Patent Application
20080020033
The following will describe one embodiment of the present invention in detail. It should be noted that the invention is not limited in any way by the following descriptions.
The embodiment below describes oil or fat compositions, a producing process thereof, and use thereof in this order.
Oil or fat compositions according to the present invention include a long-chain polyunsaturated fatty acid (LCPUFA) supply compound as a first component, and phospholipids as a second component, wherein the proportions of these components are determined according to the number of hydrolyzable fatty acid bonds. With the intake of the oil or fat compositions, the digestive system metabolizes the compounds to generate a sufficient amount of LCPUFA-PL whose constituents are LCPUFA. The LCPUFA-PL is then absorbed through the lymph vessels.
The LCPUFA supply compound used in the present invention is not particularly limited as long as it can be ingested with phospholipids and metabolized in the digestive system to generate LCPUFA-PL. The constituents of the LCPUFA-PL are LCPUFA and a phospholipids backbone structure (glycerol phosphate compound, ceramide compound, and the like). However, the LCPUFA supply compound may be any compound as long as it can supply LCPUFA.
More specifically, the LCPUFA supply compound is a compound that contains LCPUFA in its structure, and in which the LCPUFA can be separated by hydrolysis. In other words, the LCPUFA supply compound may be any compound that generates LCPUFA by being hydrolyzed by the hydrolase in the body. Note that, the LCPUFA supply compound may contain fatty acids other than LCPUFA.
Preferable examples of the LCPUFA supply compound include but are not particularly limited to: LCPUFA itself (i.e., free fatty acids); fatty acid alcohol esters such as alkylalcohol ester, aminoalcohol ester, and sterol ester; glycerolipids such as triglycerides, diglycerides, monoglycerides, glycerophospholipids, and glycoglycerolipids; sphingolipids such as sphingophospholipids and glycosphingolipids; sugars or sugar derivative esters such as sucrose fatty acid esters, sorbitan fatty acid esters, and ascorbic acid fatty acid esters; and carotenoid esters. These compounds may be used either individually or in combinations of two or more kinds.
In the present invention, the LCPUFA-PL is generated by the metabolism in the body, and therefore the only requirement for the LCPUFA supply compound is to supply LCPUFA. Nevertheless, the LCPUFA supply compound may be a phospholipid compound, i.e., a compound containing LCPUFA-PL itself. That is, the first component used in the present invention may be a non-phospholipid LCPUFA supply compound used either alone or, as required, together with the LCPUFA-PL. Alternatively, a specific kind of LCPUFA-PL may be used by itself.
The LCPUFA is not particularly limited as long as it is an unsaturated fatty acid having 20 or greater carbon atoms with the double bonds. Specific examples of such LCPUFA include: eicosadienoic acid; eicosatrienoic acids such as dihomo-γ-linolenic acid and mead acid; eicosatetraenoic acids such as arachidonic acid (AA); eicosapentaenoic acid; docosadienoic acid; docosatrienoic acid; docosatetraenoic acid; docosapentaenoic acid; docosahexaenoic acid (DHA); tetracosadienoic acid; tetracosatrienoic acid; tetracosatetraenoic acid; tetracosapentaenoic acid; tetracosahexaenoic acid. Among these examples, arachidonic acid (AA) and/or docosahexaenoic acid (DHA) are particularly preferable.
When used as a LCPUFA supply compound (free fatty acid), these fatty acids may be used either individually or in combinations of two or more kinds. When contained as constituents of the LCPUFA supply compound, these fatty acids may be contained in one kind or two or more kinds.
In the LCPUFA, at least one of the C—C double bonds (—C═C—) in its structure may be conjugated. The conjugate double bonds may include a carbonyl group (C═O), or may be formed by adjacent C—C double bonds.
The LCPUFA is abundant in fish oils and animal fats, and it is also found in algae. Thus, extracts from animals and plants containing a large amount of LCPUFA can be directly used as the free fatty acid LCPUFA. Examples of such extracts include: fish oils such as sardine oil, salmon oil, and purified fish oil; animal fats such as lard, beef tallow, and milk fat; and algae extracts such as a seaweed extract or tangle weed extract. These extracts can be produced by conventional extraction and preparation methods. The extracts may be unpurified if they are usable as the LCPUFA supply compound. If not, the extracts may be purified to a usable level. Evidently, there is no harm in purifying the extracts even when they are usable as the LCPUFA supply compound.
Microorganisms, which are also known to produce LCPUFA supply compounds, are also usable. For example, Mortierella fungi are known to produce a large amount of LCPUFA supply compound containing arachidonic acid (AA), dihomo-γ-linolenic acid, mead acid, eicosapentaenoic acid, eicosadienoic acid, and the like. Further, microorganisms that belong to genus Ulkenia or Schizochytrium are known to produce a large amount of docosapentaenoic acid or docosahexaenoic acid (DHA). These compounds may be suitably purified as required to make them usable as the LCPUFA supply compound.
Further, the LCPUFA supply compound so prepared or produced may be subjected to an enzyme or chemical treatment, so as to modify its molecular structure. Alternatively, the LCPUFA supply compound may be subjected to an alkali treatment to conjugate the double bonds. For example, the free fatty acid LCPUFA may be allowed to react with a suitable compound to synthesize a LCPUFA supply compound.
As described above, the first component may be a purified LCPUFA supply compound, or a mixture containing a LCPUFA supply compound, such as the extract or the composition containing other components. As such, the first component may contain compounds other than the LCPUFA supply compound. One example of such a compound is a fatty acid-containing compound other than the LCPUFA supply compound. Further, from the LCPUFA supply compound, fatty acids other than the LCPUFA may be separated.
When the first compound contains other compounds, or when it can separate fatty acids other than LCPUFA, the proportion of LCPUFA in all suppliable fatty acids from the total amount of the first component should be not less than 25 percent by weight, preferably not less than 33 percent by weight, or more preferably not less than 50 percent by weight.
As noted above, the LCPUFA in the present invention is preferably arachidonic acid (AA) and/or docosahexaenoic acid (DHA). In this case, the proportion of arachidonic acid (AA) in all suppliable fatty acids from the total amount of the first component should be not less than 1 percent by weight, preferably not less than 20.5 percent by weight, more preferably not less than 23 percent by weight, and even more preferably not less than 40 percent by weight. Further, the proportion of docosahexaenoic acid (DHA) in all suppliable fatty acids from the total amount of the first component should be not less than 11 percent by weight, preferably not less than 22.5 percent by weight, more preferably not less than 40 percent by weight, and even more preferably not less than 45 percent by weight.
Among all suppliable fatty acids from the total amount of the first component, it is preferable that arachidonic acid (AA) and docosahexaenoic acid (DHA) have substantially the same weight proportion. In other words, arachidonic acid (AA) and docosahexaenoic acid (DHA) should be contained in substantially equal amount with respect to all suppliable fatty acids.
By thus specifying the amount of suppliable fatty acids, the LCPUFA can be absorbed efficiently as the LCPUFA-PL. Note that, a proportion of the LCPUFA supply compound in the oil or fat composition is determined according to the number of hydrolyzable fatty acid bonds contained in the phospholipids, as will be described later in Producing Examples. In other words, the amount of suppliable fatty acids is determined based on the number of hydrolyzable fatty acid bonds contained in the phospholipids.
The phospholipids as the second component of the present invention are not particularly limited as long as they can be ingested with the LCPUFA supply compound and generate LCPUFA-PL by being metabolized in the digestive system. However, non-LCPUFA-PL whose constituents are not LCPUFA is particularly preferable. Specifically, non-LCPUFA-PL refers to those phospholipids in which the fatty acids forming the fatty acid bonds in the phospholipids are not LCPUFA.
As described above, in the present invention, the LCPUFA-PL is generated by the metabolism in the body. As such, the phospholipids contained in the oil or fat compositions according to the present invention need to supply a phospholipid backbone structure for the LCPUFA-PL. To this end, the non-LCPUFA-PL is particularly preferable as the second component. The LCPUFA-PL may be used instead of the non-LCPUFA-PL. However, the latter is more preferable because it allows the phospholipid backbone structure and the LCPUFA to be supplied from identifiable sources, which is preferable in obtaining desirable LCPUFA-PL structure in the body.
It is to be noted here that the phospholipids constitute a component for supplying a phospholipid backbone structure. Accordingly, the second component may be called “phospholipid backbone structure supply component” as opposed to the term “LCPUFA supply component” used for the first component.
The phospholipids used in the present invention are not particularly limited. Phospholipids are found in organisms of many different species, and therefore may be obtained from any organism. Namely, the phospholipids may be derived from non-animal sources as well as animal sources. The non-animal source of the phospholipids is not particularly limited, but plants are commonly used. Non-limiting examples of source plants include soy bean, corn, and rice. Among these examples, soy bean-derived phospholipids are preferable. More specifically, soy bean lecithin and/or soy bean phosphatidylserine are suitable.
Other than plants, microorganisms may be used as the non-animal source of phospholipids. Non-limiting examples of the source microbe include filamentous fungi, yeasts, and bacteria.
The animal source of the phospholipids is not particularly limited either. Commonly, phospholipids derived from various edible animals or domestic animals are preferably used. Specific examples include: marine organisms such as fish or seashells; birds such as chicken; mammals such as cows and pigs; and other vertebrates. Eggs including fish egg or chicken egg may also be used since they are rich in phospholipids. Egg yolk lecithin is one common example of a commercially available source. Other than eggs, various vertebrate organs such as cow brain or pig liver may be used.
Note that, the present invention preferably uses plant-derived phospholipids. This is to provide identifiable sources for the phospholipid backbone structure and the LCPUFA and thereby improve the LCPUFA-PL structure generated in the body, as described above. Plant-derived phospholipids are preferable for this purpose because most of plant-derived phospholipids are non-LCPUFA.
Preferable examples of non-LCPUFA used in the present invention include phospholipids in which the fatty acids forming fatty acid bonds in the phospholipids are not arachidonic acid (AA) or docosahexaenoic acid (DHA).
Specific examples of such phospholipids include: glycerophospholipids such as phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylglycerol (PG), and cardiolipin (CL); sphingophospholipids such as sphingomyelin (SP); and lysophospholipids such as lysophosphatidylcholine (LPC), lysophosphatidylserine (LPS), lysophosphatidylethanolamine (LPE), lysophosphatidylinositol (LPI), and lysophosphatidylglycerol (LPG). Among these phospholipids, PC, PS, PE, and PI are preferable, and PS is particularly preferable.
These phospholipids may be used either individually or in combinations of two or more kinds. Further, as in the case of the LCPUFA supply compound, the phospholipids may be of a purified form, or may be contained in a mixture as in extracts or compositions containing other components. That is, the second component may contain compounds other than phospholipids. Further, when the phospholipids are PS, a proportion of PS with respect to the total amount of phospholipids contained as the second component is preferably not less than 5 percent by weight, or more preferably not less than 23 percent by weight. This enables the PS to exhibit its brain function improving effect.
Oil or fat compositions according to the present invention can be produced by mixing the first and second components, i.e., the LCPUFA supply compound and the phospholipids. However, in order to increase the LCPUFA-PL level in the body, the present invention determines the respective proportions of these two components according to the number of hydrolyzable fatty acid bonds contained in the phospholipid molecules, taking into account the metabolism in the body. This enables the LCPUFA-PL to be efficiently generated in the body.
As described in the BACKGROUND ART section, orally administered fatty acids such as the LCPUFA-PL are known to reach the brain tissue without entering the liver (Non-Patent Publication 2). In the case of glycerophospholipids, as indicated by Formula (1) below, one of the fatty acid molecules (“Fatty Acid” in Formula (1)) being absorbed is hydrolyzed to generate lysophospholipids in the digestive system. In Formula (1) below, the fatty acid at the second position is hydrolyzed to become a free fatty acid. Note that, in Formula (1) below, the symbol X varies depending on the type of phospholipids. For example, X may be a hydrogen atom (H2) or a base such as choline.
The resulting lysophospholipids and fatty acids are absorbed in the small intestine, and are resynthesized in the small intestine cells to phospholipids, as represented in Formula (2) below. The phospholipids are then absorbed through the lymph vessels (Non-Patent Publication (3)).
Based on these findings, the inventors of the present invention have found that (i) the amount of LCPUFA-PL in the body can be increased by independently supplying the phospholipid backbone structure such as that of glycerophospholipids, and the LCPUFA (fatty acids in Formulae (1) and (2) above), and resynthesizing the two in the small intestine, (ii) LCPUFA-PL can be efficiently produced in the body by specifying the respective proportions of the LCPUFA supply compound (first component) and the phospholipids (second component) contained in oil or fat compositions, and (iii) the resulting LCPUFA-PL is actually absorbed through the lymph vessels. The inventors have also found that (iv) the value of the product can be increased with increased freedom of design by appropriately combining different types of first components and second components according to the target organism or required properties for the end product. The inventors have thus accomplished the present invention.
As described, in the present invention, with the intake of the oil or fat compositions containing the phospholipids and the LCPUFA supply compound, the molecules are metabolized and LCPUFA-PL is generated and absorbed in the body. Generally, LCPUFA-PL is obtained only in a small amount, supply thereof is instable, and the same quality cannot be always obtained. The present invention, on the other hand, produces oil or fat compositions using phospholipids and a LCPUFA supply compound, which are available in good supply and good quality at relatively low price. It is therefore possible to sufficiently increase the LCPUFA-PL level in the body and absorb it, without the need to directly ingest the LCPUFA-PL.
The properties and actions of the phospholipids and LCPUFA widely vary depending on types of phospholipids and LCPUFA. Conventionally, fractions of purified phospholipids from sources such as the cow brain, pig liver, or fish egg have been directly used. The present invention, on the other hand, uses the phospholipids and LCPUFA in different combinations, and thereby provides different kinds of LCPUFA-PL. By thus using the phospholipids and LCPUFA in different combinations according to the target organism or required properties for the end product, the value of the product can be increased with increased freedom of design.
In the present invention, the respective proportions of the phospholipids and LCPUFA supply compound are determined according to the number of hydrolyzable fatty acid bonds contained in the phospholipid molecules. Specifically, the respective proportions of the phospholipids and LCPUFA supply compound are determined according to the structure of the phospholipids, which may be glycerophospholipids or sphingophospholipids according to the broad classification scheme of phospholipids.
In the case of the glycerophospholipids, as indicated by Formula (3) below, a molecule of glycerophospholipid contains two fatty acids forming fatty acid bonds (“CO-Fatty Acid” in Formula (3)). The proportions of the glycerophospholipids and LCPUFA supply compound are determined in such a manner that at least one of these two fatty acids is replaced with LCPUFA.
On the other hand, in the case of the sphingophospholipids, as indicated by Formula (4) below, a molecule of sphingophospholipid contains one fatty acid forming a fatty acid bond (“O-Fatty Acid” in Formula (4)). Accordingly, the proportions of the sphingophospholipids and LCPUFA supply compound are determined in such a manner that the fatty acid forming the fatty acid bond in the sphingophospholipids is replaced with LCPUFA.
Note that, when the phospholipids are absorbed in the small intestine and resynthesized in the small intestine cells, not all but only some of the fatty acids forming the fatty acid bonds in the phospholipids are required to be replaced with LCPUFA. That is, the form or efficiency of resynthesis in the body will not be affected as long as the first and second components are suitably absorbed in the form of LCPUFA-PL.
When oil or fat compositions according to the present invention are orally ingested, the resynthesis does not always produce LCPUFA-PL according to the level calculated based on the numbers of moles or other factors, owning to the fact that the metabolism involves complex biochemical reactions and physiological activities in the body. As such, the present invention in actual practice determines the respective proportions of the first and second components according to the weight of suppliable LCPUFA in the first component, and the weight of the second component. This is done based on the results of Examples to be described later.
Specifically, the respective proportions of the first and second components are determined in such a manner that the weight ratio of all suppliable LCPUFA to all phospholipids (second component) is not less than 0.5. Preferably, the weight ratio is not less than 1, more preferably not less than 2, and even more preferably not less than 3. A weight ratio of less than 0.5 is not preferable because in this case the amount of suppliable LCPUFA becomes deficient with respect to the phospholipid backbone structure supplied by the phospholipids (second component). On the contrary, the amount of suppliable LCPUFA may be in excess of the supplied amount of phospholipid backbone structure, because the excess LCPUFA is absorbed after it is resynthesized to compounds other than phospholipids (for example, triglycerides). Accordingly, the upper limit of weight ratio is not particularly limited and is suitably set according to the intended use or applied field of the oil or fat compositions.
Oil or fat compositions according to the present invention are produced by mixing the phospholipids and LCPUFA supply compound using a conventional method, wherein the two components are mixed at specific proportions that are obtained based on calculated amounts of the phospholipids and LCPUFA. The mixing method and mixing conditions are not particularly limited.
Additionally, a substance (third component) that is miscible to the first and second components may be added. This enables the concentrations of the respective components to be adjusted, and stability of the oil or fat composition can be improved. Specifically, the third component may be diluent solvents or various kinds of additives. The type of diluent solvent is not particularly limited as long as it can dissolve the respective components. Examples of such diluent solvents include: an oil or fat containing triglycerides as the main constituents, such as soy bean oil, rape oil, and olive oil. Other examples include compounds that can be used as the LCPUFA supply compound, such as free fatty acids, fatty acid alcohol esters, glycerolipids, sphingolipids, sugars or sugar derivative esters, and carotenoid esters. These compounds may contain LCPUFA, though it is not preferable.
Non-limiting examples of additives include: vitamin E, tocotrienol, sesamin, sesaminol, sesamol, astaxanthin, astaxanthin esters, sterols, and carotenes. Oil or fat compositions according to the present invention are available as nutritional compositions for various food products. As such, any additive can be added if it is usable for food.
Oil or fat compositions according to the present invention include phospholipids. With the phospholipids (second component), the other components (first component and/or third component) may be solubilized in the form of a liposome. In this case, a dispersion liquid of an oil-in-water (O/W) type with a liposome can be used as oil or fat compositions according to the present invention (see Examples 8 below).
That is, oil or fat compositions according to the present invention can be produced by any of the techniques commonly used in the processing of fats and oils, such as dissolving in foods, or powderizing. The form of product oil or fat compositions is not particularly limited. For example, the oil or fat compositions may be in the form of a liquid, powder, or capsule (see Example 7 below). Further, the oil or fat compositions may be provided as pills or tablets.
Use of oil or fat compositions according to the present invention is not particularly limited. As a representative example, the oil or fat compositions can be suitably used as nutritional compositions for supplementing LCPUFA-PL. Such nutritional compositions can be used for any organisms, including humans as a representative example, as well as domestic animals and laboratory animals. Further, while the manner in which the nutritional compositions are ingested is not particularly limited, oral ingestion is most preferable. As such, the present invention also provides food containing the oil or fat compositions.
Food according to the present invention is not particularly limited as long as it contains the oil or fat compositions. Non-limiting examples include: bread, sweets of various kinds (including cold or frozen sweets), prepared food, dairy products, cereals, tofu, fried tofu, noodles, box lunch, seasonings, agricultural products such as wheat flour or meat, preserved food (canned food, frozen food, retort-packed food, etc.), soft drinks, milk beverage, soy milk, soups such as potage soup, and all other common foods. The method by which the oil or fat compositions are added to these common foods is not particularly limited, and any conventional method can be suitably used depending on the type of food used.
Further, food according to the present invention includes functional foods used for specific purposes, such as health foods or nutriments. Specific examples include nutraceutical foods including various supplements, and specified health foods. In the case of supplements, the oil or fat composition can be used only by processing it into appropriate form. Optionally, the third component may be added, as described in section (2-3) above.
Conventionally, animal organs such as the cow brain have been the major source of LCPUFA-PL. However, with the epidemic of the mad cow disease, these conventional sources have been avoided. The present invention, on the other hand, relies on plants and fermented microorganisms as the sources of the first and second components, which are the main components of LCPUFA-PL. This is advantageous in terms of customer acceptance, making the present invention particularly suitable when it is used in common foods or functional foods.
The applicable field of the invention is not just limited to food, but the invention is also applicable in the field of medicine. That is, oil or fat compositions according to the present invention may also be used as medicines. In this case, the invention is not just limited to a particular application, and any conventional technique may be used depending on the intended use.
The invention being thus described, it will be obvious that the same way may be varied in many ways. Such variations are not to be regarded as a departure from the spirit and scope of the invention, and all such modifications as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.
The following will describe the present invention in more detail based on Examples. It is to be noted that the invention is not limited in any way by the following Examples.
As the LCPUFA supply compound (first component), arachidonic acid (AA)-containing triglycerides (LCPUFA content of 50 percent by weight), and triglycerides with a low arachidonic acid content (LCPUFA content of 25 percent by weight) was used. As the second component, soy bean lecithin (Sigma product) was used.
These components were mixed at varying proportions as indicated in Table 1 below (all readings are in mg), so as to obtain oil or fat compositions A1 through F1, and a control composition 1.
Note that, the arachidonic acid (AA)-containing triglycerides were obtained as a fermented oil produced by arachidonic acid-producing filamentous Mortierella alpina, and it contained 40 percent by weight of arachidonic acid (AA), 5 percent by weight of dihomo-γ-linolenic acid (DGLA), and 5 percent by weight of other LCPUFA (eicosadienoic acid, eicosatrienoic acid, etc) as the constituents of the total fatty acids. The triglycerides with a low arachidonic acid content was also obtained as a fermented oil produced by arachidonic acid-producing filamentous Mortierella alpina, and it contained 23 percent by weight of arachidonic acid (AA), and 2 percent by weight of dihomo-γ-linolenic acid (DGLA) as the constituents of the total fatty acids. The soy bean lecithin contained constituent phospholipids but did not contain LCPUFA.
Further, in order to increase the absorbing efficiency of the oil or fat compositions administered to rats in the Examples below, 200 mg of taurocholic acid and 50 mg of bovine serum albumin were added to the oil or fat compositions.
Oil or fat compositions A2 through C2, and a control composition 2 were produced by mixing the first and second components at the proportions indicated in Table 2 below (all readings are in mg). The procedure of Producing Example 1 was used except that three kinds of LCPUFA supply compounds, the arachidonic acid-containing triglyceride of Producing Example 1, dihomo-γ-linolenic acid (DGLA)-containing triglycerides, and sardine oil (NISSUI) were used as the first component.
Note that, the dihomo-γ-linolenic acid (DGLA)-containing triglycerides were obtained as a fermented oil produced by Mortierella alpina, and it contained 50 percent by weight of dihomo-γ-linolenic acid (DGLA) as the constituent of the total fatty acids. The sardine oil contained 1 percent by weight of arachidonic acid, 21 percent by weight of eicosapentaenoic acid (EPA), and 11 percent by weight of docosahexaenoic acid (DHA) as the constituents of the total fatty acids.
Oil or fat compositions A3 through D3, and a control composition 3 were produced by mixing the first and second components at the proportions indicated in Table 3 below (all readings are in mg). The procedure of Producing Example 1 was used except that three kinds of LCPUFA supply compounds, the arachidonic acid-containing triglyceride of Producing Example 1, the dihomo-γ-linolenic acid-containing triglycerides of Producing Example 2, and purified fish oil (NISSUI) were used as the first component, and that soy bean phosphatidylserine (BIZEN CHEMICAL) was used as the second component.
Note that, the purified fish oil contained 1 percent by weight of arachidonic acid (AA), 4 percent by weight of eicosapentaenoic acid (EPA), and 45 percent by weight of docosahexaenoic acid (DHA) as the constituents of the total fatty acids. Further, an oil mixture containing the arachidonic acid (AA)-containing triglycerides and the purified fish oil in equal amount contained 20.5 percent by weight of arachidonic acid (AA), 2.5 percent by weight of dihomo-γ-linolenic acid (DGLA), 2 percent by weight of eicosapentaenoic acid (EPA), 22.5 percent by weight of docosahexaenoic acid (DHA), and 2.5 percent by weight of other LCPUFA (eicosadienoic acid, eicosatrienoic acid, etc.) as the constituents of the total fatty acids. As to the soy bean phosphatidylserine, 23 percent by weight of its constituent phospholipids was phosphatidylserine (PS), and the LCPUFA was not contained.
Oil or fat compositions A4 through E4, and a control composition 4 were produced by mixing the first and second components at the proportions indicated in Table 4 below (all readings are in mg). The procedure of Producing Example 1 was used except that the arachidonic acid-containing triglycerides of Producing Example 1 was used as the first component, the soy bean lecithin of Producing Example 1 as the second component, and a soy bean oil as a diluent solvent.
Note that, the soy bean oil almost entirely consisted of triglycerides, and did not contain LCPUFA.
Oil or fat compositions A5 through E5, and a control composition 5 were produced by mixing the first and second components at the proportions indicated in Table 5 below (all readings are in mg except those bracketed). The procedure of Producing Example 1 was used except that the purified fish oil of Producing Example 3 was used as the LCPUFA supply compound (first component), the soy bean lecithin of Producing Example 1 as the second component, and the soy bean oil of Producing Example 4 as a diluent solvent.
Oil or fat compositions A6 through D6, and a control composition 6 were produced by mixing the first and second components at the proportions indicated in Table 6 below (all readings are in mg except those bracketed). The procedure of Producing Example 1 was used except that the arachidonic acid-containing triglycerides of Producing Example 1 and the purified fish oil of Producing Example 3 were used as the first component, and that the soy bean lecithin of Producing Example 1 and the soy bean phosphatidylserine of Producing Example 3 were used as the second component.
Using SD male rats, 8-9 weeks of age, cannulation was carried out on the thoracic-lymph duct and stomach of anesthetized rats. The lymph flow was stabilized overnight by injecting a saline solution (Otsuka Pharmaceutical Co., Ltd.) at 3 ml/hr. In the next morning, the lymph was collected for 2 hours. The oil or fat compositions A1 through F1, and control composition 1 of Producing Example 1 were then administered through a gastric tube, and the lymph was collected over time.
The lymph was analyzed according to an ordinary method, as described below. First, lipids were collected from the lymph using the Folch method. After fractionating the PL fractions by thin-layer chromatography, transmethylation was carried out with a solvent of hydrochloric acid in methanol. By gas chromatography, the fatty acids in PL were quantified. As the internal standard, pentadecanoic acid was used.
Table 7 represents LCPUFA-PL concentrations in the lymph collected from rats administered with the respective oil or fat compositions, 2 hours (average concentration after 1 to 2 hours) and 6 hours (average concentration after 5 to 6 hours) after the administration. Note that, all readings are in mg/mL.
As can be seen from Table 7, the LCPUFA-PL concentration of the rats administered with the oil or fat composition A1 was substantially the same as that of the rats administered with the control composition 1, whereas the rats administered with the oil or fat compositions B1 through F1 had higher LCPUFA-PL concentrations than those administered with the control composition 1.
According to the procedure of Example 1, the oil or fat compositions A2 through C2, and control composition 2 were administered through a gastric tube to SD male rats, 8-9 weeks of age. The lymph was collected and analysis was made according to the procedure of Example 1.
Table 8 represents LCPUFA-PL concentrations in the lymph collected from rats administered with the respective oil or fat compositions, 2 hours (average concentration after 1 to 2 hours) and 6 hours (average concentration after 5 to 6 hours) after the administration. Note that, all readings are in mg/mL.
As can be seen from Table 8, the rats administered with the oil or fat compositions A2 through C2 had higher LCPUFA-PL concentrations than those administered with the control composition 2. Further, the LCPUFA-PL concentration in the lymph increased according to the fatty acid compositions of the LCPUFA supply compounds used for the oil or fat compositions A2 through C2.
According to the procedure of Example 1, the oil or fat compositions A3 through D3, and control composition 3 were administered through a gastric tube to SD male rats, 8-9 weeks of age. The lymph was collected and analysis was made according to the procedure of Example 1.
Table 9 represents LCPUFA-PL concentrations in the lymph collected from rats administered with the respective oil or fat compositions, 2 hours (average concentration after 1 to 2 hours) and 6 hours (average concentration after 5 to 6 hours) after the administration. Note that, all readings are in μg/mL.
As can be seen from Table 9, the rats administered with the oil or fat compositions A3 through D3 had higher (PS)-type LCPUFA-PL concentrations than those administered with the control composition 3. Further, the phosphatidylserine LCPUFA-PL concentration in the lymph increased according to the fatty acid compositions of the LCPUFA supply compounds used for the oil or fat compositions A3 through D3.
According to the procedure of Example 1, the oil or fat compositions A4 through E4, and control composition 4 were administered through a gastric tube to SD male rats, 8-9 weeks of age. The lymph was collected and analysis was made according to the procedure of Example 1.
Table 10 represents LCPUFA-PL concentrations in the lymph collected from rats administered with the respective oil or fat compositions, 2 hours (average concentration after 1 to 2 hours) and 6 hours (average concentration after 5 to 6 hours) after the administration. Note that, all readings are in mg/mL.
As can be seen from Table 10, in the rats administered with the oil or fat composition A4, the concentration of AA in PL and the LCPUFA-PL concentration were substantially the same as those of the rats administered with the control composition 4, whereas the rats administered with the oil or fat compositions B4 through E4 had higher concentrations of AA in PL and higher LCPUFA-PL concentrations than those administered with the control composition 4.
According to the procedure of Example 1, the oil or fat compositions A5 through E5, and control composition 5 were administered through a gastric tube to SD male rats, 8-9 weeks of age. The lymph was collected and analysis was made according to the procedure of Example 1.
Table 11 represents LCPUFA-PL concentrations in the lymph collected from rats administered with the respective oil or fat compositions, 2 hours (average concentration after 1 to 2 hours) and 6 hours (average concentration after 5 to 6 hours) after the administration. Note that, all readings are in mg/mL.
As can be seen from Table 11, in the rats administered with the oil or fat composition A5, the concentration of DHA in PL was substantially the same as that of the rats administered with the control composition 5, whereas the rats administered with the oil or fat compositions B5 through E5 had higher concentrations of DHA in PL and higher LCPUFA-PL concentrations than those administered with the control composition 5.
According to the procedure of Example 1, the oil or fat compositions A6 through E6, and control composition 6 were administered through a gastric tube to SD male rats, 8-9 weeks of age. The lymph was collected and analysis was made according to the procedure of Example 1.
Table 12 represents LCPUFA-PL concentrations in the lymph collected from rats administered with the respective oil or fat compositions, 2 hours (average concentration after 1 to 2 hours) and 6 hours (average concentration after 5 to 6 hours) after the administration. Note that, all readings are in μg/mL.
As can be seen from Table 12, in the rats administered with the oil or fat composition A6, the phosphatidylserine (PS)-type LCPUFA-PL concentration was substantially the same as that of the rats administered with the control composition 6, whereas the rats administered with the oil or fat compositions B6 through D6 had higher phosphatidylserine LCPUFA-PL concentrations than those administered with the control composition 6.
Three kinds of LCPUFA supply compounds were used as the first component: The arachidonic acid (AA)-containing triglycerides used in Example 1, the dihomo-γ-linolenic acid-containing triglycerides used in Example 2, and the purified fish oil used in Example 3. As the second component, the soy bean phosphatidylserine used in Example 3, and the soy bean lecithin used in Example 1 or 2 were used.
As the third component, a soy bean oil (SHOWASANGYO Co., Ltd.), a vitamin E oil (Eisai Co., Ltd.), sesamin (SUNTORY), and astaxanthin oil (SUNTORY) were used. Note here that, the soy bean oil was used as a diluent solvent. The vitamin E oil was an additive used as a stabilizer. The sesamin and astaxanthin oil were additives used as nutrients.
These components were mixed at varying proportions as indicated in Table 13 below (all readings are in weight %), and were mixed together according to the procedures of Producing Examples 1 through 6, so as to produce capsule contents 1 through 5.
A 100:35 mixture (weight ratio) of galatin (Nitta Gelatin Inc.) and food-additive glycerine (Kao Corporation) was prepared and water was added thereto. By dissolving the solution at 50° C. to 60°, a gelatin coat with a viscosity of 2,000 cp was prepared. With the contents 1 through 5, capsules were formed and dried using an ordinary method, so as to prepare soft capsules, each containing 180 mg of the content. The soft capsules are suitable for oral ingestion.
The arachidonic acid-containing triglycerides and purified fish oil used in Example 3, and the vitamin E oil and astaxanthin oil used in Example 4 were mixed together at a weight ratio of 50:50:0.05:0.01 according to the procedures of Producing Examples 1 through 3, so as to prepare an oil or fat composition G. A 1:5 mixture (weight ratio) of the oil or fat composition G and soy bean lecithin was stirred in water at 60° C. for 5 to 30 minutes, using a mixing-and-dispersing device (the Mtechnique product, CLEARMIX). As a result, the oil or fat composition G was uniformly dispersed in water in the form of a liposome, and a dispersion liquid of oil or fat composition G was obtained.
The concentration of the oil or fat composition G in the dispersion liquid was controllable in a range of 0.1 to 20 percent by weight, and the liquid was substantially transparent or opaque white. The average particle size of the liposome was 50 nm to 100 nm. A dispersion liquid containing 10% oil or fat composition G was used as an oil or fat composition of the present invention, and was added to orange juice, carbonated water, coffee, milk, soy milk, and a potage soup, each in 1/100 (v/v), so as to prepare (produce) drinks as the food of the present invention. These drinks were suitable for oral ingestion.
As described, the present invention effectively increases the LCPUFA-PL level in the body without the need to directly take LCPUFA-PL. Further, the components of the LCPUFA-PL can be suitably obtained from plant or microorganism sources. The product is therefore advantageous in terms of customer acceptance. Further, by combining these components, the value of the product can be increased with increased freedom of design. The present invention therefore greatly and efficiently enhances the effectiveness of the LCPUFA-PL.
With these and other advantages, the present invention is useful not only in industries dealing with functional food but also in industries dealing with food in general and even medicines.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2003-403630 | Dec 2004 | JP | national |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/JP04/17915 | 12/2/2004 | WO | 00 | 4/16/2007 |
