Methods and devices for streamline extraction, clean-up, and enrichment of target analytes from solid and semi-solid samples

Abstract

A method of extracting a sample, the method includes adding a sample to a disruption and dispersion (2D) vessel, the sample being solid or semi-solid; adding a collision object and a reagent to a sample in the 2D vessel; agitating the sample, collision object, and reagent in the extraction vessel to (a) break up larger material in the sample to increase available surface area for extraction and (b) mix the reagent with the sample to disperse the sample; and performing a fluidic extraction of the sample to isolate compounds of interest in a liquid phase separate from the sample matrix material.

Description

METHODS AND DEVICES FOR STREAMLINE EXTRACTION, CLEAN-UP, AND ENRICHMENT OF TARGET ANALYTES FROM SOLID AND SEMI-SOLID SAMPLESFIELD

[0001] This application claims the benefit of the U.S. Provisional Application S / N 63 / 649,305, filed May 17, 2024, which is incorporated by reference in its entirety.

[0002] The present disclosure generally relates to the field of analytical chemistry including methods and devices for streamline extraction, clean-up, and enrichment of target analytes from solid and semi-solid samples.BACKGROUND

[0003] Pressurized liquid extraction or accelerated solvent extraction (ASE) is a technique for extracting target analytes from solid and semisolid samples with liquid solvents. Pressurized liquid extraction utilizes increased temperature and pressure with common solvents to increase the efficiency of the extraction process. Pressurized liquid extraction can be used to replace more conventional Soxhlet, sonication, boiling, wrist-shaker, and other extraction methods. Several pressurized liquid systems are currently offered by the Thermo Fisher Scientific including the ASE™ 350 Accelerated Solvent Extractor and the EXTREVA ASE™ Accelerated Solvent Extractor.

[0004] Pressurized liquid extraction methods performed by the ASE™ extractors are accepted solid liquid extraction (SLE) methods useful for extraction of many types of analytes. One such method is described in U.S. Pat. No. 5,843,311 (“the '311 patent”) and in EPA Method 3545. An automated system for performing a pressurized liquid extraction method is described in U.S. Pat. No. 5,785,856 (“the '856 patent”). An apparatus for parallel pressurized liquid extraction is described in U.S. Pat. No. 11,123,655 (“the ‘655 patent”).

[0005] With pressurized liquid extraction, the sample is generally first ground and / or mixed with a dispersing agent. A weighed portion is placed in an extraction cell made ofstainless steel or other materials, which is then heated to a predetermined temperature. Initially, a conventional static valve is opened and solvent is pumped into the extraction cell until a target pressure is reached. Once the target pressure is reached, a second static valve is opened to allow the solvent with the extracted sample to flow into a collection vial.

[0006] Therefore, there is a need for a more cost-effective means of pursuing high throughput extractions without any of the above limitations and disadvantages.BRIEF SUMMARY

[0007] In a first aspect, a method of extracting a sample can include adding a sample to a disruption and dispersion (2D) vessel, the sample being solid or semi-solid; adding a collision object and a reagent to a sample in the 2D vessel; agitating the sample, collision object, and reagent in the extraction vessel to (a) break up larger material in the sample to increase available surface area for extraction and (b) mix the reagent with the sample to disperse the sample; and performing a fluidic extraction of the sample to isolate compounds of interest in a liquid phase separate from the sample matrix material.

[0008] In various embodiments of the first aspect, performing the fluidic extraction can include supplying an extraction solvent at elevated pressure, elevated temperature, or a combination thereof.

[0009] In various embodiments of the first aspect, the method can further include removing excess water from the sample. In particular embodiments, removing excess water from the sample can include centrifuging the 2D vessel.

[0010] In various embodiments of the first aspect, the method further includes adding the 2D vessel to a sample extraction vessel. In some embodiments, the method can further include adding a cleaning vessel to a sample extraction vessel in series with the 2D vessel. In particular embodiments, the cleaning vessel can include one or more reagents for drying, defatting, or discoloration. In particular embodiments, the cleaning vessel can include one or more analyte trapping reagent. In particular embodiments, the method can further include an acetone wash to remove water from the sample where the analyte trapping reagent binds at least a portion of the compounds of interest prior to performing the fluidic extraction.

[0011] In various embodiments of the first aspect, the method can further include concentrating the extracted compounds.

[0012] In various embodiments of the first aspect, the method can further include analyzing the extracted compounds to identify and / or quantify the compounds of interest.

[0013] In a second aspect, an extraction apparatus can include a disruption and dispersion (2D) vessel for containing a solid or semi-solid sample, one or more types of collision objects, and one or more reagents; a cleaning vessel containing one or more cleaning reagent; and a sample extraction vessel configured to hold the 2D vessel and at least one cleaning vessel in series.

[0014] In various embodiments of the second aspect, the 2D vessel can include titanium, stainless steel, aluminum, cellulose, polymeric material, or any combination thereof

[0015] In various embodiments of the second aspect, the cleaning vessel can include cellulose, stainless steel, aluminum, titanium, polymeric material, or any combination thereof. In particular embodiments, the cleaning vessel can contain one or more reagents for drying, defatting, or discoloration. In particular embodiments, the cleaning vessel can contain one or more analyte trapping reagents.

[0016] In various embodiments of the second aspect, the sample extraction vessel can include titanium, stainless steel, aluminum, polymeric material, or any combination thereof.

[0017] In various embodiments of the second aspect, the 2D vessel can be disposable.

[0018] In various embodiments of the second aspect, the cleaning vessel can be disposable.

[0019] In various embodiments of the second aspect, the sample extraction vessel can be disposable.

[0020] In a third aspect, a disposable extraction vessel can include a bottom cap comprised of titanium, stainless steel, aluminum, cellulose, polymeric substances, or combinations thereof; a cell body comprised of cellulose, polymeric substances, titanium, stainless steel, aluminum, or combinations thereof; and a top cap comprised of cellulose,stainless steel, aluminum, or combinations thereof, wherein the disposable extraction vessel is configured to withstand pressures of between about 100 psi and about 2000 psi.

[0021] These and other objects and advantages shall be made apparent from the accompanying drawings and the description thereof.BRIEF DESCRIPTION OF THE FIGURES

[0022] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments, and together with the general description given above, and the detailed description of the embodiments given below, serve to explain the principles of the present disclosure.

[0023] FIGs. 1 through 4 illustrate exemplary sample extraction vessel, an exemplary disruption and dispersion (2D) vessel, and an exemplary cleaning vessel, in accordance with various embodiments.

[0024] FIGs. 5 through 8 illustrate exemplary methods of sample extraction, in accordance with various embodiments.DETAILED DESCRIPTION

[0025] Embodiments of methods and devices for streamline extraction, clean-up, and enrichment of target analytes from solid and semi-solid samples are described herein.

[0026] The section headings used herein are for organizational purposes only and are not to be construed as limiting the described subject matter in any way.

[0027] In this detailed description of the various embodiments, for purposes of explanation, numerous specific details are set forth to provide a thorough understanding of the embodiments disclosed. One skilled in the art will appreciate, however, that these various embodiments may be practiced with or without these specific details. In other instances, structures and devices are shown in block diagram form. Furthermore, one skilled in the art can readily appreciate that the specific sequences in which methods are presented and performed are illustrative and it is contemplated that the sequences can be varied and still remain within the spirit and scope of the various embodiments disclosed herein.

[0028] All literature and similar materials cited in this application, including but not limited to, patents, patent applications, articles, books, treatises, and internet web pages are expressly incorporated by reference in their entirety for any purpose. Unless described otherwise, all technical and scientific terms used herein have a meaning as is commonly understood by one of ordinary skill in the art to which the various embodiments described herein belongs.

[0029] It will be appreciated that there is an implied “about” prior to the temperatures, concentrations, times, pressures, flow rates, cross-sectional areas, etc. discussed in the present teachings, such that slight and insubstantial deviations are within the scope of the present teachings. In this application, the use of the singular includes the plural unless specifically stated otherwise. Also, the use of “comprise”, “comprises”, “comprising”, “contain”, “contains”, “containing”, “include”, “includes”, and “including” are not intended to be limiting. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present teachings.

[0030] As used herein, "a" or "an" also may refer to "at least one" or "one or more." Also, the use of “or” is inclusive, such that the phrase “A or B” is true when “A” is true, “B” is true, or both “A” and “B” are true. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular.

[0031] A “system” sets forth a set of components, real or abstract, comprising a whole where each component interacts with or is related to at least one other component within the whole.

[0032] Samples containing significant amounts of water can present problems for pressurized solvent extraction techniques. In some cases, the water can be co-extracted with the sample into the solvent. This can result in an extracted sample having a different solution composition than intended, specifically containing more water than expected. This can interfere with techniques, such as evaporation, used to concentrate the sample. Additionally, in some cases, the co-extraction of the water can even be damaging to the analytical instruments, such as by damaging a chromatographic column through sudden shifts in solvent composition when a sample plug containing a high percentage of water is applied to a column. In other cases, thewater may be immiscible in the solvent and result in two distinct phases, an aqueous phase and a solvent phase. This could result in the sample compounds partitioning between the phases and interfere with quantification of the compounds. As such, dewatering the sample before solvent extraction can improve the pressurized solvent extraction process.

[0033] Described herein are systems and methods for improving pressurized liquid extraction for solid or semi solid samples. Preparing the samples for analysis can include disruption, dispersion, drying, defatting, and discoloration, which is referred to a 5D process. Disruption refers to the mechanical or chemical breakdown of sample matrices to enhance the release of target analytes. This process is vital for improving the efficiency of extraction and ensuring comprehensive analysis. Dispersion denotes the mechanical process of ensuring the sample is homogeneous to enhance extraction efficiency. This may be achieved by introducing reagents or dispersing agents that prevent sample particles from clogging or forming agglomerations. Addressing the agglomeration issue is crucial for achieving consistent distribution of particles within a solvent, leading to reliable analytical outcomes. Drying refers to the removal of moisture from samples to facilitate subsequent analytical processes. This step is crucial for preventing clogging of fluidic lines and avoiding detrimental effects on analytical instrument performance. In the case of EXTREVA Accelerated Solvent Extractor, improper drying can interfere with the level sensing mechanism, leading to inaccurate results. Defatting involves the extraction of lipids from samples to prevent interference with target analytes. This process is critical when dealing with tissue, biosolids, and other matrices where fat not only limits extraction but can cause ionization suppression and faster damage to components of the analytical instrument. Discoloration pertains to the process of removing compounds, such as pigments, that generate color in the sample. This step is essential for ensuring that the sample does not retain any color that could interfere with subsequent analytical processes or the performance of the analytical instrument. Depending on the sample and the target analytes, one or more of these steps can be omitted.

[0034] FIG. 1 illustrates a sample extraction vessel 100 capable of handling solid and semi-solid samples. Semi-solid samples are highly viscous materials with properties of both solids and liquids. The sample extraction vessel 100 includes a top cap 102, a sample cell body 104, and a bottom cap 106. Sample cell body 104 can include an interior space 108 for holdingthe sample. The interior space 108 can be surrounded by an annular wall 1 10. Tn various embodiments, the annular wall 110 can be made of titanium, stainless steel, aluminum, polymeric material, or any combination thereof. Preferentially, the interior of the annular wall 110 should be inert so as to not react with the sample. In some embodiments, the sample extraction vessel 100, or portions thereof, can be disposable. The annular wall 110 of the sample cell body 104 can be sufficiently rigid to withstand pressures of up to about 2000 psi used during extraction. In various embodiments, the sample cell body 104 can be pressured to an internal pressure of between about 100 psi and about 2000 psi during extraction, such as between about 100 psi to about 250 psi. Various techniques can be used to couple the top cap 102 and the bottom cap 106 to the sample cell body 104, such as threaded attachments, magnetic attachments, snap-fit connections, bayonet mounts or other coupling mechanisms known in the art. A particular example is disclosed in US Patent 12,134,050.

[0035] FIG. 2 illustrates a disruption and dispersion (2D) vessel 200. The 2D vessel 200 can include a top cap 202, a cell body 204, and a bottom cap 206. Cell body 204 can include an interior space 208 surrounded by an annular wall 210. In various embodiments, the annular wall 210 can be made of metal, such as titanium, stainless steel, aluminum, cellulose, polymeric material, or any combination thereof. Preferentially, the interior of the annular wall 210 should be inert so as to not react with the sample. While in some embodiments it can be advantageous for the annular wall 210 of the 2D vessel 200 to be able to withstand pressures of up to about 2000 psi used during extraction, in other embodiments, the annular wall 210 of the 2D vessel 200 can be supported by the annular wall 110 if sample extraction vessel 100 to provide sufficient rigidity to withstand the elevated pressures. In various embodiments, the 2D vessel 200 can be able to withstand the forces during centrifugation. In some embodiments, the 2D vessel 200, or portions thereof, can be disposable. In various embodiments, the top cap 202 and the bottom cap 206 can include a screen or membrane to allow the flow of gas and liquid but retain solid material, such as particles above a certain size, such as at least about 0.02 pm.

[0036] In various embodiments, the sample can be added to the interior space 208 along with collision objects and a reagent to assist with water removal (drying), disruption and dispersion

[0037] In various embodiments, the 2D vessel 200 can be sized to fit into a disposable centrifuge tube. In various embodiments, excess water can be removed from a sample by centrifuging the sample within the 2D vessel 200 in the disposable centrifuge tube.

[0038] FIG. 3 illustrates a cleaning vessel 300. The cleaning vessel 300 can include a top cap 302, a cell body 304, and a bottom cap 306. Cell body 304 can include an interior space 308 surrounded by an annular wall 310. In various embodiments, the annular wall 310 can be made of cellulose, stainless steel, aluminum, titanium, polymeric material, or any combination thereof. Preferentially, the interior of the annular wall 310 should be inert so as to not react with the sample. While in some embodiments it can be advantageous for the annular wall 310 of the cleaning vessel 300 to be able to withstand pressures of up to about 2000 psi used during extraction, in other embodiments, the annular wall 310 of the cleaning vessel 300 can be supported by the annular wall 110 if sample extraction vessel 100 to provide sufficient rigidity to withstand the elevated pressures. In some embodiments, the cleaning vessel 300 can be disposable. In various embodiments, the top cap 302 and the bottom cap 306 can include a screen or membrane filter to allow the flow of liquid but retain solid material, such as particles above a certain size, such as at least about 0.02 pm.

[0039] In various embodiments, the interior space 308 of the cleaning vessel 300 can be filled with one or more reagents that can be used for drying, defatting, and / or discoloration of the sample extract. In some embodiments, the cleaning vessel 300 can be prefilled with the one or more reagents, while in other embodiments, the one or more reagents can be provided separately and may need to be added to the cleaning vessel 300 before use.

[0040] In various embodiments, multiple cleaning vessels 300 can be used in sequence to ensure proper drying, defatting, and discoloration of the sample extract. In some embodiments, the multiple cleaning vessels 300 can contain the same mix of reagents, while in other embodiments, each cleaning vessel 300 can include different reagents, such as using a drying cleaning vessel, a defatting cleaning vessel, and a discoloration cleaning vessel.

[0041] FIG. 4 illustrates an extraction assembly 400 including a sample extraction vessel 100, a 2D vessel 200, and a cleaning vessel 300. The inner diameter of the sample extraction vessel 100 can be slightly larger than the outer diameter of the 2D vessel 200 and the cleaningvessel 300 to allow for the 2D vessel 200 and the cleaning vessel 300 to be placed inside the sample extraction vessel 100. In various embodiments, the sample extraction vessel 100 can hold at least one 2D vessel 200 and at least one cleaning vessel 300.

[0042] FIG. 5 is a flow diagram 500 illustrating a method 500 of the extraction of a sample. In various embodiments, the sample can include between about 0% and about 75% water content. In various embodiments, the sample can include between 0% and 60% lipid content. FIGs. 6 and 7 provide further illustration of the method 500. At step 502, a sample is provided in 2D vessel, such as 2D vessel 200. In various embodiments, an operator may attach a bottom cap on one end of the cell body and add solid or semi-solid material to the 2D vessel. Optionally, at step 504, the 2D vessel can be centrifuged to remove excess water, as illustrated in FIG. 7. In other embodiments, it may not be necessary to remove excess water, as illustrated in FIG. 6. At step 506, collision objects and reagent can be added to the 2D vessel. In various embodiments, the reagent can include reagents to assist with disruption and dispersion. In some embodiments, the reagents can also assist with analyte desolvation, water removal (drying), lipid removal (defatting), and pigment removal (discoloration).

[0043] At 508, the 2D vessel can be agitated to dry, disrupt, and disperse the sample. Through agitation, the collision objects can break down the sample into finer material and the reagent and the sample can be thoroughly mixed. Optionally, at 510, the 2D vessel can be placed in an extraction vessel along with a cleaning vessel to form an extraction assembly. In various embodiments, depending on the needs of the sample, the extraction assembly can include multiple cleaning vessels.

[0044] At 512, the extraction assembly can be placed in an extraction system, and at 514, a liquid extraction can be performed. The extraction system can be a liquid extraction system, pressurized liquid extraction system, supercritical extraction system, Soxhlet extraction system, microwave assisted extraction system, or combinations thereof. Generally, the extraction system acts to extract the compounds of interest from the sample into a liquid solvent. In various embodiments, the compounds of interest can include polyaromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), dioxins, per-and polyfluoroalkyl substances (PF AS), organochlorine pesticides (OCPs). The use of the reagents can improve extraction of thecompounds of interest and can reduce the background of contaminate compounds that can make analysis of the sample more challenging.

[0045] In various embodiments, at 518, the extracted compounds can optionally be concentrated, such as by evaporation of the extraction solvent. At 520, the sample can be analyzed, such as to determine the identity and concentration of the compounds of interest. Depending on the compounds of interest, the compounds can be analyzed by spectrometry, mass spectrometry, chromatography, including gas chromatography, liquid chromatography, and ion chromatography, other analytical techniques known in the art for analysis and identification of compounds, or any combination thereof.

[0046] In an alternate embodiment, illustrated in FIG. 8, water can be removed using acetone while the compounds of interest, such as PAHs, SVOC, and PFAS, are retained by an absorbing material. The absorbing material can include nonimprinted polymer (NIP), molecularly imprinted polymer (MIP), or hyper-crosslinked polymer (HCP). The absorbing material can be used to retain the analyte in the sample can also include various polymeric materials such as XAD-2, PS-DVB, HLB, polymethacrylate, polystyrene-divinylbenzene, and polyacrylate. These materials are chosen for their ability to effectively capture and hold target analytes while allowing the solvent, acetone, to carry away water and other impurities. The main purpose of this step is to place a suitable material in the cleaning vessel that will retain the target analytes, ensuring that the extraction process is efficient, and the analytes are isolated for further analysis. By using these polymeric materials, the process can achieve higher purity and concentration of the target analytes, which is crucial for accurate and reliable analytical results.

[0047] After removing the water, the retained compounds of interest can be extracted from the absorbing material with a polar solvent, such as di chloromethane, hexane, or combinations thereof.

[0048] While the present teachings are described in conjunction with various embodiments, it is not intended that the present teachings be limited to such embodiments. On the contrary, the present teachings encompass various alternatives, modifications, and equivalents, as will be appreciated by those of skill in the art.

[0049] Further, in describing various embodiments, the specification may have presented a method and / or process as a particular sequence of steps. However, to the extent that the method or process does not rely on the particular order of steps set forth herein, the method or process should not be limited to the particular sequence of steps described. As one of ordinary skill in the art would appreciate, other sequences of steps may be possible. Therefore, the particular order of the steps set forth in the specification should not be construed as limitations on the claims. In addition, the claims directed to the method and / or process should not be limited to the performance of their steps in the order written, and one skilled in the art can readily appreciate that the sequences may be varied and still remain within the spirit and scope of the various embodiments.

Claims

WHA T IS CI. AIMED IS:

1. A method of extracting a sample, the method comprising: adding a sample to a disruption and dispersion (2D) vessel, the sample being solid or semi -solid; adding a collision object and a reagent to a sample in the 2D vessel; agitating the sample, collision object, and reagent in the extraction vessel to (a) break up larger material in the sample to increase available surface area for extraction and (b) mix the reagent with the sample to disperse the sample; and performing a fluidic extraction of the sample to isolate compounds of interest in a liquid phase separate from the sample matrix material.

2. The method of claim 1, wherein performing the fluidic extraction includes supplying an extraction solvent at elevated pressure, elevated temperature, or a combination thereof.

3. The method of claim 1, further comprising removing excess water from the sample.

4. The method of claim 3, wherein removing excess water from the sample includes centrifuging the 2D vessel.

5. The method of claim 1, further comprising adding the 2D vessel to a sample extraction vessel.

6. The method of claim 5, further comprising adding a cleaning vessel to a sample extraction vessel in series with the 2D vessel.

7. The method of claim 6, wherein the cleaning vessel includes one or more reagents for drying, defatting, or discoloration.

8. The method of claim 6, wherein the cleaning vessel includes one or more analyte trapping reagent.

9. The method of claim 8, further includes an acetone wash to remove water from the sample where the analyte trapping reagent binds at least a portion of the compounds of interest prior to performing the fluidic extraction.

10. The method of claim 1, further comprising concentrating the extracted compounds.

11. The method of claim 1, further comprising analyzing the extracted compounds to identify and / or quantify the compounds of interest.

12. An extraction apparatus comprising: a disruption and dispersion (2D) vessel for containing a solid or semi-solid sample, one or more types of collision objects, and one or more reagents; a cleaning vessel containing one or more cleaning reagent; and a sample extraction vessel configured to hold the 2D vessel and at least one cleaning vessel in series.

13. The extraction apparatus of claim 12, wherein the 2D vessel includes titanium, stainless steel, aluminum, cellulose, polymeric material, or any combination thereof14. The extraction apparatus of claim 12, wherein the cleaning vessel includes cellulose, stainless steel, aluminum, titanium, polymeric material, or any combination thereof15. The extraction apparatus of claim 14, wherein the cleaning vessel contains one or more reagents for drying, defatting, or discoloration.

16. The extraction apparatus of claim 14, wherein the cleaning vessel contains one or more analyte trapping reagents.

17. The extraction apparatus of claim 12, wherein the sample extraction vessel includes titanium, stainless steel, aluminum, polymeric material, or any combination thereof.

18. The extraction apparatus of claim 12, wherein the 2D vessel is disposable.

19. The extraction apparatus of claim 12, wherein the cleaning vessel is disposable.

20. The extraction apparatus of claim 12, wherein the sample extraction vessel is disposable.

21. A disposable extraction vessel comprising: a bottom cap comprised of titanium, stainless steel, aluminum, cellulose, polymeric substances, or combinations thereof; a cell body comprised of cellulose, polymeric substances, titanium, stainless steel, aluminum, or combinations thereof; and a top cap comprised of cellulose, stainless steel, aluminum, or combinations thereof, wherein the disposable extraction vessel is configured to withstand pressures of between about 100 psi and about 2000 psi.

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