was analyzed using T-tests (Table 5) and Wilcoxon's rank-sum tests (Table 6).


Tables 5 and 6 include, for each listed biomarker and non-biomarker compound, the p-value and the q-value determined in the statistical analysis of the data concerning the biomarkers, an indication of whether the mean of a particular compound was higher in the ALS or Control samples (a “+” indicating a higher mean in ALS samples as compared to the control samples and a “−” indicating a lower mean in ALS samples as compared to the control samples), and an indication of the percentage difference in the ALS mean as compared to the control mean. Throughout the tables, names of metabolites ending with the notation “−35” indicate that the levels of those compounds were measured using LC-MS, and names ending with the notation “−9” indicate that the levels of those compounds were measured using GC-MS. The term “Isobar” as used in the tables indicates the compounds that could not be distinguished from each other on the analytical platform used in the analysis (i.e., the compounds in an isobar elute at nearly the same time and have similar (and sometimes exactly) quant ions, and thus cannot be distinguished).


Non-biomarker compounds identified in the analyses are also listed in the tables below as those compounds that having a percentage change in ALS of 0%.

TABLE 5ALS Biomarkers from blood plasma samples - T-Test Analysis of Plasmafrom Control vs. ALSIncrease(+) or%DecreasechangeCompoundp-valueq-value(−) in ALSin ALSMetabolite - 2045 - 357.39E−281.09E−25+412%Isobar: glutamine/lysine - 351.95E−271.44E−25−39%Metabolite - 2567 - 356.35E−243.13E−22+744%Glycerate - 92.23E−218.24E−20+125%Glyceric acid - 94.90E−211.45E−19+124%Metabolite - 1111-possible-7.52E−211.85E−19−38%methylnitronitrosoguanidine-or-ethyl-thiocarbamoylacetate - 35pyridoxamine - 351.59E−203.36E−19−36%Methionine - 351.85E−203.42E−19−40%Metabolite - 2033 retired: 2-4.47E−207.35E−19−46%isopropylmalic acid - 35Metabolite - 1192 - 354.30E−186.36E−17−45%Metabolite - 2648 - 96.57E−188.83E−17+1548%Glutamic acid - 97.23E−188.91E−17+580%gamma-L-glutamyl-L-tyrosine - 353.14E−153.57E−14+70%Metabolite - 2568 - 354.53E−144.78E−13+1569%Metabolite - 1850 - 95.24E−145.17E−13+377%4-hydroxyphenylacetate - 351.37E−121.27E−11−40%3-hydroxy-3-methylglutarate - 351.85E−121.61E−11−52%Metabolite - 1071 - 352.82E−122.20E−11+757%Metabolite - 1302 - 352.82E−122.20E−11+757%Metabolite - 128 - 95.44E−123.60E−1191%Metabolite - 264 - 95.44E−123.60E−11+−45%Metabolite - 1815 - 95.46E−123.60E−11−44%Metabolite - 1575 - 355.60E−123.60E−11+141%Metabolite - 499 - 94.59E−112.83E−10−44%Metabolite - 1775 - 94.80E−112.84E−10+109%Metabolite - 1261 - 359.67E−115.50E−10+496%Metabolite - 1576 - 351.40E−107.66E−10−29%Metabolite - 270 - 92.44E−101.29E−09+913%Epinephrine - 92.61E−101.33E−09+41%Metabolite - 763 - 94.94E−102.43E−09+60%Metabolite - 2608 - 356.15E−102.94E−09+80%Arginine - 351.38E−096.36E−09+53%Metabolite - 736 - 91.58E−097.10E−09+196%N-acetyl-L-glutamine - 91.89E−098.20E−09−46%Metabolite - 1286 - 352.19E−099.27E−09−23%trans-4-hydroxyproline - 355.29E−092.17E−08+241%3-phospho-l-serine - 356.03E−092.40E−08+236%Metabolite - 1457 - 356.17E−092.40E−08+136%Metabolite - 841 - 97.65E−092.90E−08−60%Metabolite - 1129 - 357.85E−092.90E−08+355%Ornithine - 351.64E−085.91E−08−30%Isobar: 5-oxoproline/glutamine - 92.18E−087.68E−08+64%Metabolite - 655 - 92.44E−088.41E−08−51%Isobar: lysine/tyramine/putrescine - 96.22E−082.04E−07+114%Metabolite - 1538 - 96.77E−082.18E−07−39%Metabolite - 1220 - 359.40E−082.94E−07+1107%Metabolite - 2237 - 359.55E−082.94E−07+398%Metabolite - 1497 - 351.64E−074.94E−07−39%Metabolite - 1092 - 352.10E−076.20E−07+5519%Thyroxine - 352.69E−077.80E−07−25%Metabolite - 1208 - 352.74E−077.80E−07+3120%Metabolite - 1073 - 356.51E−071.82E−06+536%Metabolite - 1265 - 351.06E−062.90E−06+662%Metabolite - 2591 - 351.14E−063.07E−06+265%Metabolite - 2220 - 91.49E−063.94E−06+65%Metabolite - 2051 - 351.59E−064.13E−06−38%Metabolite - 451 - 91.72E−064.38E−06+31%o-phosphoethanolamine - 93.19E−068.00E−06+37%Metabolite - 2041 - 354.84E−061.19E−05−37%Metabolite - 1114 - 355.82E−061.41E−05+47%Metabolite - 2192 - 96.61E−061.58E−05−37%Carnosine - 358.07E−061.89E−05−18%Glycerol - 98.36E−061.93E−05+34%Metabolite - 1342 - possible-9.09E−062.04E−05+49%phenylacetylglutamine-or-formyl-N-acetyl-5-methoxykynurenamine - 35Metabolite - 1616 - 359.37E−062.07E−05−36%Metabolite - 1738 - 351.42E−053.09E−05−22%4-acetamidobutyric acid - 351.50E−053.20E−05+122%Xanthine - 351.68E−053.54E−05+56%12-hydroxydodecanoic acid - 91.75E−053.64E−05−42%N-formyl-L-glycine - 352.34E−054.81E−05+130%Guanosine - 352.43E−054.93E−05−35%Metabolite - 734 - 92.47E−054.94E−05+534%Metabolite - 1322 retired: citric acid - 352.50E−054.94E−05−40%Metabolite - 2559 - 352.66E−055.18E−05+192%Phosphate - 92.91E−055.59E−05+12%Metabolite - 1824 - 92.97E−055.64E−05−42%Metabolite - 1910 retired: uric acid - 353.20E−055.98E−05−9%Metabolite - 1064 - 353.82E−057.07E−05+20%Creatinine - 354.00E−057.24E−05−12%Tartarate - 94.01E−057.24E−05+20%5-oxoproline - 354.13E−057.36E−05+25%Metabolite - 2564 - 355.74E−050.000101019+3145%Serine - 96.60E−050.000114885+51%N-formyl-L-methionine - 356.76E−050.000116223−29%alpha-keto-glutarate - 350.0001083720.000184178−33%N-6-trimethyl-l-lysine - 350.0001226580.000206088−16%Metabolite - 1264 - 350.0001250140.000207686+1018%Pantothenic acid - 350.0001392610.000228783+57%Metabolite - 557 - 90.0002321830.000377248−33%Isobar 8: anthranilic/salicylamide - 350.0002533850.000407222+25%Caffeine - 350.00028630.000455173−44%Metabolite - 1713 retired: n-acetyl-L-0.0003041530.000478413−44%aspartic acid - 35Metabolite - 1203 - 350.0003227140.000502265+1236%alpha-4-dihydroxybenzenepropanoic0.0006000040.000924105−20%acid - 35Metabolite - 1734 - 350.0007760780.001182966+100%Metabolite - 1839 - 350.0007939910.001197921−39%Metabolite - 1327 - 350.000819870.001224469−43%Metabolite - 1465 - 350.0008345110.001233873−17%Metabolite - 2592 - 350.0008767330.001283467+605%Metabolite - 1065 - 350.0011066540.001604169+386%Arabinose-3 - 90.0013845960.001987579+24%Metabolite - 2546 - 350.0016206750.002304098−25%Metabolite - 404 - 90.0018398230.002590748−45%Metabolite - 1975 - 350.0019559350.002728267+39%Metabolite - 2005 - 350.0023068330.00318765−18%Metabolite - 1560 - 90.0027261860.003732244−14%Isobar 3: hydroxybutanoic acid/butanoic0.0032399470.004394908+76%acid - 9Metabolite - 2589 - 350.0033712480.004531442+3636%Metabolite - 2105 - 350.0035335140.004706762−30%Metabolite - 1329 - 350.0037571850.004960014−39%gamma-L-glutamyl-L-glutamine - 350.0039323590.005145328−12%Metabolite - 2185 - 350.0040011130.005189366−16%Metabolite - 671 - 90.0040859810.005253356−25%Metabolite - 1289 - 350.0041544520.005295343+40%Metabolite - 1281 - 350.0048051780.006072422+55%Metabolite - 1564 - 90.0049640430.006220022+263%Metabolite - 470 - 90.0050945960.006329963+11%Metabolite - 621 - 90.0054675130.006736695+30%Metabolite - 2560 - 350.0080823680.009876239−13%Isoleucine - 90.0082363780.009936974+30%Selenocystine - 350.0083165910.009936974+24%Metabolite - 526 - 90.0083336930.009936974+52%Praline - 90.0089319790.01056516+30%Metabolite - 1346 - 350.0098186140.011521738+7%Metabolite - 613 - 90.0099516220.011585867+18%Metabolite - 423 - 90.0107580740.012426904+25%Metabolite - 1254 - 350.0113473590.013005992+776%Metabolite - 2565 - 350.0124089190.014077917+7127%Metabolite - 578 - 90.0124730120.014077917−15%1-7-dihydro-6h-purin-6-one - 350.0127267580.014173847+39%Metabolite - 273 - 90.0127497310.014173847+54%Metabolite - 485 - 90.0129103360.014245284−28%Metabolite - 1347 - 350.0133823450.014656721−35%Metabolite - 1968 - 90.0136096850.014796109+17%Metabolite - 543 - 90.0139668340.015073558+22%Alanine - 90.0140903340.01509665+20%Metabolite - 1368 - 350.0144374590.015357281+241%Biliverdin - 350.0152963240.016154644−24%Metabolite - 681 - 90.0154146890.016164193−21%Metabolite - 1829 retired: oxalic acid -0.0160386110.016700012−18%35Metabolite - 2588 - 350.0174158230.017868851+19307%Dulcitol - 350.0174555240.017868851−35%Metabolite - 1802 - 350.0175237180.017868851+69%Metabolite - 2058 - 350.0190739930.019316442+31%Metabolite - 1914 - 350.0204377980.020556783+37%Metabolite - 1127 - 350.0225482360.022526267+14%Metabolite - 2646 - 90.0227932050.022618172+8%N-acetylserotonin - 90.0230885250.022758483+100%Metabolite - 1086 - 350.0233093650.022824006+24%Metabolite - 941 - 90.0239578920.023304693+30%Uric acid - 90.0243857830.023565879−19%Metabolite - 279 - 90.0267084430.025642846+15%Oxitryptan - 90.0273439160.026083591+86%N-acetyl-D-galactosamine - 350.0289016730.027392821−22%Valine - 90.029397620.027685406+20%Succinate - 350.0311596190.029159054+46%Metabolite - 1958 - 350.0314222030.029219843−5%1-Hexadecanol - 90.0319604470.029534609+14%Oleic acid - 90.0333925440.030666344+19%Sugar6 - 90.0337817610.03083228+22%Metabolite - 1328 - 350.0365743710.033176276−21%Leucine - 90.0390035140.035163999+24%Metabolite - 1753 - 90.040019430.035861242−26%Metabolite - 709 - 90.0427008940.038033583+15%Metabolite - 2053 - 350.043419730.038442269−19%Metabolite - 706 - 90.0471369360.041484937−24%Metabolite - 664 - 90.0480560120.042043551+18%Metabolite - 2238 - 350.0485432390.042219997−46%Metabolite - 1831-possible-Cl-adduct-of-0.0509537920.04405739−10%citrulline - 35Metabolite - 523 - 90.0537217170.046180625+66%Metaboilte-1834 - 350.0551032490.047094422−28%Arachidonic acid - 90.0558926360.047494543+21%Metabolite - 1288 - 350.0578560720.048882034+22%Metabolite - 1113 - 350.0586287660.049253427−11%Metabolite - 1974 - 350.0598731670.050014661−17%Metabolite - 407 - 90.062994040.052326035−11%Uridine - 350.0639579230.052829888−7%Metabolite - 1340 - 350.0647274360.053168483+27%Metabolite - 293 - 90.0662079890.054084173−12%Metabolite - 1242 - 350.0668345620.054296031+160%Metabolite - 1979 retired: CL adduct of0.0679282950.054883019−7%Isobar 19 - 35beta-D-Iactose - 350.0689944160.055441438−13%Metabolite - 1335 - 350.0695020550.05554557−15%Metabolite - 1826 - 90.0698753520.05554557+20%Metabolite - 1735 - 350.0709765110.056119192+64%Metabolite - 614 - 90.0727109320.05718475+34%3-hydroxypropanoate - 90.0749145070.058606055−19%N-acetyl-L-leucine - 350.0759707560.059093074+45%Isobar 2: 2-amino-3amino-GABA-etc -0.0763363830.059093074−11%35Metabolite - 1849 - 90.0784690440.060427622−14%Metabolite - 2055 - 350.0830281520.063607223+10%Glutarate - 90.0936288550.071358605−7%Metabolite - 2009 - 90.0970581530.073544335+15%Metabolite - 1737 retired: 2,3-0.0974915390.073544335−37%dihydroxybenzoic acid - 35N-acetyl-L-alanine - 90.099198310.074188172+11%Metabolite - 268 - 90.099348540.074188172−10%Glucarate - 350.1002927990.074516946−7%Metabolite - 2561 - 350.1100755990.081376579−10%Metabolite - 687 - 90.1118148180.081878989+11%Metabolite - 278 - 90.1122771030.081878989−15%Metabolite - 683 - 90.1124165210.081878989−8%9,12-octadecadienoic acid-z-z- - 90.1143099440.082849943+13%Metabolite - 1835 - 350.1151928180.083082568−12%Metabolite - 1842 retired: 4-0.1163985560.083235488+18%Guanidinobutanoic acid - 35alpha-Hydroxyisobutyric acid-tms - 90.116530740.083235488+12%Octadecanoic acid - 90.1174067120.083457997+8%DL-beta-hydroxyphenylethylamine - 90.118433020.083784732−15%Metabolite - 1262 - 350.1212552280.085372804+78%Metabolite - 1332 - 350.1233276540.086420423+15%Metabolite - 1597 - 350.129151190.090074305+5%Metabolite - 1911 - 350.1302134260.090388781−18%Metabolite - 2074 - 350.1375385870.095027467+14%D-fructose-3 - 90.1421173070.097734271+11%3-phospho-d-glycerate - 350.144977910.098900438−22%Metabolite - 285 - 90.1451508490.098900438−10%Metabolite - 1414 - 90.1460445820.09905293−9%Metabolite - 1285 - 350.1479906930.099914532+14%Palmitoleic acid - 90.1506235570.101229848+17%Threonine - 90.1536288740.102636481+10%Metabolite - 562 - 90.1541048680.102636481−15%Serotonin - 350.1602519450.106251927−24%Metabolite - 341 - 90.1630564940.107628789−7%Menadione-Vitamin-K3 - 90.1666069470.109483572+17%Metabolite - 596 - 90.1686850810.110358707−2%Metabolite - 2047 - 350.1714478830.111672088−28%Metabolite - 1133 - 350.1741830860.112956051−12%Isobar: noradrenaline/normetanephrine - 90.1749631870.112966473−12%Melatonin - 90.1781551920.114527299+21%Catechol - 350.1790751290.114620332−28%Palmitate - 90.1836436480.117037839+9%Methylmalonic acid - 350.1851456490.117488662+37%alpha-2-diamino-gamma-0.1873237190.118362814+6%oxobenzenebutanoic acid - 35Metabolite - 393 - 90.1950916250.12196942−15%Metabolite - 506 - 90.1958694140.12196942+62%Raffinose - 350.1964037430.12196942−19%Metabolite - 594 - 90.1971562230.12196942−2%Metabolite - 1193-confirmed-3-indoxyl-0.1982025950.122105848−10%sulfate - 35alpha-tocopherol - 90.2038328710.125053413−18%Metabolite - 2548-possible-Cl-adduct-of-0.206784720.126340167−10%uric acid - 35Metabolite - 406 - 90.2165764590.131778131+5%Arabinose - 350.2193220410.132901787+8%D-galactose-1 - 90.2246264860.135560527+17%Isobar 4: includes-Gluconic acid-0.2265660370.136175215+8%arabinose-D-ribose - 35Ascorbic acid - 350.2343978910.140312097−28%Metabolite - 2052 retired: potassium0.2372274760.141433301+3%adduct of Isobar 1 - 35GABA - 90.2393488750.142124979+10%Metabolite - 2056 - 350.2403404590.142142924+7%Cholesterol - 90.2425439990.14287465+3%Metabolite - 1336 retired: carnitine - 350.2483153940.145693935+8%gamma-aminobutyryl-L-histidine - 350.250306410.146281639−7%Histidine - 350.2529476780.147202235+4%Saccaropine - 350.2538728250.147202235−7%Metabolite - 344 - 90.2551714820.147377281+2%Metabolite - 1818 - 350.2590814980.149053317−7%2-ethylhexanoic acid - 90.2610534510.149605687+7%Metabolite - 1087 - 350.2695211050.153862004−49%Isobar 9: includes-sucrose-beta-D-0.2711130170.154175509+7%lactose - 35Metabolite - 1830 - 350.2918170410.165313572+10%Metabolite - 669 - 90.2939973080.165913006+7%Metabolite - 1961 retired: glycocholic0.301182260.169321461+31%acid - 35D-galactose - 90.3090192030.173069249+22%Metabolite - 1108 - 350.3220918530.179709996+24%Metabolite - 618 - 90.3266872640.181588748−7%Metabolite - 2175 - 350.3297322680.182594864+23%Metabolite - 2067 retired: carnitine - 350.3333573930.183913524+5%Metabolite - 1554 - 90.3395234520.186619004−8%Metabolite - 1388 - 350.3417866460.187167179−37%N-tigloylglycine - 90.3487970660.190301364−7%N-acetyl-L-valine - 350.3657950720.198204653−7%Metabolite - 1537 - 90.3659637960.198204653+2%Metabolite - 1373 - 90.3704890910.199923217+11%Metabolite - 2587 retired: p-0.3911655890.209179781+96%acetaminophen-beta-d-glucuronide - 35Metabolite - 2173 - 350.3953165920.209179781−12%Metabolite - 2100 - 350.3959622010.209179781−5%glyoxylate - 90.3960523530.209179781+4%Metabolite - 1847 - 90.3961314890.209179781+15%Hydroorotate - 90.4008629110.210924935+5%Metabolite - 2627 - 90.4041997350.211926509+6%Aspartate - 90.4079781490.212792325−4%Metabolite - 443 - 90.4087294510.212792325−5%Inosine - 350.4123496430.213923814+12%Metabolite - 1609 - 350.4181446460.215709956−8%Mannose-6-phosphate - 90.4187103710.215709956−7%Metabolite - 1915 - 350.4219849620.216642099−15%DL-pipecolic acid - 350.4246989210.217280966+5%Metabolite - 1358 - 90.4326734240.22002834+8%Metabolite - 1926 retired: trans-2,3,4-0.4339175710.22002834+12%trimethoxycinnamic acid - 35Metabolite - 1088 - 350.4345333410.22002834−11%Metabolite - 1909 - 350.4404309360.222253477+4%Metabolite - 1379 retired: hippuric acid -0.4437488670.223166137−8%35Metabolite - 1656 - 350.4497625670.225293772−4%alpha-aminoadipic acid - 90.4510269860.225293772−4%Metabolite - 760 - 90.4565302310.227274892−7%Metabolite - 1827 - 90.4591666920.227450012−19%Metabolite - 1323-possible-p-cresol-0.4599586430.227450012+7%sulfate - 35Inositol - 90.4653062650.229327435+4%n-dodecanoate - 90.4728296060.231037316+4%Metabolite - 642 - 90.4732179140.231037316+5%Metabolite - 222 - 90.4756037810.231037316−25%Metabolite - 2628 - 90.4762231550.231037316−7%Xylitol - 350.4765885460.231037316−8%Malic acid - 90.491077080.237282991−5%3-amino-isobutyrate - 90.4987106230.240186511−4%Metabolite - 1337 - 350.5043055750.242092548−21%Metabolite - 1888 - 90.5202038590.248113414−10%Tryptophan - 90.5374507240.255515136−3%Metabolite - 702 - 90.5403110790.256051691−7%Riboflavine - 350.5541006590.261747594+5%Metabolite - 704 - 90.5641633380.265652301+3%N-5--aminocarbonyl-L-ornithine - 350.572857080.268889659−7%Isobar: adenosine-5-0.5819978650.272315698−4%diphosphoribose/glucosamine-6-phosphate - 9Metabolite - 386 - 90.5892063340.273366105−5%tetradecanoic acid - 90.590052990.273366105−3%Pyrophosphate - 350.5912028210.273366105+7%2-amino-heptanedioic acid - 90.5932495940.273366105−4%Metabolite - 2221 - 90.5934871610.273366105+2%Isobar 3: includes-inositol-1-phosphate-0.5969405350.274102861−4%mannose-6-phosphate-glucose-6-phosphate - 35Metabolite - 1353 - 90.6151588770.281526493−1%Metabolite - 1324 - 350.6169158250.281526493−3%Urea - 90.6282899020.285647975+3%Metabolite - 1372 retired: 2-0.6298112010.285647975+35%hydroxyhippuric acid- 35Isobar-1-includes-mannose-fructose-0.6340821420.286130487−3%galactose-alpha-L-sorbopyranose-Inositol - 35Hypotaurine - 90.6347454680.286130487−5%Phenylalanine - 350.6470935470.290810136−1%Cystine - 90.6506029720.291403283−4%Metabolite - 2606 - 350.6525868450.291403283+3%Metabolite - 670 - 90.6543259670.291403283+3%Metabolite - 1956 - 90.6571360730.291775918−3%4-hydroxy-3-methoxymandelate - 350.6604358810.292363103+17%Niacinamide - 350.6646823930.29336462+7%Metabolite - 1534 - 90.6820628790.300139742−2%Gulono-1-4-lactone - 350.6875409970.301608282−6%Gluconic acid- 90.6894798760.301608282+2%Metabolite - 1104 - 350.694020830.302699132−2%Isobar: 2′deoxyguanosine/adenosine0.7001526320.304475377−14%Decanoic acid - 90.7084930790.305642167+2%Metabolite - 1351 retired: urea adduct of0.7089629780.305642167+2%Isobar 6 - 35Metabolite - 1814 - 90.7090371990.305642167+3%Metabolite - 638 - 90.7143883430.306809695+2%Metabolite - 1110 - 350.7158957850.306809695+5%Metabolite - 1066 - 350.7195664410.307491539−1%N-carbamoyl-L-aspartate - 350.7228871460.30802034−4%Metabolite - 1929 - 350.7256691180.308317209−1%Metabolite - 1757 - 90.7291811590.308643713+4%Metabolite - 1836 - 350.7306125210.308643713+2%methyl-indole-3-acetate - 350.7515248350.316560675−2%Metabolite - 1349 retired: Isobar 7 - 350.7536353430.316560675+5%Metabolite - 2558 - 350.7560938320.316693652−4%Metabolite - 1551 - 90.7602093740.317517983−3%Metabolite - 1247 - 350.768706650.320051482−5%alpha-D-ribose-5-phosphate - 350.7724339080.320051482+1%Metabolite - 1331 - 350.7727689960.320051482+5%Metabolite - 645 - 90.7840523430.323201768+2%Metabolite - 1397 - 350.7864259310.323201768+2%Metabolite - 1754 - 90.7869331850.323201768−2%Histamine - 350.7893667830.323303208−1%Metabolite - 1514 - 90.7940167610.324309349+1%Metabolite - 1131 - 350.7971304630.324684196+1%Isobar: dulcitol/gluano-1,4-lactone - 90.803138610.3262327+1%Glycine - 90.805745150.32639478+3%Isobar: 3-chloro-L-tyrosine/DOPA - 90.8172328730.330143774+2%Metabolite - 2550 - 350.8614927120.347075464+13%Metabolite - 1142-possible-5-0.8865668460.356206661−2%hydroxypentanoate-or-beta-hydroxyisovaleric acid - 35Metabolite - 1068 - 350.8934933240.3570491930%Metabolite - 1181 - 350.8934935270.3570491930%Metabolite - 1981 - 350.8966394150.357340537−1%5-hydroxyindoleacetate - 350.9004141320.35788025−1%Guanidine acetic acid - 90.9083513120.359336309+1%Lactate - 90.9089381590.3593363090%Metabolite - 1736 retired: p-0.9166094010.361402713+1%hydroxybenzaldehyde - 35Metabolite - 1519 - 90.9324203870.3666589450%Metabolite - 1161 - 350.938829010.367225704−1%Metabolite - 1334 - 350.9388290150.367225704−1%Metabolite - 580 - 90.9416465870.3673559650%Metabolite - 749 - 90.9491448070.368863678+1%Metabolite - 1972 - 350.9505008270.368863678−1%Metabolite - 1673 - 90.9637967660.372339766−1%Tyrosine - 350.9644946570.3723397660%Metabolite - 1385 - 350.9696603970.373359153−1%Metabolite - 995 - 90.9747053280.374326850%Metabolite - 1383 - 350.9902158440.378318235+1%Metabolite - 1201 - 350.9966669820.3798015340%









TABLE 6










ALS Biomarkers from blood plasma samples - Wilcoxon's Rank Sum-Tests


ALS vs. Control














Increase






(+) or
%





Decrease
change


Compound
p-value
q-value
(−) in ALS
in ALS














trans-4-hydroxyproline - 35
7.95E−19
1.13E−16
+
241%


Glycerate - 9
2.13E−17
1.51E−15
+
125%


Glyceric acid - 9
1.31E−16
6.17E−15
+
124%


Metabolite - 841 - 9
4.44E−16
1.52E−14

−60%


Metabolite - 2045 - 35
5.37E−16
1.52E−14
+
412%


Metabolite - 2550 - 35
8.80E−16
2.08E−14
+
13%


Isobar: glutamine/lysine - 35
2.00E−15
4.04E−14

−39%


Metabolite - 128 - 9
2.89E−15
5.11E−14

−45%


Methionine - 35
7.55E−15
1.19E−13

−40%


Metabolite - 2033 retired: 2-
4.35E−14
6.16E−13

−46%


isopropylmalic acid - 35


Metabolite - 2568 - 35
1.24E−13
1.60E−12
+
1569%


Metabolite - 1192 - 35
5.70E−13
6.73E−12

−45%


3-hydroxy-3-methylglutarate - 35
1.31E−12
1.43E−11

−52%


Metabolite - 1111-possible-
2.54E−12
2.57E−11

−38%


methylnitronitrosoguanidine-or-ethyl


thiocarbamoylacetate - 35


Glutamic acid - 9
6.18E−12
5.47E−11
+
1548%


Metabolite - 2648 - 9
6.18E−12
5.47E−11
+
580%


Metabolite - 655 - 9
7.63E−12
6.36E−11

−51%


Metabolite - 499 - 9
1.37E−11
1.07E−10

−44%


Metabolite - 2588 - 35
1.77E−11
1.32E−10
+
19307%


N-acetyl-L-glutamine - 9
3.65E−11
2.59E−10

−46%


Pyridoxamine - 35
4.12E−11
2.78E−10

−36%


Metabolite - 1850 - 9
9.06E−11
5.83E−10
+
377%


Metabolite - 1815 - 9
1.29E−10
7.96E−10

−44%


Metabolite - 1071 - 35
2.01E−10
1.10E−09
+
757%


Metabolite - 1302 - 35
2.01E−10
1.10E−09
+
757%


Metabolite - 270 - 9
2.02E−10
1.10E−09
+
913%


Metabolite - 2567 - 35
2.74E−10
1.43E−09
+
744%


Metabolite - 1286 - 35
2.83E−10
1.43E−09

−23%


4-hydroxyphenylacetate - 35
8.59E−10
4.20E−09

−40%


Metabolite - 1220 - 35
2.16E−09
1.01E−08
+
1107%


Metabolite - 1208 - 35
2.22E−09
1.01E−08
+
3120%


Metabolite - 1327 - 35
2.41E−09
1.07E−08

−43%


Guanosine - 35
2.80E−09
1.20E−08

−35%


Metabolite - 1497 - 35
8.04E−09
3.35E−08

−39%


Metabolite - 1261 - 35
8.51E−09
3.44E−08
+
496%


Metabolite - 264 - 9
1.85E−08
7.26E−08
+
91%


alpha-keto-glutarate - 35
4.28E−08
1.61E−07

−33%


Metabolite - 1073 - 35
4.31E−08
1.61E−07
+
536%


Metabolite - 2105 - 35
4.76E−08
1.68E−07

−30%


Metabolite - 1538 - 9
5.80E−08
2.00E−07

−39%


Metabolite - 2589 - 35
7.19E−08
2.43E−07
+
3636%


Metabolite - 763 - 9
7.38E−08
2.43E−07
+
60%


Metabolite - 1265 - 35
7.77E−08
2.50E−07
+
662%


Metabolite - 1824 - 9
1.41E−07
4.45E−07

−42%


Ornithine - 35
2.57E−07
7.93E−07

−30%


Metabolite - 2051 - 35
2.70E−07
8.13E−07

−38%


Thyroxine - 35
3.68E−07
1.07E−06

−25%


Metabolite - 451 - 9
3.77E−07
1.07E−06
+
31%


Metabolite - 1576 - 35
3.79E−07
1.07E−06

−29%


gamma-L-glutamyl-L-tyrosine - 35
4.24E−07
1.18E−06
+
70%


3-phospho-I-serine - 35
4.36E−07
1.19E−06
+
236%


Metabolite - 1129 - 35
4.59E−07
1.20E−06
+
196%


Metabolite - 736 - 9
4.59E−07
1.20E−06
+
355%


Metabolite - 1738 - 35
4.75E−07
1.22E−06

−22%


Metabolite - 1329 - 35
6.52E−07
1.65E−06

−39%


Caffeine - 35
6.87E−07
1.71E−06

−44%


Isobar: 5-oxoproline/glutamine - 9
7.91E−07
1.93E−06
+
64%


Metabolite - 1203 - 35
9.88E−07
2.31E−06
+
1236%


Metabolite - 1616 - 35
9.91E−07
2.31E−06

−36%


Metabolite - 2041 - 35
9.95E−07
2.31E−06

−37%


Metabolite - 734 - 9
1.02E−06
2.32E−06
+
534%


Metabolite - 1465 - 35
1.53E−06
3.44E−06

−17%


Metabolite - 1775 - 9
1.60E−06
3.54E−06
+
109%


Dulcitol - 35
1.71E−06
3.73E−06

−35%


Metabolite - 1575 - 35
2.89E−06
6.21E−06
+
141%


Metabolite - 404 - 9
3.27E−06
6.90E−06

−45%


Metabolite - 1457 - 35
3.38E−06
7.04E−06
+
136%


Phosphate - 9
4.21E−06
8.65E−06
+
12%


Epinephrine - 9
4.53E−06
9.05E−06
+
41%


Metabolite - 2608 - 35
4.53E−06
9.05E−06
+
80%


Isobar: 2′deoxyguanosine/adenosine
5.34E−06
1.05E−05

−14%


Metabolite - 1713 retired: n-acetyl-L-
5.59E−06
1.07E−05

−44%


aspartic acid - 35


Metabolite - 2564 - 35
5.83E−06
1.10E−05
+
3145%


Metabolite - 1322 retired: citric acid- 35
9.50E−06
1.77E−05

−40%


Metabolite - 2565 - 35
1.04E−05
1.91E−05
+
7127%


Metabolite - 2192 - 9
1.24E−05
2.25E−05

−37%


o-phosphoethanolamine - 9
1.27E−05
2.28E−05
+
37%


Carnosine - 35
1.48E−05
2.62E−05

−18%


Metabolite - 470 - 9
2.01E−05
3.51E−05
+
11%


N-6-trimethyl-I-lysine - 35
2.03E−05
3.51E−05

−16%


12-hydroxydodecanoic acid - 9
2.48E−05
4.23E−05

−42%


Metabolite - 1092 - 35
2.59E−05
4.37E−05
+
5519%


N-formyl-L-methionine - 35
2.66E−05
4.39E−05

−29%


Xanthine - 35
2.67E−05
4.39E−05
+
56%


Metabolite - 2546 - 35
3.73E−05
6.07E−05

−25%


Metabolite - 2220 - 9
4.25E−05
6.85E−05
+
65%


Metabolite - 1839 - 35
6.64E−05
0.00010564

−39%


Metabolite - 485 - 9
7.28E−05
0.000114563

−28%


Metabolite - 2560 - 35
8.81E−05
0.0001372

−13%


Metabolite - 1254 - 35
9.07E−05
0.000139626
+
776%


Metabolite - 1829 retired: oxalic acid -
0.00010345
0.000157572

−18%


35


Serotonin - 35
0.000113
0.000170287

−24%


Metabolite - 2047 - 35
0.000132691
0.000197857

−28%


Metabolite - 2559 - 35
0.00014073
0.000207659
+
192%


alpha-4-dihydroxybenzenepropanoic
0.00016961
0.000247693

−20%


acid - 35


Metabolite - 2646 - 9
0.000176617
0.000255293
+
8%


Metabolite - 1968 - 9
0.000185937
0.000266049
+
17%


Creatinine - 35
0.000188809
0.000267458

−12%


Metabolite - 557 - 9
0.000217403
0.000304913

−33%


Metabolite - 1342-possible-
0.000254676
0.000353309
+
49%


phenylacetylglutamine-or-formyl-N-


acetyl-5-methoxykynurenamine - 35


gamma-L-glutamyl-L-glutamine - 35
0.000256897
0.000353309

−12%


Ascorbic acid - 35
0.000330688
0.000449733

−28%


Metabolite - 1560 - 9
0.000333359
0.000449733

−14%


Tartarate - 9
0.000371962
0.000497079
+
20%


Metabolite - 1264 - 35
0.000458966
0.000604582
+
1018%


Metabolite - 1735 - 35
0.000460942
0.000604582
+
64%


Biliverdin - 35
0.000499595
0.000649269

−24%


Metabolite - 1910 retired: uric acid - 35
0.000566772
0.000727419

−9%


Metabolite - 344 - 9
0.00057
0.000727419
+
2%


Arginine - 35
0.000589024
0.000744985
+
53%


Pantothenic acid - 35
0.000603624
0.000756696
+
57%


Serine - 9
0.000649558
0.000807134
+
51%


Metabolite - 2592 - 35
0.000685228
0.000844053
+
605%


Isobar: lysine/tyramine/putrescine- 9
0.000808049
0.000986762
+
114%


Metabolite - 1114 - 35
0.000836548
0.001012833
+
47%


Metabolite - 2005 - 35
0.000854054
0.001025266

−18%


Metabolite - 2237 - 35
0.000864524
0.001029113
+
398%


Glycerol - 9
0.001035023
0.001221804
+
34%


Metabolite - 273 - 9
0.00114569
0.001341265
+
54%


Arabinose-3 - 9
0.001173106
0.001362103
+
24%


Metabolite - 671 - 9
0.001216735
0.001401276

−25%


5-oxoproline - 35
0.001437638
0.001642331
+
25%


4-acetamidobutyric acid - 35
0.001459636
0.001654121
+
122%


Metabolite - 1734 - 35
0.001498197
0.001684345
+
100%


Metabolite - 2173 - 35
0.001525064
0.001701051

−12%


Metabolite - 1335 - 35
0.002025014
0.002241046

−15%


Metabolite - 706 - 9
0.002158019
0.002369727

−24%


Metabolite - 1064 - 35
0.002197332
0.002394336
+
20%


Metabolite - 1242 - 35
0.002388033
0.002582271
+
160%


Metabolite - 1281 - 35
0.003022562
0.003243651
+
55%


Metabolite - 1834 - 35
0.003372683
0.003592168

−28%


3-hydroxypropanoate - 9
0.003501016
0.003680208

−19%


Raffinose - 35
0.003507303
0.003680208

−19%


Catechol - 35
0.004257739
0.004434789

−28%


Metabolite - 1346 - 35
0.004482293
0.004634603
+
7%


Isobar 8: anthranilic/salicylamide - 35
0.004608619
0.00470831
+
25%


Metabolite - 1087 - 35
0.004620054
0.00470831

−49%


Metabolite - 1347 - 35
0.004696061
0.004751585

−35%


Uridine - 35
0.005661959
0.005688273

−7%


Metabolite - 1911 - 35
0.006615412
0.006578227

−18%


Uric acid - 9
0.006640673
0.006578227

−19%


Metabolite - 1065 - 35
0.007251176
0.007133107
+
386%


Metabolite - 526 - 9
0.007729522
0.007551225
+
52%


Xylitol - 35
0.008558145
0.00830347

−8%


Metabolite - 2185 - 35
0.009068095
0.008738392

−16%


Metabolite - 1849 - 9
0.009824748
0.009403564

−14%


Metabolite - 681 - 9
0.010216325
0.009712728

−21%


Metabolite - 279 - 9
0.010974852
0.010364306
+
15%


Oxitryptan - 9
0.012769081
0.01197886
+
86%


Metabolite - 1127 - 35
0.013601982
0.012676268
+
14%


Metabolite - 278 - 9
0.013824904
0.012799809

−15%


alpha-tocopherol - 9
0.01408719
0.012957954

−18%


N-acetylserotonin - 9
0.014928383
0.013643126
+
100%


Metabolite - 1974 - 35
0.015032093
0.013649843

−17%


Metabolite - 1554 - 9
0.015441641
0.013932421

−8%


Metabolite - 1088 - 35
0.015755837
0.014125935

−11%


Metabolite - 543 - 9
0.0180751
0.016038689
+
22%


Metabolite - 613 - 9
0.018115739
0.016038689
+
18%


Metabolite - 2591 - 35
0.018617915
0.016380908
+
265%


Isobar: 3-hydroxybutanoic acid/butanoic
0.019560566
0.017104059
+
76%


acid - 9


Metabolite - 578 - 9
0.01968929
0.017110995

−15%


Inosine - 35
0.023673433
0.02044797
+
12%


Metabolite - 1958 - 35
0.02610764
0.02241385

−5%


Metabolite - 407 - 9
0.026532892
0.022641714

−11%


Metabolite - 1328 - 35
0.026815296
0.02274568

−21%


Metabolite - 1379 retired: hippuric acid -
0.028105709
0.023698348

−8%


35


Praline - 9
0.028692147
0.024049671
+
30%


Metabolite - 1113 - 35
0.028962419
0.024133411

−11%


1-Hexadecanol - 9
0.029546959
0.024476509
+
14%


N-acetyl-D-galactosamine- 35
0.037024349
0.030460228

−22%


Glutarate - 9
0.037200301
0.030460228

−7%


Metabolite - 2009 - 9
0.039234333
0.031941097
+
15%


Isoleucine - 9
0.043920994
0.035552235
+
30%


4-hydroxy-3-methoxymandelate - 35
0.046820624
0.037580744
+
17%


N-formyl-L-glycine - 35
0.046957594
0.037580744
+
130%


Metabolite - 1753 - 9
0.047736353
0.037989365

−26%


Cholesterol - 9
0.051016232
0.040372733
+
3%


Isobar 2: 2-amino-3amino-GABA-etc - 35
0.052977943
0.041692255

−11%


Metabolite - 1336 retired: carnitine - 35
0.053534798
0.04189772
+
8%


Metabolite - 1826 - 9
0.056530983
0.043999521
+
20%


Metabolite - 621 - 9
0.058160055
0.044884379
+
30%


N-acetyl-L-alanine - 9
0.05830157
0.044884379
+
11%


Metabolite - 2053 - 35
0.061188067
0.046851965

−19%


Metabolite - 1915 - 35
0.061720642
0.047005675

−15%


Metabolite - 2627 - 9
0.064605585
0.048939695
+
6%


Arachidonic acid - 9
0.066466939
0.050081881
+
21%


Metabolite - 1736 retired: p-
0.067663771
0.050713921
+
1%


hydroxybenzaldehyde - 35


Metabolite - 341 - 9
0.068268247
0.050897676

−7%


DL-beta-hydroxyphenylethylamine - 9
0.070205187
0.052067729

−15%


Metabolite - 687 - 9
0.071440216
0.052707732
+
11%


Isobar 9: includes-sucrose-beta-D-
0.072457954
0.053181619
+
7%


lactose - 35


Glucarate - 35
0.073165256
0.053423946

−7%


Metabolite - 423 - 9
0.074943355
0.054441655
+
25%


Metabolite - 1975 - 35
0.078489196
0.056726581
+
39%


Metabolite - 285 - 9
0.083915392
0.060340402

−10%


Leucine - 9
0.091494377
0.065457892
+
24%


2-ethylhexanoic acid - 9
0.092707835
0.06599274
+
7%


Metabolite - 1835 - 35
0.093301471
0.066083235

−12%


Metabolite - 1133 - 35
0.094018603
0.066259864

−12%


Metabolite - 393 - 9
0.095389256
0.066653314

−15%


Metabolite - 2561 - 35
0.095517949
0.066653314

−10%


Metabolite - 1597 - 35
0.096845978
0.067248751
+
5%


Metabolite - 1414 - 9
0.097590754
0.067435349

−9%


Metabolite - 760 - 9
0.102318258
0.070331993

−7%


Metabolite - 1979 retired: CL adduct of
0.102775706
0.070331993

−7%


Isobar 19 - 35


Niacinamide - 35
0.112955903
0.076926929
+
7%


Valine - 9
0.123934053
0.08399959
+
20%


Metabolite - 709 - 9
0.125381408
0.084575904
+
15%


Metabolite - 1142 - possible-5-
0.128826455
0.08648791

−2%


hydroxypentanoate-or-beta-


hydroxyisovaleric acid - 35


N-acetyl-L-valine - 35
0.131541799
0.08758105

−7%


N-acetyl-L-leucine - 35
0.13169126
0.08758105
+
45%


Metabolite - 1262 - 35
0.132521917
0.087721639
+
78%


gamma-aminobutyryl-L-histidine - 35
0.13706282
0.090305458

−7%


Metabolite - 1551 - 9
0.140029039
0.091832659

−3%


Metabolite - 664 - 9
0.14880096
0.097135683
+
18%


Metabolite - 1737 retired: 2,3-
0.150196526
0.097596938

−37%


dihydroxybenzoic acid - 35


Alanine - 9
0.154234783
0.099763347
+
20%


D-galactose - 9
0.155488128
0.10011689
+
22%


Metabolite - 1831-possible-Cl-adduct-
0.158637911
0.101682803

−10%


of-citrulline - 35


Metabolite - 2058 - 35
0.159657851
0.101875582
+
31%


Metabolite - 596 - 9
0.161628617
0.102670622

−2%


Metabolite - 2055 - 35
0.165607853
0.104728699
+
10%


Metabolite - 2606 - 35
0.169515822
0.106723614
+
3%


Metabolite - 2100 - 35
0.172035129
0.10783047

−5%


Isobar: noradrenaline/normetanephrine - 9
0.174990745
0.109045616

−12%


Metabolite - 614 - 9
0.175513392
0.109045616
+
34%


Isobar 3: includes-inositol-1-phosphate-
0.180431829
0.111611892

−4%


mannose-6-phosphate-glucose-6-


phosphate - 35


Oleic acid - 9
0.186145422
0.114645582
+
19%


Metabolite - 1961 retired: glycocholic
0.19133582
0.11733217
+
31%


acid - 35


beta-D-lactose - 35
0.195019064
0.119075354

−13%


Metabolite - 1324 - 35
0.198740723
0.120826931

−3%


Arabinose - 35
0.204819003
0.123990151
+
8%


Isobar 4: includes-Gluconic acid-
0.206407713
0.12442019
+
8%


arabinose-D-ribose - 35


Methyl-indole-3-acetate - 35
0.214527642
0.128766845

−2%


Metabolite - 1383 - 35
0.216364677
0.12926928
+
1%


Metabolite - 1288 - 35
0.217536402
0.12926928
+
22%


Metabolite - 2238 - 35
0.218102394
0.12926928

−46%


Tryptophan - 9
0.221846663
0.13094064

−3%


Metabolite - 268 - 9
0.225783468
0.132711298

−10%


Hydroorotate - 9
0.230150861
0.134719368
+
5%


Metabolite - 1368 - 35
0.233591591
0.136170719
+
241%


Metabolite - 1289 - 35
0.236763471
0.137454092
+
40%


alpha-Hydroxyisobutyric acid-tms - 9
0.244720651
0.141493775
+
12%


Metabolite - 594 - 9
0.250759914
0.144396214

−2%


Metabolite - 1193 - confirmed-3-indoxyl-
0.252904153
0.145041341

−10%


sulfate - 35


Metabolite - 1914 - 35
0.259413238
0.147579348
+
37%


Metabolite - 2221 - 9
0.262843415
0.148932641
+
2%


Metabolite - 1981 - 35
0.266853676
0.15060253

−1%


Melatonin - 9
0.269895636
0.15137824
+
21%


D-fructose-3 - 9
0.270365439
0.15137824
+
11%


3-phospho-d-glycerate - 35
0.271439573
0.151381305

−22%


Metabolite - 293 - 9
0.272708164
0.15149237

−12%


Metabolite - 704 - 9
0.277602508
0.153608844
+
3%


1-7-dihydro-6h-purin-6-one - 35
0.282866742
0.155912726
+
39%


Metabolite - 2587 retired: p-
0.286465619
0.157284382
+
96%


acetaminophen-beta-d-glucuronide - 35


Metabolite - 1066 - 35
0.29122259
0.158593867

−1%


Metabolite - 2067 retired: carnitine - 35
0.291897238
0.158593867
+
5%


Metabolite - 683 - 9
0.292209342
0.158593867

−8%


9,12-octadecadienoic acid - 9
0.293621439
0.158752024
+
13%


Metabolite - 1929 - 35
0.296658476
0.159784193

−1%


Metabolite - 1537 - 9
0.305289189
0.163809957
+
2%


alpha-D-ribose-5-phosphate - 35
0.313376853
0.167515053
+
1%


Metabolite - 941 - 9
0.323784479
0.172427761
+
30%


Palmitoleic acid - 9
0.327398741
0.17369949
+
17%


Metabolite - 1110 - 35
0.333616326
0.176337753
+
5%


Sugar 6 - 9
0.33653011
0.17709421
+
22%


Mannose-6-phosphate - 9
0.337547834
0.17709421

−7%


Metabolite - 1814 - 9
0.339582604
0.177235683
+
3%


Metabolite - 702 - 9
0.340319839
0.177235683

−7%


Metabolite - 1201 - 35
0.347867888
0.180236718

0%


Tyrosine - 35
0.348627012
0.180236718

0%


Pyrophosphate - 35
0.351583476
0.181104217
+
7%


Metabolite - 2056 - 35
0.355678177
0.182549622
+
7%


Metabolite - 1337 - 35
0.357011191
0.182572289

−21%


Lactate - 9
0.360039201
0.183115973

0%


Isobar: adenosine-5-
0.362902611
0.183115973

−4%


diphosphoribose/glucosamine-6-


phosphate - 9


Metabolite - 1909 - 35
0.363241082
0.183115973
+
4%


Metabolite - 1086 - 35
0.363245088
0.183115973
+
24%


palmitate - 9
0.367154133
0.183808241
+
9%


Metabolite - 1888 - 9
0.367213479
0.183808241

−10%


Cystine - 9
0.38705669
0.19238115

−4%


Metabolite - 1372 retired: 2-
0.393185108
0.194743884
+
35%


hydroxyhippuric acid - 35


Inositol - 9
0.398559375
0.195469757
+
4%


Octadecanoic acid - 9
0.399174013
0.195469757
+
8%


Metabolite - 1754 - 9
0.400147965
0.195469757

−2%


Glyoxylate - 9
0.400170225
0.195469757
+
4%


Urea - 9
0.410210662
0.199391682
+
3%


Metabolite - 1656 - 35
0.411014449
0.199391682

−4%


alpha-2-diamino-gamma-
0.413581035
0.199952019
+
6%


oxobenzenebutanoic acid - 35


Metabolite - 1340 - 35
0.415523104
0.200180266
+
27%


Metabolite - 2558 - 35
0.416879441
0.200180266

−4%


Tetradecanoic acid - 9
0.421956382
0.201933628

−3%


Metabolite - 642 - 9
0.424576017
0.202416604
+
5%


Metabolite - 1802 - 35
0.425823475
0.202416604
+
69%


Metabolite - 506 - 9
0.436096446
0.206350302
+
62%


Metabolite - 1842 retired: 4-
0.437891207
0.206350302
+
18%


Guanidinobutanoic acid - 35


Metabolite - 1534 - 9
0.438920776
0.206350302

−2%


N-5-aminocarbonyl-L-ornithine - 35
0.439925617
0.206350302

−7%


Saccaropine - 35
0.443725996
0.20744599

−7%


Metabolite - 1247 - 35
0.44819327
0.208845224

−5%


Metabolite - 1358 - 9
0.455211922
0.211420253
+
8%


Metabolite - 2548-possible-Cl-adduct-
0.461217293
0.213509378

−10%


of-uric acid - 35


Metabolite - 2052 retired: potassium
0.475091649
0.218745839
+
3%


adduct of Isobar 1 - 35


Metabolite - 222 - 9
0.475617382
0.218745839

−25%


Histamine - 35
0.481435239
0.21999306

−1%


Metabolite - 406 - 9
0.484208862
0.220549026
+
5%


Metabolite - 1131 - 35
0.487971612
0.22102226
+
1%


gulono-1-4-lactone - 35
0.488368398
0.22102226

−6%


Histidine - 35
0.502924778
0.225879949
+
4%


Metabolite - 1609 - 35
0.503194155
0.225879949

−8%


Metabolite - 1373 - 9
0.50388562
0.225879949
+
11%


Isobar 1: includes-mannose-fructose-
0.520966974
0.232800404

−3%


galactose-alpha-L-sorbopyranose-


Inositol - 35


Metabolite - 1108 - 35
0.536390627
0.238867648
+
24%


n-dodecanoate - 9
0.537916934
0.238867648
+
4%


Metabolite - 995 - 9
0.543076999
0.240405407

0%


Metabolite - 1332 - 35
0.555102236
0.244963142
+
15%


Succinate - 35
0.560617086
0.246628498
+
46%


Metabolite - 1331 - 35
0.569051953
0.249564148
+
5%


N-tigloylglycine - 9
0.571240346
0.24975067

−7%


Phenylalanine - 35
0.576089348
0.251095705

−1%


Hypotaurine - 9
0.579240029
0.251694524

−5%


Metabolite - 1285 - 35
0.592320498
0.25659123
+
14%


Metabolite - 1673 - 9
0.612947221
0.264717124

−1%


Metabolite - 1323-possible-p-cresol-
0.61860681
0.265186844
+
7%


sulfate - 35


Metabolite - 2563 retired: lactate - 35
0.61912429
0.265186844
+
11%


Metabolite - 2074 - 35
0.622269085
0.265186844
+
14%


Metabolite - 523 - 9
0.624114598
0.265186844
+
66%


5-hydroxyindoleacetate - 35
0.625925876
0.265186844

−1%


Metabolite - 2175 - 35
0.626397572
0.265186844
+
23%


Metabolite - 1827 - 9
0.627139249
0.265186844

−19%


Metabolite - 1161 - 35
0.640126082
0.268828198

−1%


Metabolite - 1334 - 35
0.640126082
0.268828198

−1%


Metabolite - 1847 - 9
0.641443963
0.268828198
+
15%


Metabolite - 562 - 9
0.659500458
0.27438888

−15%


Metabolite - 1514 - 9
0.66032802
0.27438888
+
1%


N-carbamoyl-L-aspartate - 35
0.660523215
0.27438888

−4%


Metabolite - 1353 - 9
0.664614073
0.275280993

−1%


Aspartate - 9
0.679701031
0.280327428

−4%


Metabolite - 1104 - 35
0.680755621
0.280327428

−2%


Metabolite - 638 - 9
0.689784761
0.283222205
+
2%


DL-pipecolic acid - 35
0.713563633
0.292138913
+
5%


Isobar: dulcitol/gluano-1,4-lactone - 9
0.726823595
0.296710109
+
1%


alpha-aminoadipic acid - 9
0.737943315
0.299825714

−4%


Metabolite - 1818 - 35
0.740158119
0.299825714

−7%


Isobar: 3-chloro-L-tyrosine/DOPA - 9
0.740805369
0.299825714
+
2%


Selenocystine - 35
0.744615875
0.30050934
+
24%


Metabolite - 1388 - 35
0.750101919
0.301863367

−37%


Metabolite - 1972 - 35
0.759306974
0.304702125

−1%


Metabolite - 1836 - 35
0.769196748
0.30706915
+
2%


Glycine - 9
0.770801436
0.30706915
+
3%


Metabolite - 1349 retired: Isobar 7 - 35
0.773026197
0.30706915
+
5%


Metabolite - 386 - 9
0.7738764
0.30706915

−5%


Methylmalonic acid - 35
0.777999626
0.307842914
+
37%


Metabolite - 1564 - 9
0.782695577
0.308838355
+
263%


Metabolite - 645 - 9
0.789345703
0.310597209
+
2%


D-galactose-1 - 9
0.805459667
0.316059898
+
17%


Metabolite - 618 - 9
0.816440265
0.319483655

−7%


Threonine - 9
0.820620665
0.320234873
+
10%


Metabolite - 670 - 9
0.842922157
0.32739804
+
3%


Metabolite - 1926 retired: trans-2,3,4-
0.846964798
0.32739804
+
12%


trimethoxycinnamic acid - 35


Gluconic acid - 9
0.848156442
0.32739804
+
2%


Decanoic acid - 9
0.849254587
0.32739804
+
2%


Metabolite - 669 - 9
0.850532853
0.32739804
+
7%


Metabolite - 580 - 9
0.861036973
0.330543205

0%


Metabolite - 1397 - 35
0.867406129
0.33208829
+
2%


Metabolite - 1956 - 9
0.884897064
0.337871566

−3%


Metabolite - 1385 - 35
0.89021962
0.338990105

−1%


Metabolite - 749 - 9
0.901760902
0.34246435
+
1%


Menadione-Vitamin-K3 - 9
0.907156807
0.34359241
+
17%


Riboflavine - 35
0.910014277
0.343755565
+
5%


Malic acid - 9
0.92863219
0.349427329

−5%


Metabolite - 1519 - 9
0.929962462
0.349427329

0%


GABA - 9
0.949912268
0.355979097
+
10%


3-amino-isobutyrate - 9
0.952883348
0.35615031

−4%


Metabolite - 1351 retired: urea adduct
0.955825028
0.356309664
+
2%


of Isobar 6 - 35


Metabolite - 443 - 9
0.961671599
0.357548215

−5%


Metabolite - 1830 - 35
0.970420648
0.359856594
+
10%


Metabolite - 2628 - 9
0.980511879
0.362590137

−7%


2-amino-heptanedioic acid - 9
0.982911488
0.362590137

−4%


Metabolite - 1068 - 35
0.991160619
0.363738719

0%


Metabolite - 1181 - 35
0.991160619
0.363738719

0%


Guanidine acetic acid - 9
0.997630416
0.364656039
+
1%


Metabolite - 1757 - 9
0.998808745
0.364656039
+
4%









Example 3

Biomarkers were discovered by (1) analyzing plasma samples from different groups of human subjects to determine the levels of metabolites in the samples and then (2) statistically analyzing the results to determine those metabolites that were differentially present in the two groups. As listed below in Tables 7-8, biomarkers were discovered that were differentially present between samples from ALS subjects and Control subjects not diagnosed with ALS. Non-biomarker compounds identified in the analyses are also listed in the tables below as those compounds that having a percentage change in ALS of 0%.


The plasma samples used for the analysis were from 62 ALS subjects and 62 control subjects not diagnosed with ALS. After the levels of metabolites were determined, the data was analyzed using T-tests (Table 7) and Wilcoxon's rank-sum tests (Table 8).


Tables 7 and 8 include, for each listed biomarker and non-biomarker compound, the analytical method used to detect the compound, the p-value and the q-value determined in the statistical analysis of the data concerning the biomarkers, and an indication of the percentage difference in the ALS mean level as compared to the control mean level. A percentage change that is positive indicates an increase in ALS compared to control and a negative percentage change indicates a decrease in ALS compared to control. The term “Isobar” as used in the tables indicates the compounds that could not be distinguished from each other on the analytical platform used in the analysis (i.e., the compounds in an isobar elute at nearly the same time and have similar (and sometimes exactly) quant ions, and thus cannot be distinguished).

TABLE 7ALS Biomarkers from blood plasma samples - T-Test Analysis of Plasmafrom Control vs. ALS%AnalyticalchangeNamePlatformp-valueq-valuein ALSMetabolite - 2550LC-MS<0.00010.0031289%alpha-4-dihydroxybenzenepropanoic acidGC-MS0.00120.0925−29%N-6-trimethyl-l-lysineLC-MS0.00140.0925−34%CarnosineLC-MS0.00180.1048−35%Metabolite - 3033GC-MS0.00310.1342−20%glutamic acidGC-MS0.00380.134247%XanthineLC-MS0.00380.134242%Metabolite - 1114LC-MS0.00390.134230%alpha-tocopherolGC-MS0.00520.150948%Metabolite - 2256LC-MS0.00530.1509−45%Metabolite - 2139LC-MS0.00590.154230%arachidonic acidGC-MS0.00730.178147%D-lyxoseGC-MS0.0080.1815−16%Metabolite - 3498LC-MS0.00990.2081−15%Metabolite - 4019GC-MS0.01040.208126%Metabolite - 1497LC-MS0.01150.208134%6-phosphogluconic acidLC-MS0.01220.2081−22%OrnithineGC-MS0.01220.208136%Metabolite - 3030GC-MS0.01340.2162−15%CreatinineLC-MS0.01390.2162−17%Isobar-28-includes-L-threonine-L-LC-MS0.0160.2318−16%allothreonineMetabolite - 3977LC-MS0.01640.2318−27%MethionineLC-MS0.01710.2318−14%Metabolite - 3058GC-MS0.01760.2318−24%Metabolite - 3881 retired: azelaic acidLC-MS0.01880.2374−29%Metabolite - 2041LC-MS0.01960.238429%Metabolite - 3088GC-MS0.02080.2384−31%D-arabitolGC-MS0.02160.2384−14%Metabolite - 3181LC-MS0.02160.238436%Metabolite - 2407LC-MS0.02440.255853%Metabolite - 2313LC-MS0.02480.2558−52%Metabolite - 3994LC-MS0.02580.255835%L-allo-threonineGC-MS0.02620.2558−17%Isobar-27-includes-L-kynurenine-alpha-2-LC-MS0.03240.288815%diamino-gamma-oxobenzenebutanoic acidMetabolite - 3218LC-MS0.0330.288824%NormetanephrineGC-MS0.03660.304923%Metabolite - 3073GC-MS0.03810.308426%Metabolite - 1089LC-MS0.03880.3084−30%Metabolite - 2886 retired: CL adduct of p-LC-MS0.04410.337151%acetiminophen-beta-d-glucuronideThreonineGC-MS0.04440.337−16%1-methyladenineGC-MS0.04840.3578−16%Metabolite - 3056LC-MS0.04920.357817%Metabolite - 3108GC-MS0.05360.3809−15%Metabolite - 3180LC-MS0.05550.380925%anthranilic acidGC-MS0.05570.3809−15%SelenocystineLC-MS0.0570.382219%5-6-dihydroorotic acidGC-MS0.05990.3936−7%Metabolite - 3951LC-MS0.0610.39369%Metabolite - 2266 retired 4-acetominophenLC-MS0.06370.4034571%sulfate5-hydroxylysineLC-MS0.06590.4095−24%4-methyl-2-oxopentanoateGC-MS0.06750.4118−12%Isobar-19-includes-D-saccharic acid-2-deoxy-LC-MS0.06920.4149−25%D-galactose-2-deoxy-D-glucose-L-fucose-L-rhamnosealpha-methyl-L-beta-3-4-GC-MS0.07420.41524%dihydroxyphenylalanineMetabolite - 3320LC-MS0.07550.415−47%2-deoxy-D-glucoseGC-MS0.07630.41522%Metabolite - 1127LC-MS0.07640.415−18%Metabolite - 2546LC-MS0.07860.415−25%Metabolite - 2130LC-MS0.07890.41557%Metabolite - 4134GC-MS0.07890.41519%Metabolite - 2973GC-MS0.08040.4165−9%SaccharopineLC-MS0.08230.4199−20%Metabolite - 2687LC-MS0.08870.4458−8%Metabolite - 4196GC-MS0.09050.448239%Metabolite - 3534LC-MS0.09480.462965%Metabolite - 3134LC-MS0.0970.466866%Metabolite - 3183LC-MS0.09910.4704−14%Metabolite - 2316LC-MS0.10220.476158%p-acetamidophenyl-beta-D-gluguronideLC-MS0.10310.4761137%Metabolite - 1656LC-MS0.10550.4772−18%Metabolite - 3040GC-MS0.10740.477226%Metabolite - 3813LC-MS0.10760.477223%Metabolite - 3331LC-MS0.10890.4772−69%Metabolite - 1831-possible-Cl-adduct-of-LC-MS0.11470.4936−17%citrullineMetabolite - 1979 retired: CL adduct of IsobarLC-MS0.11770.4936−13%19Metabolite - 2894LC-MS0.12080.4936−63%Metabolite - 3098GC-MS0.12520.4936−13%Metabolite - 1839LC-MS0.12530.4936−39%Metabolite - 3843LC-MS0.12620.4936−21%Metabolite - 1975LC-MS0.12840.493625%Metabolite - 3078GC-MS0.12940.4936−9%4-hydroxyphenylacetateLC-MS0.13050.4936−21%Metabolite - 3313LC-MS0.13260.493618%Metabolite - 4275GC-MS0.13660.4936−11%Metabolite - 3090GC-MS0.13790.4936−51%Metabolite - 2924 retired: 2-hydroxybutanoicGC-MS0.13850.493621%acid (also called (s)-2-hydroxybutyric acid)Metabolite - 4274GC-MS0.13890.493614%beta-hydroxypyruvic acidLC-MS0.14030.493630%Metabolite - 1220LC-MS0.14220.493635%CholesterolGC-MS0.14490.493610%Metabolite - 1817LC-MS0.14660.493614%3alpha-7alpha-12alpha-trihydroxy-5beta-GC-MS0.14880.493611%cholanateMetabolite - 3022GC-MS0.15020.4936−11%octadecanoic acidGC-MS0.15040.493611%Metabolite - 3002GC-MS0.15190.493613%Metabolite - 4077GC-MS0.15260.4936−23%UreaGC-MS0.15270.493615%Isobar-5-includes-asparagine-ornithineLC-MS0.15280.4936−19%Metabolite - 2051LC-MS0.15310.493663%Metabolite - 3113GC-MS0.15840.503−33%Metabolite - 2270LC-MS0.15920.503−31%1-HexadecanolGC-MS0.16090.503−16%Metabolite - 3012GC-MS0.16190.503−11%Metabolite - 2915GC-MS0.17020.5122−7%Metabolite - 3099GC-MS0.17080.5122−15%N-carbamoyl-L-aspartateLC-MS0.17090.512254%Metabolite - 3125LC-MS0.17360.512211%MercaptopyruvateLC-MS0.1740.5122−23%Metabolite - 1085-possible-isolobinine-or-4-LC-MS0.17460.512214%aminoestra-1,3-5-10-triene-3-17beta-diolMetabolite - 2366LC-MS0.17530.512235%pantothenic acidLC-MS0.1840.532630%Metabolite - 1335LC-MS0.19010.532619%Metabolite - 3077GC-MS0.19030.5326−7%Metabolite - 3103GC-MS0.19070.5326−17%Metabolite - 2698LC-MS0.19250.532639%Metabolite - 1655LC-MS0.19310.532613%Metabolite - 2866LC-MS0.19320.5326−33%Metabolite - 3160LC-MS0.19610.5361−8%n-dodecanoateGC-MS0.20680.5567−13%Metabolite - 1104LC-MS0.20790.556712%Metabolite - 4365GC-MS0.20850.556717%Metabolite - 4272GC-MS0.21640.5695−12%Metabolite - 1251LC-MS0.21780.569549%Metabolite - 3365LC-MS0.21830.569547%Metabolite - 2392LC-MS0.22240.5746−30%Metabolite - 2329LC-MS0.22360.574623%citric acidGC-MS0.22680.5752−9%Metabolite - 2697LC-MS0.22720.575217%Metabolite - 4043 retired: lysineGC-MS0.23150.579515%Metabolite - 1346LC-MS0.23510.579520%Metabolite - 1351 retired: urea adduct ofLC-MS0.23720.579519%Isobar 6Metabolite - 3624LC-MS0.23780.5795−18%alpha-D-ribose-5-phosphateLC-MS0.23830.579522%GlyceraldehydesGC-MS0.23910.579538%Metabolite - 1988LC-MS0.2460.5892−37%ethyl-3-indoleacetateGC-MS0.25020.589216%TyramineGC-MS0.25170.589215%Metabolite - 2249LC-MS0.25250.5892−14%alpha-keto-glutarateLC-MS0.25440.589212%Metabolite - 3457LC-MS0.25510.589213%Metabolite - 2074LC-MS0.26090.589218%Metabolite - 2986GC-MS0.26120.589228%Metabolite - 2005LC-MS0.26190.589213%Metabolite - 2194LC-MS0.26190.589226%Metabolite - 1368LC-MS0.2620.589258%Metabolite - 4357GC-MS0.26480.5915−14%Metabolite - 3027 retired: arginineGC-MS0.26740.5935−8%Metabolite - 3896LC-MS0.27170.5992−19%Metabolite - 2255LC-MS0.28150.606−35%Isobar-9-includes-sucrose-beta-D-lactose-D-LC-MS0.28450.60625%trehalose-D-cellobiose-D-Maltose-palatinose-melibiose-alpha-D-lactosemethyl-indole-3-acetateLC-MS0.28580.60614%trans-4-hydroxyprolineGC-MS0.28630.60610%Metabolite - 2387 retired: gamma glu leuLC-MS0.28680.60663%diaminopimelic acidLC-MS0.28840.6067%Metabolite - 3517LC-MS0.29010.60652%Metabolite - 3131 retired: N4 adduct ofLC-MS0.29020.60619%indole-3-acetic acidMetabolite - 4276GC-MS0.29080.606−18%Metabolite - 1961 retired: glycocholic acidLC-MS0.29490.610939%Metabolite - 3100GC-MS0.30480.62222%Metabolite - 2212LC-MS0.30580.622−16%Metabolite - 4364GC-MS0.3080.622−9%Metabolite - 2052 retired: potassium adductLC-MS0.30850.6225%of Isobar 1adenosine-inosine-uridine-xanthosine-5-GC-MS0.31120.622−17%monophosphateMetabolite - 3067GC-MS0.31120.622−10%4-Guanidinobutanoic acidLC-MS0.3170.6293−7%Metabolite - 2347LC-MS0.31850.629326%Metabolite - 3143LC-MS0.32050.6295−12%Metabolite - 2306 retired: gamma glu leuLC-MS0.32660.63136%benzoic acidGC-MS0.32780.6313−4%HistidineGC-MS0.33640.6313−7%Metabolite - 3317LC-MS0.33930.6313222%2-deoxyinosineLC-MS0.33980.6313293%Metabolite - 2386LC-MS0.33980.63138%Metabolite - 1834LC-MS0.34310.6313−27%Metabolite - 2806LC-MS0.34320.63135%1-7-dihydro-6h-purin-6-oneLC-MS0.34530.6313−15%Metabolite - 3075GC-MS0.34660.6313−5%Metabolite - 2111LC-MS0.34720.631324%Metabolite - 3334LC-MS0.34840.631315%Metabolite - 4084GC-MS0.34950.6313−9%Metabolite - 3830LC-MS0.35080.6313−15%Metabolite - 1389LC-MS0.35230.6313227%pyridoxamine-phosphateLC-MS0.35250.631312%Metabolite - 2287LC-MS0.35280.631389%3-amino-isobutyrateGC-MS0.3590.6351−7%Metabolite - 1086LC-MS0.36170.635117%Metabolite - 2506LC-MS0.36230.635129%Metabolite - 3074GC-MS0.37430.6473−27%N-acetylserotoninGC-MS0.37490.6473−8%Metabolite - 2389LC-MS0.3810.6544−12%Metabolite - 3093GC-MS0.38660.6585−23%ValineGC-MS0.38720.65857%GlutamineGC-MS0.39410.6615−8%Metabolite - 3097GC-MS0.39510.6615−12%4-hydroxy-2-quinolinecarboxylic acidLC-MS0.39560.66155%shikimic acidGC-MS0.39720.661532%Metabolite - 4031LC-MS0.40.661511%Metabolite - 1350LC-MS0.40360.6615−17%PyridoxamineLC-MS0.40380.6615−3%PhosphateGC-MS0.40450.66154%3-methoxy-L-tyrosineGC-MS0.40840.662115%Metabolite - 2285LC-MS0.40870.66218%tartaric acidLC-MS0.41190.6641−15%Metabolite - 3023GC-MS0.41860.666−6%Metabolite - 3653LC-MS0.41880.666−22%Metabolite - 1960 retired: adduct of EDTALC-MS0.41990.666−10%palmitoleic acidGC-MS0.42250.666−17%OxitryptanLC-MS0.42280.666−7%Metabolite - 4361GC-MS0.43260.6709−22%Metabolite - 3017GC-MS0.43930.6709−8%Possible-Isobar-DL-aspartic acid--aspartate-GC-MS0.43970.6709−10%tetradecanoic acidGC-MS0.44360.6709−11%decanoic acidGC-MS0.44460.670910%Metabolite - 4080GC-MS0.44720.6709−11%Metabolite - 3003GC-MS0.44760.6709−6%AsparaginesGC-MS0.44830.6709−9%NonanateGC-MS0.44870.6709−4%Metabolite - 4032 retired: lysineGC-MS0.44940.6709−16%Metabolite - 3019GC-MS0.44950.6709−4%Metabolite - 2151LC-MS0.45190.670925%Metabolite - 4198GC-MS0.45350.6709−4%Metabolite - 1216LC-MS0.45560.670916%Metabolite - 1349 retired: Isobar 7LC-MS0.45570.6709−6%GlycineGC-MS0.46290.67099%MannitolGC-MS0.46330.670938%D-alanyl-D-alanineLC-MS0.46380.67097%Metabolite - 3102GC-MS0.46470.6709−7%3-methyl-2-oxovaleric acidGC-MS0.46520.6709−9%Metabolite - 4133GC-MS0.47240.678−7%Metabolite - 1736 retired: p-LC-MS0.47420.67812%hydroxybenzaldehydeMetabolite - 2989GC-MS0.47610.678−12%NiacinamideLC-MS0.480.680814%Metabolite - 4003LC-MS0.48730.6878−15%Metabolite - 2100LC-MS0.4890.6878−3%Isobar-6-includes-valine-betaineLC-MS0.49180.68815%25-hydroxycholesterolGC-MS0.49320.68813%Metabolite - 4096LC-MS0.49990.68967%Metabolite - 1342-possible-LC-MS0.50150.689614%phenylacetylglutamine-or-formyl-N-acetyl-5-methoxykynurenamineIsobar-21-includes-gamma-aminobutyryl-L-LC-MS0.50430.6896−12%histidine-L-anserineMetabolite - 1116LC-MS0.50430.689615%3-methyl-L-histidineLC-MS0.50730.689612%Metabolite - 4020GC-MS0.50840.68966%Metabolite - 4362GC-MS0.50970.689610%PhenylalanineLC-MS0.51050.6896−2%Metabolite - 1323-possible-4-sulfobenzyl-LC-MS0.51350.69116%alcohol-possible-p-cresol-sulfateMetabolite - 4360GC-MS0.51980.6927−14%Metabolite - 2046LC-MS0.52020.692713%Metabolite - 4055GC-MS0.52290.692711%Metabolite - 3165LC-MS0.53430.7026−4%Metabolite - 1283LC-MS0.53540.7026−14%MelatoninGC-MS0.53850.7026−4%Metabolite - 1911LC-MS0.53860.702616%N-acetylneuraminateGC-MS0.54410.706−8%creatine-creatinineGC-MS0.54810.70645%SerineGC-MS0.54940.706−4%Metabolite - 3025GC-MS0.55150.706−5%GlycerolGC-MS0.55270.706−7%uric acidGC-MS0.55530.706−5%Metabolite - 3085 retired: inositolGC-MS0.55560.7064%Metabolite - 3668LC-MS0.55850.70632%Metabolite - 3707LC-MS0.55980.70620%Metabolite - 1301LC-MS0.56840.71163%Metabolite - 2388LC-MS0.57230.713716%Metabolite - 3065 retired: 1,5-anhydro-d-GC-MS0.57580.713710%glucitolmalic acidLC-MS0.57630.7137−8%GalactoseGC-MS0.58010.7158−2%CarnitineLC-MS0.58620.72039%glucono-gamma-lactoneGC-MS0.59090.72037%elaidic acidGC-MS0.59210.72039%Isobar-1-includes-mannose-fructose-glucose-LC-MS0.59220.72035%galactose-alpha-L-sorbopyranose-Inositol-D-alloseMetabolite - 3044LC-MS0.59520.72148%Metabolite - 3370LC-MS0.59830.7227−3%Metabolite - 2047LC-MS0.60050.72278%Metabolite - 1843LC-MS0.60860.72948%Metabolite - 3781 retired: Na adduct of IsobarLC-MS0.61030.72945%21Metabolite - 1498LC-MS0.61570.73146%Metabolite - 4271GC-MS0.62080.7314−6%Metabolite - 2055LC-MS0.62110.7314−10%Metabolite - 4147GC-MS0.6230.73147%Metabolite - 3138LC-MS0.62790.731411%3-nitro-L-tyrosineGC-MS0.62940.7314−5%Metabolite - 1113LC-MS0.63070.73146%Metabolite - 3807LC-MS0.63120.7314−4%Metabolite - 3698LC-MS0.63850.7373−10%DL-homocysteineLC-MS0.64160.73848%Metabolite - 3166LC-MS0.65080.7434−14%TryptamineGC-MS0.6550.74345%ascorbic acidLC-MS0.65620.7434−12%n-hexadecanoic acidGC-MS0.65740.74345%Metabolite - 3832LC-MS0.65860.7434−14%Metabolite - 2026LC-MS0.6590.7434−5%Metabolite - 1915LC-MS0.66370.746215%lactateLC-MS0.66890.7479−4%AcetylphosphateLC-MS0.67550.7479−10%SerotoninLC-MS0.67880.747911%Isobar-2-includes-3-amino-isobutyrate-2-LC-MS0.67960.74795%amino-butyrate-4-aminobutanoic acid-dimethylglycine-choline5-hydroxyindoleacetateGC-MS0.68110.747911%Metabolite - 4012GC-MS0.68170.74795%Metabolite - 3489LC-MS0.68360.7479−10%Metabolite - 3402LC-MS0.68560.747921%N-N-dimethylarginineLC-MS0.68730.7479−8%Metabolite - 3708LC-MS0.69230.74794%ThyroxineLC-MS0.69830.7479−4%Metabolite - 1829 retired: oxalic acidLC-MS0.69940.74798%Metabolite - 3615LC-MS0.69980.7479−6%vitamin-B6GC-MS0.70250.74795%Metabolite - 3055LC-MS0.70270.7479−9%Metabolite - 3081GC-MS0.70340.74794%Isobar-8-includes-anthranilic acid-LC-MS0.70410.74795%salicylamideMetabolite - 2774LC-MS0.70450.7479−6%EpinephrineGC-MS0.70890.75013%Metabolite - 4044GC-MS0.71260.75023%FructoseGC-MS0.71480.750214%OctopamineGC-MS0.71590.7502−2%guanidineacetic acidLC-MS0.71770.7502−5%HistamineLC-MS0.73210.76152%hydroxyacetic acidGC-MS0.74020.7615−2%Metabolite - 1329LC-MS0.74320.76155%LactateGC-MS0.74510.76153%BiliverdinLC-MS0.74730.76154%4-hydroxyphenylpyruvateGC-MS0.74740.7615−5%Metabolite - 1914LC-MS0.74740.761511%gulono-1-4-lactoneGC-MS0.74930.7615−3%Metabolite - 3952LC-MS0.75080.76153%AlanineGC-MS0.7530.7615−3%Metabolite - 2898LC-MS0.75310.761521%L-beta-imidazolelactic acidGC-MS0.76340.76194%Metabolite - 2242LC-MS0.76610.761928%N-5-aminocarbonyl-L-ornithineLC-MS0.76690.7619−3%GlucarateGC-MS0.76780.76194%LeucineGC-MS0.76960.76193%N-acetyl-L-alanineGC-MS0.76960.76193%InositolGC-MS0.77230.76192%Metabolite - 3314LC-MS0.77940.76193%XylitolLC-MS0.780.76197%Metabolite - 3014 retired: meso-erythritolGC-MS0.78010.7619−2%AspartateLC-MS0.78330.76193%Metabolite - 2567LC-MS0.78790.76192%2-keto-L-gulonic acidGC-MS0.78870.76196%Metabolite - 3129LC-MS0.78970.7619−1%Metabolite - 2486LC-MS0.79040.76195%Metabolite - 3235 retired: DL-indole-3-lacticLC-MS0.79250.76193%acidDopamineGC-MS0.79630.7619−3%Metabolite - 3139LC-MS0.79740.76194%Metabolite - 2694 retired: lactateLC-MS0.80240.7619−3%Metabolite - 3430LC-MS0.80430.7619−3%5-oxoprolineGC-MS0.80690.7619−2%Metabolite - 3894 retired: pyroglutamic acidLC-MS0.80840.76194%(5-oxoproline)-pyroglutamic acidMetabolite - 1974LC-MS0.80860.7619−3%Metabolite - 2370LC-MS0.81270.76194%PralineGC-MS0.81310.76193%Metabolite - 3696LC-MS0.81370.76198%Metabolite - 3758LC-MS0.81840.7638−10%Metabolite - 3020 retired: threonic acidGC-MS0.82970.77062%Metabolite - 3327LC-MS0.83540.77136%9-12-octadecadienoic acid-z-zGC-MS0.83720.77133%glyceric acidGC-MS0.83770.7713−2%SuccinateGC-MS0.84040.77131%Metabolite - 1458 retired: hypoxanthineLC-MS0.84290.77133%GlycerateLC-MS0.8440.77133%hippuric acidLC-MS0.85150.77616%Metabolite - 2056LC-MS0.8580.77823%AllantoinLC-MS0.8590.7782−3%IsoleucineGC-MS0.86060.77822%Metabolite - 1926 retired: trans-2,3,4-LC-MS0.86990.7845−5%trimethoxycinnamic acidIsocitrateLC-MS0.87510.78571%Metabolite - 1597LC-MS0.87850.78571%Metabolite - 1209LC-MS0.88010.78574%Metabolite - 2269LC-MS0.88040.78574%Metabolite - 3992LC-MS0.88360.7865−1%MannoseGC-MS0.89220.78910%2-amino-butyrateGC-MS0.8930.78911%Metabolite - 3900LC-MS0.89770.78910%Metabolite - 4354GC-MS0.90060.78911%sn-Glycerol-3-phosphateLC-MS0.90130.78911%Metabolite - 3450 retired: 1-LC-MS0.90190.78912%methylnicotinamideTryptophanLC-MS0.90270.78910%Metabolite - 2279LC-MS0.90660.79024%5-6-Dimethylbenzimidazole-GC-MS0.91060.79022%3-hydroxybutanoic acidGC-MS0.91090.7902−4%Metabolite - 1465LC-MS0.91460.79073%Metabolite - 2469LC-MS0.91610.7907−1%Metabolite - 3101GC-MS0.92220.7914−1%alpha-L-sorbopyranoseGC-MS0.92640.79142%TyrosineGC-MS0.92690.7914−1%Metabolite - 2563 retired: lactateLC-MS0.92760.7914−1%3-chloro-L-tyrosineGC-MS0.93410.79141%DOPAGC-MS0.93410.79141%Metabolite - 4251GC-MS0.93440.79141%Metabolite - 1063LC-MS0.93540.79141%Metabolite - 1286LC-MS0.95250.79670%Metabolite - 2250LC-MS0.95390.79672%3-phospho-d-glycerateLC-MS0.9540.7967−1%Metabolite - 1110LC-MS0.95880.79671%DethiobiotinGC-MS0.95950.79670%glucose-6-phosphateGC-MS0.96420.79670%Metabolite - 3783LC-MS0.96470.79670%Metabolite - 3837 retired: 3-indoxylsulfateLC-MS0.96470.79670%Metabolite - 1111-possible-LC-MS0.96490.79670%methylnitronitrosoguanidine-or-ethyl-thiocarbamoylacetateMetabolite - 1573 retired: glycerol-2-LC-MS0.96490.7967−1%phosphateMetabolite - 3178 retired: NH3 adduct of citricLC-MS0.97350.8002−1%acidMetabolite - 2027LC-MS0.97490.80020%Metabolite - 3094GC-MS0.97910.80020%Metabolite - 3604 retired: CL adduct ofLC-MS0.97990.80021%hippuric acidL-alpha-glycerophosphorylcholineLC-MS0.98290.80020%Metabolite - 4148GC-MS0.98330.80020%ArabinoseGC-MS0.99380.80680%Metabolite - 2292LC-MS0.99850.80730%Metabolite - 3132LC-MS0.9990.80730%









TABLE 8










ALS Biomarkers from blood plasma samples- Wilcoxon's Rank Sum-Tests ALS vs. Control












Analytical


% change


Name
Platform
p-value
q-value
in ALS














Metabolite - 2550
LC-MS
<0.0001
1.00E−04
289%


creatine-creatinine
GC-MS
1.00E−04
0.0053
45%


alpha-4-dihydroxybenzenepropanoic acid
GC-MS
0.001
0.0494
−29%


Metabolite - 2256
LC-MS
0.0011
0.0494
−45%


Metabolite - 3033
GC-MS
0.0012
0.0502
−20%


Carnosine
LC-MS
0.0017
0.0627
−35%


N-6-trimethyl-l-lysine
LC-MS
0.002
0.0699
−34%


Metabolite - 1114
LC-MS
0.0024
0.0738
30%


alpha-tocopherol
GC-MS
0.0028
0.0757
48%


glutamic acid
GC-MS
0.0029
0.0757
47%


Metabolite - 3881 retired: azelaic acid
LC-MS
0.0031
0.0757
−29%


Xanthine
LC-MS
0.0032
0.0757
42%


Metabolite - 3218
LC-MS
0.0048
0.101
24%


Metabolite - 3977
LC-MS
0.0048
0.101
−27%


Metabolite - 1368
LC-MS
0.0052
0.102
58%


Metabolite - 2139
LC-MS
0.0054
0.102
30%


Metabolite - 3088
GC-MS
0.006
0.107
−31%


arachidonic acid
GC-MS
0.0063
0.107
47%


6-phosphogluconic acid
LC-MS
0.0096
0.1564
−22%


Metabolite - 3058
GC-MS
0.0106
0.1661
−24%


Ornithine
GC-MS
0.012
0.1801
36%


D-lyxose
GC-MS
0.013
0.1873
−16%


Metabolite - 3498
LC-MS
0.0136
0.1888
−15%


Isobar-28-includes-L-threonine-L-
LC-MS
0.0149
0.1995
−16%


allothreonine


Metabolite - 2313
LC-MS
0.0164
0.2122
−52%


5-6-dihydroorotic acid
GC-MS
0.0173
0.2172
−7%


Metabolite - 2041
LC-MS
0.0187
0.2189
29%


Creatinine
LC-MS
0.0198
0.2189
−17%


D-arabitol
GC-MS
0.0198
0.2189
−14%


Metabolite - 3073
GC-MS
0.0206
0.2207
26%


Metabolite - 2407
LC-MS
0.022
0.2298
53%


Metabolite - 3331
LC-MS
0.0234
0.2328
−69%


Metabolite - 4019
GC-MS
0.0235
0.2328
26%


Metabolite - 3090
GC-MS
0.0252
0.2385
−51%


Metabolite - 3653
LC-MS
0.0254
0.2385
−22%


Metabolite - 2266 retired 4-acetominophen
LC-MS
0.0262
0.2403
571%


sulfate


Metabolite - 1497
LC-MS
0.0276
0.2415
34%


Metabolite - 3030
GC-MS
0.0276
0.2415
−15%


L-allo-threonine
GC-MS
0.0291
0.2485
−17%


Metabolite - 3758
LC-MS
0.0321
0.2683
−10%


Isobar-19-includes-D-saccharic acid-2-deoxy-
LC-MS
0.0329
0.269
−25%


D-galactose-2-deoxy-D-glucose-L-fucose-L-


rhamnose


Metabolite - 3994
LC-MS
0.0353
0.2813
35%


Isobar-27-includes-L-kynurenine-alpha-2-
LC-MS
0.0359
0.2813
15%


diamino-gamma-oxobenzenebutanoic acid


Metabolite - 3183
LC-MS
0.0392
0.3003
−14%


octadecanoic acid
GC-MS
0.0405
0.3045
11%


Metabolite - 3951
LC-MS
0.0434
0.3171
9%


Metabolite - 1089
LC-MS
0.0454
0.3171
−30%


Metabolite - 2387 retired: gamma glu leu
LC-MS
0.0454
0.3171
63%


Threonine
GC-MS
0.0456
0.3171
−16%


Metabolite - 3181
LC-MS
0.0465
0.3171
36%


2-deoxy-D-glucose
GC-MS
0.0479
0.3171
22%


Metabolite - 2130
LC-MS
0.0481
0.3171
57%


Metabolite - 3108
GC-MS
0.0499
0.3202
−15%


Metabolite - 2316
LC-MS
0.0503
0.3202
58%


Methionine
LC-MS
0.0546
0.3422
−14%


anthranilic acid
GC-MS
0.057
0.3509
−15%


pantothenic acid
LC-MS
0.0588
0.3566
30%


Metabolite - 3012
GC-MS
0.0602
0.3589
−11%


alpha-methyl-L-beta-3-4-
GC-MS
0.0621
0.3591
24%


dihydroxyphenylalanine


Metabolite - 2973
GC-MS
0.0621
0.3591
−9%


Metabolite - 3813
LC-MS
0.0632
0.3596
23%


Metabolite - 2886 retired: CL adduct of p-
LC-MS
0.066
0.3699
151%


acetiminophen-beta-d-glucuronide


Isobar-21-includes-gamma-aminobutyryl-L-
LC-MS
0.0693
0.3829
−12%


histidine-L-anserine


5-hydroxylysine
LC-MS
0.0703
0.3829
−24%


Metabolite - 3002
GC-MS
0.0715
0.384
13%


Metabolite - 1656
LC-MS
0.0738
0.385
−18%


Metabolite - 3098
GC-MS
0.0738
0.385
−13%


Metabolite - 3180
LC-MS
0.0749
0.3856
25%


Metabolite - 2392
LC-MS
0.078
0.3897
−30%


Metabolite - 3313
LC-MS
0.0818
0.3897
18%


Saccharopine
LC-MS
0.0821
0.3897
−20%


Metabolite - 1127
LC-MS
0.0846
0.3897
−18%


Metabolite - 4043 retired: lysine
GC-MS
0.0846
0.3897
15%


Normetanephrine
GC-MS
0.0859
0.3897
23%


Metabolite - 3134
LC-MS
0.0871
0.3897
66%


Metabolite - 2924 retired: 2-hydroxybutanoic
GC-MS
0.0872
0.3897
21%


acid (also called (s)-2-hydroxybutyric acid)


Selenocystine
LC-MS
0.0882
0.3897
19%


Metabolite - 2866
LC-MS
0.0883
0.3897
−33%


Metabolite - 1351 retired: urea adduct of
LC-MS
0.0898
0.3897
19%


Isobar 6


3alpha-7alpha-12alpha-trihydroxy-5beta-
GC-MS
0.0911
0.3897
11%


cholanate


Metabolite - 2687
LC-MS
0.0911
0.3897
−8%


Metabolite - 3056
LC-MS
0.0913
0.3897
17%


3-amino-isobutyrate
GC-MS
0.0925
0.3905
−7%


Metabolite - 3843
LC-MS
0.094
0.3923
−21%


Metabolite - 2074
LC-MS
0.0995
0.4109
18%


Metabolite - 4196
GC-MS
0.1049
0.4286
39%


4-methyl-2-oxopentanoate
GC-MS
0.1085
0.4337
−12%


Metabolite - 1086
LC-MS
0.1085
0.4337
17%


Metabolite - 1817
LC-MS
0.1116
0.4415
14%


Mercaptopyruvate
LC-MS
0.1156
0.448
−23%


Metabolite - 2546
LC-MS
0.1165
0.448
−25%


Metabolite - 2255
LC-MS
0.1175
0.448
−35%


beta-hydroxypyruvic acid
LC-MS
0.118
0.448
30%


Metabolite - 4134
GC-MS
0.1215
0.4537
19%


Metabolite - 1975
LC-MS
0.122
0.4537
25%


Metabolite - 3160
LC-MS
0.1232
0.4537
−8%


1-methyladenine
GC-MS
0.1319
0.4813
−16%


Metabolite - 3067
GC-MS
0.1392
0.5031
−10%


Isobar-9-includes-sucrose-beta-D-lactose-D-
LC-MS
0.1444
0.5068
25%


trehalose-D-cellobiose-D-Maltose-palatinose-


melibiose-alpha-D-lactose


Tyramine
GC-MS
0.1451
0.5068
15%


Metabolite - 3365
LC-MS
0.1456
0.5068
47%


Metabolite - 1220
LC-MS
0.1467
0.5068
35%


Metabolite - 3143
LC-MS
0.147
0.5068
−12%


N-carbamoyl-L-aspartate
LC-MS
0.1496
0.5111
54%


Metabolite - 1979 retired: CL adduct of Isobar
LC-MS
0.155
0.521
−13%


19


Metabolite - 3077
GC-MS
0.1571
0.521
−7%


Metabolite - 4275
GC-MS
0.1571
0.521
−11%


Metabolite - 3113
GC-MS
0.1581
0.521
−33%


Metabolite - 1839
LC-MS
0.1627
0.5318
−39%


Metabolite - 1085-possible-isolobinine-or-4-
LC-MS
0.1655
0.5361
14%


aminoestra-1,3-5-10-triene-3-17beta-diol


Metabolite - 3022
GC-MS
0.1699
0.5363
−11%


Metabolite - 3078
GC-MS
0.1699
0.5363
−9%


Metabolite - 4361
GC-MS
0.1699
0.5363
−22%


pyridoxamine-phosphate
LC-MS
0.1743
0.5413
12%


Metabolite - 4077
GC-MS
0.1743
0.5413
−23%


3-methyl-2-oxovaleric acid
GC-MS
0.1786
0.5442
−9%


Metabolite - 1335
LC-MS
0.1788
0.5442
19%


Metabolite - 3320
LC-MS
0.1796
0.5442
−47%


Metabolite - 1831-possible-Cl-adduct-of-
LC-MS
0.188
0.5608
−17%


citrulline


Carnitine
LC-MS
0.1881
0.5608
9%


Metabolite - 2249
LC-MS
0.1928
0.5668
−14%


Metabolite - 1961 retired: glycocholic acid
LC-MS
0.1942
0.5668
39%


p-acetamidophenyl-beta-D-gluguronide
LC-MS
0.1946
0.5668
137%


Metabolite - 1283
LC-MS
0.1989
0.5682
−14%


Cholesterol
GC-MS
0.2001
0.5682
10%


Metabolite - 4272
GC-MS
0.2001
0.5682
−12%


Metabolite - 2915
GC-MS
0.2026
0.5682
−7%


Metabolite - 4357
GC-MS
0.2026
0.5682
−14%


Metabolite - 4276
GC-MS
0.2055
0.5719
−18%


Metabolite - 3832
LC-MS
0.2077
0.572
−14%


Metabolite - 3103
GC-MS
0.2086
0.572
−17%


Metabolite - 3027 retired: arginine
GC-MS
0.2102
0.5723
−8%


Metabolite - 2697
LC-MS
0.2128
0.5752
17%


Metabolite - 3402
LC-MS
0.2154
0.578
21%


Metabolite - 2806
LC-MS
0.218
0.5809
5%


Metabolite - 3166
LC-MS
0.2201
0.5825
−14%


Metabolite - 1113
LC-MS
0.2233
0.5827
6%


Metabolite - 3040
GC-MS
0.2233
0.5827
26%


4-hydroxyphenylacetate
LC-MS
0.2277
0.5884
−21%


Metabolite - 3624
LC-MS
0.2286
0.5884
−18%


Metabolite - 3457
LC-MS
0.2369
0.6057
13%


Metaboiite-1655
LC-MS
0.2426
0.6114
13%


Metabolite - 2051
LC-MS
0.2436
0.6114
63%


adenosine-inosine-uridine-xanthosine-5-
GC-MS
0.2456
0.6114
−17%


monophosphate


Metabolite - 3003
GC-MS
0.2503
0.6114
−6%


citric acid
GC-MS
0.2512
0.6114
−9%


alpha-D-ribose-5-phosphate
LC-MS
0.2527
0.6114
22%


Metabolite - 2366
LC-MS
0.2541
0.6114
35%


Metabolite - 2388
LC-MS
0.2559
0.6114
16%


Metabolite - 3099
GC-MS
0.2571
0.6114
−15%


Metabolite - 3830
LC-MS
0.2571
0.6114
−15%


Metabolite - 4274
GC-MS
0.2571
0.6114
14%


methyl-indole-3-acetate
LC-MS
0.2587
0.6114
14%


Metabolite - 2329
LC-MS
0.2621
0.6155
23%


L-beta-imidazolelactic acid
GC-MS
0.266
0.6201
4%


Metabolite - 1251
LC-MS
0.2673
0.6201
49%


Metabolite - 1104
LC-MS
0.2743
0.6323
12%


Metabolite - 1960 retired: adduct of EDTA
LC-MS
0.2782
0.6328
−10%


trans-4-hydroxyproline
GC-MS
0.2783
0.6328
10%


benzoic acid
GC-MS
0.2815
0.6328
−4%


vitamin-B6
GC-MS
0.2815
0.6328
5%


elaidic acid
GC-MS
0.2846
0.6328
9%


Mannose
GC-MS
0.2846
0.6328
0%


Metabolite - 1988
LC-MS
0.2868
0.634
−37%


4-hydroxy-2-quinolinecarboxylic acid
LC-MS
0.291
0.6357
5%


Metabolite - 1911
LC-MS
0.291
0.6357
16%


Metabolite - 2212
LC-MS
0.2942
0.6391
−16%


Urea
GC-MS
0.3041
0.6566
15%


Metabolite - 4096
LC-MS
0.3073
0.6599
7%


25-hydroxycholesterol
GC-MS
0.3175
0.6779
3%


Metabolite - 1389
LC-MS
0.3206
0.6805
227%


Phosphate
GC-MS
0.3244
0.6847
4%


1-Hexadecanol
GC-MS
0.3291
0.6909
−16%


n-hexadecanoic acid
GC-MS
0.3348
0.6986
5%


Biliverdin
LC-MS
0.3384
0.6986
4%


diaminopimelic acid
LC-MS
0.3384
0.6986
7%


Metabolite - 4365
GC-MS
0.3421
0.6986
17%


Isobar-5-includes-asparagine-ornithine
LC-MS
0.3454
0.6986
−19%


Metabolite - 1116
LC-MS
0.3455
0.6986
15%


Metabolite - 1350
LC-MS
0.3474
0.6986
−17%


Metabolite - 4364
GC-MS
0.3492
0.6986
−9%


Metabolite - 3125
LC-MS
0.3523
0.6986
11%


Metabolite - 3334
LC-MS
0.3523
0.6986
15%


Metabolite - 3696
LC-MS
0.3532
0.6986
8%


Metabolite - 3075
GC-MS
0.3601
0.7011
−5%


Metabolite - 1346
LC-MS
0.3621
0.7011
20%


Metabolite - 3896
LC-MS
0.3635
0.7011
−19%


Metabolite - 3131 retired: N4 adduct of
LC-MS
0.3638
0.7011
19%


indole-3-acetic acid


Metabolite - 3837 retired: 3-indoxylsulfate
LC-MS
0.3638
0.7011
0%


tartaric acid
LC-MS
0.3662
0.7011
−15%


Inositol
GC-MS
0.3676
0.7011
2%


Metabolite - 3017
GC-MS
0.3713
0.7047
−8%


Serotonin
LC-MS
0.3748
0.7077
11%


alpha-keto-glutarate
LC-MS
0.3827
0.717
12%


Metabolite - 4031
LC-MS
0.3835
0.717
11%


Metabolite - 3430
LC-MS
0.3904
0.7227
−3%


Metabolite - 3370
LC-MS
0.3931
0.7228
−3%


Metabolite - 2100
LC-MS
0.3943
0.7228
−3%


Metabolite - 3097
GC-MS
0.3973
0.7229
−12%


Phenylalanine
LC-MS
0.3983
0.7229
−2%


Metabolite - 1843
LC-MS
0.4009
0.7243
8%


Metabolite - 2270
LC-MS
0.408
0.7336
−31%


9-12-octadecadienoic acid-z-z-
GC-MS
0.4142
0.7343
3%


4-hydroxyphenylpyruvate
GC-MS
0.4155
0.7343
−5%


Galactose
GC-MS
0.4182
0.7343
−2%


Tyrosine
GC-MS
0.4182
0.7343
−1%


Metabolite - 3894 retired: pyroglutamic acid
LC-MS
0.4182
0.7343
4%


(5-oxoproline)-pyroglutamic acid


Thyroxine
LC-MS
0.4223
0.738
−4%


Metabolite - 2005
LC-MS
0.425
0.7393
13%


gulono-1-4-lactone
GC-MS
0.4305
0.7454
−3%


Metabolite - 3534
LC-MS
0.4325
0.7455
65%


Metabolite - 3489
LC-MS
0.4425
0.7525
−10%


Metabolite - 2986
GC-MS
0.4429
0.7525
28%


5-6-Dimethylbenzimidazole-
GC-MS
0.4447
0.7525
2%


Metabolite - 3327
LC-MS
0.4447
0.7525
6%


4-Guanidinobutanoic acid
LC-MS
0.4471
0.7525
−7%


Metabolite - 3074
GC-MS
0.4486
0.7525
−27%


glucose-6-phosphate
GC-MS
0.4556
0.7541
0%


Nonanate
GC-MS
0.4556
0.7541
−4%


Metabolite - 3783
LC-MS
0.4556
0.7541
0%


Metabolite - 2285
LC-MS
0.4598
0.7578
8%


Metabolite - 1498
LC-MS
0.4721
0.7657
6%


3-hydroxybutanoic acid
GC-MS
0.4727
0.7657
−4%


Metabolite - 3139
LC-MS
0.4727
0.7657
4%


Metabolite - 2306 retired: gamma glu leu
LC-MS
0.4771
0.7661
6%


Metabolite - 3044
LC-MS
0.4771
0.7661
8%


Metabolite - 3023
GC
0.4859
0.7746
−6%


Metabolite - 1829 retired: oxalic acid
LC-MS
0.4937
0.7827
8%


Oxitryptan
LC-MS
0.4992
0.7853
−7%


Metabolite - 2506
LC-MS
0.5035
0.7853
29%


Lactate
GC-MS
0.5037
0.7853
3%


Metabolite - 4044
GC-MS
0.5037
0.7853
3%


Metabolite - 4080
GC-MS
0.507
0.7853
−11%


Metabolite - 2194
LC-MS
0.5079
0.7853
26%


Metabolite - 1974
LC-MS
0.5126
0.7894
−3%


Metabolite - 2386
LC-MS
0.5172
0.7933
8%


Epinephrine
GC-MS
0.5218
0.797
3%


Metabolite - 2347
LC-MS
0.5286
0.8033
26%


Metabolite - 4133
GC-MS
0.531
0.8033
−7%


3-methyl-L-histidine
LC-MS
0.5333
0.8033
12%


Metabolite - 3093
GC-MS
0.5345
0.8033
−23%


Metabolite - 3019
GC-MS
0.5449
0.8098
−4%


Asparagines
GC-MS
0.5496
0.8098
−9%


glucono-gamma-lactone
GC-MS
0.5496
0.8098
7%


Metabolite - 2052 retired: potassium adduct
LC-MS
0.5543
0.8098
5%


of Isobar 1


Alanine
GC-MS
0.559
0.8098
−3%


Metabolite - 4147
GC-MS
0.5627
0.8098
7%


3-nitro-L-tyrosine
GC-MS
0.5685
0.8098
−5%


sn-Glycerol-3-phosphate
LC-MS
0.5685
0.8098
1%


Metabolite - 3615
LC-MS
0.5685
0.8098
−6%


Metabolite - 2698
LC-MS
0.5773
0.8098
39%


Metabolite - 3698
LC-MS
0.5776
0.8098
−10%


Tryptamine
GC-MS
0.5781
0.8098
5%


uric acid
GC-MS
0.5781
0.8098
−5%


N-acetylneuraminate
GC-MS
0.5803
0.8098
−8%


Metabolite - 1915
LC-MS
0.5944
0.8098
15%


Metabolite - 2055
LC-MS
0.5967
0.8098
−10%


Metabolite - 1301
LC-MS
0.5975
0.8098
3%


Metabolite - 4354
GC-MS
0.5975
0.8098
1%


2-deoxyinosine
LC-MS
0.5986
0.8098
293%


Xylitol
LC-MS
0.6023
0.8098
7%


N-5-aminocarbonyl-L-ornithine
LC-MS
0.6024
0.8098
−3%


Metabolite - 3604 retired: CL adduct of
LC-MS
0.607
0.8098
1%


hippuric acid


Praline
GC-MS
0.6073
0.8098
3%


Metabolite - 2279
LC-MS
0.6087
0.8098
4%


Isobar-8-includes-anthranilic acid-
LC-MS
0.6106
0.8098
5%


salicylamide


Melatonin
GC-MS
0.6123
0.8098
−4%


Metabolite - 1216
LC-MS
0.6123
0.8098
16%


Metabolite - 3316 retired: lactate
LC-MS
0.6123
0.8098
−2%


shikimic acid
GC-MS
0.6172
0.8098
32%


Metabolite - 3025
GC-MS
0.6172
0.8098
−5%


Niacinamide
LC-MS
0.6186
0.8098
14%


Acetylphosphate
LC-MS
0.6194
0.8098
−10%


Metabolite - 1573 retired: glycerol-2-
LC-MS
0.6232
0.8098
−1%


phosphate


Metabolite - 4148
GC-MS
0.6245
0.8098
0%


Metabolite - 1323-possible-4-sulfobenzyl-
LC-MS
0.6254
0.8098
16%


alcohol-possible-p-cresol-sulfate


Metabolite - 4020
GC-MS
0.6272
0.8098
6%


Metabolite - 3517
LC-MS
0.6301
0.8098
52%


2-keto-L-gulonic acid
GC-MS
0.6321
0.8098
6%


1-7-dihydro-6h-purin-6-one
LC-MS
0.6322
0.8098
−15%


D-alanyl-D-alanine
LC-MS
0.6345
0.8098
7%


Metabolite - 4271
GC-MS
0.6345
0.8098
−6%


Metabolite - 2151
LC-MS
0.6356
0.8098
25%


Glyceraldehydes
GC-MS
0.6399
0.8098
38%


Metabolite - 1063
LC-MS
0.6407
0.8098
1%


Glycine
GC-MS
0.6473
0.8098
9%


Serine
GC-MS
0.6473
0.8098
−4%


Metabolite - 2287
LC-MS
0.6516
0.8098
89%


n-dodecanoate
GC-MS
0.6523
0.8098
−13%


Metabolite - 1349 retired: Isobar 7
LC-MS
0.6523
0.8098
−6%


Metabolite - 3165
LC-MS
0.6523
0.8098
−4%


Metabolite - 2242
LC-MS
0.6538
0.8098
28%


Fructose
GC-MS
0.6562
0.8098
14%


Mannitol
GC-MS
0.6571
0.8098
38%


Metabolite - 2292
LC-MS
0.6574
0.8098
0%


Metabolite - 3100
GC-MS
0.6598
0.8098
22%


Metabolite - 3055
LC-MS
0.6611
0.8098
−9%


N-acetylserotonin
GC-MS
0.6625
0.8098
−8%


Metabolite - 3101
GC-MS
0.6647
0.8098
−1%


malic acid
LC-MS
0.6728
0.8098
−8%


Pyridoxamine
LC-MS
0.6779
0.8098
−3%


Lactate
LC-MS
0.6779
0.8098
−4%


Metabolite - 3317
LC-MS
0.6813
0.8098
222%


ethyl-3-indoleacetate
GC-MS
0.683
0.8098
16%


2-amino-butyrate
GC-MS
0.6831
0.8098
1%


Isobar-1-includes-mannose-fructose-glucose-
LC-MS
0.6831
0.8098
5%


galactose-alpha-L-sorbopyranose-Inositol-D-


allose


Glutamine
GC-MS
0.6882
0.8098
−8%


Metabolite - 2469
LC-MS
0.6882
0.8098
−1%


3-chloro-L-tyrosine
GC-MS
0.6933
0.8098
1%


DOPA
GC-MS
0.6933
0.8098
1%


Metabolite - 3132
LC-MS
0.6955
0.8098
0%


Metabolite - 2486
LC-MS
0.6986
0.8098
5%


Metabolite - 3707
LC-MS
0.6986
0.8098
20%


DL-homocysteine
LC-MS
0.7031
0.8098
8%


Metabolite - 2567
LC-MS
0.7058
0.8098
2%


Metabolite - 2894
LC-MS
0.7086
0.8098
−63%


Glycerol
GC-MS
0.7091
0.8098
−7%


Isobar-6-includes-valine-betaine
LC-MS
0.7091
0.8098
5%


Metabolite - 4198
GC-MS
0.7091
0.8098
−4%


Metabolite - 2046
LC-MS
0.7143
0.8133
13%


Isobar-2-includes-3-amino-isobutyrate-2-
LC-MS
0.7267
0.8227
5%


amino-butyrate-4-aminobutanoic acid-


dimethyiglycine-choline


3-methoxy-L-tyrosine
GC-MS
0.7269
0.8227
15%


hydroxyacetic acid
GC-MS
0.7301
0.8238
−2%


Dethiobiotin
GC-MS
0.7354
0.827
0%


Metabolite - 2269
LC-MS
0.7407
0.827
4%


Metabolite - 2111
LC-MS
0.7425
0.827
24%


Metabolite - 3235 retired: DL-indole-3-lactic
LC-MS
0.7459
0.827
3%


acid


Possible-Isobar-DL-aspartic acid--aspartate
GC-MS
0.7461
0.827
−10%


Metabolite - 2250
LC-MS
0.7547
0.8306
2%


5-oxoproline
GC-MS
0.7568
0.8306
−2%


Metabolite - 3952
LC-MS
0.7568
0.8306
3%


Metabolite - 3138
LC-MS
0.7582
0.8306
11%


Metabolite - 4012
GC-MS
0.7674
0.8377
5%


Metabolite - 3085 retired: inositol
GC-MS
0.7691
0.8377
4%


Allantoin
LC-MS
0.7726
0.8391
−3%


Histidine
GC-MS
0.7783
0.8428
−7%


Octopamine
GC-MS
0.7837
0.8439
−2%


N-N-dimethylarginine
LC-MS
0.7869
0.8439
−8%


Metabolite - 3102
GC-MS
0.789
0.8439
−7%


Metabolite - 2047
LC-MS
0.7891
0.8439
8%


alpha-L-sorbopyranose
GC-MS
0.7906
0.8439
2%


Arabinose
GC-MS
0.7966
0.8479
0%


decanoic acid
GC-MS
0.7999
0.8482
10%


Metabolite - 2989
GC-MS
0.8014
0.8482
−12%


Metabolite - 3992
LC-MS
0.8163
0.8616
−1%


Valine
GC-MS
0.8218
0.8649
7%


Metabolite - 1926 retired: trans-2,3,4-
LC-MS
0.8246
0.8655
−5%


trimethoxycinnamic acid


Metabolite - 2563 retired: lactate
LC-MS
0.8327
0.8698
−1%


Metabolite - 3781 retired: Na adduct of Isobar
LC-MS
0.8382
0.8698
5%


21


Glucarate
GC-MS
0.8435
0.8698
4%


Leucine
GC-MS
0.8437
0.8698
3%


N-acetyl-L-alanine
GC-MS
0.8437
0.8698
3%


Metabolite - 1834
LC-MS
0.8449
0.8698
−27%


Metabolite - 3081
GC-MS
0.8533
0.8738
4%


Isocitrate
LC-MS
0.8558
0.8738
1%


Metabolite - 1342-possible-
LC-MS
0.8558
0.8738
14%


phenylacetylglutamine-or-formyl-N-acetyl-5-


methoxykynurenamine


Glycerate
LC-MS
0.8603
0.8746
3%


guanidineacetic acid
LC-MS
0.8645
0.8746
−5%


Metabolite - 1110
LC-MS
0.8657
0.8746
1%


Metabolite - 3020 retired: threonic acid
GC-MS
0.8658
0.8746
2%


Tryptophan
LC-MS
0.8714
0.8778
0%


Metabolite - 4360
GC-MS
0.8767
0.8796
−14%


Metabolite - 1465
LC-MS
0.8778
0.8796
3%


Succinate
GC-MS
0.8821
0.8796
1%


Metabolite - 2774
LC-MS
0.8825
0.8796
−6%


hippuric acid
LC-MS
0.888
0.8828
6%


Metabolite - 1329
LC-MS
0.8965
0.8845
5%


Metabolite - 4032 retired: lysine
GC-MS
0.8981
0.8845
−16%


Metabolite - 3129
LC-MS
0.8992
0.8845
−1%


palmitoleic acid
GC-MS
0.9047
0.8861
−17%


Dopamine
GC-MS
0.9102
0.8861
−3%


Metabolite - 4251
GC-MS
0.9159
0.8861
1%


Metabolite - 4003
LC-MS
0.9167
0.8861
−15%


5-hydroxyindoleacetate
GC-MS
0.9172
0.8861
11%


Metabolite - 1736 retired: p-
LC-MS
0.9207
0.8861
12%


hydroxybenzaldehyde


Isoleucine
GC-MS
0.9215
0.8861
2%


Metabolite - 1286
LC-MS
0.9215
0.8861
0%


Metabolite - 3178 retired: NH3 adduct of
LC-MS
0.9221
0.8861
−1%


citric acid


Metabolite - 4084
GC-MS
0.9271
0.8867
−9%


Metabolite - 3450 retired: 1-
LC-MS
0.929
0.8867
2%


methylnicotinamide


Metabolite - 1914
LC-MS
0.9359
0.8867
11%


Metabolite - 3708
LC-MS
0.9383
0.8867
4%


Metabolite - 4055
GC-MS
0.9383
0.8867
11%


Metabolite - 3014 retired: meso-erythritol
GC-MS
0.9439
0.8867
−2%


glyceric acid
GC-MS
0.9495
0.8867
−2%


Metabolite - 4362
GC-MS
0.9495
0.8867
10%


3-phospho-d-glycerate
LC-MS
0.9549
0.8867
−1%


Metabolite - 3807
LC-MS
0.9551
0.8867
−4%


Metabolite - 2026
LC-MS
0.9554
0.8867
−5%


Aspartate
LC-MS
0.9663
0.8867
3%


Metabolite - 3065 retired: 1,5-anhydro-d-
GC-MS
0.9663
0.8867
10%


glucitol


Histamine
LC-MS
0.9719
0.8867
2%


Metabolite - 3900
LC-MS
0.9719
0.8867
0%


Metabolite - 3668
LC-MS
0.9761
0.8867
32%


tetradecanoic acid
GC-MS
0.9775
0.8867
−11%


Metabolite - 2056
LC-MS
0.9775
0.8867
3%


Metabolite - 2370
LC-MS
0.9775
0.8867
4%


Metabolite - 2694 retired: lactate
LC-MS
0.9775
0.8867
−3%


Metabolite - 1209
LC-MS
0.9823
0.8867
4%


ascorbic acid
LC-MS
0.9831
0.8867
−12%


L-alpha-glycerophosphorylcholine
LC-MS
0.9832
0.8867
0%


Metabolite - 3094
GC-MS
0.9832
0.8867
0%


Metabolite - 1458 retired: hypoxanthine
LC-MS
0.988
0.8867
3%


Metabolite - 1111-possible-
LC-MS
0.9888
0.8867
0%


methylnitronitrosoguanidine-or-ethyl-


thiocarbamoylacetate


Metabolite - 2389
LC-MS
0.9888
0.8867
−12%


Metabolite - 1597
LC-MS
0.9944
0.8883
1%


Metabolite - 2027
LC-MS
1
0.8883
0%


Metabolite - 2898
LC-MS
1
0.8883
21%


Metabolite - 3314
LC-MS
1
0.8883
3%









Example 4
Caffeine and Caffeine Metabolites in ALS Patients and in Healthy Control Individuals

Metabolomic analysis of two independent ALS studies was carried out to assess the changes of metabolite levels in plasma from ALS patients relative to healthy control subjects. Thirty-one healthy control volunteers and 31 participants with ALS enrolled in Study 1. Study 2 was comprised of ninety-nine participants with ALS and 94 healthy volunteers. All research participants provided informed consent. A board-certified neurologist reviewed and confirmed diagnosis for all participants. Data on gender, age, weight, and medications was collected. For participants with ALS, date and site of symptom onset, date of diagnosis, and family history of ALS was recorded. Healthy control subjects were defined by the absence of a known neurological disorder.


Table 9 lists the results of T-test analysis between levels of caffeine and caffeine metabolites (i.e., 1,7-dimethylxanthine, theobromine, and theophylline) in plasma samples from ALS patients as compared to plasma samples from healthy control subjects. The mean levels of caffeine and caffeine metabolites were lower in the ALS patients compared to the healthy control volunteers (Table 9).


The difference in levels is highly statistically significant with the p-value of 0.001 or lower, with the exception of theobromine where the p-value is 0.0591. In addition, the q-value, a measure of the false discovery rate, is also significant, with all values lower than 0.1 with the exception of theobromine where the q-value is 0.3478.

TABLE 9Levels of Caffeine and Caffeine Metabolites inALS Patients Compared to Control Subjects.Higher% ChangeCOMPOUNDLibraryp-valueq-valueMeanin ALSCaffeineLC-MS0.0010.0349Control−73%1-7-LC-MS0.0010.0349Control−50%dimethylxanthineTheobromineLC-MS 0.05910.3478Control−42%TheophyllineLC-MS4.00E−040.0245Control−58%


Example 5
Xenobiotic Metabolism in ALS Patients

As shown in Table 9, ALS patients have lower levels of caffeine than control subjects. This may be due to lower caffeine intake in ALS patients. Alternatively it may indicate increased activity of enzymes that metabolize xenobiotics (e.g. CYP1A2).


A measure of caffeine clearance rate is reflected in the ratio of caffeine metabolites to caffeine. A higher ratio is observed in both the ratios of P/C and T/C for ALS patients than for control individuals (Table 10). This indicates that ALS patients may metabolize caffeine more rapidly than healthy control subjects (rather than indicating lower caffeine intake in ALS patients). Further, subjects with ALS may generally metabolize xenobiotics differently than healthy individuals or individuals suffering from illnesses that have symptoms similar to ALS symptoms.


For example, the rate of caffeine clearance is used to measure the activity of CYP1A2, an inducible Cytochrome P450 enzyme in the liver that metabolizes xenobiotics, including caffeine. Rapid metabolism of caffeine is associated with “pathological detoxifiers” because the products of the P450s include toxins and free radicals. The Phase II conjugation reactions further metabolize these products turning them into less toxic, water soluble compounds that can be excreted. Possibly an overactive xenobiotic metabolizing process (e.g. hepatic P450 enzyme system) and/or an under active Phase II detoxification process may contribute to the etiology of ALS and provide a target for drug discovery and therapeutic intervention.

TABLE 10Ratios of Caffeine Metabolites to Caffeine in plasma fromControl subjects and ALS subjectsSubjectParaxanthine/CaffeineTheophylline/CaffeineGroup(P/C) Ratio(T/C) RatioControl0.82140.8857ALS1.50001.3684


In a metabolomics demographic study of plasma from 270 healthy volunteers the levels of both caffeine and the metabolite paraxanthine increase with age (Table 11). However, the P/C ratio is essentially the same in the younger 2 age groups and is lower in the oldest age group (Table 11). This older age group is the group that is similar in age to the ALS patients and control subjects for the ALS studies.

TABLE 11Ratios of Caffeine Metabolites to Caffeine in Plasma fromNormal Adults of Various Ages.25-3536-5051-65Paraxanthine0.320.690.81Caffeine0.280.671.15Paraxanthine/Caffeine1.14671.03580.7105(P/C) Ratio


Example 6
Caffeine and Caffeine Metabolites in ALS Patients and in Patients with Symptoms Similar to ALS

Metabolomic analysis was carried out to assess the changes of metabolite levels in plasma from ALS patients relative to patients with peripheral neuropathy or myopathy, two neurodegenerative diseases that have symptoms similar to those of ALS. The study was comprised of 99 participants with ALS, 36 participants with myopathy, and 52 participants with peripheral neuropathy. All research participants provided informed consent. A board-certified neurologist reviewed and confirmed diagnosis for all participants. Data on gender, age, weight, and medications was collected. For participants with ALS, date and site of symptom onset, date of diagnosis, and family history of ALS was recorded.


Table 12 lists the results of T-test analysis between levels of caffeine and paraxanthine in plasma samples from (1) ALS patients as compared to plasma samples from peripheral neuropathy patients as well as (2) ALS patients as compared to plasma samples from myopathy patients. The mean levels of caffeine and caffeine metabolites (Table 12) were shown to be lower in the ALS patients compared to the patients with peripheral neuropathy (PN) or myopathy. The difference in levels is statistically significant with the p-value of 0.05. In addition, the q-value, a measure of the false discovery rate, is also significant, especially for the individuals with PN.

TABLE 12Lower Caffeine and Paraxanthine Levels in Plasmafrom ALS Patients Compared to Plasma fromPatients with Myopathy or Peripheral Neuropathy.ALS vs. PeripheralNeuropathyALS vs. MyopathyALS/ALS/Compoundp-valueq-valuePNp-valueq-valuemyopathyCaffeine0.01930.08160.600.00660.14890.38Paraxanthine0.03480.10170.680.04270.45190.57


Example 7
Xenobiotic Metabolism in Neurodegenerative Diseases with Symptoms Similar to Symptoms of ALS

Lower caffeine and caffeine metabolite levels in ALS than in myopathy or PN are possibly due to lower caffeine intake, but may also be the result of a higher rate of caffeine metabolism. One well-characterized system for caffeine metabolism is the Phase I detoxification system carried out in the liver by cytochrome P450 enzymes.


Table 13 lists the mean levels of paraxanthine and caffeine as well as the paraxanthine/caffeine ratio in plasma samples from subjects having ALS, subjects having peripheral neuropathy, and subjects having myopathy. Ratios of caffeine metabolites to caffeine are higher in ALS patients compared to the subjects with diseases with similar symptoms (i.e., PN and myopathy) (Table 13). The difference in ratios is more pronounced when ALS is compared to PN. Thus, the data indicates higher levels and/or higher activity of CYP1A2 and/or similar xenobiotic metabolizing enzymes in ALS patients (rather than suggesting lower caffeine intake) as compared to normal levels and/or activity of CYP1A2 and/or similar xenobiotic metabolizing enzymes in patients having diseases with related symptoms. This result supports the idea that caffeine and/or xenobiotic metabolism (e.g. enzyme levels and/or activity of CYP1A2 and/or other similar enzyme systems) is altered in ALS. Thus, the metabolism of caffeine provides a useful way to distinguish ALS patients from patients with similar symptoms to ALS.

TABLE 13Ratios of the Caffeine Metabolite Paraxanthine to Caffeine in Plasmafrom Patients with ALS, Myopathy or Peripheral Neuropathy.ALSPNMyopathyParaxanthine0.680.021.20Caffeine0.920.722.44Paraxanthine/Caffeine Ratio0.73890.02940.4900


Example 8e
Biomarkers for Differentiating ALS from Other Neurodegenerative Diseases

Metabolomic analysis was carried out to assess the changes of metabolite levels in ALS patients relative to patients with peripheral neuropathy or myopathy, two neurodegenerative diseases that have symptoms similar to those of ALS.


Table 14 below lists the biomarkers that were discovered that distinguish ALS subjects from subjects with peripheral neuropathy.


Table 15 below lists the biomarkers that were discovered that distinguish ALS subjects from myopathy subjects.


Tables 14 and 15 include, for each listed biomarker, the p-value and the q-value determined in the statistical analysis of the data concerning the biomarkers and an indication of the median level of each biomarker in the control, ALS, peripheral neuropathy (Table 14), and myopathy (Table 15) samples. The term “Isobar” as used in the tables indicates the compounds that could not be distinguished from each other on the analytical platform used in the analysis (i.e., the compounds in an isobar elute at nearly the same time and have similar (and sometimes exactly) quant ions, and thus cannot be distinguished).

TABLE 14Biomarkers that distinguish ALS subjects from subjectswith Peripheral Neuropathy (PN).Metabolitep-valueq-valueControlALSPNAspartate2.36E−050.00090.970.971.18Isobar citric acid-isocitrate0.00010.00250.980.851.08p-hydroxyphenyllactate0.00010.00251.020.801.12Alanine0.00010.00250.980.841.09Isobar lysine-tyramine-0.00010.00270.830.701.00putrescineHistamine0.00030.00581.000.891.06Creatinine0.00080.01141.030.911.033-hydroxybutanoic acid0.0010.01230.931.530.83N-6-trimethyl-1-lysine0.00100.01231.050.871.05Inosine0.00150.01501.091.080.83Proline0.00150.01500.950.811.15Adenosine0.00210.01690.920.930.63Arachidonic acid0.00220.01690.200.680.20Paraxanthine0.00320.02271.140.270.98Guanosine0.00510.02851.001.000.73Malic acid0.00510.02850.900.911.11Threonine0.00530.02850.990.821.04Oleic acid0.00620.03151.001.230.91Tetradecanoic acid0.00830.03510.991.090.82Caffeine0.00840.03511.340.430.933-phospho-d-glycerate0.01020.03920.800.360.98Citrulline0.01140.04130.380.010.81Palmitoleic acid0.01200.04281.021.230.90Palmitate0.01280.04450.991.100.96alpha-Hydroxyisobutyric0.01340.04510.991.090.93acidArginine0.01760.05571.020.780.923-hydroxypropanoate0.01800.05500.840.980.86Valine0.01940.05680.970.831.00Uric acid0.01970.05681.100.991.29N-formyl-L-glycine0.02180.05940.120.120.56Glutarate0.03150.07831.040.871.01Kynurenine0.03720.08600.970.981.05









TABLE 15










Biomarkers that distinguish ALS from Myopathy















Median
Median
Median


Metabolite
p-value
q-value
Control
ALS
Myopathy















Isobar citric acid-
0.0005
0.0354
0.98
0.85
1.16


isocitrate


Caffeine
0.0055
0.2132
1.34
0.43
1.04


Aspartate
0.0090
0.2425
0.97
0.97
1.06


Kynurenine
0.0132
0.3076
0.97
0.98
1.12


3-hydroxybutanoic
0.0173
0.3377
0.93
1.53
0.92


acid


Glutarate
0.0249
0.3377
1.04
0.87
1.07


uric acid
0.0290
0.3605
1.10
0.99
1.26


Dulcitol
0.0318
0.3729
0.94
0.40
1.00


Palmitate
0.0337
0.3729
0.99
1.10
0.89


selenocystine
0.0375
0.3729
0.99
0.92
1.06


N-acetyl-L-glutamine
0.0389
0.3764
1.03
1.05
1.35


Threonine
0.0496
0.4298
0.99
0.82
0.99









Example 9
Biomarkers for Disease Progression

As listed below in Tables 16, 17 and 18, biomarkers were discovered that were differentially present among samples from ALS subjects over the course of the disease that indicate the progression of the disease. Tables 16, 17, and 18 include, for each listed biomarker and non-biomarker compound, the p-value and the q-value determined in the statistical analysis of the data concerning the biomarkers. Throughout the tables, the column heading “LIB_ID” indicates the analytical platform used to measure the level of the compound. The number “61” indicates that the levels of those compounds were measured using LC-MS, and the number “50” indicates that the levels of those compounds were measured using GC-MS. The term “Isobar” as used in the tables indicates the compounds that could not be distinguished from each other on the analytical platform used in the analysis (i.e., the compounds in an isobar elute at nearly the same time and have similar (and sometimes exactly the same) quant ions, and thus cannot be distinguished).


Non-biomarker compounds identified in the analyses are also listed in the Tables 16, 17 and 18 below as those compounds having a slope of 0 and/or a percentage change over time of 0%.


Biomarkers were discovered by (1) analyzing plasma samples from human subjects with ALS at various times to determine the levels of metabolites in the samples and then (2) statistically analyzing the results to determine those metabolites that are differentially present at various time points. As listed below in Tables 16-18, biomarkers were discovered that were differentially present in samples from ALS subjects early in the disease compared to samples collected at later times when the disease severity had progressed. The metabolite changes were also evaluated relative to changes in the Forced Vital Capacity (FVC), a clinical measurement that indicates disease severity. As the disease progresses the FVC measurement decreases. The most severely affected ALS patients have the lowest FVC scores.


The plasma samples used for the analysis were collected from 40 ALS subjects at several times during the course of the disease. Samples were collected early (screening/month 0 and/or month 1), at 6 months after screening, and at 12 months after screening. After the levels of metabolites were determined, the data from the subjects was analyzed using random coefficient regression analysis and T-tests.


For the initial random coefficient regression analysis, a random coefficient regression (Little, R., Milliken G., Stroup, W., Wolfinger, R. (1996) SAS System for Mixed Models, Chapter 7, SAS® Institute, Cary, N.C.) was first performed for each compound. This model allows for different intercepts and different slopes for each subject with a common intercept and slope across all subjects. The analysis tests whether or not the common slope is zero. The x-variable was the month, which had possible values of 0, 1, 6, and 12. The results of this analysis are shown in Table 16. If the slope is positive, it means the level of the compound increased over time; likewise, if the slope is negative, it means the level of the compound decreased over time.


Another random coefficient regression was performed for each compound on the FVC (forced vital capacity). The x-values were the average FVC values since the original FVC values were measured in triplicate per time point. The 40 patients used in the initial analysis were also used for this random coefficient regression analysis. The results are shown in Table 17. A positive slope means that the compound was positively correlated with FVC (i.e., as FVC increased, the compound increased or as FVC decreased, the compound decreased). A negative slope indicates that as the FVC increased, the compound decreased (or vice-versa).


A T-test was performed by comparing the levels of the compounds in the samples collected at month 1 to the levels of the compounds in the samples at month 12. All patients with both the 1 month and 12 month time points were included in the analysis (37 patients total). The standard matched-pairs T-test was used to perform this analysis. The results are shown in Table 18. The table includes, for each listed biomarker and non-biomarker compound, an indication of the percentage difference in the mean level at 1 month after screening as compared to the mean level at 12 months after screening, where a positive percentage change indicates that there was an increase in the metabolite level as the disease progressed and a negative percentage change indicates that there was a decrease in the metabolite level as the disease progressed.

TABLE 16Random Coefficient Regression Analysis Over TimeCOMPOUNDLIB_IDp-valueq-valueSLOPEMetabolite - 3073505.93E−081.09E−05−0.020gamma-L-glutamyl-L-tyrosine611.15E−050.0011−0.019Metabolite - 3183-possible-gamma-L-glutamyl-L-613.21E−050.0020−0.017phenylalanineMetabolite - 6246505.36E−050.0025−0.011inositol-1-phosphate509.40E−050.0035−0.026Metabolite - 1208610.00050.0139−0.049methionine-sulfoxide610.00070.0139−0.023Metabolite - 4275500.00070.01390.029Metabolite - 3078500.00080.01390.028Metabolite - 3088500.00080.0139−0.027Metabolite - 3026500.00080.0139−0.015O-acetyl-L-carnitine-hydrochloride610.00090.01390.027Isobar-22-includes-glutamic acid-O-acetyl-L-serine610.00110.0151−0.012Metabolite - 5233610.00120.0151−0.043gamma-glu-leu610.00140.0173−0.015Metabolite - 3098500.00150.0174−0.014Metabolite - 3022500.00180.0194−0.010Metabolite - 2567610.00210.0214−0.014Arginine610.00240.0231−0.019Metabolite - 3143610.00260.0238−0.049hippuric acid610.00290.0251−0.040Metabolite - 3114500.00320.0267−0.035Metabolite - 5976610.00360.0278−0.043Metabolite - 3019500.00370.0278−0.008Metabolite - 3025500.00380.0278−0.012Metabolite - 7050610.00470.0331−0.024N-6-trimethyl-l-lysine610.00530.0362−0.022Metabolite - 4769500.00580.03840.021Glutamine500.00620.03960.049Metabolite - 4627610.00650.0400−0.0414-Guanidinobutanoic acid610.00730.0431−0.016Metabolite - 5346500.00750.0431−0.015Metabolite - 2688610.00850.0473−0.017Metabolite - 5907500.00890.0481−0.013Metabolite - 7009610.00960.0503−0.017Metabolite - 3830610.01040.0503−0.037Thyroxine610.01050.05030.025Isobar-8-includes-anthranilic acid-salicylamide610.01050.0503−0.021Metabolite - 3077500.01090.0503−0.012Metabolite - 2546610.01090.0503−0.061Metabolite - 3109500.01210.0531−0.027Metabolite - 6326500.01210.0531−0.007glyceric acid500.01370.0581−0.014Metabolite - 2005610.01400.05810.028Metabolite - 2319610.01420.0581−0.022Methionine610.01490.05940.0461-7-dimethylxanthine610.01520.0594−0.025Isobar-19-includes-D-saccharic acid-1,5-anhydro-D-610.01750.06710.014glucitol-2′-deoxy-D-galactose-2′-deoxy-D-glucose-L-fucose-L-rhamnoseCreatinine610.01940.0730−0.016Metabolite - 1988610.02020.0743−0.0203-methyl-L-histidine610.02170.0765−0.013Tyrosine610.02190.0765−0.008Metabolite - 6907500.02210.0765−0.012Biotin610.02340.0797−0.028Metabolite - 4511500.02550.0854−0.011Metabolite - 5887610.02650.0871−0.094Arabinose500.02730.0881−0.016Metabolite - 3012500.02800.0887−0.005Metabolite - 1206-possible-methyltestosterone-and-610.02940.0917−0.029othersMetabolite - 7089610.03230.0990−0.012Metabolite - 4252500.03390.1002−0.024D-quinic acid500.03430.1002−0.045Metabolite - 5349500.03430.1002−0.005Metabolite - 3138610.03600.1033−0.017Metabolite - 3094500.03750.1060−0.012DL-pipecolic acid610.03850.1071−0.015Metabolite - 5086610.04560.12500.022Metabolite - 1111-possible-610.04890.13230.019methylnitronitrosoguanidine-or-ethyl-thiocarbamoylacetateMetabolite - 3783610.05210.13870.013Metabolite - 5728610.05300.1391−0.027Metabolite - 1092610.05570.1442−0.047Metabolite - 6955500.05930.1480−0.011Metabolite - 3653-Possible-stachydrine610.05940.1480−0.072Mannose500.06030.14800.009Metabolite - 4611500.06070.1480−0.008Metabolite - 1911610.06120.1480−0.022Oleic-Acid500.06830.16290.019beta-hydroxypyruvic acid500.07430.17340.010Metabolite - 7846500.07450.1734−0.014Metabolite - 3951610.07720.17730.0091-methyl-guanidine500.08960.1969−0.006Histamine610.09030.19690.017Metabolite - 6272500.09050.1969−0.014Metabolite - 1656610.09070.19690.018Metabolite - 1127610.09290.19690.0121,5-anhydro-D-glucitol500.09310.1969−0.005Lactate500.09320.19690.018Metabolite - 6346500.09600.2004−0.006(p-Hydroxyphenyl)lactic acid500.10240.2115−0.012Metabolite - 1286610.10380.21190.009Metabolite - 4586610.11440.22830.009Metabolite - Metabolite - 5982 retired: 1-oleoyl-rac500.11530.2283−0.018glycerolMetabolite - 2139610.11550.22830.011Hypoxanthine610.11960.23390.141Metabolite - 4351610.12610.24390.011L-alpha-glycerophosphorylcholine610.12900.2469−0.028Metabolite - 1345610.13120.2487−0.016alpha-tocopherol500.13320.2499−0.046Metabolite - 4767500.13820.2552−0.005Metabolite - 1086610.13980.2552−0.036Metabolite - 3707610.14020.25520.047Metabolite - 1819610.14160.2553−0.011Metabolite - 3030500.14660.2603−0.005Metabolite - 6711610.14730.26030.011Inosine610.15810.27680.060Caffeine610.16150.2778−0.030Metabolite - 2168610.16170.27780.005N-5-aminocarbonyl-L-ornithine500.17020.2873−0.010Isobar-21-includes-gamma-aminobutyryl-L-histidine-610.17170.28730.005L-anserineGlycine500.17200.28730.070Metabolite - 2568610.17370.2876−0.019Metabolite - 3033-possible-threonine-deriv500.18200.2987−0.004Metabolite - 3044610.18560.29930.018Metabolite - 5247610.18810.29930.010Metabolite - 4020500.18870.2993−0.007Metabolite - 2390610.19110.2993−0.011Tryptophan610.19160.2993−0.004Eythrose500.19210.2993−0.010pantothenic acid610.19470.30000.057Serine500.19590.30000.026Metabolite - 1975610.19950.3031−0.038Metabolite - 2074610.21650.3236−0.025Riboflavine610.22250.3249−0.024Metabolite - 3131 retired: N4 adduct of indole-3-610.22300.3249−0.007acetic acidMetabolite - 3052610.22410.3249−0.010n-dodecanoate500.22450.3249−0.018Cholesterol500.23220.3334−0.004arachidonic acid500.23570.3358−0.006Praline610.23960.3376−0.004Metabolite - 2109610.24060.3376−0.007Metabolite - 5791610.25430.35410.033Metabolite - 6347500.25800.3546−0.009Metabolite - 5788610.26150.3546−0.038Inositol500.26320.3546−0.005Metabolite - 6126610.26480.3546−0.016Metabolite - 1193-confirmed-3-indoxyl-sulfate610.26580.3546−0.009Metabolite - 4732610.27150.35460.013Metabolite - 3708610.27160.3546−0.023Metabolite - 7888500.27190.3546−0.005Metabolite - 2027610.27580.35460.007Threonine500.27870.35460.017Metabolite - 1835610.27880.3546−0.006palmitoleic acid500.27980.35460.015Metabolite - 1831-possible-Cl-adduct-of-citrulline610.28070.3546−0.005Alanine500.28170.35460.017Biliverdin610.28510.3551−0.015Metabolite - 6226500.28720.3551−0.010Metabolite - 3093500.28910.3551−0.007Metabolite - 7765610.29090.3551−0.042Metabolite - 3017500.29200.3551−0.005Metabolite - 7008610.29670.3551−0.006Metabolite - 4274500.30210.35510.026Metabolite - 1597610.30410.3551−0.003Metabolite - 1142-possible-5-hydroxypentanoate-or-610.30520.3551−0.010beta-hydroxyisovaleric acidMetabolite - 6488500.30650.3551−0.014Glycerol500.30670.35510.007Metabolite - 4612610.31000.3551−0.010Metabolite - 5847500.31170.3551−0.019Metabolite - 6467500.31350.3551−0.010Carnitine610.31390.3551−0.007Metabolite - 5231610.31400.3551−0.007Metabolite - 3603610.31490.3551−0.012L-kynurenine610.32080.3574−0.004Metabolite - 2053610.32080.35740.011Metabolite - 1114610.32430.35750.017Metabolite - 3040500.32490.35750.004Metabolite - 2321610.32670.3575−0.011Metabolite - 3099500.33080.3598−0.008citric acid500.33960.3672−0.005Metabolite - 4055500.34920.3753−0.013Bicine610.35320.37750.008phosphoenolpyruvate610.36130.37970.007Uridine610.36260.37970.009Metabolite - 4523500.36430.3797−0.006benzoic acid610.36600.37970.013glycochenodeoxycholic acid610.36610.3797−0.017Metabolite - 3377610.36770.37970.026Metabolite - 2506610.36980.3797−0.0083-phospho-d-glycerate610.38020.3883−0.009malic acid500.38270.38870.019Metabolite - 4522500.38610.38990.003Metabolite - 4547610.38970.3915−0.011Metabolite - 5730610.39260.39220.006hydroxyacetic acid500.39740.3947−0.004Metabolite - 5366500.40160.3947−0.013methyl-indole-3-acetate610.40210.3947−0.009Metabolite - 3002500.40390.3947−0.014Metabolite - 2249610.40750.3947−0.006Metabolite - 4806500.40800.3947−0.006Metabolite - 4658610.41290.3973−0.006Theobromine610.42460.40100.011Isobar-28-includes-L-threonine-L-allothreonine-L-610.42650.4010−0.005homoserine-S-4-amino-2-hydroxybutyric acidMetabolite - 6269500.42710.4010−0.003Metabolite - 6227500.42910.4010−0.009Lysine500.43010.40100.011Metabolite - 2594610.43030.40100.161Metabolite - 2370610.43200.4010−0.008Metabolite - 3100500.43610.4028−0.012Ornithine500.44190.40430.023Metabolite - 2559610.44390.4043−0.015Metabolite - 3772610.44850.40430.015Metabolite - 7706610.44880.40430.005Metabolite - 2469610.44900.40430.009Metabolite - 6869500.45080.4043−0.008Metabolite - 4624500.45520.4048−0.007Valine500.45800.40480.011Metabolite - 6963500.45810.4048−0.008Metabolite - 4362500.46230.4066−0.0072-hydroxybutyric acid500.46760.40930.005Metabolite - 1335610.47060.4097−0.013heptadecanoic acid500.47250.4097−0.005Isobar-1-includes-mannose-fructose-glucose-610.47490.4098−0.003galactose-alpha-L-sorbopyranose-Inositol-D-allose-D-altrose-D-psicone2-keto-L-gulonic acid500.48570.4172−0.006n-hexadecanoic acid500.49160.42030.004Metabolite - 2269610.49590.42200.021Metabolite - 3832-possible-phenol-sulfate610.50440.4231−0.018Metabolite - 7807610.50830.4231−0.003Metabolite - 3056610.50950.42310.003Metabolite - 7815610.50980.4231−0.002Metabolite - 5848610.51090.4231−0.005Metabolite - 7707610.51100.42310.003Metabolite - 2973500.52820.43420.001Metabolite - 4931610.53230.43420.010Metabolite - 4986500.53520.4342−0.004Urea500.53550.4342−0.002Metabolite - 3087500.53620.43420.020Isobar-6-includes-valine-betaine610.54220.4352−0.002Metabolite - 1914610.54220.43520.016p-hydroxybenzaldehyde610.54740.4375−0.002Metabolite - 1254610.55200.4392−0.033Metabolite - 1834610.55900.44290.008Metabolite - 2825-possible-Riluzole610.56320.44400.006Metabolite - 1085-possible-isolobinine-or-4-610.56760.4440−0.003aminoestra-1,3-5-10-triene-3-17beta-diolglycocholic acid610.56770.44400.015Metabolite - 2548-possible-Cl-adduct-of-uric acid610.57410.44710.005Metabolite - 2395610.57660.4472−0.006Metabolite - 5983500.58170.4492−0.004Isobar-2-includes-2-aminoisobutyric acid-3-amino-610.58610.45080.008isobutyrate-2-amino-butyrate-4-aminobutanoic acid-dimethylglycine-cholineMetabolite - 4510500.58880.4510−0.012Leucine500.59610.45470.010Saccharopine610.60590.4602−0.004dehydroisoandrosterone-3-sulfate-sodium-salt-610.61080.4621−0.005hydrateDL-homocysteine610.61650.4645−0.005Metabolite - 2100610.63410.47500.003meso-erythritol500.63570.4750−0.002oxalic acid610.64400.4765−0.004Metabolite - 7336610.64440.4765−0.008Metabolite - 3097500.64820.4765−0.005Metabolite - 1323-possible-p-cresol-sulfate610.65050.47650.004octadecanoic acid500.65240.4765−0.002Metabolite - 2386610.65810.47650.003Metabolite - 1110610.66010.47650.016Histidine500.66170.47650.006DL-indole-3-lactic acid610.66500.4765−0.002glutamic acid500.66630.47650.006Phosphate500.66630.47650.002Creatine610.66890.4765−0.0114-O-beta-galactopyranosyl-D-mannopyranose610.67590.47970.004Acetylpyrazine610.68050.4800−0.002Metabolite - 2952500.68520.48000.030Metabolite - 3441610.68610.4800−0.003Metabolite - 5769610.68680.48000.002Metabolite - 6551610.69050.48080.006Metabolite - 4470610.69600.4811−0.003Phenylalanine610.69630.4811−0.001Metabolite - 5403500.71960.4954−0.001Metabolite - 3003500.74080.50810.002Metabolite - 3972610.74530.50920.002Metabolite - 2056610.76140.51550.001Metabolite - 7146610.76260.5155−0.0023-hydroxybutanoic acid500.76280.51550.010decanoic acid500.76870.5174−0.001Metabolite - 1836610.77500.51740.002Nonanate500.77570.5174−0.001Metabolite - 4428610.77680.5174−0.002Metabolite - 6270500.78120.5182−0.004D-glucose500.78730.51820.001Metabolite - 4167610.78830.5182−0.002Fructose500.79290.51820.007Metabolite - 1342-possible-phenylacetylglutamine-or-610.79450.51820.002formyl-N-acetyl-5-methoxykynurenaminetetradecanoic acid500.79490.51820.0035-oxoproline500.79930.5192−0.003Metabolite - 4873610.80470.51930.004Metabolite - 2753610.80690.5193−0.003Metabolite - 4363610.80800.51930.007Isobar-13-includes-5-keto-D-gluconic acid-2-keto-L-610.81710.52200.004gulonic acid-D-glucuronic acid-D-galacturonic acidMetabolite - 3808610.81790.52200.002Metabolite - 1264610.84020.53440.014Metabolite - 3218610.84560.53540.003Isoleucine500.84760.53540.003Allantoin500.85660.53920.0055-6-dihydrouracil500.86060.53990.002Metabolite - 2986500.86560.5412−0.001Aspartate500.86960.5419−0.002Metabolite - 1498610.87830.5454−0.001Metabolite - 2561610.88500.54770.001Metabolite - 2981500.89340.54780.001Metabolite - 1215610.89520.5478−0.001Metabolite - 4364500.89640.5478−0.001Metabolite - 1261610.89710.54780.002Metabolite - 2915500.92750.56460.001Metabolite - 7147610.93380.56460.000Metabolite - 4795500.93710.56460.001Metabolite - 7890500.94700.5646−0.001Metabolite - 3125610.94970.5646−0.001sn-Glycerol-3-phosphate500.95240.56460.001Metabolite - 2185610.95440.56460.000Aldosterone610.95640.56460.000Metabolite - 4906610.96120.5646−0.001Metabolite - 6931500.96190.56460.000Metabolite - 3129610.96310.56460.000Metabolite - 5234610.96380.56460.001Linoleic acid500.96710.56460.000Metabolite - 7762610.96750.5646−0.001Metabolite - 5673610.98140.57090.000uric acid500.98900.57150.000Metabolite - 6827610.99180.57150.000Metabolite - 7889500.99190.57150.000Metabolite - 7177610.99750.57300.000









TABLE 17










Random Coefficient Regression Analysis Over Forced Vital Capacity.











COMPOUND
LIB_ID
p-value
q-value
slope














Creatinine
61
0.0003
0.0782
0.109


Metabolite - 7089
61
0.0032
0.2716
0.099


Metabolite - 4769
50
0.0036
0.2716
−0.148


Metabolite - 3951
61
0.0048
0.2716
−0.078


Metabolite - 1988
61
0.0068
0.2716
0.138


Aspartate
50
0.0072
0.2716
0.155


glutamic acid
50
0.0080
0.2716
0.135


3-methyl-L-histidine
61
0.0081
0.2716
0.064


Metabolite - 3073
50
0.0106
0.3157
0.066


Metabolite - 4275
50
0.0127
0.3293
−0.121


Metabolite - 2249
61
0.0135
0.3293
0.152


Metabolite - 2973
50
0.0156
0.3458
−0.029


Isobar-22-includes-glutamic acid-O-acetyl-L-serine
61
0.0204
0.3458
0.051


4-Guanidinobutanoic acid
61
0.0206
0.3458
0.088


Metabolite - 3183-possible-gamma-L-glutamyl-L-
61
0.0219
0.3458
0.073


phenylalanine


1-7-dimethylxanthine
61
0.0225
0.3458
0.220


Metabolite - 2688
61
0.0240
0.3458
0.068


gamma-glu-leu
61
0.0253
0.3458
0.100


Metabolite - 1142-possible-5-hydroxypentanoate-or-
61
0.0271
0.3458
0.163


beta-hydroxyisovaleric acid


Metabolite - 1345
61
0.0286
0.3458
0.201


Metabolite - 3078
50
0.0286
0.3458
−0.123


Metabolite - 7889
50
0.0295
0.3458
−0.084


Metabolite - 2567
61
0.0297
0.3458
0.074


Metabolite - 4364
50
0.0326
0.3645
−0.072


Metabolite - 4547
61
0.0353
0.3787
−0.114


Metabolite - 5976
61
0.0417
0.4297
0.180


Metabolite - 4511
50
0.0454
0.4412
0.078


Metabolite - 7846
50
0.0463
0.4412
0.114


Metabolite - 1208
61
0.0486
0.4412
0.128


Leucine
50
0.0517
0.4412
0.123


Metabolite - 1254
61
0.0528
0.4412
0.308


Metabolite - 2005
61
0.0561
0.4412
−0.107


Metabolite - 1834
61
0.0563
0.4412
0.246


Arginine
61
0.0564
0.4412
0.055


Metabolite - 3143
61
0.0576
0.4412
0.236


Metabolite - 7050
61
0.0612
0.4522
0.101


DL-indole-3-lactic acid
61
0.0639
0.4522
0.079


Isoleucine
50
0.0681
0.4522
0.108


Metabolite - 1597
61
0.0713
0.4522
0.033


Metabolite - 4767
50
0.0718
0.4522
−0.041


Metabolite - 6907
50
0.0729
0.4522
−0.056


Metabolite - 5788
61
0.0731
0.4522
0.272


Tryptophan
61
0.0742
0.4522
0.042


Metabolite - 1911
61
0.0744
0.4522
0.211


Metabolite - 1206-possible-methyltestosterone-and-
61
0.0759
0.4522
0.108


others


Creatine
61
0.0781
0.4550
−0.181


Metabolite - 3783
61
0.0868
0.4948
−0.062


Metabolite - 5848
61
0.0889
0.4966
0.089


1,5-anhydro-D-glucitol
50
0.0965
0.5251
0.042


Acetylpyrazine
61
0.0979
0.5251
−0.033


Metabolite - 4732
61
0.1037
0.5450
−0.123


N-5-aminocarbonyl-L-ornithine
50
0.1076
0.5493
0.059


inositol-1-phosphate
50
0.1086
0.5493
0.064


Valine
50
0.1159
0.5756
0.084


Metabolite - 2395
61
0.1197
0.5835
0.108


Metabolite - 1835
61
0.1291
0.5916
0.046


5-6-dihydrouracil
50
0.1318
0.5916
−0.071


Metabolite - 7147
61
0.1319
0.5916
−0.040


O-acetyl-L-carnitine-hydrochloride
61
0.1343
0.5916
−0.067


Biotin
61
0.1364
0.5916
0.113


Metabolite - 4522
50
0.1369
0.5916
−0.032


Metabolite - 4873
61
0.1415
0.5916
−0.118


Isobar-8-includes-anthranilic acid-salicylamide
61
0.1432
0.5916
0.104


Metabolite - 2981
50
0.1451
0.5916
−0.035


D-quinic acid
50
0.1467
0.5916
0.166


4-O-beta-galactopyranosyl-D-mannopyranose
61
0.1480
0.5916
−0.051


Glycerol
50
0.1497
0.5916
−0.074


Histamine
61
0.1501
0.5916
−0.051


citric acid
50
0.1546
0.6009
−0.048


Metabolite - 7888
50
0.1571
0.6016
−0.039


Metabolite - 6270
50
0.1600
0.6043
−0.108


Metabolite - 5234
61
0.1624
0.6049
−0.113


Metabolite - 6467
50
0.1695
0.6069
−0.082


Metabolite - 1819
61
0.1705
0.6069
0.045


Metabolite - 7765
61
0.1712
0.6069
0.200


Metabolite - 1193-confirmed-3-indoxyl-sulfate
61
0.1720
0.6069
0.076


Metabolite - 2185
61
0.1774
0.6088
0.050


Metabolite - 5086
61
0.1812
0.6088
−0.231


Metabolite - 3044
61
0.1862
0.6088
−0.065


Metabolite - 1127
61
0.1869
0.6088
−0.044


Caffeine
61
0.1910
0.6088
0.232


1-oleoyl-rac glycerol
50
0.1920
0.6088
0.073


Metabolite - 3114
50
0.1977
0.6088
0.063


Metabolite - 5728
61
0.2012
0.6088
0.082


Lysine
50
0.2036
0.6088
0.071


malic acid
50
0.2055
0.6088
0.096


Glutamine
50
0.2099
0.6088
−0.088


Metabolite - 5247
61
0.2145
0.6088
−0.078


Phosphate
50
0.2183
0.6088
−0.028


Metabolite - 1831-possible-Cl-adduct-of-citrulline
61
0.2233
0.6088
−0.032


Isobar-6-includes-valine-betaine
61
0.2240
0.6088
0.022


Metabolite - 3077
50
0.2250
0.6088
0.031


Metabolite - 7815
61
0.2254
0.6088
0.014


Metabolite - 1656
61
0.2265
0.6088
−0.043


Metabolite - 5730
61
0.2268
0.6088
−0.035


Metabolite - 3087
50
0.2277
0.6088
0.116


Allantoin
50
0.2292
0.6088
−0.135


Metabolite - 3040
50
0.2305
0.6088
0.005


(p-Hydroxyphenyl)lactic acid
50
0.2311
0.6088
0.049


Metabolite - 2056
61
0.2316
0.6088
−0.028


Phenylalanine
61
0.2348
0.6088
0.015


Metabolite - 2825-possible-Riluzole
61
0.2389
0.6088
−0.079


Metabolite - 3097
50
0.2394
0.6088
0.060


Metabolite - 6488
50
0.2426
0.6088
0.059


glyceric acid
50
0.2438
0.6088
0.039


gamma-L-glutamyl-L-tyrosine
61
0.2464
0.6088
0.034


Metabolite - 1092
61
0.2481
0.6088
0.140


Metabolite - 3025
50
0.2493
0.6088
0.022


Oleic-Acid
50
0.2497
0.6088
−0.070


Metabolite - 3109
50
0.2498
0.6088
0.052


Metabolite - 1086
61
0.2539
0.6133
0.118


glycochenodeoxycholic acid
61
0.2640
0.6230
0.111


Alanine
50
0.2648
0.6230
0.063


Metabolite - 2561
61
0.2659
0.6230
−0.053


Metabolite - 2952
50
0.2672
0.6230
−0.216


Biliverdin
61
0.2778
0.6387
0.068


Metabolite - 3088
50
0.2787
0.6387
0.047


methyl-indole-3-acetate
61
0.2994
0.6803
0.072


Metabolite - 5791
61
0.3028
0.6812
−0.333


Metabolite - 3138
61
0.3091
0.6812
0.038


Metabolite - 1335
61
0.3102
0.6812
0.079


phosphoenolpyruvate
61
0.3110
0.6812
−0.030


alpha-tocopherol
50
0.3127
0.6812
0.245


Metabolite - 4906
61
0.3179
0.6812
0.074


Metabolite - 4363
61
0.3244
0.6812
−0.214


2-keto-L-gulonic acid
50
0.3248
0.6812
−0.051


Mannose
50
0.3267
0.6812
−0.043


Metabolite - 5403
50
0.3345
0.6812
0.023


Histidine
50
0.3346
0.6812
0.065


DL-homocysteine
61
0.3391
0.6812
0.056


Metabolite - 3832-possible-phenol-sulfate
61
0.3398
0.6812
0.116


beta-hydroxypyruvic acid
50
0.3419
0.6812
−0.033


Metabolite - 4627
61
0.3423
0.6812
0.080


sn-Glycerol-3-phosphate
50
0.3487
0.6812
−0.053


p-hydroxybenzaldehyde
61
0.3548
0.6812
−0.018


Metabolite - 1286
61
0.3578
0.6812
−0.025


Metabolite - 2168
61
0.3598
0.6812
−0.015


Metabolite - 5233
61
0.3612
0.6812
0.100


Metabolite - 2321
61
0.3624
0.6812
−0.068


hippuric acid
61
0.3627
0.6812
0.067


Metabolite - 4586
61
0.3636
0.6812
−0.027


Metabolite - 4510
50
0.3641
0.6812
−0.091


decanoic acid
50
0.3663
0.6812
−0.032


Metabolite - 4986
50
0.3674
0.6812
0.060


DL-pipecolic acid
61
0.3714
0.6812
−0.042


Metabolite - 2594
61
0.3719
0.6812
−1.133


Metabolite - 2074
61
0.3748
0.6812
0.072


Metabolite - 3653-Possible-stachydrine
61
0.3760
0.6812
−0.332


Isobar-21-includes-gamma-aminobutyryl-L-histidine-L-
61
0.3801
0.6839
−0.021


anserine


Ornithine
50
0.3874
0.6851
0.066


Metabolite - 2506
61
0.3875
0.6851
0.078


Uridine
61
0.3884
0.6851
−0.043


Isobar-19-includes-D-saccharic acid-1,5-anhydro-D-
61
0.3941
0.6897
−0.018


glucitol-2′-deoxy-D-galactose-2′-deoxy-D-glucose-L-


fucose-L-rhamnose


Metabolite - 2386
61
0.3983
0.6897
−0.032


Metabolite - 4362
50
0.3988
0.6897
−0.050


Metabolite - 3377
61
0.4022
0.6912
−0.188


benzoic acid
61
0.4136
0.6976
0.013


Thyroxine
61
0.4143
0.6976
0.036


Metabolite - 3003
50
0.4162
0.6976
−0.018


Metabolite - 6347
50
0.4186
0.6976
−0.031


Metabolite - 5349
50
0.4193
0.6976
−0.010


Metabolite - 7008
61
0.4255
0.6976
−0.035


Metabolite - 7177
61
0.4266
0.6976
0.075


Metabolite - 3830
61
0.4267
0.6976
0.083


Metabolite - 2053
61
0.4344
0.7059
−0.031


Metabolite - 6827
61
0.4388
0.7084
0.027


Metabolite - 5231
61
0.4413
0.7084
0.030


Metabolite - 3002
50
0.4508
0.7165
−0.111


heptadecanoic acid
50
0.4537
0.7165
0.023


Metabolite - 4806
50
0.4544
0.7165
0.033


Metabolite - 6272
50
0.4588
0.7193
−0.035


Metabolite - 2027
61
0.4715
0.7287
−0.024


Isobar-28-includes-L-threonine-L-allothreonine-L-
61
0.4748
0.7287
−0.021


homoserine-S-4-amino-2-hydroxybutyric acid


Metabolite - 2370
61
0.4757
0.7287
0.036


uric acid
50
0.4764
0.7287
0.038


Metabolite - 6246
50
0.4829
0.7287
−0.015


Metabolite - 3026
50
0.4832
0.7287
0.023


Metabolite - 3093
50
0.4838
0.7287
0.039


Metabolite - 5487 retired: eythrose
50
0.4913
0.7352
−0.023


Metabolite - 3019
50
0.4936
0.7352
0.012


Metabolite - 1498
61
0.5012
0.7364
0.027


arachidonic acid
50
0.5045
0.7364
−0.019


Fructose
50
0.5050
0.7364
−0.068


Metabolite - 4167
61
0.5054
0.7364
−0.028


Metabolite - 4428
61
0.5152
0.7447
0.025


2-hydroxybutyric acid
50
0.5167
0.7447
0.024


Metabolite - 3708
61
0.5199
0.7453
−0.093


Metabolite - 5983
50
0.5394
0.7508
0.020


Metabolite - 5346
50
0.5423
0.7508
−0.009


Lactate
50
0.5525
0.7508
−0.040


Metabolite - 2915
50
0.5540
0.7508
−0.019


Metabolite - 6551
61
0.5542
0.7508
−0.019


Isobar-1-includes-mannose-fructose-glucose-
61
0.5551
0.7508
0.013


galactose-alpha-L-sorbopyranose-Inositol-D-allose-D-


altrose-D-psicone


Urea
50
0.5563
0.7508
0.015


Metabolite - 3056
61
0.5570
0.7508
0.019


Metabolite - 6227
50
0.5590
0.7508
−0.041


Metabolite - 2986 retired: CL adduct of p-
50
0.5611
0.7508
−0.015


acetiminophen-beta-d-glucuronide


methionine-sulfoxide
61
0.5674
0.7508
0.020


1-methyl-guanidine
50
0.5729
0.7508
0.012


Metabolite - 4274
50
0.5752
0.7508
0.042


Metabolite - 3218
61
0.5796
0.7508
0.027


Isobar-13-includes-5-keto-D-gluconic acid-2-keto-L-
61
0.5816
0.7508
−0.035


gulonic acid-D-glucuronic acid-D-galacturonic acid


Hypoxanthine
61
0.5818
0.7508
−0.136


Metabolite - 3033-possible-threonine-deriv
50
0.5827
0.7508
−0.011


Metabolite - 1342-possible-phenylacetylglutamine-or-
61
0.5835
0.7508
0.035


formyl-N-acetyl-5-methoxykynurenamine


Metabolite - 3100
50
0.5865
0.7508
0.021


Glycine
50
0.5887
0.7508
0.048


3-phospho-d-glycerate
61
0.5903
0.7508
−0.026


Metabolite - 7009
61
0.5968
0.7508
0.015


Saccharopine
61
0.5975
0.7508
−0.022


L-alpha-glycerophosphorylcholine
61
0.5988
0.7508
−0.030


Metabolite - 3603
61
0.6010
0.7508
−0.033


Isobar-2-includes-2-aminoisobutyric acid-3-amino-
61
0.6019
0.7508
0.027


isobutyrate-2-amino-butyrate-4-aminobutanoic acid-


dimethylglycine-choline


Metabolite - 3707
61
0.6149
0.7508
−0.099


glycocholic acid
61
0.6167
0.7508
0.071


Metabolite - 2753
61
0.6211
0.7508
−0.031


Metabolite - 1085-possible-isolobinine-or-4-
61
0.6276
0.7508
−0.010


aminoestra-1,3-5-10-triene-3-17beta-diol


n-hexadecanoic acid
50
0.6314
0.7508
−0.016


Methionine
61
0.6324
0.7508
−0.046


Metabolite - 6711
61
0.6374
0.7508
0.016


Metabolite - 2568
61
0.6399
0.7508
0.053


Metabolite - 1261
61
0.6407
0.7508
0.033


Metabolite - 3030
50
0.6409
0.7508
0.010


Metabolite - 7146
61
0.6459
0.7508
0.013


Metabolite - 4252
50
0.6468
0.7508
−0.027


Metabolite - 1836
61
0.6472
0.7508
−0.024


Cholesterol
50
0.6482
0.7508
−0.010


Metabolite - 4611
50
0.6496
0.7508
0.013


L-kynurenine
61
0.6505
0.7508
−0.019


Metabolite - 6226
50
0.6528
0.7508
−0.027


Tyrosine
61
0.6534
0.7508
0.013


Metabolite - 5769
61
0.6540
0.7508
0.016


Metabolite - 7890
50
0.6553
0.7508
0.025


palmitoleic acid
50
0.6635
0.7570
−0.035


Metabolite - 7706
61
0.6695
0.7588
0.024


Metabolite - 1323-possible-p-cresol-sulfate
61
0.6708
0.7588
0.026


D-glucose
50
0.6847
0.7698
−0.007


Metabolite - 1111-possible-
61
0.6882
0.7698
−0.022


methylnitronitrosoguanidine-or-ethyl-


thiocarbamoylacetate


Metabolite - 6869
50
0.6891
0.7698
−0.026


hydroxyacetic acid
50
0.6927
0.7705
0.009


Metabolite - 3052
61
0.6990
0.7734
−0.016


Metabolite - 1975
61
0.7010
0.7734
−0.045


Metabolite - 2469
61
0.7106
0.7775
−0.017


Metabolite - 3772 pieces of lactate
61
0.7116
0.7775
0.038


Metabolite - 2319
61
0.7134
0.7775
−0.027


Bicine
61
0.7201
0.7816
0.016


Metabolite - 3972
61
0.7338
0.7891
−0.014


Aldosterone
61
0.7368
0.7891
0.021


Linoleic acid
50
0.7382
0.7891
−0.012


Metabolite - 4470
61
0.7388
0.7891
−0.009


Metabolite - 5907
50
0.7417
0.7891
−0.012


Metabolite - 3808
61
0.7467
0.7913
−0.018


Metabolite - 6346
50
0.7514
0.7929
−0.008


Threonine
50
0.7581
0.7929
−0.013


Metabolite - 4055
50
0.7602
0.7929
0.017


Metabolite - 2139
61
0.7638
0.7929
−0.013


Riboflavine
61
0.7643
0.7929
0.005


Metabolite - 7707
61
0.7680
0.7929
−0.009


Metabolite - 3129
61
0.7774
0.7929
0.014


Metabolite - 6126
61
0.7775
0.7929
0.028


Metabolite - 6931
50
0.7792
0.7929
0.008


Metabolite - 2269
61
0.7804
0.7929
0.112


Metabolite - 3441
61
0.7808
0.7929
−0.009


3-hydroxybutanoic acid
50
0.7864
0.7956
0.065


Metabolite - 3098
50
0.7970
0.8024
−0.006


Metabolite - 4931
61
0.8000
0.8024
0.015


Metabolite - 4624
50
0.8041
0.8024
−0.012


Metabolite - 1114
61
0.8051
0.8024
0.008


Metabolite - 2109
61
0.8094
0.8037
−0.004


Metabolite - 3017
50
0.8198
0.8077
0.007


Metabolite - 7762
61
0.8223
0.8077
0.024


Theobromine
61
0.8271
0.8077
0.016


Metabolite - 1914
61
0.8278
0.8077
0.055


Metabolite - 2548-possible-CI-adduct-of-uric acid
61
0.8285
0.8077
−0.008


Metabolite - 5887
61
0.8423
0.8181
0.026


Metabolite - 2100
61
0.8454
0.8181
−0.007


Metabolite - 3012
50
0.8486
0.8181
−0.004


5-oxoproline
50
0.8520
0.8181
0.005


Inosine
61
0.8544
0.8181
−0.028


Arabinose
50
0.8663
0.8230
−0.006


Carnitine
61
0.8690
0.8230
0.005


Metabolite - 7336
61
0.8716
0.8230
0.013


Metabolite - 2390
61
0.8719
0.8230
−0.006


Nonanate
50
0.8793
0.8230
0.002


Metabolite - 4658
61
0.8866
0.8230
−0.004


tetradecanoic acid
50
0.8897
0.8230
−0.007


Metabolite - 3125
61
0.8931
0.8230
0.008


Metabolite - 1110
61
0.8945
0.8230
0.066


Metabolite - 3131 retired: N4 adduct of indole-3-acetic
61
0.8955
0.8230
0.005


acid


Metabolite - 4795
50
0.8961
0.8230
−0.011


Metabolite - 6963
50
0.8987
0.8230
0.007


Metabolite - 3094
50
0.8995
0.8230
0.004


octadecanoic acid
50
0.9048
0.8230
0.003


Metabolite - 6269
50
0.9056
0.8230
0.003


Metabolite - 6326
50
0.9087
0.8230
0.002


Metabolite - 4523
50
0.9126
0.8238
0.002


Metabolite - 6955
50
0.9190
0.8268
−0.003


Metabolite - 2546
61
0.9264
0.8278
−0.024


Metabolite - 1215
61
0.9296
0.8278
0.005


Metabolite - 3099
50
0.9354
0.8278
0.004


Metabolite - 2559
61
0.9423
0.8278
0.009


Metabolite - 5847
50
0.9441
0.8278
0.003


Metabolite - 4612
61
0.9484
0.8278
0.005


pantothenic acid
61
0.9493
0.8278
−0.026


Metabolite - 3022
50
0.9526
0.8278
−0.001


dehydroisoandrosterone-3-sulfate-sodium-salt-hydrate
61
0.9536
0.8278
0.019


Metabolite - 5673
61
0.9538
0.8278
−0.004


n-dodecanoate
50
0.9541
0.8278
0.005


Metabolite - 4351
61
0.9632
0.8291
0.005


Metabolite - 1264
61
0.9643
0.8291
0.009


Metabolite - 4020
50
0.9691
0.8291
−0.001


Metabolite - 7807
61
0.9695
0.8291
0.001


N-6-trimethyl-l-lysine
61
0.9737
0.8291
0.038


Metabolite - 5366
50
0.9760
0.8291
−0.002


Inositol
50
0.9787
0.8291
0.001


meso-erythritol
50
0.9803
0.8291
−0.001


oxalic acid
61
0.9900
0.8346
0.000


Praline
61
0.9964
0.8374
0.000


Serine
50
1.0000
0.8378
0.104
















TABLE 18










T-tests comparing month 1 with month 12.















% Change






with disease


COMPOUND
LIB_ID
p-value
q-value
progression














Metabolite - 3073
50
0
1.00E−04
−19%


Metabolite - 5233
61
0
0.0022
−46%


Metabolite - 3183-possible-gamma-L-glutamyl-L-
61
0
0.0022
−22%


phenylalanine


gamma-L-glutamyl-L-tyrosine
61
1.00E−04
0.0056
−22%


Metabolite - 6246
50
1.00E−04
0.0056
−17%


Metabolite - 3022
50
4.00E−04
0.0172
−14%


O-acetyl-L-carnitine-hydrochloride
61
7.00E−04
0.0234
39%


Metabolite - 5887
61
8.00E−04
0.0234
−49%


Metabolite - 4275
50
0.0014
0.0396
49%


Metabolite - 6907
50
0.0018
0.0435
−19%


Metabolite - 1208
61
0.002
0.0435
−45%


Metabolite - 3088
50
0.0022
0.0435
−26%


Metabolite - 4611
50
0.0026
0.0435
−15%


Isobar-19-includes-D-saccharic acid-1-5-anhydro-
61
0.0027
0.0435
28%


D-glucitol-2-deoxy-D-galactose-2-deoxy-D-


glucose-L-fucose-L-rhamnose


Metabolite - 7009
61
0.0027
0.0435
−23%


Metabolite - 2567
61
0.0028
0.0435
−17%


gamma-glu-leu
61
0.0035
0.0486
−16%


Metabolite - 3012
50
0.0036
0.0486
−9%


hippuric acid
61
0.0037
0.0486
−37%


L-alpha-glycerophosphorylcholine
61
0.0055
0.0668
−43%


Metabolite - 3026
50
0.0059
0.0668
−14%


glyceric acid
50
0.006
0.0668
−16%


Metabolite - 4769
50
0.0064
0.0668
29%


Metabolite - 3830
61
0.0064
0.0668
−37%


inositol-1-phosphate
50
0.0076
0.076
−25%


Metabolite - 3078
50
0.0081
0.0772
37%


Metabolite - 3109
50
0.0087
0.0772
−33%


Isobar-22-includes-glutamic acid-O-acetyl-L-
61
0.0094
0.0772
−11%


serine


Metabolite - 3098
50
0.0095
0.0772
−14%


Metabolite - 3114
50
0.0096
0.0772
−34%


Biotin
61
0.0096
0.0772
−32%


Metabolite - 7177
61
0.0109
0.0846
36%


Glutamine
50
0.0133
0.0987
65%


Metabolite - 4020
50
0.0136
0.0987
−14%


glycochenodeoxycholic acid
61
0.0139
0.0987
−33%


4-Guanidinobutanoic acid
61
0.0144
0.0999
−16%


Metabolite - 2319
61
0.0156
0.1048
−33%


Metabolite - 3019
50
0.0189
0.1235
−8%


Metabolite - 3094
50
0.0202
0.1261
−14%


Metabolite - 2688
61
0.0203
0.1261
−16%


Metabolite - 3138
61
0.022
0.1335
−20%


Arabinose
50
0.0278
0.1619
−22%


Metabolite - 1988
61
0.0286
0.1619
−21%


1-7-dimethylxanthine
61
0.0295
0.1619
−27%


Metabolite - 5346
50
0.0305
0.1619
−12%


Metabolite - 3143
61
0.0305
0.1619
−27%


Metabolite - 3025
50
0.0311
0.1619
−9%


Metabolite - 3077
50
0.0312
0.1619
−15%


methionine-sulfoxide
61
0.0335
0.1702
−19%


Metabolite - 4627
61
0.0357
0.1749
−44%


3-methyl-L-histidine
61
0.0359
0.1749
−15%


Metabolite - 7050
61
0.0375
0.1782
−23%


D-quinic acid
50
0.0385
0.1782
−43%


Metabolite - 2546
61
0.0387
0.1782
−37%


Metabolite - 5907
50
0.0394
0.1782
−11%


Metabolite - 7846
50
0.0438
0.1945
−13%


Metabolite - 2568
61
0.0459
0.1953
−26%


N-6-trimethyl-l-lysine
61
0.0459
0.1953
−22%


1-5-anhydro-D-glucitol
50
0.0471
0.1953
−8%


Metabolite - 6272
50
0.0477
0.1953
−15%


Metabolite - 7089
61
0.0485
0.1953
−13%


Methionine
61
0.0487
0.1953
48%


Metabolite - 2005
61
0.051
0.1981
39%


Metabolite - 4986
50
0.0511
0.1981
−14%


Metabolite - 4511
50
0.0517
0.1981
−12%


Metabolite - 1286
61
0.0568
0.214
13%


Metabolite - 5231
61
0.0603
0.2241
−18%


Oleic-Acid
50
0.0615
0.2249
31%


Metabolite - 3783
61
0.068
0.2397
18%


Metabolite - 5349
50
0.0707
0.2397
−6%


Tyrosine
61
0.0709
0.2397
−8%


Metabolite - 3707
61
0.0712
0.2397
43%


Metabolite - 5976
61
0.0712
0.2397
−34%


palmitoleic acid
50
0.0713
0.2397
29%


3-phospho-d-glycerate
61
0.0745
0.2473
−19%


Arginine
61
0.0763
0.2499
−12%


Metabolite - 3653-Possible-stachydrine
61
0.0809
0.2566
−34%


Metabolite - 2386
61
0.0814
0.2566
13%


Metabolite - 3951
61
0.0815
0.2566
11%


Metabolite - 4767
50
0.0858
0.2631
−7%


Metabolite - 3033-possible-threonine-deriv
50
0.0866
0.2631
−6%


Metabolite - 3603
61
0.0867
0.2631
−21%


Metabolite - 4612
61
0.0881
0.2642
−19%


Caffeine
61
0.0912
0.2701
−36%


Metabolite - 6126
61
0.0926
0.2711
−21%


Metabolite - 1345
61
0.0948
0.2717
−35%


Metabolite - 6711
61
0.095
0.2717
23%


Histamine
61
0.0968
0.2737
24%


Inositol
50
0.1018
0.2828
−10%


Mannose
50
0.1023
0.2828
12%


Metabolite - 7008
61
0.1061
0.2903
−11%


Metabolite - 6326
50
0.1075
0.2907
−7%


Metabolite - 1142-possible-5-hydroxypentanoate-
61
0.1096
0.2933
−14%


or-beta-hydroxyisovaleric acid


DL-indole-3-lactic acid
61
0.1113
0.2947
−9%


meso-erythritol
50
0.1215
0.3165
−7%


Metabolite - 5728
61
0.1221
0.3165
−32%


Fructose
50
0.1265
0.3219
−32%


alpha-tocopherol
50
0.1268
0.3219
−21%


Bicine
61
0.1298
0.3237
17%


Isobar-8-includes-anthranilic acid-salicylamide
61
0.1301
0.3237
−15%


Metabolite - 1092
61
0.1361
0.3339
−36%


Metabolite - 4252
50
0.1368
0.3339
−23%


Metabolite - 1111-possible-
61
0.1389
0.3339
19%


methylnitronitrosoguanidine-or-ethyl-


thiocarbamoylacetate


Metabolite - 3052
61
0.1396
0.3339
−21%


Metabolite - 3044
61
0.142
0.3364
24%


Metabolite - 7707
61
0.1433
0.3364
−8%


Metabolite - 7807
61
0.1483
0.342
−13%


Metabolite - 6346
50
0.1484
0.342
−6%


Metabolite - 3030
50
0.1541
0.3519
−6%


Metabolite - 1127
61
0.1569
0.355
13%


Metabolite - 5487 retired: eythrose
50
0.1652
0.3699
−13%


Aldosterone
61
0.1665
0.3699
10%


Metabolite - 3131 retired: N4 adduct of indole-3-
61
0.1733
0.3817
−12%


acetic acid


Metabolite - 2168
61
0.1783
0.3885
9%


Metabolite - 2561
61
0.1795
0.3885
22%


(p-Hydroxyphenyl)lactic acid
50
0.1874
0.402
−15%


Metabolite - 6269
50
0.1919
0.4082
−7%


Metabolite - 2027
61
0.1969
0.4153
18%


Metabolite - 2139
61
0.2034
0.4255
12%


L-kynurenine
61
0.2054
0.426
−6%


Metabolite - 1911
61
0.2116
0.4351
−16%


Glycine
50
0.2148
0.4383
69%


Hypoxanthine
61
0.22
0.4429
73%


Metabolite - 6955
50
0.2213
0.4429
−11%


Tryptophan
61
0.2237
0.4429
−5%


DL-pipecolic acid
61
0.2258
0.4429
−9%


Metabolite - 3002
50
0.226
0.4429
−27%


Isobar-1-includes-mannose-fructose-glucose-
61
0.2326
0.4523
−6%


galactose-alpha-L-sorbopyranose-Inositol-D-


allose-D-altrose-D-psicone


1-methyl-guanidine
50
0.2407
0.4644
−5%


Glycerol
50
0.2485
0.4757
12%


Metabolite - 3097
50
0.2611
0.496
−12%


2-hydroxybutyric acid
50
0.2663
0.5021
11%


Metabolite - 1193-confirmed-3-indoxyl-sulfate
61
0.2685
0.5024
−11%


n-hexadecanoic acid
50
0.2754
0.5115
9%


pantothenic acid
61
0.2801
0.5165
33%


Metabolite - 6226
50
0.2843
0.5167
−9%


Riboflavine
61
0.2844
0.5167
−24%


Serine
50
0.2881
0.5196
28%


Metabolite - 5847
50
0.2914
0.5218
−21%


n-dodecanoate
50
0.2959
0.526
−22%


Creatinine
61
0.3005
0.528
−10%


Metabolite - 6347
50
0.303
0.528
−8%


Praline
61
0.3049
0.528
−6%


Metabolite - 1835
61
0.3055
0.528
−7%


Metabolite - 1831-possible-Cl-adduct-of-citrulline
61
0.3104
0.5328
−6%


Metabolite - 2506
61
0.3135
0.5331
−12%


Metabolite - 6963
50
0.3198
0.5331
−14%


Metabolite - 2469
61
0.3199
0.5331
12%


Metabolite - 1086
61
0.32
0.5331
−18%


Metabolite - 5086
61
0.3213
0.5331
9%


Metabolite - 4364
50
0.3352
0.5511
8%


Metabolite - Metabolite - 5982 retired: 1-oleoyl-
50
0.3366
0.5511
−13%


rac glycerol


Metabolite - 4931
61
0.3422
0.5567
22%


Metabolite - 7765
61
0.3468
0.5586
−21%


Metabolite - 1261
61
0.3479
0.5586
13%


Metabolite - 2053
61
0.3527
0.5626
15%


Metabolite - 6931
50
0.3581
0.566
−5%


Metabolite - 6227
50
0.3601
0.566
−11%


Metabolite - 3708
61
0.3616
0.566
−15%


Metabolite - 3093
50
0.3655
0.5686
−9%


Metabolite - 4523
50
0.3698
0.5716
−7%


Metabolite - 2269
61
0.375
0.5761
17%


2-keto-L-gulonic acid
50
0.3795
0.5795
−8%


dehydroisoandrosterone-3-sulfate-sodium-salt-
61
0.3853
0.5847
−5%


hydrate


Metabolite - 4624
50
0.3923
0.5885
−9%


Linoleic acid
50
0.3925
0.5885
6%


Metabolite - 1656
61
0.4005
0.5945
9%


Inosine
61
0.4021
0.5945
41%


Metabolite - 5366
50
0.4037
0.5945
−14%


Metabolite - 4510
50
0.4134
0.5952
−17%


Cholesterol
50
0.4137
0.5952
−3%


Isobar-28-includes-L-threonine-L-allothreonine-L-
61
0.4184
0.5952
−6%


homoserine-S-4-amino-2-hydroxybutyric acid


Metabolite - 4586
61
0.4212
0.5952
6%


Metabolite - 1206-possible-methyltestosterone-
61
0.426
0.5952
−14%


and-others


Metabolite - 1975
61
0.4288
0.5952
−11%


Metabolite - 6869
50
0.4294
0.5952
−8%


Metabolite - 4362
50
0.4312
0.5952
−8%


Metabolite - 2390
61
0.4321
0.5952
−7%


Metabolite - 2074
61
0.4326
0.5952
−14%


Metabolite - 3100
50
0.4338
0.5952
−12%


arachidonic acid
50
0.4369
0.5952
−5%


Threonine
50
0.4376
0.5952
16%


Metabolite - 4658
61
0.4377
0.5952
−7%


Metabolite - 4522
50
0.4516
0.61
4%


Alanine
50
0.4534
0.61
16%


Thyroxine
61
0.4594
0.6135
10%


hydroxyacetic acid
50
0.461
0.6135
−5%


Metabolite - 7815
61
0.4664
0.6148
−3%


N-5-aminocarbonyl-L-ornithine
50
0.4669
0.6148
−7%


Metabolite - 2594
61
0.4827
0.6323
146%


glycocholic acid
61
0.4869
0.6342
−16%


Metabolite - 2395
61
0.4893
0.6342
−8%


Metabolite - 1215
61
0.4933
0.6359
9%


Metabolite - 2548-possible-Cl-adduct-of-uric acid
61
0.5035
0.6359
8%


Valine
50
0.5043
0.6359
12%


Metabolite - 7890
50
0.505
0.6359
−8%


Metabolite - 5848
61
0.5063
0.6359
6%


Metabolite - 2559
61
0.5077
0.6359
−11%


Lysine
50
0.5103
0.6359
12%


Metabolite - 3377
61
0.512
0.6359
15%


Acetylpyrazine
61
0.5136
0.6359
−4%


Metabolite - 3832-possible-phenol-sulfate
61
0.5245
0.6438
−13%


Metabolite - 4363
61
0.5294
0.6438
13%


Ornithine
50
0.5303
0.6438
23%


Metabolite - 7888
50
0.5303
0.6438
−3%


Metabolite - 7336
61
0.5421
0.6479
−15%


Metabolite - 3056
61
0.5457
0.6479
4%


Metabolite - 7706
61
0.5477
0.6479
7%


Metabolite - 4732
61
0.5482
0.6479
12%


Isobar-13-includes-5-keto-D-gluconic acid-2-keto-
61
0.5509
0.6479
−11%


L-gulonic acid-D-glucuronic acid-D-galacturonic


acid


Metabolite - 4274
50
0.5512
0.6479
18%


Metabolite - 1834
61
0.5526
0.6479
4%


decanoic acid
50
0.5545
0.6479
3%


Metabolite - 3099
50
0.5574
0.6483
−6%


Saccharopine
61
0.5656
0.6547
−7%


Metabolite - 5983
50
0.569
0.6556
8%


Phenylalanine
61
0.5746
0.6581
−3%


oxalic acid
61
0.5765
0.6581
−6%


Uridine
61
0.592
0.6704
5%


5-oxoproline
50
0.5956
0.6704
−7%


malic acid
50
0.5961
0.6704
13%


Metabolite - 1264
61
0.598
0.6704
32%


Metabolite - 4806
50
0.6192
0.6911
−5%


Metabolite - 3441
61
0.6232
0.6925
5%


Metabolite - 1914
61
0.637
0.6993
9%


Leucine
50
0.6405
0.6993
10%


Metabolite - 1836
61
0.6407
0.6993
−6%


Metabolite - 1085-possible-isolobinine-or-4-
61
0.6509
0.7035
−4%


aminoestra-1-3-5-10-triene-3-17beta-diol


Metabolite - 2825-possible-Riluzole
61
0.6519
0.7035
−7%


Metabolite - 6551
61
0.6584
0.7035
13%


Metabolite - 1498
61
0.6621
0.7035
−4%


Metabolite - 5791
61
0.6626
0.7035
4%


Metabolite - 1335
61
0.6691
0.7035
−9%


citric acid
50
0.6694
0.7035
−3%


Metabolite - 3129
61
0.6711
0.7035
−5%


beta-hydroxypyruvic acid
50
0.679
0.7035
3%


Isobar-6-includes-valine-betaine
61
0.679
0.7035
−2%


methyl-indole-3-acetate
61
0.6795
0.7035
−6%


Metabolite - 6488
50
0.6842
0.7035
−8%


D-glucose
50
0.6866
0.7035
−1%


Metabolite - 2249
61
0.6868
0.7035
−3%


Metabolite - 4055
50
0.6869
0.7035
−8%


Metabolite - 4470
61
0.6946
0.7038
−2%


tetradecanoic acid
50
0.6972
0.7038
5%


Metabolite - 2056
61
0.6983
0.7038
3%


Metabolite - 6270
50
0.7005
0.7038
6%


Metabolite - 4906
61
0.7013
0.7038
11%


Metabolite - 3003
50
0.7059
0.7055
−3%


Metabolite - 7889
50
0.7108
0.7076
−3%


Metabolite - 1254
61
0.7149
0.7088
−15%


Metabolite - 3040
50
0.7211
0.7104
1%


DL-homocysteine
61
0.7222
0.7104
−5%


Metabolite - 5730
61
0.7295
0.7148
3%


Metabolite - 4167
61
0.7332
0.7156
−3%


Metabolite - 1342-possible-
61
0.7373
0.7168
3%


phenylacetylglutamine-or-formyl-N-acetyl-5-


methoxykynurenamine


glutamic acid
50
0.7453
0.7182
5%


Metabolite - 1114
61
0.749
0.7182
7%


Metabolite - 2370
61
0.7509
0.7182
−4%


Metabolite - 7762
61
0.755
0.7182
6%


Phosphate
50
0.7557
0.7182
2%


Metabolite - 4795
50
0.7587
0.7182
−4%


Isobar-21-includes-gamma-aminobutyryl-L-
61
0.759
0.7182
2%


histidine-L-anserine


Metabolite - 2986
50
0.7649
0.7211
2%


Metabolite - 5403
50
0.7693
0.7225
2%


Metabolite - 5788
61
0.7751
0.7229
−10%


Nonanate
50
0.7809
0.7229
0%


Metabolite - 1819
61
0.7832
0.7229
−2%


benzoic acid
61
0.7845
0.7229
2%


Metabolite - 6467
50
0.7846
0.7229
−4%


Metabolite - 2981
50
0.7884
0.7229
−1%


Metabolite - 3017
50
0.7901
0.7229
−2%


Histidine
50
0.7975
0.7246
4%


Allantoin
50
0.7998
0.7246
−8%


Metabolite - 2753
61
0.8025
0.7246
−3%


Metabolite - 2973
50
0.8036
0.7246
1%


Metabolite - 5247
61
0.8088
0.7266
2%


Metabolite - 1323-possible-p-cresol-sulfate
61
0.8124
0.7266
2%


Metabolite - 7146
61
0.8146
0.7266
−2%


Metabolite - 3087
50
0.8213
0.7271
8%


Metabolite - 4547
61
0.8235
0.7271
2%


4-O-beta-galactopyranosyl-D-mannopyranose
61
0.8239
0.7271
3%


Isoleucine
50
0.829
0.7277
4%


Metabolite - 3218
61
0.8304
0.7277
4%


Metabolite - 5673
61
0.8384
0.7321
3%


Metabolite - 4428
61
0.8469
0.737
2%


uric acid
50
0.8635
0.7488
−2%


Metabolite - 2100
61
0.8676
0.7492
1%


octadecanoic acid
50
0.87
0.7492
−1%


sn-Glycerol-3-phosphate
50
0.8783
0.7538
−2%


Metabolite - 3125
61
0.8898
0.761
−2%


Metabolite - 2321
61
0.8933
0.7613
2%


phosphoenolpyruvate
61
0.9033
0.7635
1%


Metabolite - 6827
61
0.906
0.7635
−1%


Metabolite - 1597
61
0.9085
0.7635
0%


heptadecanoic acid
50
0.9106
0.7635
−1%


Metabolite - 2185
61
0.9112
0.7635
1%


Metabolite - 3808
61
0.9235
0.766
−1%


Biliverdin
61
0.9263
0.766
1%


p-hydroxybenzaldehyde
61
0.9273
0.766
0%


5-6-dihydrouracil
50
0.9289
0.766
1%


Metabolite - 2109
61
0.9316
0.766
−1%


Lactate
50
0.9331
0.766
2%


Metabolite - 7147
61
0.9393
0.766
−1%


Isobar-2-includes-2-aminoisobutyric acid-3-amino-
61
0.9407
0.766
−2%


isobutyrate-2-amino-butyrate-4-aminobutanoic


acid-dimethylglycine-choline


Metabolite - 2711
61
0.9442
0.766
−2%


Theobromine
61
0.9449
0.766
2%


Aspartate
50
0.9491
0.7662
1%


Metabolite - 1110
61
0.9513
0.7662
−1%


Metabolite - 2952
50
0.9725
0.7739
2%


Urea
50
0.9731
0.7739
1%


Carnitine
61
0.9734
0.7739
1%


Metabolite - 4873
61
0.9752
0.7739
1%


3-hydroxybutanoic acid
50
0.9785
0.7739
1%


Metabolite - 2915
50
0.9841
0.7739
−1%


Creatine
61
0.986
0.7739
−1%


Metabolite - 5769
61
0.9874
0.7739
0%


Metabolite - 3972
61
0.9891
0.7739
0%


Metabolite - 5234
61
0.9932
0.7739
0%


Metabolite - 4351
61
0.9951
0.7739
0%









Example 10
Analytical Characterization of Isobars, Unnamed Biomarkers, and Unnamed Non-Biomarker Compounds

Table 19 below includes analytical characteristics of each of the Isobars and the unnamed metabolites listed in Tables 3, 4, 5, 6, 7, 8, 16, 17, and 18 above. The table includes, for each listed Isobar and Metabolite, the retention time (RT), retention index (RI), mass, quant mass, and polarity obtained using the analytical methods described above. “Mass” refers to the mass of the C12 isotope of the parent ion used in quantification of the compound. The values for “Quant Mass” give an indication of the analytical method used for quantification: “Y” indicates GC-MS and “1” indicates LC-MS. “Polarity” indicates the polarity of the quantitative ion as being either positive (+) or negative (−).

TABLE 19Analytical Characteristics of Isobars and Unnamed MetabolitesNamePlatformRTRIMASSQUANT_MASSPolarityIsobar-13-includes-5-keto-D-gluconic acid; 2-LC1.41530193.11−iketo-L-gulonic acid-D-glucuronic acid; D-galacturonic acidIsobar-19-includes-D-saccharic acid; 2-deoxy-D-LC1.5517002091galactose; 2-deoxy-D-glucose; L-fucose; L-rhamnoseIsobar-1-includes-mannose-fructose-glucose-LC1.451481224.91galactose-alpha-L-sorbopyranose-Inositol-D-alloseIsobar-21-includes-gamma-aminobutyryl-L-LC1.5916202411+histidine-L-anserineIsobar-22-includes-glutamic acid; O-acetyl-L-LC1.551635148.01+iserineIsobar-27-includes-L-kynurenine-alpha-2-LC8.238470209.11+diamino-gamma-oxobenzenebutanoic acidIsobar-28-includes-L-threonine-L-allothreonineLC1.4615251201+Isobar-2-includes-3-amino-isobutyrate-2-amino-LC1.61671104.11+butyrate-4-aminobutanoic acid-dimethylglycine-choline-Isobar-5-includes-asparagine-ornithineLC1.51395133.11+Isobar-6-includes-valine-betaineLC2.132160118.11+Isobar-8-includes-anthranilic acid-salicylamideLC1010116138.11+Isobar-9-includes-sucrose-beta-D-lactose-D-LC1.61605386.91trehalose-D-cellobiose-D-Maltose-palatinose-melibiose-alpha-D-lactoseMetabolite - 1063LC8.58845391.41+Metabolite - 1064LC1.91950258.11+Metabolite - 1065LC9.6698707691+Metabolite - 1066LC1.571520215.11Metabolite - 1068LC1.441490203.11+Metabolite - 1071LC15.215445279.31+Metabolite - 1073LC15.4156303381+Metabolite - 1085 isolobinine or 4-aminoestra-LC15.815964288.11+1,3-5-10-triene-3-17beta-diolMetabolite - 1086LC4.564811294.11+Metabolite - 1087LC9.29440371.71+Metabolite - 1088LC13.113298369.11Metabolite - 1089LC2.012017346.91+Metabolite - 1092LC11.51168410061+Metabolite - 1104LC2.4324102011Metabolite - 1108LC4.154369144.11+Metabolite - 1110LC11.711841269.11Metabolite - 1111-methylnitronitrosoguanidineLC2.692782148.11+or ethyl-thiocarbamoylacetateMetabolite - 1113LC4.915190204.21+Metabolite - 1114LC2.192198104.11+Metabolite - 1116LC4.24480103.41Metabolite - 1126LC3.043188175.11+Metabolite - 1127LC12.212369363.11+Metabolite - 1129LC5.165419260.11+Metabolite - 1131LC1.6216491211+Metabolite - 1132LC1.661689291.11Metabolite - 1133LC1.6316363151+Metabolite - 1142-5-hydroxypentanoat or beta-LC8.5487391171hydroxyisovaleric acidMetabolite - 1161LC2.3123501351Metabolite - 1181LC1.441486203.11+Metabolite - 1192LC8.788983129.11Metabolite - 1193 3-indoxyl-sulfateLC8.859031212.11Metabolite - 1201LC5.7559991821+Metabolite - 1203LC9.119288510.21+Metabolite - 1206 methyltestosteroneLC15.315475303.21+iMetabolite - 1208LC15.315494319.41Metabolite - 1209LC8.899077.8426.91+Metabolite - 121GC5.541161.4102.079Y+Metabolite - 1215LC8.969390550.11+iMetabolite - 1216LC1.61631.4343.91Metabolite - 1220LC15.215403319.21+Metabolite - 1242LC8.438627.6355.91+Metabolite - 1247LC14.814959448.31Metabolite - 1251LC16.316406718.21+Metabolite - 1252LC8.128326229.91+Metabolite - 1254LC9.89987.5733.41+Metabolite - 1261LC10.710905528.41+Metabolite - 1262LC9.9710163808.91+Metabolite - 1264LC10.710879617.81Metabolite - 1265LC15.315440361.91+Metabolite - 128GC10.11697.1227.171Y+Metabolite - 1281LC8.398591.3328.11+Metabolite - 1283LC9.049244.5434.81+Metabolite - 1285LC2.332342280.11+Metabolite - 1286LC14.4145802291+Metabolite - 1288LC2.112120.53021Metabolite - 1289LC8.969139.7338.41+Metabolite - 1301LC8.628819243.31+Metabolite - 1302LC15.215391279.21+Metabolite - 1304LC9.749930.5729.51+Metabolite - 1322 retired: citric acidLC2.782892190.91Metabolite - 1323LC9.319719.81871Metabolite - 1323 p-cresol-sulfateLC9.319719.81871Metabolite - 1324LC3.193393.91911Metabolite - 1327LC13.213706585.41+Metabolite - 1328LC3.213430.7210.11+Metabolite - 1329LC2.692791210.11+Metabolite - 1331LC12.913343239.21Metabolite - 1332LC2.272400280.11+Metabolite - 1333 retired: citric acidLC3.053194.6321.91+Metabolite - 1334LC2.062217.71351Metabolite - 1335LC8.749162.2367.21+Metabolite - 1336 retired: carnitineLC1.872039162.21+Metabolite - 1337LC8.2886961731Metabolite - 1340LC3.764214.5173.11Metabolite - 1342 phenylacetylglutamine-or-LC9.049459.4265.21+formyl-N-acetyl-5-methoxykynurenamineMetabolite - 1343 retired: p-hydroxyphenyllacticLC8.919327181.21+acidMetabolite - 1344 retired: Na adduct of citricLC3.293511.8406.71+acidMetabolite - 1345LC13.314600369.31−iMetabolite - 1346LC1.271449.51131Metabolite - 1347LC9.379777.7247.21+Metabolite - 1349 retired: Isobar 7LC3.53876323.91+Metabolite - 1350LC13.814249909.81+Metabolite - 1351 retired: urea adduct of Isobar 6LC1.771936.5177.91+Metabolite - 1353GC13.22104.2371.94Y+Metabolite - 1358GC12.72038.3288.015Y+Metabolite - 136GC6.071221.7211.034Y+Metabolite - 1368LC8.188607.4184.11+Metabolite - 1372 retired: 2-hydroxyhippuricLC9.6310038194.11acidMetabolite - 1373GC10.31749.6218.014Y+Metabolite - 1379 retired: hippuric acidLC9.069454.8180.11+Metabolite - 138GC10.81770.3156.123Y+Metabolite - 1383LC8.669077.9370.11Metabolite - 1385LC11.912303225.11Metabolite - 1388LC12.512940456.91+Metabolite - 1389LC13.614111425.31Metabolite - 1397LC15.716277425.51+Metabolite - 141GC6.131228.9221.119Y+Metabolite - 1414GC10.61788.9259.012Y+Metabolite - 145GC12.71991.9204.129Y+Metabolite - 1457LC1.591675188.21+Metabolite - 1458 retired: hypoxanthineLC6.56815.7137.11+Metabolite - 146GC6.581279.5231.108Y+Metabolite - 1465LC3.453600162.11+Metabolite - 147GC8.581513.2217.127Y+Metabolite - 1497LC13.914032332.21+Metabolite - 1498LC1.561650143.11Metabolite - 150GC6.211237.9174.076Y+Metabolite - 1500LC1.721760.4391.11+Metabolite - 1514GC6.241239.8148.068Y+Metabolite - 1519GC11.21812.1204.088Y+Metabolite - 1534GC11.31824.3246.08Y+Metabolite - 1537GC6.141220.1171.055Y+Metabolite - 1538GC10.61730.7156.074Y+Metabolite - 1551GC8.731526.9205.077Y+Metabolite - 1554GC8.881541.3292.084Y+Metabolite - 1559GC10.21683.3217.071Y+Metabolite - 1564GC9.41592.8115.081Y+Metabolite - 1573 retired: glycerol-2-phosphateLC1.631669170.91Metabolite - 1574LC1.9219392331+Metabolite - 1575LC2.252243.5219.11+Metabolite - 1576LC2.512530247.11+Metabolite - 1597LC3.663894265.91+Metabolite - 1608LC8.0882533501+Metabolite - 1609LC8.3185293781+Metabolite - 1612LC8.648850.3230.91+Metabolite - 1616LC12.712910331.21+Metabolite - 1655LC1.3113741071+Metabolite - 1656LC1.461509154.91Metabolite - 1673GC12.51982.2217.019Y+Metabolite - 1713 retired: n-acetyl-L-asparticLC2.7327701741acidMetabolite - 1734LC8.7289234751Metabolite - 1735LC8.758929644.11Metabolite - 1736 retired: p-LC9.789961.5121.11hydroxybenzaldehydeMetabolite - 1737 retired: 2,3-dihydroxybenzoicLC9.8410036153.11acidMetabolite - 1738LC14.414574277.91+Metabolite - 1753GC8.161446.9356.938Y+Metabolite - 1754GC12.61981.6204.056Y+Metabolite - 1757GC11.91889.9318.169Y+Metabolite - 1775GC11.21809.3518.181Y+Metabolite - 1802LC8.959328486.91+Metabolite - 1814GC12.82004.7290.098Y+Metabolite - 1815GC11.81886.8289.079Y+Metabolite - 1817LC1.371552.32521+Metabolite - 1818LC1.431608.7126.91Metabolite - 1819LC1.361540244.81−iMetabolite - 1820 retired: glycerol-2-phosphateLC1.451626.8342.81Metabolite - 1824GC8.551509.8126.046Y+Metabolite - 1826GC5.51168101.993Y+Metabolite - 1827GC11.31835.6205.014Y+Metabolite - 1829 retired: oxalic acidLC1.4316001351Metabolite - 1830LC1.491661.3192.91Metabolite - 1831 Cl-adduct-of-citrullineLC1.461638.7209.91Metabolite - 1834LC1.641794.51041Metabolite - 1835LC1.861999.3152.11Metabolite - 1836LC2.12215.5205.91Metabolite - 1839LC2.572624138.11+Metabolite - 1842 retired: 4-GuanidinobutanoicLC3.213259146.11+acidMetabolite - 1843LC3.253295288.71Metabolite - 1847GC11.61854.5373.02Y+Metabolite - 1849GC13.12041.3361.035Y+Metabolite - 1850GC13.72110.6392.092Y+Metabolite - 1888GC10.11675.2173.077Y+Metabolite - 1909LC1.661803.5162.11+Metabolite - 1910 retired: uric acidLC5.856058.1169.31+Metabolite - 1911LC11.411800464.11+Metabolite - 1912LC9.229570.4181.21+Metabolite - 1914LC10.410720239.11+Metabolite - 1915LC14.414799507.21Metabolite - 1926 retired: trans-2,3,4-LC11.511840239.21+trimethoxycinnamic acidMetabolite - 1927 retired: Metabolite - 1323LC9.229581.5186.91aboveMetabolite - 1929LC1.331496.3215.11Metabolite - 1956GC8.41482.3218.037Y+Metabolite - 1958LC8.929282.3188.21+Metabolite - 1960LC1.421592337.11+Metabolite - 1961 retired: glycocholic acidLC1414431466.11+Metabolite - 1968GC10.91778.2414.045Y+Metabolite - 1972LC9.649958.6531.71+Metabolite - 1974LC5.936077160.21+Metabolite - 1975LC5.9560933441+Metabolite - 1977 5-oxoprolineLC3.563815260.91+Metabolite - 1979 retired: CL adduct of Isobar 19LC1.521690.31991Metabolite - 1981LC7.948266.8158.11+Metabolite - 1988LC11.111515190.11+Metabolite - 2005LC8.629048232.11+Metabolite - 2009GC11.91905.9217.024Y+Metabolite - 2026LC1.361556.2239.21+Metabolite - 2027LC1.561729.3184.11+Metabolite - 2033 retired: 2-isopropylmalic acidLC9.039332.51751Metabolite - 2038LC7.868139.34221+Metabolite - 2041LC13.814198246.31+Metabolite - 2045LC1.371548.91481+Metabolite - 2046LC15.816200502.51+Metabolite - 2047 retired: Metabolite - 4328LC8.829150.3828.11+Metabolite - 2051LC1.4516343091+Metabolite - 2052 retired: potassium adduct ofLC1.31429.8219.11+Isobar 1Metabolite - 2053LC1.351482.3324.91Metabolite - 2055LC1.371502269.91+Metabolite - 2056LC1.371499165.11Metabolite - 2058LC1.411538.42821+Metabolite - 2067 retired: carnitineLC1.671750162.11+Metabolite - 2074LC2.242380.9280.11+Metabolite - 2100LC1.331532.94991+Metabolite - 2105LC8.158442433.61+Metabolite - 2109LC8.999266321.11+Metabolite - 2111LC9.199442.3365.11+Metabolite - 2130LC16.316626792.41+Metabolite - 2139LC8.098416.7218.11+Metabolite - 2141LC9.399605409.11+Metabolite - 2151LC14.414722531.31+Metabolite - 2168LC1.361549261.11+iMetabolite - 2173LC2.682748.2230.11+Metabolite - 2174LC2.52569250.11+Metabolite - 2175LC3.844148.41441+Metabolite - 2181LC8.378715.52981+Metabolite - 2185LC9.229499.4246.21+Metabolite - 2192GC8.881538.7174.05Y+Metabolite - 2194LC13.713961544.21+Metabolite - 221GC111829205.106Y+Metabolite - 2212LC1616271478.21+Metabolite - 222GC111835.7319.092Y+Metabolite - 2220GC9.751639.3450.927Y+Metabolite - 2221GC11.61862.5308.112Y+Metabolite - 223GC9.651677.7217.137Y+Metabolite - 2231LC14.314629278.11+Metabolite - 2237LC10.110454453.11+Metabolite - 2238LC10.510817792.21+Metabolite - 2242LC11.611926254.31+Metabolite - 2249LC14.214571267.21Metabolite - 2250LC14.314668286.31+Metabolite - 2254LC1.531687.6217.21+Metabolite - 2255LC9.089394539.11+Metabolite - 2256LC9.9310232460.81+Metabolite - 226GC3.871011200.137Y+Metabolite - 2266 retired 4-acetominophenLC8.318713.4229.91sulfateMetabolite - 2269LC10.410727255.11Metabolite - 2270LC1111402495.21Metabolite - 2272LC7.968377189.11Metabolite - 2279LC12.412781490.11+Metabolite - 2285LC22146699.61Metabolite - 2287LC1313336502.81+Metabolite - 2292LC2.42513.6343.91Metabolite - 2306 retired: gamma glu leuLC8.899246261.11+Metabolite - 2313LC1.561685.6352.91Metabolite - 2316LC8.829163.6100.11+Metabolite - 2319LC12.212626367.21−iMetabolite - 2321LC13.412940314.31+iMetabolite - 2329LC11.812178541.21Metabolite - 2347LC13.714091450.11+Metabolite - 2366LC8.478870.22711+Metabolite - 2370LC16.116561476.41Metabolite - 2386LC11.912320539.21Metabolite - 2387 retired: gamma glu leuLC8.558838.5182.11Metabolite - 2388LC16.216567259.11Metabolite - 2389LC1.491641.5314.91Metabolite - 2390LC6.096144.9517.41+Metabolite - 2392LC13.1134603791Metabolite - 2395LC10.110015471.91+iMetabolite - 2407LC15.716128637.31+Metabolite - 2469LC1616436502.31+Metabolite - 2486LC1.521667635.71Metabolite - 2506LC14.114438624.41Metabolite - 2526LC1.3815162151Metabolite - 2527 retired: citrullineLC1.621742175.81+Metabolite - 2546LC1.631747.3129.11+Metabolite - 2546LC1.631747.3129.11+Metabolite - 2548 Cl-adduct-of-uric acidLC5.976016202.91Metabolite - 2550LC11.111719411.11+Metabolite - 2558LC8.148674153.11+Metabolite - 2559LC13.814151539.21Metabolite - 2560LC14.414754235.21+Metabolite - 2561LC10.210481352.11+Metabolite - 2563 retired: lactateLC2.182302178.91Metabolite - 2564LC10.510903677.11+Metabolite - 2565LC1111341886.61+Metabolite - 2567LC7.798164.7247.11+Metabolite - 2568LC8.548790.8342.11+Metabolite - 2586 retired: pieces of Isobar 1LC1.31489394.31Metabolite - 2587 retired: p-acetaminophen-beta-LC8.278613.4371.11d-glucuronideMetabolite - 2588LC8.8791086081+Metabolite - 2589LC9.049334.7472.21+Metabolite - 2591LC9.9910189279.31+Metabolite - 2592LC10.610778697.21Metabolite - 2593LC9.8110139627.21+Metabolite - 2594LC9.7810112332.11+iMetabolite - 2606LC1.581755.5114.11+Metabolite - 2607LC1010354578.21+Metabolite - 2608LC1111326191.91+Metabolite - 2627GC9.191601.6334.058Y+Metabolite - 2628GC12.52000.2308.035Y+Metabolite - 263GC5.351185215.178Y+Metabolite - 264GC14.42244.3299.11Y+Metabolite - 2646GC12.92043265.127Y+Metabolite - 2647GC14.12190.6295.028Y+Metabolite - 2648GC15.42351.6309.096Y+Metabolite - 268GC8.11507.6144.159Y+Metabolite - 2686LC1.415932171Metabolite - 2687 retired: CL adduct of Isobar 10LC1.41593181.11Metabolite - 2688LC1.421614182.01−iMetabolite - 2694 retired: lactateLC2.2323211351Metabolite - 2696LC3.383455.51051+Metabolite - 2697LC3.774241.2209.91+Metabolite - 2698LC3.884338.51571+Metabolite - 270GC10.91834.9362.22Y+Metabolite - 2703LC8.869054.8384.11+Metabolite - 2711 retired: pieces of lactateLC2.2223001231+Metabolite - 2726LC8.38550.6375.21+Metabolite - 273GC10.41781.9312.126Y+Metabolite - 274GC11.81940.6221.128Y+Metabolite - 2750 retired: Metabolite 3316LC2.172260125.61Metabolite - 2752LC2.922802.3189.11+Metabolite - 2753LC3.3833581471+Metabolite - 276GC4.831124.8269.159Y+Metabolite - 2774LC3.533796230.91+Metabolite - 278GC9.11624.2117.074Y+Metabolite - 279GC101731.2274.171Y+Metabolite - 2806LC1.381491185.11+Metabolite - 2821LC6.86913119.11+Metabolite - 2822LC8.658838607.61+Metabolite - 2825LC13.1131552351+Metabolite - 2825 retired: RiluzoleLCMetabolite - 283GC6.131278.4100.125Y+Metabolite - 285GC9.61681.9191.07Y+Metabolite - 2854LC8.6889337671+Metabolite - 286GC10.51789217.145Y+Metabolite - 2866LC9.539795.32311Metabolite - 2867LC9.659908235.31+Metabolite - 2886 retired: CL adduct of p-LC8.348637361.91acetiminophen-beta-d-glucuronideMetabolite - 289GC7.581446.1540.205Y+Metabolite - 2890 retired: pieces of tryptophanLC9.2395022031Metabolite - 2894LC9.9410320226.11Metabolite - 2898LC11.211463213.11Metabolite - 2915GC3.771099174Y+Metabolite - 2924 retired: 2-hydroxybutanoicGC4.381170.7130.9Y+acid (also called (s)-2-hydroxybutyric acid)Metabolite - 293GC8.641570.5158.16Y+Metabolite - 2952GC3.151025281.0Y+LMetabolite - 2973GC4.741213.4281Y+Metabolite - 2981GC5.211265210.9Y+LMetabolite - 2986GC5.561304.3201.1Y+Metabolite - 2989GC5.871340341Y+Metabolite - 3002GC6.741440.8296.1Y+Metabolite - 3003GC6.791446.6218.1Y+Metabolite - 3012GC7.171489.8232Y+Metabolite - 3014 retired: meso-erythritolGC7.431520.6217.1Y+Metabolite - 3017GC7.611541.4246.1Y+Metabolite - 3019GC7.741556.4260.1Y+Metabolite - 3020 retired: threonic acidGC7.811564.1292Y+Metabolite - 3022GC7.981584.9142Y+Metabolite - 3023GC8.041590.9274.1Y+Metabolite - 3025GC8.111600.3274.1Y+Metabolite - 3026GC8.171606274.1Y+LMetabolite - 3027 retired: arginineGC8.211610.6142Y+Metabolite - 303 threonine-derivGC8.881689.4116.9Y+Metabolite - 3030GC8.621659.7320Y+Metabolite - 3040GC9.271735.7274.1Y+Metabolite - 3044LC1.521615.3150.11+Metabolite - 3045 retired: Isobar 1LC1.511601.5180.61+Metabolite - 3052LC8.79035426.21+iMetabolite - 3055LC9.29443196.81+Metabolite - 3056LC9.199432185.21+Metabolite - 3058GC9.71786.9335.1Y+Metabolite - 3065 retired 1,5-anhdro-d-glucitolGC9.741790.8217.1Y+Metabolite - 3067GC101824.2132Y+Metabolite - 3073GC10.21838.8362.1Y+Metabolite - 3074GC10.21844.5204.1Y+Metabolite - 3075GC10.41857.9204Y+Metabolite - 3077GC10.41866.2308.1Y+Metabolite - 3078GC10.71887203.1Y+Metabolite - 3081GC10.91911.5204Y+Metabolite - 3085 retired inositolGC111926.1217Y+Metabolite - 3087GC11.21942174.1Y+LMetabolite - 3088GC11.21946.1372.2Y+Metabolite - 3090GC11.31955243.1Y+Metabolite - 3093GC11.51975.6204Y+Metabolite - 3094GC11.61980.6299Y+Metabolite - 3097GC11.61990.4204Y+Metabolite - 3098GC11.82003307.8Y+Metabolite - 3099GC11.82005.2204Y+Metabolite - 3100GC11.92013.2204Y+Metabolite - 3101GC11.92022.2290Y+Metabolite - 3102GC122028.2217.1Y+Metabolite - 3103GC12.12039.8290.1Y+Metabolite - 3108GC12.22056.5246Y+Metabolite - 3109GC12.62093202.1Y+LMetabolite - 3113GC12.72113.5406.2Y+Metabolite - 3114GC12.82121204.0Y+LMetabolite - 3125LC11.912095187.11+Metabolite - 3127LC8.618812260.11Metabolite - 3129LC8.89012337.11+Metabolite - 3130LC9.099328158.21+Metabolite - 3131 retired: NH4 adduct of indole-LC10.510770192.91+3-acetic acidMetabolite - 3132LC10.110392260.21+Metabolite - 3134LC14.314487483.11+Metabolite - 3138LC8.638749229.21+Metabolite - 3139LC8.828934.5176.11+Metabolite - 3143LC9.8110070160.11+Metabolite - 3160LC12.1122473611+Metabolite - 3162 retired: N,N-dimethylarginineLC2.412444203.11+Metabolite - 3165LC8.388472.22651+Metabolite - 3166LC8.698746.5394.21+Metabolite - 3178 retired NH3 adduct of citricLC3.1532802101+acidMetabolite - 3179 retired 4-guanidnobutanoicLC3.453603146.11+acidMetabolite - 3180LC4.1443561391+Metabolite - 3181LC8.598621.4165.11+Metabolite - 3182LC8.838971332.71+Metabolite - 3183-gamma-L-glutamyl-L-LC9.379441295.21+phenylalanineMetabolite - 3218LC2.22257148.11+Metabolite - 3230LC3.13043.22451+Metabolite - 3231LC3.083026104.11+Metabolite - 3235 retired DL-indole-3-lactic acidLC10.5105812061+Metabolite - 3249LC3.283298.31411+Metabolite - 3305 retired: duplicate of MetaboliteLC13.814129369.311088Metabolite - 3313LC8.18529.6196.91Metabolite - 3314LC8.929143.5264.81+Metabolite - 3316 retired: lacateLC2.092308125.11Metabolite - 3317LC8.428702.3429.61+Metabolite - 3320LC10.7109852451Metabolite - 3327LC11.611784385.31Metabolite - 3331LC9.3895305311+Metabolite - 3334LC3.153371.54091+Metabolite - 3365LC1.872068.3115.11+Metabolite - 3370LC8.118529.1226.21+Metabolite - 3377LC8.868964270.21+iMetabolite - 3401LC1.731863.3131.11+Metabolite - 3402LC8.99052.3343.21+Metabolite - 341GC6.571278.3100.098Y+Metabolite - 3416LC2.252582.31271+Metabolite - 3430LC2.783319.7189.11+Metabolite - 3441LC1.511565515.01+iMetabolite - 3450 retired: 1-methlynicotinamideLC1.681767.81371+Metabolite - 3457LC3.814193.3212.91+Metabolite - 3468 retired: Metabolite 1498LC1.751813.51431Metabolite - 3489LC3.2636052261+Metabolite - 3498LC7.88368.7279.11+Metabolite - 3517LC10.310892382.31+Metabolite - 3534LC10.511174426.31+Metabolite - 3603LC8.418971313.61+iMetabolite - 3604 retired CL adduct of hippuricLC8.999551.9214.21acidMetabolite - 3615LC13.6143448681+Metabolite - 3624LC10.410984205.11+Metabolite - 3653-stachydrineLC4.054500144.11+Metabolite - 3668LC9.639536379.11+Metabolite - 3696LC1515200450.31+Metabolite - 3698LC8.318640.2273.11+Metabolite - 3707LC13.1133402411+Metabolite - 3708LC1.661625.3159.91+Metabolite - 3758LC12.412714309.11Metabolite - 3772LC2.2222741091+Metabolite - 3781LC1.451544262.91+Metabolite - 3783LC1.371464271.11+Metabolite - 3807LC33398.52451+Metabolite - 3808LC3.283719288.81−iMetabolite - 381GC4.251012101.043Y+Metabolite - 3813LC3.814312212.11+Metabolite - 382GC5.121106.8221.063Y+Metabolite - 383GC5.431141.4198.035Y+Metabolite - 3830LC8.4287251891Metabolite - 3832-phenol-sulfateLC8.738995.81731Metabolite - 3837 retired: 3-incoxylsulfateLC9.269466.8212.11Metabolite - 3843LC9.549721.9263.11+Metabolite - 386GC9.31580.1142.092Y+Metabolite - 387GC10.71746.5204.077Y+Metabolite - 388GC4.621052.7121.065Y+Metabolite - 3881: retired: azelaic acidLC11.111318188.91+Metabolite - 3894 retired: pyroglutamic acid (5-LC3.393883.5130.11+oxoproline)Metabolite - 3896LC3.383868245.21+Metabolite - 3900LC4.534871.7173.11Metabolite - 393GC10.71750.9217.073Y+Metabolite - 394GC4.731065.2157.087Y+Metabolite - 3951LC8.418705.4367.11+Metabolite - 3952LC8.78941.3297.21+Metabolite - 3972LC6.166304432.61−iMetabolite - 3977LC1111312187.11Metabolite - 398GC5.151110.7103.026Y+Metabolite - 3992LC1.41400127.21−iMetabolite - 3994GC1.631640.44271+Metabolite - 4003GC3.9443972051+Metabolite - 4012GC7.021458.2357Y+Metabolite - 4019GC7.681534.5174Y+Metabolite - 4020GC7.911561.5220.1Y+Metabolite - 4031GC14.2614607244.21+Metabolite - 4032 retired: lysineGC8.951682.6156.1Y+Metabolite - 404GC9.341584.2227.098Y+Metabolite - 4044GC10.531863.1149Y+Metabolite - 4055GC12.042022.2304.1Y+Metabolite - 406GC11.81883.2204.074Y+Metabolite - 407GC16.72483.2283.2Y+Metabolite - 4077GC142266.5227Y+Metabolite - 4080GC14.022270.2299Y+Metabolite - 4084GC14.982393.9441.3Y+Metabolite - 4096GC8.68763.6318.21+Metabolite - 413GC11.11806342.146Y+Metabolite - 4133GC4.351108.9198Y+Metabolite - 4134GC5.51123960.9Y+Metabolite - 4147GC10.071767.1290.2Y+Metabolite - 4148GC10.231786.3249.2Y+Metabolite - 4167LC1110920286.21+iMetabolite - 4196GC12.142000.4290.2Y+Metabolite - 4198GC13.572173.7218.2Y+Metabolite - 421GC9.181567.4243.176Y+Metabolite - 423GC132031.5204.087Y+Metabolite - 4251GC4.091130.7217Y+Metabolite - 4252GC6.021348282.1Y+LMetabolite - 4271GC9.691777.4419.2Y+Metabolite - 4272GC10.281840.2669.3Y+Metabolite - 4274GC10.371857.0158.1Y+Metabolite - 4275GC10.681887.0271.1Y+Metabolite - 4276GC13.922262.9223.1Y+Metabolite - 4351LC11.811937427.11+iMetabolite - 4354GC3.901074.3110Y+Metabolite - 4357GC8.001541.1216Y+Metabolite - 4360GC9.151678.2347.2Y+Metabolite - 4361GC9.401706.2232.2Y+Metabolite - 4362GC10.021779.9319.2Y+Metabolite - 4363LC13.713709830.11+iMetabolite - 4364GC10.661852.4232Y+Metabolite - 4365GC11.051892.9204Y+Metabolite - 441GC3.81964.273.0775Y+Metabolite - 442GC4.11997.9176.069Y+Metabolite - 4428LC7.928237229.21+iMetabolite - 443GC8.661506.6205.092Y+Metabolite - 4470LC10.811037271.21+iMetabolite - 451GC5.211118.4173.075Y+Metabolite - 4510GC9.71740254.0Y+LMetabolite - 4511GC10.11788206.0Y+LMetabolite - 4522GC12.32025217.1Y+LMetabolite - 4523GC12.52047258.1Y+LMetabolite - 4547LC11.411464310.31+iMetabolite - 458GC11.91897204.089Y+Metabolite - 4586LC7.147487260.01−iMetabolite - 4611GC8.071547292.1Y+LMetabolite - 4612LC11.711784453.21−iMetabolite - 4624GC101779342.2Y+LMetabolite - 4627LC10.811035591.31+iMetabolite - 465GC10.81757.9204.088Y+Metabolite - 4658LC10.110321194.11+iMetabolite - 470GC15.42329.7299.05Y+Metabolite - 472GC3.96980.2160.082Y+Metabolite - 4732LC5.846102221.21+iMetabolite - 4767GC8.771626116.9Y+LMetabolite - 4769GC11.31916156.0Y+LMetabolite - 4795GC14.82350309.0Y+LMetabolite - 4806GC4.21123104.9Y+LMetabolite - 482GC7.711396.1117.055Y+Metabolite - 485GC5.311128.8217.029Y+Metabolite - 4873LC12.111838288.01+iMetabolite - 490GC4.371026.7191.028Y+Metabolite - 4906LC10.310119313.21+iMetabolite - 4931LC1.51660431.01+iMetabolite - 4986GC11.61956204.1Y+LMetabolite - 499GC6.641276.1259.062Y+Metabolite - 501GC5.551155.4143.084Y+Metabolite - 502GC13.82125.7361.127Y+Metabolite - 504GC11.61864.2156.068Y+Metabolite - 505GC10.61737.5454.125Y+Metabolite - 506GC11.81885.8204.075Y+Metabolite - 508GC7.251343.8466.076Y+Metabolite - 5086LC9.519738388.21+iMetabolite - 511GC7.511375247.045Y+Metabolite - 522GC6.351256.3211.009Y+Metabolite - 523GC10.71762.7265.075Y+Metabolite - 5231LC2.092271113.21+iMetabolite - 5233LC2.562928138.11+iMetabolite - 5234LC13.513823316.41+iMetabolite - 5247LC9.9410272525.31+iMetabolite - 526GC11.71886.7353.125Y+Metabolite - 5346GC8.331573202.0Y+LMetabolite - 5349GC10.11782312.1Y+LMetabolite - 5366GC12.52045204.0Y+LMetabolite - 5403GC5.921300319.0Y+LMetabolite - 543GC12.72006.8117.082Y+Metabolite - 547GC5.261131.8113.055Y+Metabolite - 549GC6.51273.6189.101Y+Metabolite - 553GC4.161005.6244.035Y+Metabolite - 554GC10.71771.6221.093Y+Metabolite - 557GC10.91784.5245.094Y+Metabolite - 562GC6.11227.1102.061Y+Metabolite - 5673LC12.812402383.31−iMetabolite - 568GC5.821196.6281.047Y+Metabolite - 571GC4.621057.8105.063Y+Metabolite - 5728LC1.591733148.01−iMetabolite - 5730LC8.918739290.31+iMetabolite - 5769LC11.110753485.21−iMetabolite - 577GC11.51865.4272.14Y+Metabolite - 578GC12.51979.2498.188Y+Metabolite - 5788LC9.249026267.11+iMetabolite - 5791LC9.799529327.31+iMetabolite - 580GC11.61876.8102.025Y+Metabolite - 5847GC12.42040288.2Y+LMetabolite - 5848LC10.410571601.21+iMetabolite - 5887LC9.629786547.21+iMetabolite - 5907GC8.691643229.1Y+LMetabolite - 591GC12.21957217.117Y+Metabolite - 593GC10.61765.3196.11Y+Metabolite - 594GC11.51871.3205.116Y+Metabolite - 595GC3.8968.8100.003Y+Metabolite - 596GC11.71878.8217.115Y+Metabolite - 597GC7.911439218.11Y+Metabolite - 5976LC11.611366671.31+iMetabolite - 5983GC14.72309207.0Y+LMetabolite - 601GC5.191131.3201.139Y+Metabolite - 606GC6.161242131.105Y+Metabolite - 609GC5.041113.3190.09Y+Metabolite - 6126LC9.769577202.91−iMetabolite - 613GC9.251599.6156.088Y+Metabolite - 614GC5.81196.7233.092Y+Metabolite - 618GC8.461507.3103.045Y+Metabolite - 621GC12.92040.3217.114Y+Metabolite - 6226GC4.381137154.0Y+LMetabolite - 6227GC51211196.1Y+LMetabolite - 6246GC6.941428160.1Y+LMetabolite - 6269GC10.91881217.1Y+LMetabolite - 6270GC11.41930320.2Y+LMetabolite - 6272GC12.62070131.0Y+LMetabolite - 6326GC7.661511144.1Y+LMetabolite - 6346GC81551263.2Y+LMetabolite - 6347GC8.161569244.1Y+LMetabolite - 638GC7.861437.9218.112Y+Metabolite - 642GC10.31724.5103.045Y+Metabolite - 645GC11.61873.5203.109Y+Metabolite - 6467GC11.11894320.1Y+LMetabolite - 6488GC12.32022204.1Y+LMetabolite - 651GC4.551059.6175.108Y+Metabolite - 653GC6.571291192.106Y+Metabolite - 655GC11.71912.8156.085Y+Metabolite - 6551LC9.249020492.11+iMetabolite - 664GC12.41961.4174.095Y+Metabolite - 669GC12.72017.7361.164Y+Metabolite - 670GC12.92039.7258.125Y+Metabolite - 671GC13.72144.9204.123Y+Metabolite - 6711LC1.541647381.11+iMetabolite - 681GC12.31955.4243.083Y+Metabolite - 6827LC4.595000139.01−iMetabolite - 683GC8.461502.7188.09Y+Metabolite - 6869GC81546257.1Y+LMetabolite - 687GC9.261591.8210.977Y+Metabolite - 688GC9.961674.9156.111Y+Metabolite - 6907GC9.221687337.1Y+LMetabolite - 691GC10.31711274.083Y+Metabolite - 6931GC10.41820267.1Y+LMetabolite - 6955GC11.81979306.1Y+LMetabolite - 6963GC11.91989103.0Y+LMetabolite - 7008LC1.471616125.11−iMetabolite - 7009LC1.621577349.91+iMetabolite - 702GC10.81784218.077Y+Metabolite - 704GC6.511275.1171.059Y+Metabolite - 7050LC9.449626242.91−iMetabolite - 706GC10.61751.4437.153Y+Metabolite - 7089LC9.239391297.01−iMetabolite - 709GC12.51979.8109.065Y+Metabolite - 7146LC2.132248229.71+iMetabolite - 7147LC3.133467245.01+iMetabolite - 7177LC12.112101405.11−iMetabolite - 721GC7.541421.2373.062Y+Metabolite - 7336LC8.789007470.81+iMetabolite - 734GC8.421524.1232.108Y+Metabolite - 736GC13.52133.5506.129Y+Metabolite - 749GC12.82030.8101.143Y+Metabolite - 753GC7.921465.7218.137Y+Metabolite - 760GC8.671552.4220.099Y+Metabolite - 761GC10.61777.9217.104Y+Metabolite - 763GC8.451528.5230.135Y+Metabolite - 770GC6.871341.4247.075Y+Metabolite - 7706LC2.322457222.71+iMetabolite - 7707LC2.292371250.81+iMetabolite - 771GC11.21854.4246.092Y+Metabolite - 7762LC8.928776197.01−iMetabolite - 7765LC11.210919245.01−iMetabolite - 777GC10.31741.6299.052Y+Metabolite - 780GC6.131233.1280.996Y+Metabolite - 7807LC11.310966256.71+iMetabolite - 7815LC8.438450381.91+iMetabolite - 782GC10.81776287.056Y+Metabolite - 783GC5.161121.3180.969Y+Metabolite - 7846GC5.11208145.1Y+LMetabolite - 7888GC162513311.3Y+LMetabolite - 7889GC16.82629311.3Y+LMetabolite - 7890GC17.82752129.0Y+LMetabolite - 841GC152261.1488.377Y+Metabolite - 861GC7.641401.8247.111Y+Metabolite - 863GC9.981675347.161Y+Metabolite - 941GC11.61912.5204.1Y+Metabolite - 961GC6.311296.1319.078Y+Metabolite - 982GC6.121274.8175.039Y+Metabolite - 984GC8.391538.8314.083Y+Metabolite - 985GC6.351301.3174.11Y+Metabolite - 987GC8.861595.2244.121Y+Metabolite - 988GC9.771701.3519.157Y+Metabolite - 990GC7.241404.6405.107Y+Metabolite - 991GC4.141051.9117.094Y+Metabolite - 992GC7.531434296.155Y+Metabolite - 995GC11.41890.9305.152Y+Metabolite - 996GC12.21984.3357.151Y+Metabolite - 998GC7.361415.4241.171Y+


While the invention has been described in detail and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made without departing from the spirit and scope of the invention.

Claims
  • 1. A method of diagnosing whether a subject has amyotrophic lateral sclerosis (ALS), comprising: analyzing a biological sample from a subject to determine the level(s) of one or more biomarkers for amyotrophic lateral sclerosis in the sample, wherein the one or more biomarkers are selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, and 8, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; and comparing the level(s) of the one or more biomarkers in the sample to ALS-positive and/or ALS-negative reference levels of the one or more biomarkers in order to diagnose whether the subject has amyotrophic lateral sclerosis.
  • 2. The method of claim 1, wherein the ALS-negative reference levels of the one or more biomarkers comprise levels of the one or more biomarkers in one or more samples from one or more subjects not having ALS and the ALS-positive reference levels of the one or more biomarkers comprise levels of the one or more biomarkers in one or more samples from one or more subjects diagnosed with ALS.
  • 3. The method of claim 2, wherein differential levels of the one or more biomarkers between the sample and the ALS-negative reference levels are indicative of a diagnosis of ALS in the subject.
  • 4. The method of claim 2, wherein differential levels of the one or more biomarkers between the sample and the ALS-positive reference levels are indicative of a diagnosis of no ALS in the subject.
  • 5. The method of claim 2, wherein levels of the one or more biomarkers in the sample corresponding to the ALS-positive reference levels are indicative of a diagnosis of ALS in the subject.
  • 6. The method of claim 2, wherein levels of the one or more biomarkers in the sample corresponding to the ALS-negative reference levels are indicative of a diagnosis of no ALS in the subject.
  • 7. The method of claim 1, wherein the one or more biomarkers are selected from those biomarkers in Tables 3, 4, 5, 6, 7, and 8 having p values of less than 0.05 and/or those biomarkers in Tables 3, 4, 5, 6, 7, and 8 having q values of less than 0.10.
  • 8. The method of claim 1, wherein the method comprises analyzing the biological sample to determine the level of two or more biomarkers selected from Tables 3, 4, 5, 6, 7, and 8.
  • 9. The method of claim 1, wherein the method comprises analyzing the biological sample to determine the level of three or more biomarkers selected from Tables 3, 4, 5, 6, 7, and 8.
  • 10. The method of claim 1, wherein the method comprises analyzing the biological sample to determine the level of four or more biomarkers selected from Tables 3, 4, 5, 6, 7, and 8.
  • 11. The method of claim 1, wherein the method comprises analyzing the biological sample to determine the level of five or more biomarkers selected from Tables 3, 4, 5, 6, 8, and 9.
  • 12. The method of claim 1, wherein the method comprises analyzing the biological sample to determine the level of ten or more biomarkers selected from Tables 3, 4, 5, 6, 7, and 8.
  • 13. The method of claim 1, wherein the method comprises analyzing the biological sample to determine the level of fifteen or more biomarkers selected from Tables 3, 4, 5, 6, 7, and 8.
  • 14. The method of claim 1, wherein the biological sample is cerebral spinal fluid and the one or more biomarkers comprise one or more biomarkers listed in Tables 3 and 4.
  • 15. The method of claim 1, wherein the biological sample is blood plasma and the one or more biomarkers comprise one or more biomarkers listed in Tables 5, 6, 7, and 8.
  • 16. The method of claim 1, wherein the sample is analyzed using one or more techniques selected from the group consisting of mass spectrometry, ELISA, and antibody linkage.
  • 17. The method of claim 1, wherein the one or more biomarkers comprise one or more xenobiotics and/or phase I metabolites of xenobiotics.
  • 18. The method of claim 1, wherein the one or more biomarkers comprise caffeine and/or one or more metabolites of caffeine.
  • 19. The method of claim 18, wherein the one or more metabolites of caffeine are selected from the group consisting of paraxanthine, theophylline, and theobromine.
  • 20. The method of claim 1, wherein the one or more biomarkers comprise caffeine and one or more metabolites of caffeine.
  • 21. The method of claim 20, wherein the ALS-positive and/or ALS-negative reference levels of the one or more biomarkers comprise ALS-positive and/or ALS-negative reference ratios of the levels of caffeine and a metabolite of caffeine.
  • 22. The method of claim 1, wherein the one or more biomarkers comprise at least one biomarker selected from Tables 3, 4, 5, 6, 7, and 8.
  • 23. The method of claim 1, wherein the one or more biomarkers comprise at least one biomarker selected from Tables 3, 4, 5, 6, 7, and 8 having a p value of less than 0.05 and/or having a q value of less than 0.10.
  • 24. A method of determining whether a subject is predisposed to developing amyotrophic lateral sclerosis (ALS), comprising: analyzing a biological sample from a subject to determine the level(s) of one or more biomarkers for amyotrophic lateral sclerosis in the sample, wherein the one or more biomarkers are selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, and 8, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; and comparing the level(s) of the one or more biomarkers in the sample to ALS-positive and/or ALS-negative reference levels of the one or more biomarkers in order to determine whether the subject is predisposed to developing amyotrophic lateral sclerosis.
  • 25. A method of monitoring progression/regression of amyotrophic lateral sclerosis (ALS) in a subject comprising: analyzing a first biological sample from a subject to determine the level(s) of one or more biomarkers for amyotrophic lateral sclerosis in the sample, wherein the first sample is obtained from the subject at a first time point and the one or more biomarkers are selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, 8, 16, 17, and 18, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; analyzing a second biological sample from a subject to determine the level(s) of the one or more biomarkers, wherein the second sample is obtained from the subject at a second time point; and comparing the level(s) of one or more biomarkers in the first sample to the level(s) of the one or more biomarkers in the second sample in order to monitor the progression/regression of ALS in the subject.
  • 26. The method of claim 25, wherein the method further comprises comparing the level(s) of one or more biomarkers in the first sample, the level(s) of one or more biomarkers in the second sample, and/or the results of the comparison of the level(s) of the one or more biomarkers in the first and second samples to ALS-positive reference levels, ALS-negative reference levels, ALS-progression-positive reference levels, and/or ALS-regression-positive reference levels of the one or more biomarkers.
  • 27. The method of claim 25, wherein the one or more biomarkers are selected from Tables 16, 17, and 18.
  • 28. The method of claim 25, wherein the one or more biomarkers are selected from those biomarkers in Tables 16, 17, and 18 having p values of less than 0.05 and/or those biomarkers in Tables 16, 17, and 18 having q values of less than 0.10.
  • 29. A method of assessing the efficacy of a composition for treating amyotrophic lateral sclerosis (ALS) comprising: analyzing, from a subject having amyotrophic lateral sclerosis and currently or previously being treated with a composition, a biological sample to determine the level(s) of one or more biomarkers for ALS selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, 8, 16, 17, and 18, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; and comparing the level(s) of the one or more biomarkers in the sample to (a) levels of the one or more biomarkers in a previously-taken biological sample from the subject, wherein the previously-taken biological sample was obtained from the subject before being treated with the composition, (b) ALS-positive reference levels of the one or more biomarkers, (c) ALS-negative reference levels of the one or more biomarkers, (d) ALS-progression-positive reference levels of the one or more biomarkers, and/or (e) ALS-regression-positive reference levels of the one or more biomarkers.
  • 30. The method of claim 29, wherein the one or more biomarkers are selected from Tables 16, 17, and 18.
  • 31. The method of claim 29, wherein the one or more biomarkers are selected from those biomarkers in Tables 16, 17, and 18 having p values of less than 0.05 and/or those biomarkers in Tables 16, 17, and 18 having q values of less than 0.10.
  • 32. A method for assessing the efficacy of a composition in treating amyotrophic lateral sclerosis (ALS), comprising: analyzing a first biological sample from a subject to determine the level(s) of one or more biomarkers for ALS, the first sample obtained from the subject at a first time point wherein the one or more biomarkers are selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, 8, 16, 17, and 18, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; administering the composition to the subject; analyzing a second biological sample from the subject to determine the level(s) of the one or more biomarkers, the second sample obtained from the subject at a second time point after administration of the composition; comparing the level(s) of one or more biomarkers in the first sample to the level(s) of the one or more biomarkers in the second sample in order to assess the efficacy of the composition for treating amyotrophic lateral sclerosis.
  • 33. The method of claim 32, wherein the one or more biomarkers are selected from Tables 16, 17, and 18.
  • 34. The method of claim 32, wherein the one or more biomarkers are selected from those biomarkers in Tables 16, 17, and 18 having p values of less than 0.05 and/or those biomarkers in Tables 16, 17, and 18 having q values of less than 0.10.
  • 35. A method of assessing the relative efficacy of two or more compositions for treating amyotrophic lateral sclerosis (ALS) comprising: analyzing, from a first subject having ALS and currently or previously being treated with a first composition, a first biological sample to determine the level(s) of one or more biomarkers selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, 8, 16, 17, and 18, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; analyzing, from a second subject having ALS and currently or previously being treated with a second composition, a second biological sample to determine the level(s) of the one or more biomarkers; and comparing the level(s) of one or more biomarkers in the first sample to the level(s) of the one or more biomarkers in the second sample in order to assess the relative efficacy of the first and second compositions for treating amyotrophic lateral sclerosis.
  • 36. The method of claim 35, further comprising analyzing, from a third subject having ALS and currently or previously being treated with a third composition, a second biological sample to determine the level(s) of the one or more biomarkers; and comparing the level(s) of one or more biomarkers in the third sample to the level(s) of the one or more biomarkers in the first and second samples in order to assess the relative efficacy of the first, second, and third compositions for treating amyotrophic lateral sclerosis.
  • 37. The method of claim 35, wherein the one or more biomarkers are selected from Tables 16, 17, and 18.
  • 38. The method of claim 35, wherein the one or more biomarkers are selected from those biomarkers in Tables 16, 17, and 18 having p values of less than 0.05 and/or those biomarkers in Tables 16, 17, and 18 having q values of less than 0.10.
  • 39. A method for screening a composition for activity in modulating one or more biomarkers of amyotrophic lateral sclerosis, comprising: contacting one or more cells with a composition; analyzing at least a portion of the one or more cells or a biological sample associated with the cells to determine the level(s) of one or more biomarkers of amyotrophic lateral sclerosis selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, and 8, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; and comparing the level(s) of the one or more biomarkers with predetermined standard levels for the biomarkers to determine whether the composition modulated the level(s) of the one or more biomarkers.
  • 40. The method of claim 39, wherein the predetermined standard levels for the biomarkers are level(s) of the one or more biomarkers in the one or more cells in the absence of the composition.
  • 41. The method of claim 39, wherein the predetermined standard levels for the biomarkers are level(s) of the one or more biomarkers in one or more control cells not contacted with the composition.
  • 42. The method of claim 39, wherein the method is conducted in vivo.
  • 43. The method of claim 39, wherein the method is conducted in vitro.
  • 44. A method for identifying a potential drug target for amyotrophic lateral sclerosis (ALS) comprising: identifying one or more biochemical pathways associated with one or more biomarkers for ALS, wherein the one or more biomarkers are selected from (a) one or more biomarkers listed in Tables 3, 4, 5, 6, 7, and 8, (b) one or more xenobiotics, (c) one or more metabolites of xenobiotics, and (d) combinations thereof; and identifying a protein affecting at least one of the one or more identified biochemical pathways, the protein being a potential drug target for amyotrophic lateral sclerosis.
  • 45. A method for treating a subject having amyotrophic lateral sclerosis (ALS) comprising administering to the subject an effective amount of one or more biomarkers selected from Tables 3, 4, 5, 6, 7, and 8 that are decreased in subjects having ALS as compared to subjects not having ALS.
  • 46. A method of distinguishing whether a subject has amyotrophic lateral sclerosis (ALS) or has peripheral neuropathy or myopathy, comprising: analyzing a biological sample from a subject to determine the level(s) of one or more biomarkers for amyotrophic lateral sclerosis in the sample, wherein the one or more biomarkers comprise one or more xenobiotics, metabolites of xenobiotics, and/or biomarkers listed in Tables 14 and 15; and comparing the level(s) of the one or more biomarkers in the sample to ALS-positive and/or ALS-negative reference levels of the one or more biomarkers in order to determine whether a subject has ALS or has peripheral neuropathy or myopathy.
CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No. 60/777,338, filed Feb. 28, 2006, U.S. Provisional Application No. 60/789,392, filed Apr. 5, 2006, and U.S. Provisional Application No. 60/851,144, filed Oct. 12, 2006; the entire contents of these applications are hereby incorporated herein by reference.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

This invention was made, in part, with Government support under Grant No. 1 R43 ES013646-01 from the National Institutes of Health. The U.S. Government may have certain rights in this invention.

Provisional Applications (3)
Number Date Country
60777338 Feb 2006 US
60789392 Apr 2006 US
60851144 Oct 2006 US