The present disclosure relates to supercritical fluid chromatography (SFC) and/or a carbon dioxide based chromatography system. More specifically, the present disclosure relates to methods and systems for controlling the density of the mobile phase in the region of interest of a chromatographic system.
Developing a successful chromatographic separation method usually requires extensive experimentation. Such method development often involves the evaluation and optimization of numerous variables. These variables may include the choice of chromatographic system (e.g., carbon dioxide based chromatography, SFC, high pressure liquid chromatography (HPLC), gas chromatography (GC)), the choice of mobile phase and mobile phase compositions, the choice of column chemistry and column dimensions, the choice of detector, etc. Once a successful chromatographic separation method has been developed, it often needs to be transferred and performed on different chromatographic systems. For example, separation on an analytical scale SFC system may need to be transferred and performed on a preparative scale SFC system. Similarly, a preparative scale SFC system may be modified thereby requiring the new separation method to be transferred and performed on a different preparative scale SFC system.
For liquid chromatography, the theory and understanding for transferring methods between different system or column configurations is generally well understood. Guidelines for transferring LC methods are straightforward and typically do not need additional optimization.
When employing a SFC and/or a carbon dioxide based chromatography system, however, effective separation method transfer between different chromatography systems requires special consideration. Chromatographic separations using a mobile phase comprising carbon dioxide that are transferred from one chromatographic system to another chromatographic system typically may need to be re-developed to achieve the same successful separation as achieved on the original chromatographic system.
In WO2014/201222 A1, researchers at Waters Technologies Corporation disclosed a methodology for scaling SFC and/or carbon dioxide based chromatography methods between different systems and/or column configurations. The methodology includes measuring an average mobile phase density from the density profile along the system during a first separation utilizing carbon dioxide as a mobile phase component and substantially duplicating the average density for a second separation to produce similar selectivity and retention factors. The researchers at Waters Technologies Corporation also disclosed that the average of the pressure profile may be used as a close approximation to duplicate average of the density profiles between separations.
In WO2015/023533 A1, researchers at Waters Technologies Corporation disclosed apparatus for regulating the average mobile phase density or pressure in a carbon dioxide based chromatographic system. The disclosed apparatus includes a controller, a set of pressure or density sensors and a set of instructions capable of determining the pressure drop across a column and adjusting at least one system component or parameter to achieve a predetermined average mobile phase density or pressure in the system. But since filing WO2015/023533 A1, researchers at Waters Technologies Corporation have discovered specific new ways to efficiently transfer a carbon dioxide-based separation procedure from a first chromatographic system to a second system.
The present disclosure relates to methods and systems for efficiently transferring a carbon dioxide based separation procedure from a first chromatographic system to a second chromatographic system. The methods involve identifying an average column pressure for the carbon dioxide based separation in the first chromatographic system; determining a measured average column pressure for the carbon dioxide based separation in the second chromatographic system; and comparing the measured average column pressure for the carbon dioxide based separation in the second chromatographic system with the identified average column pressure for the carbon dioxide based separation in the first chromatographic system.
In some embodiments, determining a measured average column pressure for the carbon dioxide based separation in the second chromatographic system comprises calculating the measured average column pressure from a plurality of measurements proximate to the column in the second chromatographic system. In some embodiments, determining a measured average column pressure for the carbon dioxide based separation in the second chromatographic system comprises calculating the measured average column pressure from a plurality of measurements at an end of the column in the second chromatographic system.
In some embodiments, methods of the present invention involve altering a cross-sectional area of a column packed with media in the second chromatographic system along a length of the column to more closely match the identified average column pressure for the carbon dioxide based separation in the first chromatographic system. Altering the flow rate may comprise using a column in the second chromatography system comprising a column jacket comprising a thickness that increases along the length of the column and packed with media within the inner surface of the column jacket such that a cross-sectional area packed with media within the inner surface of the column jacket decreases along the length of the column. Altering the flow rate may comprise using a column in the second chromatography system comprising an insert comprising a thickness that increases along the length of the column, wherein an outer surface of the insert is proximate to the inner surface of the column jacket and wherein media is packed within an inner surface of the insert such that a cross-sectional area packed with media within the column jacket decreases along the length of the column. Altering the flow rate may comprise using a column in the second chromatography system comprising an insert comprising an annular cone, wherein media is packed between an inner surface of the column jacket and an outer surface of the insert such that a cross-sectional area within the column jacket comprising packed media decreases along the length of the column.
Some embodiments involve repeating the steps of determining a measured average column pressure for the carbon dioxide based separation in the second chromatographic system; comparing the measured average column pressure for the carbon dioxide based separation in the second chromatographic system with the identified average column pressure for the carbon dioxide based separation in the first chromatographic system; and altering a cross-sectional area of a column packed with media in the second chromatographic system along a length of the column to more closely match the identified average column pressure for the carbon dioxide based separation in the first chromatographic system. Some embodiments involve, iteratively or continually, repeatedly altering a cross-sectional area of a column packed with media in the second chromatographic system along a length of the column until the measured average column pressure for the carbon dioxide based separation in the second chromatographic system substantially matches the identified average column pressure for the carbon dioxide based separation in the first chromatographic system.
In some embodiments, the present invention comprises a column for a carbon dioxide based separation procedure in a chromatography system. In some embodiments, systems of the present invention include a column for a carbon dioxide based separation procedure in a chromatography system. The column includes a column jacket and media packed within the column jacket, wherein a cross-sectional area of media packed within the column jacket decreases along the length of the column. In some such columns, the column jacket comprises a thickness that increases along the length of the column such that a cross-sectional area of media packed within the column jacket decreases along the length of the column. In some such columns, the column comprises an insert having a thickness that increases along the length of the column, wherein an outer surface of the insert is proximate to the inner surface of the column jacket and wherein media is packed within an inner surface of the insert such that a cross-sectional area packed with media within the column jacket decreases along the length of the column. In some such columns, the column comprises an insert comprising an annular cone, wherein media is packed between an inner surface of the column jacket and an outer surface of the insert such that a cross-sectional area within the column jacket comprising packed media decreases along the length of the column.
In some embodiments, methods of the present invention involve adding makeup fluid along the length of a column in the second chromatography system to more closely match the identified average column pressure for the carbon dioxide based separation in the first chromatographic system. Adding makeup fluid along the length of a column in the second chromatography system may comprise allowing makeup fluid to flow, from a channel of makeup fluid within the column in the second chromatography system, through a porous material and into packed media along the length of the column. Adding makeup fluid along the length of a column in the second chromatography system may comprise allowing makeup fluid to flow, from a channel of makeup fluid within the column in the second chromatography system, through discrete apertures and into packed media along the length of the column. The channel of makeup fluid may be formed between an inner surface of the column jacket and an outer surface of a cylinder of porous material within the column. The channel of makeup fluid may be formed within an inner surface of the cylinder of porous material within the column.
Some embodiments involve repeating the steps of determining a measured average column pressure for the carbon dioxide based separation in the second chromatographic system; comparing the measured average column pressure for the carbon dioxide based separation in the second chromatographic system with the identified average column pressure for the carbon dioxide based separation in the first chromatographic system; and adding makeup fluid along the length of a column in the second chromatography system to more closely match the identified average column pressure for the carbon dioxide based separation in the first chromatographic system. Some embodiments involve, iteratively or continually, repeatedly adding makeup fluid along the length of a column in the second chromatography system until the measured average column pressure for the carbon dioxide based separation in the second chromatographic system substantially matches the identified average column pressure for the carbon dioxide based separation in the first chromatographic system.
In some embodiments, the present invention comprises a column for a carbon dioxide based separation procedure in a chromatography system. In some embodiments, systems of the present invention include a column for a carbon dioxide based separation procedure in a chromatography system. The column includes a column jacket, media packed within the column jacket, and an annular insert (e.g., a cylindrical annular insert) within the column jacket, wherein the annular insert allows makeup fluid to flow from a channel of makeup fluid within the column jacket, through the annular insert, and into the packed media along the length of the column. In some such columns, a porosity of the annular insert allows makeup fluid to flow, from a channel of makeup fluid within the column jacket, through the cylindrical annular insert and into the packed media along the length of the column. In some such columns, a plurality of discrete apertures allow makeup fluid to flow, from a channel of makeup fluid within the column in the second chromatography system, through the plurality of discrete apertures and into the packed media along the length of the column. In some such columns, the channel of makeup fluid is formed between an inner surface of the column jacket and an outer surface of the annular insert within the column. In some such columns, the channel of makeup fluid is formed within an inner surface of the annular insert within the column.
The foregoing and other features provided by embodiments of the present invention will be more fully understood from the following description when read together with the accompanying drawings.
As used herein, the phrase “chromatographic system” refers to a combination of instruments or equipment, e.g., a pump, a column, a detector, and accompanying accessories that may be used to perform a separation to detect target analytes.
In some embodiments, the present disclosure relates to carbon dioxide based separation in a chromatographic system having a pump, a column located downstream of the pump, a detector located downstream of the column, a back pressure regulator located downstream of the detector, and a first sensor and a second sensor. In some such embodiments, the sensors may be pressure sensors for measuring mobile phase pressure in the system. Mobile phase pressure measurements may be used, along with measured or estimated mobile phase temperatures, to estimate the mobile phase density. The first sensor may be contained in or connected to an outlet of a pump, may be contained in or connected to an inlet of a column, or positioned anywhere in between. The second sensor may be contained in or connected to an inlet of a back pressure regulator, may be contained in or connected to an outlet of the column, or positioned anywhere in between. In some embodiments, the mobile phase density or pressure in the system may be at equilibrium when the first and second mobile phase density or pressure measurements are measured by the first and second sensors, or when the cross-sectional area of a column packed with media in the second system is altered or makeup fluid is added along the length of a column in a second system. In other embodiments, the mobile phase density or pressure in the system is not at equilibrium when the first and second mobile phase density or pressure measurements are measured by the first and second sensors, or when the cross-sectional area of a column packed with media in the second system is altered or makeup fluid is added along the length of a column in a second system.
In some embodiments, the present disclosure relates to carbon dioxide based separation in a chromatographic system having a controller, a first sensor and a second sensor both in signal communication with the controller, and a set of instructions utilized by the controller. The controller is capable of averaging the first and the second mobile phase pressure measurements to determine a measured average mobile phase pressure value. In some embodiments, the controller is capable of determining a measured average column pressure from the measured mobile phase pressure values. In some embodiments, the measured average mobile phase pressure value determined by the controller is a measured average column pressure or at least a close approximation thereof. In some such embodiments, the controller is capable of comparing the measured average column pressure with an identified average column pressure. In some such embodiments, the controller suggests altering a cross-sectional area of a column packed with media along the length of the column or adding makeup fluid along the length of the column to more closely match an identified average column pressure. In some such embodiments, the controller is capable of suggesting a column featuring a cross-sectional area that changes along the length of the column or a column that enable makeup fluid to be added along the length of the column to more closely match an identified average column pressure.
The present disclosure may be useful for transferring separations between analytical scale chromatographic systems, preparative scale chromatographic systems, and combinations thereof. For example, the present disclosure may be useful in transferring a separation from an analytical scale chromatographic system to a preparative scale chromatographic system, or a preparative scale chromatographic system to an analytical scale chromatographic system. The present disclosure may also be useful in transferring a separation from one analytical scale chromatographic system to another analytical scale chromatographic system, or from one preparative scale chromatographic system to another preparative scale chromatographic system. A list of chromatographic systems for which the present disclosure may be applicable include, but is not limited to, carbon dioxide-based chromatographic systems commercially available from Waters Technologies Corporation (Milford, Mass.) and branded as ACQUITY® UPC2, Method Station SFC, Resolution SFC MS, Preparative SFC Instruments (e.g., Investigator SFC, Prep 100 SFC, SFC 80/200/350 Preparative Systems). Chromatographic systems for which the present disclosure may be applicable may comprise columns designed for use with a mobile phase including carbon dioxide. In some embodiments, columns designed for use with a carbon dioxide containing mobile phase are branded as Waters Technologies Corporation (Milford, Mass.) UPC2 and/or SFC columns including both chiral and achiral stationary phases.
The distinction between different chromatographic systems, e.g., a first chromatographic system and a second chromatographic system, may include any change in the system configuration that results in a change in the overall operating average mobile phase density or average column pressure. For example, the distinction between different chromatographic systems may be the use of different instruments such as a carbon dioxide based analytical chromatographic system, for example a system commercially available from Waters Technologies Corporation (Milford, Mass.) and branded as an ACQUITY® UPC2 system versus a carbon dioxide based preparative chromatography system, for example a system commercially available from Waters Technologies Corporation (Milford, Mass.) and branded as a Prep 100 SFC system. The distinction may also be a change in one or more components on the same instrument, e.g., a change in system configuration. For example, the distinction may be a change in column configuration, e.g. length, internal diameter or particle size, or a change in tubing, e.g., length or internal diameter, a change in a valve, e.g., the addition or removal of a valve, or the addition or removal of system components such as detectors, column ovens, etc.
Preferably, the present disclosure may be applied to any change or distinction, e.g. instrument, column particle size, column length, etc., between different chromatographic systems which results in greater than about a 10% change in overall operating average mobile phase density or average column pressure. More preferably, the present disclosure may be applied to any change or distinction which results in greater than about a 5% change in overall operating average mobile phase density or average column pressure. Even more preferably, the present disclosure may be applied to any change or distinction which results in greater than about a 1% change in overall operating average mobile phase density or average column pressure.
The present disclosure relates to efficiently transferring carbon dioxide based separations between systems. As used herein, the phrase “efficiently transferring” of a carbon dioxide based separation refers to the concept of transferring a carbon dioxide based separation, methodology, or method parameters between chromatographic systems while maintaining the chromatographic integrity of the separation, e.g., preserving retention factors and selectivity of at least one target analyte, preferably two or more target analytes. An efficiently transferred separation is one that substantially reproduces the chromatographic integrity of the separation obtained on the first chromatographic system on the second chromatographic system. For example, an efficiently transferred carbon dioxide based separation is one wherein the second carbon dioxide based separation performed on the second chromatographic system has a target analyte, or target analytes, having substantially the same retention factor (k′) or selectivity as the first carbon dioxide based separation performed on the first system.
As used herein, the term “retention factor” or “k′” refers to the ratio of time an analyte is retained in the stationary phase to the time it is retained in the mobile phase under either isocratic or gradient conditions. For an efficiently transferred carbon dioxide-based separation method, the difference in retention factor for any given target analyte between a first and a second separation should be minimized. Preferably, the difference in retention factor for a target analyte between a first and a second separation is less than about 10%. More preferably, the difference in retention factor for a target analyte between a first and a second separation is less than about 5%. Even more preferably, the difference in retention factor for a target analyte between a first and a second separation is less than about 1%.
For multiple target analytes, the difference in retention factor for each target analyte, respectively, between a first and a second separation should also be minimized. Multiple target analytes may include 2 or more target analytes, e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, etc. Preferably, all or a majority of the target analytes have substantially the same retention factor between the first and second separations. Because all analytes respond differently to system changes, not all of the target analytes may have substantially the same retention factor between the first and second separations. Preferably, the difference in retention factor for each multiple target analyte, respectively, between a first and a second separation is less than about 10%. More preferably, the difference in retention factor for each multiple target analyte, respectively, between a first and a second separation is less than about 5%. Even more preferably, the difference in retention factors for each multiple target analyte, respectively, between a first and a second separation is less than about 1%.
As used herein, the term “selectivity” or “separation factor” or “a” refers to the degree of separation of two analytes in a separation. For example, the separation factor for two analytes, A and B, is the ratio of their respective retention factors, provided A elutes before B, e.g., α=k′B/k ′A. The selectivity between two target analytes between a first and a second separation should be maintained. Preferably, the change in selectivity for two target analytes between a first and a second separation is less than about 10%. More preferably, the change in selectivity for two target analytes between a first and a second separation is less than about 5%. Even more preferably, the change in selectivity for two target analytes between a first and a second separation is less than about 1%.
As used herein, the phrases “carbon dioxide-based separation” and “carbon dioxide-based separation (procedure” refer to method parameters and/or settings used with a particular carbon dioxide based chromatographic system to control or effect a separation of target analytes. The mobile phase in a carbon dioxide-based separation includes at least, in part, carbon dioxide.
Similar to
The difference between system 1000 of
Columns differences between chromatographic system are not limited to differences in particle diameter. Among other ways, columns may differ in length and internal diameter. Column stationary phases may differ in regard to chemistry, base particle, ligand, bonding density, endcapping, pore size, etc. Column manufacturers typically produce columns having the same stationary phase, e.g., same chemistry, same base particle, same ligand, same bonding density, same endcapping and same pore size, in several different particle size and column dimension configurations. In one embodiment, the two different separation systems have a first and a second respective column, wherein the first and second columns have similar stationary phases. The similar stationary phases may have, at least, same chemistry, same base particle, same ligand, same bonding density, same endcapping or same pore size. The present invention is applicable where the columns in two different chromatographic systems have the same stationary phase.
Due to the compressible nature of the carbon dioxide based mobile phase at or near supercritical conditions, the mobile phase density must be managed from the sample introduction to detection. More specifically, the average density of the mobile phase across the column must be conserved in order to match retention characteristics of the analytes.
As disclosed in the prior art, the average column pressure of the mobile phase can be adjusted by adjusting the set point of the BPR. For example, the set point of the BPR 3080 may be selected to address the pressure difference that may be caused by differences between the column particle diameters of column 1100 and column 3300. In particular, the set point of BPR 3080 may be decreased in
The inventors of the present disclosure recognized that, using average pressure as a close approximation for average density, the effect of mobile phase density on solubility and analyte retention can be normalized by substantially duplicating the average column pressure from a separation method in a first chromatographic system in a separation method in a second chromatographic system. The inventors were aware that settings related to the back pressure regulator (BPR) may be changed to substantially match an average column pressure in another chromatographic system. But the inventors further recognized that an average column pressure may be achieved in different SFC and/or a carbon dioxide based chromatographic systems without changing the settings related to BPR. The inventors further recognized that an average column pressure may be achieved in different SFC and/or a carbon dioxide based chromatographic systems without solely relying on changing the settings related to post-detector BPR.
The inventors recognized that cross-sectional area of a column packed with media can be altered along the length of the column to adjust the average column pressure in a chromatographic system.
Step 222 of
In step 222 of
In step 232 of
In step 242 of
Step 242 of method 202 may comprise selecting a different column for the second chromatographic system.
Additionally or alternatively, the diameter of a column jacket may vary along the length of the column such that the cross-sectional area within the column jacket packed with media varies along the length of column. The diameter of the jacket may uniformly and continuously vary from one end of the jacket to the other. Nonetheless, the diameter of the jacket need not vary uniformly from one end of the jacket to the other. For example, the diameter may vary incrementally from one thickness to another. Similarly, the diameter of the jacket need not vary continuously from one end of the jacket to the other. For example, the diameter may vary for only a portion of the length of the jacket. Due to the diameter variation of the jacket, the cross-sectional area packed with media varies along the length of the column.
Step 242 of method 202 may comprise modifying the column in the second chromatographic system, such as by adding an insert.
In step 242 of
As illustrated in
As illustrated in
Despite the fact that the particle diameter of the column 3300 of
The inventors also recognized that makeup fluid may be added along the length of a column in a second chromatography system to more closely match an identified average column pressure for carbon dioxide based separation in a first chromatographic system.
Like step 222 of
In step 722 of
In step 732 of
In step 742 of
The makeup fluid added in step 742 is preferably the same composition as that of the mobile phase. Nonetheless, the inventors recognized that the makeup fluid could have a different composition that was miscible with that of the mobile phase. The inventors similarly recognized that the makeup fluid could have a composition that was immiscible with that of the mobile phase to block portions of the column. The inventors further recognized that the composition of the makeup fluid could be altered over time to substantially match the composition of the mobile phase when the mobile phase is undergoing a composition program gradient separation. Alternatively, the inventors further recognized that a gradient in the makeup fluid could be introduced such that the composition of the makeup fluid does not substantially match the composition of the mobile phase when the mobile phase is undergoing a composition program gradient separation.
Step 742 of method 702 may comprise selecting a different column for the second chromatographic system. Step 742 of method 702 may comprise modifying a column in the second chromatographic system, such as by adding an insert to the column.
As illustrated in
In operation of a second chromatographic system including column 800, mobile phase fluid may be introduced into channel 840 axially and/or radially. For example, mobile phase fluid may be introduced through a single port that allows it to flow axially into channel 840. To the extent mobile phase fluid is introduced to channel 840 radially, it may be introduced only at one or more portions of the length of column 800. Similarly, fluid may be introduced into packed media 820 axially and/or radially through porous insert 830 from channel 840. The mobile phase fluid that has been introduced flows through packed media 820 and also more-freely, through channel 840.
As the pressure drops along portion of column 800 packed with media 820, some mobile phase fluid from channel 840 migrates through the pores in porous insert 830 into the packed stationary phase media 820. The amount of fluid that migrates through porous insert 830 depends on the porosity, thickness, and diameter of insert 830. The amount of fluid that migrates through porous insert 830 further depends on the pressure differential between channel 840 and media 820. The amount of fluid that migrates through porous insert 830 further depends on the pressure and temperature of the mobile phase fluid in channel 840. The fluid that migrates into the stationary phase media 820 in column 800 may be called makeup fluid and have features described above with respect to step 742. And its migration would decrease the pressure drop along column 800.
As further explained below, column 800 may alternatively feature an insert with discrete apertures. Like porous insert 830 illustrated in
As illustrated in
In operation of a second chromatographic system including column 900, mobile phase fluid may be introduced into packed media 920 axially and/or radially through the porous insert 930 from channel 940. For example, mobile phase fluid may be introduced through a single port that allows it to flow axially into packed media 920. To the extent mobile phase fluid is introduced to packed media 920 radially, it may be introduced only at one or more portions of the length of column 900. The mobile phase fluid that has been introduced flows through packed media 920 and also more-freely, through channel 940.
As the pressure drops along portion of column 900 packed with media 920, some mobile phase fluid from channel 940 migrates through the pores in porous insert 930 into packed stationary phase media 920. The amount of fluid that migrates through the pores in porous insert 930 depends on the porosity, thickness, and diameter of insert 930. The amount of fluid that migrates through porous insert 930 further depends on the pressure differential between channel 940 and media 920. The amount of fluid that migrates through porous insert 930 further depends on the pressure and temperature of the mobile phase fluid in channel 940. The fluid that migrates into the stationary phase media 920 in column 900 may be called makeup fluid and have features described above with respect to step 742. And its migration would decrease the pressure drop along column 900.
As further explained below, column 900 may alternatively feature an insert with discrete apertures. Like porous insert 930 illustrated in
As illustrated in
In operation of a second chromatographic system including column 10000, mobile phase fluid flows through packed media 1020 and also more-freely, through inner channel 1040A and outer channel 1040B. As the pressure drops along the packed media 1020 portion of column 10000, some mobile phase fluid from inner channel 1040A and/or outer channel 1040B migrates through the apertures in inserts 1030A, 1030B into the packed stationary phase media 1020. As illustrated by the arrows in
The amount of fluid that migrates through apertures in inserts 1030A, 1030B depends on the size and spacing of the apertures. The amount of fluid that migrates through apertures in inserts 1030A, 1030B further depends on the pressure differential between the packed media 1020 and inner channel 1040A or outer channel 1040B. The amount of fluid that migrates through apertures in inserts 1030A, 1030B further depends on the pressure and temperature of the mobile phase fluid in channels 1040A, 1040B. The fluid that migrates into the stationary phase media 820 in column 800 may be called makeup fluid and have features described above with respect to step 742. And its migration decreases the pressure drop within the packed media portion 1020 along column 10000.
In step 742 of
As illustrated in
As illustrated in
Despite the fact that the particle diameter of the column 3300 of
The inventors further recognized that the disclosed methods for efficiently transferring a carbon dioxide based separation from a first chromatographic system to a second chromatographic system may be combined. For example, if the comparison of the measured average column pressure for carbon dioxide based separation in the second chromatographic system with the identified average column pressure for carbon dioxide based separation in the first chromatographic system (step 232/step 732) indicates that the difference is not acceptable, step 242 of method 202 and step 742 of method 702 may be combined. Accordingly, the cross-sectional area of the column packed with media may be altered and makeup fluid may be added along the length of the column in the second chromatographic system. A combined step may include selecting a new column for the second chromatographic system or modifying the column in the second chromatographic system. For example, the combined step may include modifying a column such as columns 400, 500, or 600 by adding a porous insert such as described with respect to
This application is a National Stage Application of International Application No. PCT/US2017/051293, filed Sep. 13, 2017, which claims the benefit of and priority to U.S. Provisional Application No. 62/396,739, filed Sep. 19, 2016, and entitled “Method and Apparatus for Linearizing and Mitigating Density Differences Across Multiple Chromatographic Systems”. Each of the foregoing applications is incorporated herein by reference in its entirety.
Filing Document | Filing Date | Country | Kind |
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PCT/US2017/051293 | 9/13/2017 | WO | 00 |
Number | Date | Country | |
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62396739 | Sep 2016 | US |