Systems and methods for thin-layer chromatography

Abstract

Provided herein are systems, articles, and methods for automatically performing thin-layer chromatography. The method includes automatically extracting an analyte from a sample in a cartridge comprising a fluid tip and a phase-transfer assembly; automatically depositing the analyte onto a thin-layer chromatography plate and unfolding the plate; and automatically detecting the analyte on the unfolded plate. The articles, systems, and methods can be carried out in any setting using the required materials and can provide quantitative results for a variety of different compounds or substances of interest.

Description

[Background technology]

[0001] Description of research and development funded by the federal government. none

[0002] Cross-reference of related applications This application claims the benefit of the priority date of International Patent Application PCT / US23 / 15329, filed on 15 March 2023, and U.S. Provisional Application 63 / 582,501, filed on 13 September 2023, the contents of which are incorporated herein by reference in their entirety.

[0003] Parties in a joint research agreement none

[0004] Sequence List none

[0005] Thin-layer chromatography (TLC) is an analytical tool that separates analytes based on the different migration rates of analytes through thin layers of a chromatography medium. The analytes migrate in the direction of the separation solvent moving through the medium by capillary action.

[0006] TLC is used to detect drug analytes in human bodily fluids. For example, Meuller et al. used TLC to detect benzoylecgonine in human urine (Meuller et al., “Detection of benzoylecgonine in human urine,” Journal of Chromatography A, Volume 144, Issue 1, 1977, Pages 101-107). See also U.S. Patent No. 10,458,963 dated October 29, 2019, “Quantitative HPTLC cannabinoid field testing device and method.”

[0007] The detection of analytes by TLC presents challenges including the exchange and / or transfer of one or more analytes from a first liquid to a second liquid, fragmentation of some samples to remove contaminants before application to a TLC plate, and automation of the process to initiate chromatographic analysis using raw samples. [Prior art documents] [Patent Documents]

[0008] [Patent Document 1] U.S. Patent No. 10,458,963 [Non-patent literature]

[0009] [Non-Patent Document 1] Meuller et al., “Detection of benzoylecgonine in human urine,” Journal of Chromatography A, Volume 144, Issue 1, 1977, Pages 101-107) [Overview of the Initiative] [Means for solving the problem]

[0010] In one aspect, what is provided herein is a system for automatically performing thin-layer chromatography. The system automatically performs and integrates the following operations: (1) extraction of analytes from a sample and delivery of the analytes to the loading area of ​​a thin-layer chromatography plate; (2) deployment of the thin-layer chromatography plate to separate the analytes from other molecules in the sample; and (3) detection of the analytes and analysis of the results. The system also automatically delivers a separation medium to the chromatography plate and separates the analytes in the sample by capillary action. The system also automatically compresses the sample bands by delivering a concentration medium to the chromatography plate on one or both sides of the sample bands. The system also automatically delivers a detection chemical to the chromatography plate, enabling detection and measurement, such as visualization of the analytes on the plate. The system also automatically detects the analytes using a detector such as a camera. The system further includes a computer that measures the analytes on the plate from the signal or image provided by the detector. The system also includes auxiliary elements such as fans and heaters to evaporate the liquid from the chromatography plate.

[0011] In addition, or alternatively, the systems provided herein automatically perform and integrate the following operations: (1) exchange and / or transfer of one or more analyte molecules from a liquid sample to a second liquid, optional extraction of analytes from the liquid sample, and delivery of analytes to the loading area of ​​a thin-layer chromatography plate; (2) deployment of the thin-layer chromatography plate to separate the analytes from other molecules in the sample; and (3) detection of the analytes and analysis of the results.

[0012] In one embodiment, the system comprises an instrument configured to engage with a cartridge. The cartridge includes a thin-layer chromatography plate and a fluid tip, the fluid tip comprising a fluid circuit including fluid channels and ports that engage with a sample container and ports in the fluid assembly of the instrument. The instrument further comprises a pump and valves that, when operated, move the sample fluid from the sample container into the fluid tip and from the fluid tip onto the chromatography plate. Other pumps and valves in the instrument move the liquid from the liquid reservoir into the fluid channels in the fluid tip and from the fluid tip onto the chromatography plate. The cartridge may also comprise a phase transfer medium, which allows for the separation of contaminants from the analyte and allows the analyte to pass through the chromatography plate. The chromatography plate and fluid tip are fabricated within the cartridge such that a combination of openings in the fluid tip and the surface of the chromatography plate forms a barrier that generates wells or reservoirs. The liquid deposited in the wells is contained by the wells but can migrate through the chromatography material under the barrier. The chromatography chip also has one or more openings, one or more of which are optionally covered with a transparent material, and one or more openings expose the detection area of ​​the chromatography plate, allowing the analytes within the detection area to be visualized.

[0013] The methods described herein enable the detection of cannabinoids, particularly analytes such as THC, in samples such as saliva at remarkably low levels. For example, the methods described herein can detect trace amounts of 50 ng, 20 ng, 5 ng, 2 ng, 1 ng, or 0.2 ng in approximately 400 microliters of saliva sample. That is, the methods enable the detection of cannabinoids such as THC at any low concentration of 125 nanograms / milliliter, 50 nanograms / milliliter, 12.5 nanograms / milliliter, 2.5 nanograms / milliliter, or 0.5 nanograms / milliliter.

[0014] Furthermore, the detection process can be carried out rapidly. For example, the process of loading the sample onto the cartridge, extracting the analyte, and loading the analyte onto the chromatography plate can be carried out within 30 minutes, 10 minutes, 5 minutes, 1 minute, or 30 seconds. The total time required from loading the sample onto the cartridge, extracting the analyte, loading the analyte onto the chromatography plate, unfolding the plate, and detecting the analyte can be carried out within 1 hour, 45 minutes, 30 minutes, 15 minutes, or 5 minutes. [Brief explanation of the drawing]

[0015] The accompanying drawings incorporated herein and forming part of this specification illustrate exemplary embodiments and, together with the description, serve to enable those skilled in the art to construct and use these embodiments and others that will be obvious to those skilled in the art. The present invention will be described more specifically in conjunction with the following drawings.

[0016] [Figure 1-1] Figures 1A-1E show an exemplary chromatography cartridge. The cartridge has dimensions of approximately 11.5 cm × 11.5 cm. Figure 1A shows the fluid network. Figure 1B shows a top view of the cartridge, and Figure 1C shows a bottom view of the cartridge. Figures 1D and 1E show two sides of the cartridge with the phase-transfer assembly. [Figure 1-2] Figures 1A-1E show an exemplary chromatography cartridge. The cartridge has dimensions of approximately 11.5 cm × 11.5 cm. Figure 1A shows the fluid network. Figure 1B shows a top view of the cartridge, and Figure 1C shows a bottom view of the cartridge. Figures 1D and 1E show two sides of the cartridge with the phase-transfer assembly. [Figure 1-3]Figures 1A-1E show an exemplary chromatography cartridge. The cartridge has dimensions of approximately 11.5 cm x 11.5 cm. Figure 1A shows the fluid circuit network. Figure 1B shows a top view of the cartridge, and Figure 1C shows a bottom view of the cartridge. Figures 1D and 1E show two sides of the cartridge with a phase transfer assembly.

[0017] [Figure 2] Figure 2 shows an exemplary chromatography plate seated within a cartridge support.

[0018] [Figure 3A] Figures 3A and 3B show an exemplary reservoir formed from the mating of a fluid chip and a TLC plate. [Figure 3B] Figures 3A and 3B show an exemplary reservoir formed from the mating of a fluid chip and a TLC plate.

[0019] [Figure 4] Figure 4 shows an exemplary layout of the reservoir on the chromatography plate.

[0020] [Figure 5] Figures 5A and 5B show an exemplary phase transfer assembly.

[0021] [Figure 6] Figure 6 shows a schematic of an exemplary fluid assembly.

[0022] [Figure 7] Figure 7 shows a side view of an exemplary fluid manifold.

[0023] [Figure 8] Figure 8 shows an exemplary cartridge interface.

[0024] [Figure 9] Figure 9 shows the elements of an exemplary system.

[0025] [Figure 10] Figure 10 shows the exemplary Peltier device position below the cartridge stage.

[0026] [Figure 11] Figures 11A-11C show exemplary ball valves within a fluid tip. Figure 11A shows the fluid tip with the ball valve seated within the fluid channel. Figure 11B shows the valve in the closed position, with the ball seated within the O-ring closing the channel. Figure 11C shows the valve in the open position, with the ball pushed out of the O-ring.

[0027] [Figure 12] Figure 12 shows an exemplary fluid chip 100 with a sample container 1010, which is engaged with a port 101 of the chip. A phase transfer assembly 130 located outside the fluid chip is further shown.

[0028] [Figure 13] Figure 13 shows an exemplary syringe assembly 1304, which comprises a syringe 1301 with a notch 1302 and a measuring clip 1303. The syringe assembly is configured to limit the amount of sample loaded into the syringe by the user. [Modes for carrying out the invention]

[0029] Detailed explanation I. Introduction Disclosed herein are articles, systems, and methods for rapidly testing and precisely quantifying a diluted analyte in a biological sample, such as a mixture in a sample, also referred to herein as a drug. The articles, systems, and methods described herein can be carried out in any setting using the materials required and can provide quantitative results for a variety of different compounds or substances of interest. The systems can be configured for use in situ for in-situ testing or in an office or laboratory setting.

[0030] A sample may comprise one or more analytes of interest. In some embodiments, a sample may be, for example, a biological sample, a water sample, a food sample, an agricultural sample, an industrial sample, a research sample, or any other suitable sample that provisionally comprises an analyte of interest. Typically, a sample is a liquid sample, such as an aqueous liquid sample comprising an aqueous solvent and provisionally one or more analytes.

[0031] The articles, systems, and methods disclosed herein can provide accurate and precise quantitative analysis of one or more analytes rapidly, for example, within 20 minutes or less. The analytes may be present in the sample at a diluted concentration, and the apparatus, systems, and methods are configured to enable the quantification of the diluted analytes. The articles, systems, and methods enable the analysis of samples comprising an analyte, for example, in any volume up to about 5 mL, 4 mL, 3 mL, 2 mL, 1 mL, 0.5 mL, 0.25 mL, or 0.1 mL, or samples comprising an analyte, for example, in a volume up to about 2 mL.

[0032] The methods disclosed herein may include any preferred method for carrying out the separation and / or concentration of at least one analyte in a sample. The methods disclosed herein may include any preferred method for detecting at least one separated and / or concentrated analyte. The methods disclosed herein may include any preferred method for applying an indicator, such as a dye, to a chromatographic medium. As used herein, the terms “indicator” and / or “dye” include any compound that alters the properties of the analyte. Typically, indicators and / or dyes alter the properties of the analyte, for example, by attaching a detectable label, to enable and / or improve the detection of the analyte. The methods disclosed herein may include any preferred method for quantifying at least one separated and / or concentrated analyte.

[0033] Articles disclosed herein may comprise, at least partially, a flat solid substrate coated with a chromatographic medium. The articles may include any number of fluid barriers and / or gates to direct the flow of a solvent, and consequently, the flow of analytes and non-analytes in a sample across the chromatographic medium. The articles may comprise one or more reference standards to assist in the quantification of one or more analytes.

[0034] The systems disclosed herein may comprise a receiving area for a disclosed article, at least one solvent chamber configured to be in fluid communication with at least a portion of a chromatographic medium, and / or a detector. The system and article may be configured such that the article is in the form of a replaceable cartridge, and the system may be configured in such a manner as to receive a replaceable cartridge. The system may be configured such that the cartridge is oriented horizontally or vertically.

[0035] II. Cartridges The cartridges of this disclosure are configured to receive samples and various solutions into fluid channels of the cartridge and deliver them to various locations on a chromatography plate in order to perform thin-layer chromatography ("TLC"). Thus, such a cartridge comprises a TLC plate having a surface comprising a chromatography medium, and a fluid tip engaged therewith, the fluid tip comprising a fluid circuit including fluid channels communicating with ports. The cartridge may further comprise an interphase transfer assembly. The interphase transfer assembly may comprise channels and / or columns comprising an interphase transfer medium. The interphase transfer assembly may be located outside or inside the fluid tip. The long axis of the interphase transfer assembly may be located at any preferred angle, such as parallel to the plane of the cartridge (as shown, for example, in Figure 1) or perpendicular to the plane of the cartridge (as shown, for example, in Figure 12). The interphase transfer medium separates the analyte in the sample from other materials so that a more purified sample can be delivered to the chromatography plate.

[0036] A.TLC plate A TLC plate may comprise a solid substrate coated with a chromatographic medium.

[0037] 1. Circuit board The substrate may be made of any suitable material, such as glass, quartz, metal, aluminum, plastic, or a suitable substitute. The substrate may be made of any suitable shape. The substrate may be triangular, rectangular, square, trapezoidal, rhombus, pentagonal, hexagonal, heptagonal, octagonal, circular, elliptical, or equivalent. The substrate can have a thickness of at least about 0.1 mm, 0.25 mm, 0.5 mm, 0.75 mm, 1 mm, 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, or 10 mm and / or any thickness of about 0.1 mm, 0.25 mm, 0.5 mm, 0.75 mm, 1 mm, 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, or 20 mm. For example, the substrate can have a thickness of about 0.1 to about 20 mm, preferably about 0.1 to about 5 mm, about 0.1 μm to about 2 mm, or about 0.1 μm to about 1 mm. The substrate can have a thickness of 1 mm to 2 mm. A substrate can have three geometric axes: a first axis which constitutes the thickness of the substrate (referred to herein as the z-axis), and two additional coplanar axes which are typically substantially perpendicular to the first axis. A second axis which refers to the length of the substrate (referred to herein as the "x-axis" or "first dimension" of the substrate), and a third axis which refers to the width of the substrate (referred to herein as the "y-axis" or "second dimension" of the substrate). The first and second dimensions of the substrate can be at any angle to each other. The second dimension is at an angle of about 45 to about 135 degrees, about 75 to about 105 degrees, about 80 to about 100 degrees, about 85 to about 95 degrees, or about 90 degrees of the first dimension. The second dimension is substantially orthogonal to the first dimension. In some embodiments, the second dimension is substantially orthogonal to the first dimension.The substrate may have a first dimension and / or a second dimension of any of the following lengths up to about 50 cm, 45 cm, 40 cm, 35 cm, 30 cm, 25 cm, 20 cm, 15 cm, 10 cm, 5 cm, 4 cm, 3 cm, 2 cm, 1 cm, or 0.5 cm, and / or at least about 45 cm, 40 cm, 35 cm, 30 cm, 25 cm, 20 cm, 15 cm, 10 cm, 5 cm, 4 cm, 3 cm, 2 cm, 1 cm, 0.5 cm, or 0.25 cm, for example, about 50 cm to about 0.25 cm, preferably about 30 cm to about 5 cm, more preferably about 30 cm to about 20 cm, and even more preferably up to about 25 cm. The lengths of the first dimension and the second dimension are not the same. The lengths of the first dimension and the second dimension are the same.

[0038] 2. Chromatography media The substrate is coated at least partially with a chromatographic medium. The upper surface of the substrate is coated with the chromatographic medium to at least about 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 99.5%, and / or less than or equal to about 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100%, for example, about 70% to about 100% coated with the chromatographic medium, preferably about 85% to about 100% coated with the chromatographic medium. The chromatographic medium can be any suitable medium recognized by those skilled in the art. The chromatographic medium can be silica, alumina, cellulose, polyamide, or equivalent. In some cases, the chromatographic medium may have one or more chemical modifications such as C2, C8, C10, C18, phenol, amine, and / or chiral. The chromatographic medium can be functionalized with any suitable chemical modification. The chromatographic medium can be functionalized with C2, C8, C18, or any other suitable substitute recognized by those skilled in the art. The chromatographic medium may contain fluorescent molecules (e.g., compounds such as tin-activated strontium compounds, uranyl acetate, magnesium-activated zinc silicate, zinc cadmium sulfide, and equivalents that fluoresce in response to exposure to short-wavelength UV, e.g., 254 nm), thereby causing the chromatographic medium to fluoresce in response to exposure to electromagnetic radiation. The chromatographic medium may have an average particle size ranging from 1 μm to 50 μm. The chromatographic medium may have any suitable thickness for the application. The thickness of the chromatography medium can be any of the following: up to approximately 500 μm, 400 μm, 300 μm, 200 μm, 100 μm, 75 μm, 50 μm, or 25 μm, and / or at least approximately 400 μm, 300 μm, 200 μm, 100 μm, 75 μm, 50 μm, 25 μm, or 10 μm. For example, the thickness of the chromatography medium can range from approximately 500 μm to approximately 10 μm.

[0039] The chromatography medium is attached to the substrate using a binder. The binder can be any suitable binder, such as an organic binder, an inorganic binder, and / or gypsum.

[0040] 3. Substrate equipped with calibration material In some embodiments, the article comprises a calibrator. As used herein, “calibrator” includes any molecules that are the same as or similar to the analyte, applied in one or more amounts onto the chromatographic medium. The calibrator can provide an expected signal in response to detection. In some embodiments, one or more calibrators can be fluidically isolated from the rest of the plate. In other embodiments, one or more calibrators can be adjacent to the sample but not in contact with it, and the calibrators, as well as one or more analytes in the sample, travel through the chromatographic medium. In some embodiments, the calibrators and analytes are fluidically isolated during the analysis. In some embodiments, one or more calibrators are added to the chromatographic medium during preparation, before use, during use, or after use. In some embodiments, the calibrators are positioned within a gate such that the separation and / or concentration solvent cannot interfere with the calibrators during the separation and / or concentration step. The calibrators can be placed at any preferred location on the chromatographic medium. Typically, the calibration material is preferably placed outside the pathway of any component of the sample during the separation and / or concentration step to prevent interruption of the solvent flow pathway. In some embodiments, the calibration material is placed in an area that is fluidly isolated from the rest of the chromatographic medium.

[0041] Any suitable number of calibration substances can be placed on the substrate at any of the following concentrations of the analyte to be quantified: 10, 9, 8, 7, 6, 5, 4, 3, 2, or one or fewer different concentrations.

[0042] B. Fluid Plate 1. Plate structure A fluid plate may comprise one or more networks of internal channels that open over ports. The plate may be formed from a single piece in which the fluid channels are introduced, for example, by laser etching. More typically, the plate may comprise two interlocking pieces. One or both of the pieces may be etched to introduce grooves or valleys onto the surface of the piece. Interlocking the two pieces covers and closes the channels so that they are inside the fluid plate. One or both of the pieces may also comprise holes, for example, openings or vias, such that a first side of the plate communicates with a second side of the plate through the openings. The openings may be positioned to communicate with the fluid channels. For example, the openings may be positioned at the ends of the channels. The openings may be contained on a piece comprising grooves, or positioned on an opposing piece and aligned with a group on the opposite piece.

[0043] The components can be constructed from any material known to those skilled in the art. For example, the cartridge can be constructed from plastic, glass, or metal. The plastic material may include any plastic known to those skilled in the art, such as polypropylene, polystyrene, polyethylene, polyethylene terephthalate, polyester, polyamide, poly(vinyl chloride), polycarbonate, polyurethane, polyvinyldiene chloride, cyclic olefin copolymer, or any combination thereof. The fluid plate can be formed using any technique known to those skilled in the art, such as soft lithography, hard lithography, cutting, embossing, ablation, drilling, etching, injection molding, or any combination thereof.

[0044] 2.Fluid circuit A fluid plate may comprise one or more fluid circuits. Each fluid circuit has ports within the fluid plate, which are connected by one or more internal, and optionally external, fluid channels.

[0045] Referring to Figures 1A-C, the fluid tip 100 may comprise a first fluid circuit comprising a sample inlet port 101, a first pressure port 103, a second pressure port 105, and a fluid delivery port 108, all connected through fluid channels 120a-d. Port 103 and fluid channel 120a optionally assist in sample positioning from a sample collector, if any. The ports may communicate with two or more other ports through joints or intersections 122a-b between fluid channels that form branches within the circuit.

[0046] Referring to Figure 12, the sample container 1010 engages with the sample inlet port 101 and can supply the sample fluid to the fluid tip.

[0047] The fluid plate may also include a second fluid circuit comprising a separation solvent intake port 108 and a separation solvent delivery port 109. The separation solvent delivery port is positioned to deliver the separation solvent onto the sample loading zone of the chromatography plate. In some embodiments, the second fluid circuit may share ports and / or fluid channels with the first fluid circuit.

[0048] The fluid plate may also comprise one or more third fluid circuits. The circuits may comprise one or more concentrated solvent intake ports 111 and one or more concentrated solvent delivery ports 113a and 113b, the one or more concentrated solvent intake ports 111 and the one or more concentrated solvent delivery ports 113a and 113b being positioned to deliver concentrated solvent to one or more concentrated solvent loading zones.

[0049] The fluid plate may also comprise one or more fourth fluid circuits, each comprising a port connected by one or more fluid channels, including a dye intake port 115 and a dye delivery port 117, the dye delivery port being positioned to deliver the dye onto the separation zone of the chromatography plate. In addition, or alternatively, the cartridge may contain the dye in the fluid conduit between ports 115 and / or ports 115 and 117, for example, dried dye forming a dye film on the surface of the port or fluid conduit. The dye may also be added to the cartridge during manufacturing by any preferred method, for example, by introducing solvated dye into the fluid conduit between ports 115 and / or ports 115 and 117, the dye being dried and forming a film in the port or fluid conduit. In some cases, adding dye to the cartridge during manufacturing may be beneficial to increase the shelf life of the dye, as some dyes begin to decompose after solvation. The dried dyes are resuspended by introducing a suitable solvent from a solvent reservoir via a pressurized system, allowing the dyes to be solvated in the solvent and, if desired, homogeneously mixed. An exemplary method for homogeneously mixing the dyes is to draw the solution containing the solvated dyes into the pressurized system once or multiple times, through a fluid conduit into port 115 and out of port 117, before applying the homogeneous solution to the TLC plate for use.

[0050] The fluid plate may also include a valve positioned between the phase-movement assembly and the sample loading zone. The valve may be, for example, a ball valve seated within an O-ring and held in place by a magnet (see Figures 11A-C). When a vacuum is drawn to introduce the sample, the valve prevents air from being introduced from the sample loading zone. However, the positive pressure required to move the sample through the phase-movement assembly displaces the ball, allowing fluid to pass into the sample chamber. Any suitable valve can be used. These include, but are not limited to, check valves, diaphragm valves, Quake valves, pinch valves, and solenoid valves.

[0051] C. Sample preparation and phase transfer assembly The first fluid circuit may further include a phase-transfer assembly. The phase-transfer assembly may comprise a fluid channel and / or column, the fluid channel and / or column comprising a filtration and chromatography ("phase-transfer") medium located downstream of the sample inlet port and two pressure ports and upstream of the sample delivery port. In one embodiment, the phase-transfer assembly comprises a fluid channel located inside a fluid tip, the fluid channel comprising a phase-transfer medium. In an alternative embodiment depicted in Figures 1D and 1E, the phase-transfer assembly comprises a column 130, the column 130 contained within an external housing 621a of the fluid tip, comprising a phase-transfer medium communicating with the first fluid circuit through phase-transfer connection ports 132a and 132b, for example, through tubing. The long axis of the phase-transfer assembly can be positioned at any preferred angle to the plane of the cartridge, such as parallel to the plane of the cartridge (as shown, for example, in Figure 1) or perpendicular to the plane (as shown, for example, in Figure 12). In some cases, positioning the long axis of the inter-phase moving assembly perpendicular to the plane of the cartridge is advantageous compared to parallel placement and may provide a lower detection limit.

[0052] The phase transfer assembly performs three main functions. First, it filters out particulate matter from the sample. In the case of saliva, this includes, for example, food particles. Second, it transfers and / or exchanges one or more analytes in a liquid sample, such as an aqueous sample containing an aqueous solvent, e.g., water, to an organic solvent for thin-layer chromatography, thereby transferring one or more analytes from the liquid sample to the organic solvent. Third, it acts as a steady-state chromatographic medium, using different mobile phases to selectively bind and elute molecules, e.g., analytes. Any suitable mobile phase, such as an aqueous phase, e.g., an aqueous solvent, and / or an organic phase, e.g., an organic solvent, can be used. In some cases, the steady-state chromatographic medium selectively binds to one or more undesirable molecules in the sample, e.g., non-analytes, thereby removing them from downstream analysis. Removal of one or more undesirable molecules can improve the performance of the downstream analysis of one or more analytes, e.g., detection limits, sensitivity, resolution, and equivalents. In addition, or alternatively, a steady-state chromatographic medium selectively binds to one or more desirable molecules in the sample, such as the analyte, which are then eluted for downstream analysis.

[0053] These three functions are typically performed by three material layers within a phase-transfer assembly: a filter layer, a selective coupling layer, and a phase-transfer layer. Each layer may contribute to more than one function.

[0054] The solvents that can be used include, for example, methanol, hexane, water, acetonitrile, dichloromethane, dibromomethane, chloroform, toluene, acetone, methyl THF, pentane, cyclopentyl methyl ether, ethyl acetate, and 1-octanol. The choice of concentration or separation solvent may depend on the steady phase and the analyte. For THC detection, dichloromethane can be used to load and concentrate the analyte. Dibromomethane can be used as a separation solvent for THC as the analyte. Any organic analyte can be concentrated and separated using the correct mixture of solvents. Trinitrotoluene (high explosive) and malathion (organic pesticide) have been demonstrated to be detectable using this technique.

[0055] In this configuration, moisture from the saliva sample is captured on the phase transfer medium within the phase transfer assembly, while the analyte, dissolved in the organic solvent, passes through and is deposited on the plate.

[0056] For example, the phase transfer assembly may include a housing having a layered medium, such as beads, within it. The phase transfer assembly may include a filter layer that can remove solid particles from the sample. It may also include a hydrophilic adsorbent that can absorb water. It may also include an sorbent for solid-phase extraction of an analyte, such as THC. The isolation of one or more compounds of interest can be carried out by first loading the sample into the phase transfer assembly, and then flowing a solvent through the phase transfer assembly from the proximal end to the distal end. The fluid can therefore move through the filter, then through the medium in the phase transfer layer, such as beads, and finally through the sorbent in the layer (e.g., via force from a pumping system). The extracted compound resulting from exiting from the distal end can then be sent to a detector for analysis.

[0057] 1. First screen layer The first screen 610a holds the layer in place within the housing. Any suitable screen, such as a mesh or an aluminum screen, can be used.

[0058] 2. Filter layer Referring to Figures 5A and 5B, the filter layer 611a can be used for samples that are heterogeneous mixtures such as biological fluids, colloids, emulsions, or powdered / porous solids, in order to prevent clogging of the interphase transfer interface 664a. A layer of fibrous material, in this case cotton, further plays a role in trapping particles. Cotton has a variable pore size and prevents the passage of particles in the size range of 20 to 100 μm. Alternative materials include glass wool, filter paper, or cloth. Cotton can also absorb water.

[0059] In some embodiments, the filter layer 611a may include beads made from one or more inert materials such as polystyrene, sand, or quartz sand. In other embodiments, the beads in the filter layer 611a may consist of chemically active materials (e.g., beads with C-18 coatings, chiral coatings, derivatizers or chemical modifiers, or biologically active compounds such as those used in immunoassays) designed to pre-treat samples for the remainder of the cartridge / analysis. The overall pore size of the filter layer 611a can be controlled by the size of the beads used, which can be spherical, molded, or irregular, as long as they exhibit a narrow dispersion size. The beads in the filter layer 611a may have a diameter of 40 to 600 microns, and the heterogeneity may be 20% or less. In some embodiments, the beads in the filter layer 611a may have a diameter of 40 to 60 microns. In other embodiments, the particles may be 300 to 600 microns.

[0060] In one embodiment, the phase transfer layer comprises a single filter layer 611 that accommodates any biological sample without prior preparation (for example, the filter may have internal pores that trap interfering substances and / or collect compounds by trapping large particles in open pores and adhering them to a large surface area).

[0061] The pore size of the filter layer and subsequent options depend on the sample to be extracted. For homogeneous samples, a filter layer is not required; however, for heterogeneous (i.e., colloidal or emulsion) samples, one or more layers may be employed. For example, many biological samples contain both colloidal and emulsified components with widely varying sizes. Therefore, employing a single filter would either provide poor filtration or become clogged. The choice of material depends on the desired extraction solvent analyte, as some filters may be chemically incompatible with or bind to the analyte.

[0062] The filter layer may comprise a medium for filtering out particles in a sample that have a size greater than 0.001 microns, 0.01 microns, 0.1 microns, 1 micron, 10 microns, or 100 microns.

[0063] 3. First selective coupling layer In some embodiments, the selective binding layer 612a may contain a material to which a chemical or protein may bind, potentially interfering with the analysis and / or preventing purification. For example, if thin-layer chromatography is used as part of a detector, water may interfere with the analysis. In such cases, the particles of the binding layer 612a may be formed from a desiccant material.

[0064] The layer 612A of hydrophilic particles such as sand functions for both particle trapping and water adsorption. For example, the layer may consist of white quartz sand with particle sizes ranging from 200 to 300 μm. The pore size is 10 to 30 μm. Alternative materials include silica spheres or glass beads.

[0065] In other embodiments, silica or molecular sieves can be used to capture water (and thus remove it from the extracted compound). In another embodiment, if the collected fluid contains biological material, the biological matrix may interfere with the lower limit of detection for various analyses. In such cases, beads with functional groups such as thiols, COOH, NH3, or CHO can be used to remove the biological matrix from the fluid (and thus from the compound ultimately extracted) by the binding layer 612a. In other embodiments, the beads of layer 612a can be functionalized to bond to specific chemical groups or to break down compounds. In some embodiments, the selective binding layer 612a can be combined with the phase transfer layer 613a.

[0066] 4.Phase transfer layer The phase transfer layer 613a may include a chromatographic medium and / or sorbent medium, such as particles or beads, that reversibly binds to the compound through physical or chemical interactions with different affinities, depending on the choice of steady-state and mobile phases. Examples of phase transfer media include quartz, alumina, polystyrene, silica, and beads, which are functionalized with moieties such as C18, NH3, or COOH.

[0067] The phase transfer medium acts as the steady-state phase. Depending on the solvent used, the phase transfer medium selectively binds to the analyte and elutes it. For THC and other cannabinoids, normal-phase materials such as silica, alumina, diatomaceous earth, and magnesium silicate (e.g., Florisil) can also be used. Silica has irregular particles. They can range in size from, for example, 40 to 63 μm, and have a pore size of about 60 angstroms. When an intermediate polar solvent such as dichloromethane (DCM) is used, delta-9-tetrahydrocannabinol (Δ 9 -THC or THC) and cannabidiol (CBD) are leached from silica, while the metabolite (11-hydroxy-Δ) 9 -Tetrahydrocannabinol (11-OH-THC) and 11-NOR-9-carboxy-Δ 9 -Tetrahydrocannabinol (11-COOH-THC), etc., remains bound.

[0068] The affinity of a compound depends primarily on the polarity of the solvent, and therefore, by changing the solvent from polar (methanol) to nonpolar (hexane), it is possible to obtain or isolate multiple different sets of compounds (this can be done several times). For example, THC can bind to a reverse-phase sorbent when the mobile phase is a nonpolar solvent such as water. However, when the mobile phase is nonpolar, such as dichloromethane (DCM), the analyte is released from the sorbent along with the mobile phase. For example, flowing hexane through a sample can release nonpolar compounds (e.g., THC) from the phase-transfer interface, while polar compounds remain. Alternatively, or in addition, polar compounds (e.g., THCA) can be collected by flowing a polar solvent such as methanol through the phase-transfer assembly. Furthermore, the affinity of a compound with respect to both the solvent and the phase-transfer medium can be altered to protonate or deprotonate the compound, which can be controlled through the pH of the solvent or the inherent acid-base properties of the solvent, such as triethylamine. This removes impurities while allowing the compound of interest to be collected. Furthermore, the amount of solvent required to remove the compound also varies depending on the affinity of the compound to the phase transfer medium, e.g., beads. The compound of interest can further be targeted by altering the affinity of the compound to the surface of the phase transfer medium. Moreover, the phase transfer medium can be used to control pH and act as a buffer, which can also protonate or deprotonate the compound, thereby affecting the affinity of the compound to the solvent and medium, e.g., beads. In some embodiments, the phase transfer medium of layer 613a can provide a surface area-to-volume ratio of at least 500,000 or greater than 500,000, which provides a state in which at least about 1% of the total molecules are located on the surface (solvent interface) with respect to an immiscible solvent. The large number of surface molecules can, advantageously, enable rapid isolation of the compound using solvents that are miscible, partially miscible, or completely immiscible.

[0069] Exemplary sorbents with affinity for THC include, for example, reverse-phase sorbents such as C18 (Octadecy|silane). Reverse-phase sorbents are commercially available and include, for example, StrataX™ (Phenomex, Torrance CA), Oasis™ HLB (Waters, Milford, MA) (consisting of hydrophilic N-vinylpyrrolidene and lipid-soluble divinylbenzene), and Chromabond HR-X (Macherey-Nagel, Duren, Germany) (spherical hydrophobic polystyrene-divinylbenzene resin).

[0070] In some embodiments, the materials used for the media of the filter layer, binding layer, and phase transfer layer described herein may consist of various sands such as diatomaceous earth, silica, quartz, glass, alumina, polystyrene, sea sand, or loamy sand, or other ground materials such as powdered metal (metal powder), ceramic, cellulose, or wood. In some embodiments, these media may also be functionalized to carry out chemical reactions (carbodiimide), buffer solutions (control pH (e.g., triethylamine)), or prepare compounds for subsequent layers in the phase transfer assembly 664 (sodium cyanoborate, which can protonate or break down unwanted materials / transform compounds into known ones). Thus, the phase transfer assembly 664a may also be used to carry out synthetic chemical reactions in addition to purifying / isolating compounds. The functionalization of these materials may be carried out through well-known chemical reactions, which vary from material to material.

[0071] In some embodiments, a selective binding layer, such as an aqueous solvent or a desiccant material that absorbs water, also functions as a phase transfer layer. An exemplary selective binding layer that also functions as a phase transfer layer is an aqueous solvent or a desiccant material that absorbs water from a liquid sample as the liquid sample passes through the selective binding layer. As the organic solvent passes through the selective binding layer, analytes soluble in the organic solvent are transferred from the aqueous phase, e.g., the aqueous solvent, to the organic phase, e.g., the organic solvent. In some cases, analytes in a liquid sample can bind to the selective binding layer as the organic solvent passes through it, and then be extracted from the selective binding layer.

[0072] 5. Second selective coupling layer A second selective binding layer, such as a layer of sand 614 placed after the silica, can prevent the silica particles from being loaded onto the plate.

[0073] 6. Second filter layer A second filter layer, for example, cotton 615a, positioned after the chromatographic medium layer, may contain cannabichromene (CBC), which is used as a control.

[0074] 7. Second screen layer The final screen 616a holds the layers in place within the housing. Any suitable screen, such as a mesh or an aluminum screen, can be used.

[0075] D. Assembly and opening When assembled, the fluid chip can contact the surface of a chromatography plate containing a chromatography medium, forming a reservoir on the plate's surface. In such a configuration, chromatography can be performed horizontally.

[0076] One method for interlocking a chromatography plate with a fluid tip is to provide a support, such as a tray or holder, which comprises a surface for supporting the fluid tip and a recess within the surface, into which the chromatography plate can be inserted so that the fluid tip rests across the chromatography plate. An exemplary support 202 with a chromatography plate is depicted in Figure 2.

[0077] One or more openings within the fluid chip are positioned to expose the separation area of ​​the chromatography plate so that the separation area can be visualized from above, for example, using a camera.

[0078] An exemplary area for a large opening is depicted by a thick dashed line in Figure 1. A cross-sectional view of the reservoir is shown in Figure 3. The fluid tip 100 with reservoir 107 is meshed with a chromatography plate comprising a substrate 150 and a layer of chromatography medium 151. The liquid, designated by the diagonal lines, is contained within the reservoir and can migrate across the plate only through the chromatography medium (movement is depicted by lines with arrows).

[0079] Ports within the fluid chip (through which the fluid exits the chip and is deposited on the plate) can be configured as openings through the plate. One or more openings can be positioned to expose the surface of the chromatography plate, which contains the chromatography medium, when the plate, fluid chip, and chromatography plate are engaged. In this configuration, the combination of openings within the exposed plate surface forms a reservoir, in which the openings form the walls of the reservoir and the plate surface forms the floor of the reservoir. Thus, the fluid deposited in the reservoir cannot flow freely across the surface of the chromatography plate because the fluid chip, pressed against the plate surface, forms a physical barrier. However, the liquid deposited in the reservoir can travel through the chromatography medium beneath the barrier and thus migrate across the plate.

[0080] 1. Fluid reservoir The surface of a fluid chip can be positioned in contact with the surface of a chromatography plate containing a chromatography medium. An opening through the chip exposes the plate surface and can function as a well or reservoir. Liquid deposited within such a well can migrate through the chromatography medium along the underside of the well wall, contacting the chromatography plate.

[0081] In one embodiment, the apparatus further comprises one or more fluid reservoirs in contact with the surface of the chromatographic medium, and the liquid phase applied to the fluid reservoir is in fluid communication with the chromatographic medium such that the liquid phase moves through the chromatographic medium via capillary action. In one embodiment, the fluid reservoir does not contact the edge of the chromatographic medium or the edge of the solid substrate, and / or the liquid phase does not travel over the upper part of the surface of the chromatographic medium.

[0082] In one embodiment, the article comprises (a) a solid substrate, (b) a chromatography medium on the surface of the substrate through which a liquid phase can travel, and (c) one or more fluid reservoirs in contact with the surface of the chromatography medium, the fluid reservoirs enabling the application of the liquid phase to the surface of the chromatography medium so that the liquid phase can travel through the chromatography medium. In one embodiment, the fluid reservoirs do not come into contact with the edges of the chromatography medium or the solid substrate, and / or the liquid phase does not travel over the top of the surface of the chromatography medium.

[0083] The fluid reservoir can hold any suitable volume, such as at least 100 μL, 200 μL, 300 μL, 400 μL, 500 μL, 600 μL, 700 μL, 800 μL, 900 μL, 1,000 μL, 1,500 μL, or 2,000 μL, and / or 5 mL or less, for example, any of 100 μL to 5 mL. Typically, the fluid reservoir is manufactured from a material that is solvent-resistant and / or compatible with the solvent. Suitable materials will be readily identifiable by those skilled in the art.

[0084] In one embodiment, the fluid reservoir comprises one or more conduits configured to operate toward a solvent source.

[0085] Therefore, the reservoir can be configured as an open well, such as an open-end cylinder or block. These can be positioned on the chromatography plate in contact with the chromatography medium. In this way, the chromatography medium acts as the floor of the well, and the fluid deposited in the open upper part of the reservoir will come into contact with the chromatography medium.

[0086] The reservoir can be any preferred shape, such as a triangle, rectangle, square, trapezoid, rhombus, pentagon, hexagon, heptagon, octagon, circle, ellipse, or equivalent. Typically, the reservoir has an elongated shape so that the solvent generates a linear solvent front along the length of the elongated reservoir.

[0087] The reservoir can be any suitable length, such as at least about 2.5 mm, 5 mm, 7.5 mm, 10 mm, 15 mm, 20 mm, 25 mm, 30 mm, 35 mm, 40 mm, 45 mm, 50 mm, 100 mm, 150 mm, 200 mm, or 250 mm, and / or about 5 mm, 7.5 mm, 10 mm, 15 mm, 20 mm, 25 mm, 30 mm, 35 mm, 40 mm, 45 mm, 50 mm, 100 mm, 150 mm, 200 mm, 250 mm, or 500 mm in length or less, for example, about 2.5 mm to about 500 mm in length, preferably any 50 mm to 500 mm in length. The reservoir can be any suitable width. In one embodiment, the ratio of reservoir length to reservoir width is less than or equal to approximately 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, or 20:1.

[0088] The reservoir can be any preferred height, such as at least about 1 mm, 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 11 mm, 12 mm, 13 mm, 14 mm, 15 mm, 16 mm, 17 mm, 18 mm, or 19 mm, and / or about 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 11 mm, 12 mm, 13 mm, 14 mm, 15 mm, 16 mm, 17 mm, 18 mm, 19 mm, or 20 mm, for example, any of 1 to 20 mm or less.

[0089] An exemplary article, for example, a TLC plate with a fluid reservoir, is shown in Figures 3A and 3B. The exemplary article comprises a substrate (150) with a chromatographic medium (151) coated at least partially on the substrate. A fluid tip (100) forming a reservoir (107) is placed on top of the chromatographic medium. The fluid reservoir (107) creates a seal to the chromatographic medium (151) so that any fluid placed in the fluid reservoir (107) does not pass through the joint between the fluid reservoir (107) and the chromatographic medium (151). A side view of the article is shown in Figure 3A, and an angled view is shown in Figure 3B. The fluid pathway (Figure 3A) of the liquid phase (109) installed in the fluid reservoir (107) is indicated by an arrow, illustrating that the liquid phase moves radially outward through capillary action from the fluid reservoir to the chromatography medium (151), but not through the joint (111).

[0090] Another exemplary article is shown in Figure 4. The exemplary article comprises a TLC plate (1407), which comprises a substrate coated at least partially with a chromatographic medium. The TLC plate (1407) comprises at least two, optionally three, fluid reservoirs (1401, 1403, and 1405), wherein the first fluid reservoir (1401) is configured to apply a solvent in a first dimension (1402), the second fluid reservoir (1403) is configured to apply a solvent in a second dimension (1404), and the optional third fluid reservoir (1405) is configured to apply a solvent in a second dimension (1406) from the opposite direction to the second fluid reservoir (1403). The second (1403) and third (1405) fluid reservoirs can be filled with fluid either sequentially or simultaneously. In one embodiment, the TLC plate (1407) includes a fourth fluid reservoir (1408) configured to apply the solvent in the first dimension (1409) in the opposite direction to the first fluid reservoir (1401). In one embodiment, the first fluid reservoir also serves as a sample loading zone.

[0091] 2. Pump The fluid assembly comprises one or more pumps for moving a liquid between a container and a cartridge through the application of positive or negative pressure. Any pump known in the art may be used in the apparatus of this disclosure, including, without exception, syringe pumps, piston pumps, peristaltic pumps, and diaphragm pumps.

[0092] Syringe pumps and piston pumps move liquid by drawing it into a chamber through the movement of a syringe or piston, typically through a first valve, and by pushing the liquid out of the chamber, typically through a second different valve, using the syringe or piston. These pumps require a motor to move the piston or syringe.

[0093] A peristaltic pump uses rollers to compress and release a flexible tube; the compressive action generates a positive displacement, which propels the liquid through the tube.

[0094] A diaphragm pump includes a flexible diaphragm. For example, the movement of the diaphragm by vacuum generates negative pressure above the diaphragm in the chamber. For example, moving the diaphragm using positive pressure generates positive pressure above the diaphragm in the chamber. By positioning valves above both sides of the diaphragm, when negative pressure is applied to the diaphragm, the first valve is opened; when the second valve is opened, the first valve is closed; and by applying positive pressure and moving the liquid through the second valve, the liquid can be moved into the chamber.

[0095] III. Apparatus Also provided herein is an instrument configured to engage with a cartridge and automatically perform thin-layer chromatography on the cartridge. The instrument may comprise a cartridge interface for engaging with the cartridge, a fluid assembly for moving a liquid through the cartridge onto a chromatography plate, a detector for detecting signals from the cartridge, such as signals from an analyte or calibration substance molecules, a fan positioned to direct air onto the surface of the TLC plate, a temperature controller positioned to regulate the temperature of the cartridge engaged with the interface, and a computer with operating software for operating the instrument when performing thin-layer chromatography.

[0096] A. Cartridge Interface The cartridge interface may comprise a stage for supporting the cartridge and a fluid line configured to engage with a port in the cartridge. The fluid line may be independent. Alternatively, the fluid line may be assembled within a fluid manifold. The fluid manifold may have ports that are adapted to engage with the ports in the cartridge.

[0097] 1. Stage The stage may include a platform configured to position and hold the cartridge. The platform may include guides for proper positioning of the cartridge.

[0098] 2. Fluid manifold A fluid manifold can be configured as a block having multiple inlet points for fluid lines exiting a pump, multiple through conduits, and multiple nozzles for engaging with fluid tips. When a cartridge is loaded onto a stage, the fluid manifold can be configured to move toward the cartridge, engage with the ports, move away from the cartridge, and disengage from the ports.

[0099] Referring to Figure 8, the exemplary fluid manifold 340 includes fluid connectors 341, 342, 343, 344, and 345 configured to connect to fluid lines from a pump. The stage 710 includes alignment begs 721, 722, and 723 that align with holes in the fluid cartridge and properly position the cartridge within the interface. The interface further includes a mechanical bearing screw-driven lift 360 that moves the fluid manifold along the Y (vertical) axis to engage with the fluid tip.

[0100] B. Fluid Assembly The fluid assembly comprises a container, fluid lines, a pump, and typically a valve (e.g., a switchable valve) for moving liquid into, around, and out of the cartridge once the cartridge is engaged with the instrument. Typically, positive and / or negative pressure is delivered through a fluid manifold engaged with ports in the engaged cartridge. This includes, for example, vacuum to draw a sample from the sample cartridge into the fluid channel of the fluid tip, and the application of positive pressure to pump liquids such as separation solvents into the fluid channel and out of ports on the chromatography plate.

[0101] 1. Container The fluid assembly comprises one or more containers for containing the fluid. These include, for example, glass or plastic containers. The containers or reservoirs may have a volume of about 10 mL to about 1 L, for example, about 50 mL to about 500 mL, or about 100 mL.

[0102] 2. Fluid lines For example, a fluid line consisting of plastic tubing provides a fluid communication path between a container and a port in a cartridge interface that engages with a port in the cartridge.

[0103] 3. Exemplary Fluid Assembly Figures 6 and 7 show an exemplary fluid assembly. The fluid assembly comprises several pumps (301, 302, 303, and 304), which are connected to several fluid reservoirs (321, 322, 323) and ports (341, 342, 343, 344, and 345) in a fluid manifold (340) through fluid conduits, e.g., pipes, and several switchable valves (361, 362, 363, and 364). A syringe pump 330 is fluidly connected to a valve assembly 332. The valve assembly 332 comprises three valves that fluidly communicate with a DCM reservoir 335, and ports 340 and 341 of the fluid manifold. The pumps pump fluid from the reservoir 332 to the fluid manifold ports.

[0104] The syringe pump 303, driven by the motor 353, draws fluid from the DCM reservoir 323 through the fluid line 3333 and valve 333 of the valve assembly 3. The syringe pump 303 can deliver the drawn fluid to port 342 of the fluid manifold 340 through valve 323 and fluid line 3323. It can also deliver the drawn fluid to port 344 of the fluid manifold 340 through valve 313 and fluid line 3313.

[0105] The syringe pump 302, powered by the motor 352, can draw fluid from the 5:1 reservoir 322 through the fluid line 3242 and the valve 342 of the valve assembly. The syringe pump 302 can deliver the drawn fluid to the port 343 through the valve 332 and the fluid line 3232.

[0106] A syringe pump 301, driven by motor 351, can apply vacuum and draw the sample fluid into the cartridge through valve 341 of valve assembly 1, through port 101 of the cartridge which is in fluid communication with port 341 of the fluid manifold and fluid line 3141. Alternatively, the sample collector can be a syringe attached to the cartridge via a sample collector port, and the system can include a device configured to apply positive pressure to the syringe, for example, the plunger of the syringe pump, thereby applying the sample to the cartridge. The sample collector syringe can include a measuring clip attached to the barrel of the syringe, which limits the amount of sample loaded into the syringe by the user. This can improve the performance of the measurement, e.g., reproducibility and reliability, by providing a means for reproducible loading of the sample volume. An exemplary syringe with a measuring clip is shown in Figure 13. In this embodiment, syringe 1301 includes a notch 1302 configured to receive a measuring clip 1303. Syringe assembly 1304 may be used to collect a liquid sample and then engage with a cartridge for analysis of one or more analytes in the collected liquid sample. Syringe pump 301 can also draw liquid from Fast Blue reservoir 321 through fluid line 3152 and valve 351. Pump 301 can deliver the drawn fluid to port 345 of a fluid manifold through valve 361 and fluid line 3161. In this embodiment, a dye may be applied to the cartridge during manufacturing, and Fast Blue reservoir (321) may be replaced with a solvent reservoir.

[0107] The peristaltic pump 304 can pump fluid from the methanol reservoir 324 through fluid lines 3404 and 3151, through valves 351 and 361 of valve assembly 1 and through fluid line 3161, into port 345 of the fluid manifold 340. Fluid from the fastblue reservoir 321 passes through fluid line 3152 and valve 351 of valve assembly 1.

[0108] C. Fan The apparatus may also be equipped with a fan. When engaged with the cartridge, the fan can be configured to blow air through an opening in a fluid tip on the surface of the chromatography plate, for example, toward the interface stage. This can function to evaporate the liquid from the chromatography medium.

[0109] An exemplary fan 350 is depicted in Figures 7 and 9.

[0110] D. Temperature control assembly The apparatus may include a temperature control assembly configured to control the temperature of the chromatography plate. Therefore, the temperature control element, when engaged with the cartridge, can direct heat or cold toward the interface stage.

[0111] The temperature control assembly can be selected from any heating or cooling element suitable for this purpose. These include, but are not limited to, Peltier devices, resistance heaters, hot air heaters, and infrared heaters.

[0112] Peltier devices, also known as thermoelectric coolers or heaters, generate heat using the Peltier effect. These devices consist of semiconductor elements that create a temperature difference when an electric current passes through them. By controlling the direction of the current, a Peltier device can generate heat on one side of a plate while absorbing heat on the other side.

[0113] Resistive heating involves passing an electric current through a resistive material, such as a wire or heating element that generates heat. The heated element then transfers thermal energy to a plate, raising its temperature.

[0114] Hot air heating involves blowing heated air directly onto the plate to raise its temperature. In this method, a heating element such as an electric coil or gas burner heats the air near the plate. The heated air is then directed towards the plate using a fan or other means.

[0115] Infrared heating involves directly heating the surface of a plate using infrared radiation. Infrared heaters emit electromagnetic waves within the infrared spectrum, which are absorbed by the plate and converted into heat.

[0116] An exemplary Peltier device 1010 is depicted in Figure 10.

[0117] E. Detector The apparatus may further include a detector that can detect signals from the unfolded chromatographic plate. The detector may include, for example, a camera that detects light emitted by a label.

[0118] An exemplary camera 360 is depicted in Figures 7 and 9.

[0119] F. Computer The instrument may also be equipped with a computer capable of operating a system. The computer may include one or more of the following elements: A central processing unit ("CPU") performs calculations and data processing and executes instructions. Random access memory (RAM) is a temporary storage area where the computer stores data and instructions. A hard drive or solid-state drive (SSD) provides permanent storage for the operating system, software, and data files. Input / output (I / O) devices enable interaction between the laboratory instrument and the computer. Examples include keyboards, mice, touchscreens, and various types of sensors or probes for collecting data from the instrument. A display monitor provides a visual interface for interacting with the computer and viewing data and results from the laboratory instrument. Interface cards can connect to the computer using various protocols such as USB, Ethernet®, serial ports, or special communication standards. An operating system (OS) is software that manages the computer's resources and provides a user-friendly interface. It controls the hardware, launches software applications, and facilitates communication between the computer and the laboratory instrument. Instrument-specific software controls the instrument's operation, data acquisition, analysis, and visualization. Computers may also be connected to a network for data sharing, remote access, or collaborative purposes. Network cards or wireless adapters enable communication with other devices or systems within the laboratory network.

[0120] Therefore, for example, a computer may have code that, when executed, operates the instrument and performs steps to analyze the sample. This could include, for example, instructing a pump to draw the solvent from the solvent container and push the solvent into the channel of the fluid tip, to draw the separation solvent from the separation solvent container and push it through the fluid tip into the sample loading zone, to move the sample through the phase transfer assembly onto the loading zone of the chromatography plate, and to move the concentrate solvent from the concentrate solvent container through the fluid tip into the concentrate zone on the chromatography plate.

[0121] G.Electricity The electrical elements of the system can operate through power supplied from a power source. The power source may be a battery mounted on the system. Alternatively, the system may have an external power connector, such as a plug or AC power connector, configured to access an external power source, such as an electric wall socket.

[0122] IV. How to Use Provided herein is a step for automatically performing thin-layer chromatography on an analyte from a sample.

[0123] A. Sample The sample may comprise a biological sample, for example, a sample obtained from a biological organism. Biological organisms may include, for example, viruses, bacteria, protists, eukaryotes, animals, humans, or plants. The biological sample may comprise any suitable biological sample comprising saliva, sputum, blood, plasma, serum, urine, feces, cerebrospinal fluid, bile, lymph, or provisionally the analyte of interest.

[0124] The sample may include environmental samples, such as samples obtained from the physical environment. Environmental samples may include soil samples, water samples, air samples, or waste samples. In embodiments in which the sample includes a water sample, the water sample may include a reservoir sample, well water sample, lake water sample, river water sample, seawater sample, wetland sample, ice sample, or any suitable environmental sample that provisionally includes the analyte of interest.

[0125] The sample may comprise industrial samples, such as those obtained from industrial equipment or plants, industrial waste, waste flows from industrial equipment or plants, or environments directly used for industrial processes. The industrial sample may comprise industrial products, precursors, or intermediates thereof, such as product chemicals derived from one or more synthesis steps, and the purity and / or yield of the sample will be determined with respect to any suitable industrial sample, such as dyes, raw materials, preservatives, sweeteners, foods, and cosmetics, or provisionally comprising the analyte of interest.

[0126] The sample may include agricultural samples. Agricultural samples are obtained from agricultural locations such as farms, fields, animal farms, farms for marine organisms or marine plants, or agricultural water bodies such as paddy fields. The agricultural sample may include soil samples, plant samples, livestock samples, water samples, food samples, or any suitable agricultural sample that provisionally includes the analyte of interest.

[0127] Typically, the sample comprises a liquid sample, such as an aqueous sample comprising an aqueous solvent and, provisionally, one or more analytes.

[0128] B. Analyte The analyte of interest may comprise molecules, for example, molecules with a size of approximately 50,000 Da, 20,000 Da, 10,000 Da, 5,000 Da, 2,000 Da, or 1,000 Da, for example, molecules ranging in size from 50 Da to 50,000 Da. The analyte may be any suitable molecule, for example, an organic molecule or an inorganic molecule. The analyte may comprise drugs with small organic molecules, for example, drugs with a size smaller than 5,000 Da, 2,000 Da, 1,000 Da, or 500 Da.

[0129] One or more analytes, for example, the compound of interest, may be, for example, cocaine, opioids, ayahuasca, central nervous system depressants, DMT, GHB, hallucinogens, heroin, ketamine, KHAT, LSD, MDMA (ecstasy / Molly), mescaline (peyote), methamphetamine, over-the-counter drugs (including, but not limited to, dextromethorphan, loperamide, and equivalents), PCP, prescription drugs, stimulants, psilocybin, rohypnol (flunitrazepam), saliva, steroids (anabolic), cannabinoids or their variants, cathinone (bath salt), or any other suitable analyte of interest.

[0130] The analyte may be one or more controlled substances. The analyte may be a drug. The analyte may be a cannabinoid. Cannabinoids include 10-ethoxy-9-hydroxy-delta-6a-tetrahydrocannabinol, 10-oxo-delta-6a-tetrahydrocannabinol (OTCH), 2-arachidonylglycerol (2AG), 2-arachidonylglyceryl ether, 8,9-dihydroxy-delta-6a-tetrahydrocannabinol, anandamide (AEA), cannabichromanone (CBCN), cannabichromene (CBC), cannabichromemenin (CBCV), and Cannabichromenic acid (CBCA), cannabiclomevalic acid (CBCVA), cannabicitran (CBT-C), cannabicyclool (CBL), cannabicycloalic acid (CBLA), cannabicyclovaline (CBLV), cannabidiol (CBD), cannabidiol monomethyl ether (CBDM), cannabidiolic acid (CBDA), cannabidiol chol (CBDC1), cannabidiol chol (CBN-C1), cannabidivaline (CBDV), cannabidiva Phosphate (CBDVA), Cannabiersoic acid A (CBEA-A), Cannabiersoic acid B (CBEA-B), Cannabiersoin (CBE), Cannabifuran (CBF), Cannabigerol (CBG), Cannabigerol monomethyl ether (CBGM), Cannabigerol acid (CBGA), Cannabigerol acid monomethyl ether (CBGAM), Cannabigerovaline (CBGV), Cannabigerovalic acid (CBGVA), Cannabiglendol-C3, Cannabinodiol ( Cannabinodivaline (CBND), Cannabinodivarate (CBV), Cannabinodivarate (CBVD), Cannabinol (CBN), Cannabinol Methyl Ether (CBNM), Cannabinol-C2 (CBN-C2), Cannabinol-C4 (CBN-C4), Cannabinol Acid (CBNA), Cannabilipsol (CBR), Cannabitriol (CBT), Cannabitriol Valine (CBTV), Dehydrocannabifuran (CBFD), Delta-8-Tetrahydrocannabinol (Δ 8 -THC), delta-8-tetrahydrocannabinolic acid (Δ 8-THCA), delta-9-cis-tetrahydrocannabinol (CIS-THC), delta-9-tetrahydrocannabinol (Δ 9 -THC), delta-9-tetrahydrocannabinol-C4 (Δ 9 -THC-C4), delta-9-tetrahydrocannabinolic acid A (Δ 9 -THCA-A), delta-9-tetrahydrocannabinolic acid B (Δ 9 -THCA-B), delta-9-tetrahydrocannabinolic acid C4 (Δ 9 -THCA-C4), delta-9-tetrahydrocannabionol (Δ 9 -THCA-C1), delta-9-tetrahydrocannabionolic acid C1 (Δ 9 -THCA-C1), delta-9-tetrahydrocannabivarin (THCV), delta-9-tetrahydrocannabivarinic acid (THCVA), lysophosphatidylinositol (LPI), N-arachidonoyl dopamine (NADA), tetrahydrocannabinol (THC), trihydroxy-delta-9-tetrahydrocannabinol (TRIOH-THC), virodhamine (OAE), or a variant thereof. The variant can comprise either a natural or a synthetic variant.

[0131] C. Method Provided herein is a method for isolating an analyte from a sample matrix and measuring the analyte by thin layer chromatography. "Sample matrix" refers to a sample that, in addition to the analyte, comprises many components, impurities, and interfering compounds. Exemplary sample matrices are biological samples (e.g., blood, plasma, serum, urine, saliva, tissue extracts, or cell culture media), environmental samples (e.g., water (including drinking water, surface water, wastewater), soil, sediment, airborne particulate matter, or biota), industrial samples (e.g., samples from chemical manufacturing, oil and gas industries, polymers, cosmetics, or consumer products), and food samples (fruits, vegetables, meat, dairy products, grains, or processed foods).

[0132] The sample matrix of a biological sample may comprise, for example, particulate matter, undigested food, proteins, cells, and mucin. The method disclosed herein can separate analytes such as small organic molecules from these matrix components.

[0133] A method for isolating an analyte from a sample matrix and measuring the analyte by thin-layer chromatography includes the steps of: (1) automatically separating the analyte from a first interfering substance by passing the sample through an interphase transfer assembly that captures or delays the first interfering substance; (2) automatically separating the analyte and the remaining interfering substance by thin-layer chromatography by depositing the light remaining interfering substance onto a thin-layer chromatography plate and unfolding the plate using a separation medium; and (3) automatically detecting the separated analyte.

[0134] The step of separating the analyte from the first interfering material can be performed optionally by using a pump to move the sample matrix into an interphase transfer assembly having a fluid channel with an interphase transfer medium, by moving the sample matrix into the first fluid channel by vacuum, and then moving the sample into the interphase transfer assembly using a liquid under pressure.

[0135] The step of separating the analyte from the remaining interfering substances by thin-layer chromatography can be carried out by, firstly, using a pump to move the liquid phase across the phase transfer assembly to elute the analyte and deposit the analyte and remaining interfering substances onto the loading zone of the thin-layer chromatography plate, and secondly, using a pump to move the separation solvent onto the loading zone to elute the analyte and remaining interfering substances from the phase transfer assembly. This will result in the migration of the separation solvent across the thin-layer chromatography plate through the chromatography medium, and the analyte will be separated from the other interfering substances by chromatography.

[0136] In an optional step, the analyte may move across the chromatographic medium as a band. In this case, the analyte band can be compressed or concentrated by using a pump to deposit the concentrating solvent laterally onto the chromatographic plate relative to the band. The concentrating solvent will migrate toward the center of the band, moving the analyte molecules with it.

[0137] The steps for automatically measuring the analyte may include using a computer-controlled camera to capture an image of the chromatographic plate and using software to determine the amount of the analyte based on the intensity of the image of the analyte.

[0138] These steps can be both integrated and automated. They can be integrated in the sense that all steps can be performed within a single system using a single cartridge. They can be automated in the sense that after the sample is provided to the cartridge, the system performs the remaining steps of detection without external intervention, for example, without human intervention. Fluid deposition can occur to move liquid from one place to another without using a pipette; that is, the liquid can move in a continuous fluid line from a container to a chromatography plate. They can be automated by using mechanical and electrical devices controlled by software products on a computer, such as motorized pumps, heaters, fans, and cameras; that is, once started, the steps can be carried out to completion without human intervention.

[0139] The exemplary method involves the following steps:

[0140] The sample is collected in a sample container, which has an outlet that is fitted to engage with the sample port of the cartridge. The sample may be, for example, a saliva sample collected by spitting into the sample container. The sample contains an analyte to be detected, such as a controlled substance such as THC. The sample container is then engaged with the sample port in the cartridge through any suitable connection, such as a Luer connector.

[0141] A cartridge containing a sample container is then engaged with the system's cartridge interface. A fluid manifold is then engaged with various ports within the fluid cartridge.

[0142] The system uses the vacuum generated by the fluid assembly to draw a sample, such as saliva, from the sample container into the fluid channel of the fluid cartridge.

[0143] An organic solvent such as dichloromethane is drawn from a liquid reservoir by a pump and pressurized into a fluid cartridge. The fluid channel connected to the first solvent reservoir intersects with the channel containing the sample. As a result, the specific volume of the sample, represented by the volume from the intersection to the intersection with the channel connected to the sample inlet, is pushed into the phase transfer assembly.

[0144] The phase transfer assembly filters out particulate matter, captures water from the sample, and binds the analyte to the phase transfer medium. Sustained pumping of the organic solvent through the phase transfer assembly elutes the analyte from the phase transfer medium and deposits it in the sample loading reservoir of the chromatography plate.

[0145] The separation solvent is then deposited into the sample reservoir. This is done by using a pump in the fluid assembly to draw the separation solvent from the fluid reservoir and move the liquid into the solvent port of the fluid tip. The separation solvent is then moved back into the sample reservoir through the fluid channel in the fluid tip.

[0146] The separation solvent diffuses across the sample reservoir and, acting as a front, migrates through the chromatographic medium by capillary action, separating the molecules containing the analyte from one another.

[0147] After separation, the analyte bands are compressed by the application of a concentrating solvent on both sides of the bands. This is done by using a pump in the fluid assembly to draw the liquid from the solvent reservoir and move the drawn liquid through fluid channels in the tip into one or more concentrating ports in the fluid tip, depositing the concentrating solvent into the concentrating wells on the chromatography plate. The concentrating solvents move toward each other, concentrating the analyte by compressing it into a narrower band or spot.

[0148] A suitable detector, such as a camera, can be used to monitor the extent to which a solvent, such as a separation and / or concentration solvent, has progressed. This can be done by taking images of the TLC plate, identifying the solvent front, and measuring the distance between the sample's starting position and the solvent front's position. In addition, or alternatively, multiple images can be taken, and the velocity of the solvent front can be calculated using both the distance and time between image acquisitions. Distance and / or velocity measurements of the solvent front can be used as part of a control system to optimize the separation and / or concentration step. This can help improve the system's reproducibility and performance.

[0149] An exemplary method includes a step of using a camera and control system to determine whether an additional solvent is required. In this case, images can be taken during the analysis to determine the extent to which the solvent has advanced. The additional solvent is added until the solvent front reaches a predetermined location, optimizing, for example, the separation and / or concentration step. Reproducible positioning can help prevent variations in the manufacture of TLC plates from affecting system performance. The step of measuring the solvent front position and / or rate can improve inter-assay and / or inter-system reproducibility. In some embodiments, the use of systems and methods for measuring the solvent front position and / or rate reduces the coefficient of variation in measurements between assays and / or systems for one or more analytes to less than 40, 30, 20, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1%.

[0150] Another exemplary method involves measuring the velocity of the solvent front using a camera and control system. In this case, the system can take a photograph of the TLC plate at defined intervals, for example, every 5 seconds. The system will calculate the rate at which the solvent moves across the TLC plate and, if necessary, adjust the solvent addition rate to control the velocity of the solvent front. Controlling the velocity of the solvent front can improve performance in both the separation and concentration steps and improve the reproducibility of the assay. This can further help to improve performance in light of variations in the TLC plate, temperature, altitude, and equivalents.

[0151] In addition, or alternatively, a reference chemical may be added to the sample as an internal standard. The exemplary method uses a camera and control system to measure the position of the internal standard and thus monitor the progress of the separation and / or concentration steps. In one embodiment, the internal standard is added to the phase transfer assembly during preparation. The exemplary cartridge may contain a suitable amount of any of the following reference chemicals in the phase transfer assembly: for example, about 5, 10, 25, 50, 100, 150, 200, 300, 400, or 500 μg. For example, about 150 μg of the reference chemical is added to the cotton of the phase transfer assembly in the cartridge. In addition, the internal standard can provide an indication that the method has been completed correctly; for example, improper final placement of the internal standard on the TLC plate may indicate a failure of one or more steps in the method and suggest that the sample needs to be repeated.

[0152] The analyte is stained using a dye for detection. For example, Fast Blue B binds to cannabinoids. It also absorbs light in the range of 500 nm to 600 nm, with an absorption peak at approximately 550 nm. As a result, any suitable amount of the dye can be deposited on a chromatography plate and allowed to bind to the analyte. The dye then becomes visible when illuminated with an LED emitting light in the range of 500 nm to 600 nm. Alternative dyes include Fast Blue BB salt, Fast Black K, Fast Red TR salt, and Fast Garnet GBC sulfate. Solvents for the dyes include, for example, methanol, hexane, water, acetonitrile, dichloromethane, dibromomethane, chloroform, toluene, acetone, methyl THF, pentane, cyclopentyl methyl ether, ethyl acetate, and 1-octanol.

[0153] The analyte spot is then detected using a detector such as a camera. The camera captures images of the spot and the calibration substance. The images are uploaded to a computer. The computer uses software to determine the intensity of the signal from the analyte, which is a function of the amount of analyte on the chromatography plate. Then, based on the intensity of the signal from the calibration substance, the software generates a measurement of the amount of analyte on the plate. Spots of known concentrations of the analyte are added to the plate as a calibration substance before the test is performed. For example, spots of 10 ng, 50 ng, and 100 ng are added to the plate. Once the dye is added to the plate, each known spot will have a different intensity. The intensity and correlation concentration of the spots are fitted to a line. The unknown substance is compared to the line, and its intensity is determined from this analysis.

[0154] Exemplary Embodiments 1. A method, (a) Performing thin-layer chromatography ("TLC") on one or more analyte molecules, (i) Moving a liquid sample containing analyte molecules into the fluid channel of a fluid tip, (ii) Moving a liquid sample through an interphase transfer assembly contained within a fluid channel inside or outside a fluid tip to separate analyte molecules from contaminants, (iii) Delivering the separated analyte molecules from a port in the fluid chip to the sample loading zone of a TLC plate having a chromatography medium on a solid substrate, (iv) Moving the separation solvent through the fluid channel in the fluid tip, and delivering the separation solvent from the fluid tip to the sample loading zone, (v) Developing the TLC plate by allowing the separation solvent, along with the analyte molecules, to migrate from the sample loading zone into the separation zone extending to the first dimension. by, (b) to concentrate the analyte as desired, (i) Moving a concentrated solvent through one or more fluid channels in a fluid tip, and delivering the concentrated solvent from the fluid tip to one or more concentrated solvent loading zones on a TLC plate, wherein the one or more concentrated solvent loading zones are positioned laterally with respect to the analyte molecules within the separation zone, (ii) Further development of the TLC plate by enabling the concentrated solvent to migrate through the chromatographic medium in an orthogonal or oblique dimension with respect to the first dimension. by, (c)Optionally, tagging the analyte molecules with detectable tags, (i) Moving a fluid equipped with a detectable tag through one or more fluid channels in a fluid chip, and delivering the tagged fluid to a separation zone along the tag-detectable analyte molecule. by, (d) Using a detector, detect a signal from the TLC plate indicating the presence of analyte molecules. Methods that include...

[0155] 2. (i) The fluid chip comprises a sample inlet port, a negative pressure port, a positive pressure port, and a liquid delivery port, which are fluidically connected to each other through a network of fluid channels, wherein the channel between the sample inlet port and the positive pressure port is intersected by the channel from the negative pressure port and the channel from the liquid delivery port, thereby defining the channel compartment, (ii) Using the negative pressure applied to the negative pressure port, draw out a volume of liquid sample sufficient to fill the channel compartment. (iii) Using the positive pressure applied to the positive pressure port, the volume is pushed through the phase-transfer assembly and the analyte is extracted from the volume. The method according to Embodiment 1.

[0156] 3. The method according to Embodiment 1, wherein moving a liquid sample into a fluid channel involves engaging a sample container containing the sample liquid with a port in the fluid tip.

[0157] 4. The method according to Embodiment 1, wherein moving a liquid sample into a fluid channel involves drawing the sample through the sample port in the fluid tip by applying negative pressure to a pressure port in the fluid tip that communicates with the sample port.

[0158] 5. The method according to Embodiment 1, wherein moving the liquid through the fluid channel includes pumping the liquid using one or more pumps that apply positive pressure to the channel.

[0159] 6. The method according to Embodiment 1, wherein delivering the analyte molecule to the sample loading zone includes, for example, moving a polar organic solvent through a fluid channel in an amount of about 0.1 mL to about 1 mL.

[0160] 7. The method according to Embodiment 1, wherein developing the TLC plate includes using a fan to evaporate the solvent from the surface of the TLC plate.

[0161] 8. The method according to Embodiment 1, wherein unfolding the TLC plate includes heating the TLC plate using a temperature control device.

[0162] 9. The method according to Embodiment 1, wherein the sample comprises an environmental sample.

[0163] 10. The method according to Embodiment 1, wherein the sample comprises a biological sample, such as saliva.

[0164] 11. The method according to Embodiment 1, wherein the analyte comprises a psychoactive drug, such as THC.

[0165] 12. The method according to Embodiment 1, wherein the analyte is present in the mixture at a concentration of 1% or less.

[0166] 13. The method according to Embodiment 1, wherein the analyte comprises THC and the detectable tag comprises Fast Blue.

[0167] 14. The method according to Embodiment 1, wherein detection includes using a camera to detect an image from a TLC plate.

[0168] 15. The method according to Embodiment 1, wherein detection includes quantifying the amount of the detected analyte.

[0169] 16. The method according to Embodiment 1, comprising detecting images from TLC plates using cameras, digitizing the images, and using computer software to quantify the amount of analytes in the digitized images.

[0170] 17. The method according to Embodiment 1, comprising depositing a trace amount of any of 50 ng, 20 ng, 5 ng, 2 ng, 1 ng, or 0.2 ng of an analyte (e.g., THC) onto a thin-layer chromatography plate and detecting the analyte.

[0171] The method according to Embodiment 1, comprising passing a 200-1,000 microliter sample containing an analyte at any low concentration of 18.125 nanograms / milliliter, 50 nanograms / milliliter, 12.5 nanograms / milliliter, 2.5 nanograms / milliliter, or 0.5 nanograms / milliliter through a phase transfer assembly to detect the analyte.

[0172] 19. The method according to Embodiment 1, which is carried out within one of the following periods: 19.1 hours, 45 minutes, 30 minutes, 15 minutes, or 5 minutes.

[0173] 20. The method according to Embodiment 1, as implemented by the system described in Embodiment 48.

[0174] 21. A cartridge for performing thin-layer chromatography ("TLC"), comprising a fluid tip that can be engaged with a thin-layer chromatography ("TLC") plate, (a) The TLC plate comprises a surface having a chromatographic medium, the surface comprising a sample loading zone, a separation zone extending from the sample loading zone to a first dimension, and optionally one or more concentrated solvent loading zones located within the separation zone, (b) The fluid tip is (i) A first fluid circuit comprising one or more internal and optionally externally connected ports, including a sample inlet port, at least one pressure port, and a liquid delivery port, wherein the first fluid circuit is configured to receive a sample from the sample inlet port into the fluid channel by negative pressure delivered through the pressure port, and to deliver at least a portion of the sample from the liquid delivery port out onto the sample loading zone by positive pressure delivered through the pressure port, through a fluid channel comprising a phase-transfer medium, (ii) A second fluid circuit comprising a port connected by one or more fluid channels, the second fluid circuit comprising a separation solvent intake port and a separation solvent delivery port, the separation solvent delivery port being positioned to deliver the separation solvent onto the sample loading zone, and the second fluid circuit optionally sharing ports and / or fluid channels with the first fluid circuit, (iii) optionally, one or more third fluid circuits, each comprising a port connected by a fluid channel, including one or more concentrated solvent intake ports and one or more concentrated solvent delivery ports, the one or more concentrated solvent delivery ports being positioned to deliver concentrated solvent to one or more concentrated solvent loading zones, (iv) Optionally, one or more fourth fluid circuits, each having a port connected by one or more fluid channels, including a dye intake port and a dye delivery port, the dye delivery port being positioned to deliver the dye onto the separation zone, (v) An opening through a fluid tip that exposes the isolation zone A cartridge equipped with these features.

[0175] 22. The cartridge according to Embodiment 21, wherein one or more concentrated solvent loading zones are located laterally to the sample loading zone.

[0176] 23. The TLC plate is a cartridge according to Embodiment 21, comprising a calibration substance that is dropped onto a chromatography medium.

[0177] 24. The cartridge according to Embodiment 21, wherein the TLC plate comprises a solid substrate selected from glass, glass, quartz, metal, aluminum, and plastic.

[0178] 25. The cartridge according to Embodiment 21, wherein the chromatography medium is selected from silica, alumina, cellulose, and polyamide.

[0179] 26. The cartridge according to Embodiment 21, wherein the first fluid circuit comprises a branched fluid channel connected to a sample inlet port, the first branch connecting to a first pressure port through the branched channel, and the second branch connecting to a liquid delivery port.

[0180] 27. The cartridge according to Embodiment 21, wherein the fluid channel, which includes a phase-transfer medium, is located outside the fluid chip and connected to the internal channel through a port.

[0181] 28. A cartridge according to Embodiment 21, wherein the fluid channel comprising the interphase transfer medium is located inside the fluid tip.

[0182] 29. The cartridge according to Embodiment 21, wherein the phase transfer assembly comprises a filter for filtering particulate matter, hydrophilic resin, and solid-phase extract.

[0183] 30. The cartridge according to Embodiment 29, wherein the solid-phase extract medium comprises a reverse-phase resin.

[0184] 31. The cartridge according to Embodiment 21, wherein the second fluid circuit is not in fluid communication with the first fluid circuit.

[0185] 32. The cartridge according to embodiment 21, wherein the fluid chip comprises one or more third fluid circuits.

[0186] 33. The cartridge according to embodiment 32, wherein one or more third fluid circuits are provided with first and second concentrated solvent delivery ports.

[0187] 34. The cartridge according to embodiment 33, wherein the fluid chip comprises a single third fluid circuit.

[0188] 35. The cartridge according to embodiment 21, wherein the fluid chip comprises one or more fourth fluid circuits.

[0189] 36. The cartridge according to embodiment 35, wherein the fluid chip comprises a single fourth fluid circuit.

[0190] 37. The cartridge according to Embodiment 21, comprising one or more loading reservoirs in contact with a surface extending over a sample loading zone, and a liquid delivery port and a solvent delivery port in fluid communication with one or more loading reservoirs.

[0191] 38. The cartridge according to embodiment 37, wherein the liquid delivery port and solvent delivery port communicate with different loading reservoirs.

[0192] 39. The cartridge according to Embodiment 21, comprising one or more concentrate reservoirs in contact with a surface spanning one or more concentrate solvent loading zones, one or more concentrate solvent delivery ports in fluid communication with one or more loading reservoirs, and optionally, the concentrate reservoirs having an elongated shape oriented substantially parallel to a first dimension.

[0193] 40. The cartridge according to Embodiment 39, wherein the concentration reservoir has an aspect ratio of at least 5:1 or 10:1 in elongated dimensions.

[0194] 41. The cartridge according to Embodiment 37 or 39, wherein one or more reservoirs are formed at least partially within a fluid tip, and contact between the fluid tip and the TLC plate surface forms a barrier, thereby allowing the liquid in the reservoir to migrate through the chromatography medium below the reservoir.

[0195] 42. The cartridge according to embodiment 41, wherein the reservoir has elongated, for example, linear sides that allow the separating medium to proceed as a substantially linear front.

[0196] 43. The cartridge according to embodiment 21, comprising a holder configured to support a fluid tip and a TLC plate.

[0197] 44. The cartridge according to embodiment 43, wherein the holder has a recessed portion configured to support a fluid tip and a portion configured to support a TLC tip.

[0198] 45. The cartridge according to embodiment 21, further comprising a sample collector that engages with a sample inlet port.

[0199] 46. ​​The fluid tip comprises interlocked first and second pieces, one or both of the pieces having an open channel on the surface of the piece and / or an opening through the piece, the first and second pieces being required to close the channel and to communicate the channel with a plurality of openings, as described in Embodiment 21 of the cartridge.

[0200] 47. The cartridge according to embodiment 46, wherein at least one opening through a part communicates with an open channel on the surface of the part, or at least one opening through a part communicates with an open channel on the other part when the parts are engaged.

[0201] 48. It is a system, (a) A cartridge interface for engaging with a cartridge, (i) A stage for holding the cartridge, (ii) A fluid manifold having multiple fluid ports that, when held by a holder, are adapted to engage with ports in a cartridge. A cartridge interface equipped with, (b) A fluid assembly, (i) Multiple containers for containing fluid, (ii) Multiple fluid lines, (iii) One or more pumps configured to pump liquid from a container through a fluid line to a fluid port, and to apply positive or negative pressure to at least one fluid port through at least one fluid line, (iv) Optionally, one or more valves configured to switch the fluid connection between the container and the fluid line leading to the fluid manifold. A fluid assembly comprising, (c) A detector configured to detect a signal from a cartridge, (d) optionally, when the cartridge is engaged with the cartridge interface, a fan is positioned to direct air onto the surface of the cartridge's thin-layer chromatography plate, (e) A temperature controller, optionally positioned to adjust the temperature of a cartridge that is engaged with an interface, (f) A computer equipped with operating software, the operating software being for controlling the operation of the cartridge interface, fluid assembly, and detector, and, if present, the fan and temperature controller, (g) A cartridge that can be optionally engaged with the cartridge interface A system equipped with these features.

[0202] 49. The system according to embodiment 48, wherein the stage is slidably mounted to the system chassis, and when the stage is extended from the interface, it allows for the installation of a cartridge on the stage, and when the stage is retracted into the system, it allows for the engagement of a fluid manifold.

[0203] 50. The system according to Embodiment 48, wherein the fluid manifold comprises at least three, four, five, six, seven, or eight fluid ports.

[0204] 51. The system according to Embodiment 48, wherein the multiple containers include containers each containing one, two, three, four, or five liquids selected from a separation solvent, an organic polar solvent, a concentration solvent, and a dye in a different manner.

[0205] 52. The system according to Embodiment 51, wherein the separation solvent is selected from dichloromethane, methanol, hexane, water, acetonitrile, dibromomethane, chloroform, toluene, acetone, methyl THF, pentane, cyclopentyl methyl ether, ethyl acetate, and 1-octanol.

[0206] 53. The system according to Embodiment 51, wherein the organic polar solvent is selected from dichloromethane, methanol, hexane, water, acetonitrile, dibromomethane, chloroform, toluene, acetone, methyl THF, pentane, cyclopentyl methyl ether, ethyl acetate, and 1-octanol.

[0207] 54. The system according to Embodiment 51, wherein the concentrated solvent is selected from dichloromethane, methanol, hexane, water, acetonitrile, dibromomethane, chloroform, toluene, acetone, methyl THF, pentane, cyclopentyl methyl ether, ethyl acetate, and 1-octanol.

[0208] 55. The system according to Embodiment 51, wherein the pigment is selected from Fast Blue, Fast Blue BB salt, Fast Black K, Fast Red TR salt, and Fast Garnet GBC sulfate.

[0209] 56. The system according to embodiment 48, wherein one or more of the pumps are syringe pumps.

[0210] 57. The system according to embodiment 48, wherein one of the pumps is a peristaltic pump.

[0211] 58. The system according to Embodiment 48, wherein the one or more pumps are at least two, three, or four pumps.

[0212] 59. The system according to embodiment 48, wherein the fluid assembly comprises one or more valves.

[0213] 60. The system according to embodiment 48, wherein the detector comprises a camera.

[0214] 61. The system according to embodiment 48, wherein the detector comprises a light source in the range of 500 nm to 600 nm.

[0215] 62. The system according to embodiment 48, comprising a fan.

[0216] 63. The system according to embodiment 48, comprising a temperature controller.

[0217] 64. The system according to embodiment 63, wherein the temperature controller comprises a Peltier element.

[0218] 65. The system according to embodiment 48, wherein the cartridge is engaged with the cartridge interface.

[0219] 66. The system according to embodiment 65, wherein the cartridge comprises the cartridge described in embodiment 21.

[0220] 67. The system according to Embodiment 65, wherein the cartridge comprises a fluid tip that meshes with a thin-layer chromatography plate, the tip having an opening that exposes the chromatographic surface of the TLC plate, and the system comprises a detector configured to detect a signal from the surface.

[0221] 68. A method comprising performing thin-layer chromatography ("TLC") by moving a sample from a fluid channel in a fluid tip onto a sample loading zone of a thin-layer chromatography plate, moving a separation solvent from a fluid channel in a fluid tip onto the sample loading zone, and unfolding a TLC plate by allowing the separation solvent to separate the analyte within the separation zone and a first dimension of the TLC plate.

[0222] 69. The method of Embodiment 68, further comprising moving a concentrated solvent from one or more fluid channels in a fluid tip onto one or more concentrated solvent loading zones positioned laterally in the separation zone relative to the sample loading zone, and further unfolding a TLC plate by allowing the concentrated solvent to concentrate the analyte in a second dimension within the separation zone.

[0223] 70. The method according to embodiment 68 or 69, further comprising moving the detection dye from a channel in a fluid tip onto a separation zone, thereby enabling the dye to tag the analyte.

[0224] 71. It is a system, (a) A computer, (i) Processor and (II) Memory connected to the processor, the memory is (1) A module comprising computer executable instructions for implementing the method described in Embodiment 20 It has memory and A system that includes a computer.

[0225] 72. It is a cartridge, (1) A fluid tip, (a) A first fluid channel that is in fluid communication with a first inlet and a first outlet, (b) A second fluid channel that is in fluid communication with the second inlet and the second outlet, and optionally, (c) A third fluid channel that is in fluid communication with the third inlet and third outlet The first, second, and third fluid channels are located within a substantially planar substrate between the first and second layers, and include a fluid chip. (2) A chromatography plate attached to at least a portion of the fluid chip, wherein the first, second, and optionally third outlets are in fluid communication with the surface of the chromatography plate and A cartridge equipped with these features.

[0226] 73. It is a cartridge, (1) A fluid tip, (a) Multiple entrances, (b) Multiple exits, (c) Multiple channels in the substrate that are in fluid communication with at least one inlet and at least one outlet A fluid chip equipped with, (2) A thin-layer chromatography plate in contact with at least a portion of the fluid chip such that at least two of the multiple outlets are in fluid communication with the plate. A cartridge equipped with these features.

[0227] 74. It is a cartridge, (1) A fluid tip, (a) A plurality of reservoirs, each reservoir configured to receive a liquid phase, (b) Multiple reservoirs and multiple channels that communicate fluidly A fluid chip equipped with, (2) A thin-layer chromatography plate in contact with at least a portion of the fluid chip so that multiple reservoirs are in fluid communication with the plate. A cartridge equipped with these features.

[0228] 75. It is a cartridge, (1) A conduit comprising at least one chromatographic resin, (2) Thin-layer chromatography plate and (3) A fluid chip having at least one channel, wherein the chromatographic resin and plate are in fluid communication through the channel, A cartridge equipped with these features.

[0229] 76. It is a system, (A) A manifold for inserting the cartridge, (B) A cartridge inserted into the manifold, (1) A fluid tip, (a) A first fluid channel that is in fluid communication with a first inlet and a first outlet, (b) A second fluid channel that is in fluid communication with the second inlet and the second outlet, and optionally, (c) A third fluid channel that is in fluid communication with the third inlet and third outlet The first, second, and third fluid channels are located within a substantially planar substrate between the first and second layers, and include a fluid chip. (2) A chromatography plate attached to at least a portion of a substrate, wherein the first, second, and optionally third outlets are in fluid communication with the surface of the chromatography plate and A cartridge equipped with, (C) A first conduit, a second inlet, and a second valve, which are in fluid communication with the first inlet and the first valve, and optionally a second conduit, which are in fluid communication with the third inlet. A system equipped with these features.

[0230] 77. A method, (A) Delivering a sample comprising at least one analyte to a first inlet of a fluid tip, the fluid tip is (a) A first fluid channel in fluid communication with a first inlet and a first outlet; (b) A second fluid channel in fluid communication with a second inlet and a second outlet, and optionally, (c) A third fluid channel in fluid communication with a third inlet and a third outlet comprising, the first, second, and third fluid channels being disposed within a substantially planar substrate disposed between a first layer and a second layer, and further a chromatography plate being attached to at least a portion of the substrate, the first, second, and optionally, third outlets being in fluid communication with the surface of the chromatography plate; and (B) dispensing a first solvent through the first channel of the fluid chip such that the solvent conveys the sample through the first channel to the first outlet and further onto the chromatography plate; (C) dispensing a second solvent through the first channel of the fluid chip to the first outlet and further onto the chromatography plate such that at least one analyte in the sample is separated along a first length of the chromatography plate, and optionally, (D) dispensing a third solvent through the second channel of the fluid chip to the second outlet and further onto the chromatography plate such that at least one analyte is concentrated along a second length of the chromatography plate comprising a method.

[0231] 78. A method for automatically performing thin layer chromatography, comprising: (a) engaging a fluid cartridge comprising a sample with a cartridge interface of a system; (b) automatically using the system to (i) separate analytes in the sample from contaminating materials; (ii) deliver the separated analytes to the surface of a thin layer chromatography plate; (iii) develop the plate and move the analytes into separation zones of the plate; (iv) optionally, concentrating the analyte with a concentrating solvent, and (v) optionally, detecting the concentrated analyte on a plate performing the steps of A method comprising

[0232] 79. A method comprising (a) performing thin layer chromatography ("TLC") on one or more analyte molecules, (i) moving a liquid sample comprising analyte molecules into a fluid channel of a fluidic chip, (ii) moving the liquid sample through a phase transfer assembly contained within an internal or external fluid channel of the fluidic chip to exchange and / or transfer analyte molecules from the liquid sample into a second liquid, (iii) delivering the second liquid from a port within the fluidic chip to a sample loading zone of a TLC plate having a chromatographic medium on a solid substrate, (iv) moving a separation solvent through a fluid channel within the fluidic chip and delivering the separation solvent from the fluidic chip to the sample loading zone, (v) developing the TLC plate by enabling the separation solvent, together with the analyte molecules, to migrate from the sample loading zone into a separation zone extending in a first dimension by (b) optionally, concentrating the analyte, (i) moving a concentrating solvent through one or more fluid channels within the fluidic chip and delivering the concentrating solvent from the fluidic chip to one or more concentrating solvent loading zones on the TLC plate, wherein the one or more concentrating solvent loading zones are positioned laterally with respect to the analyte molecules within the separation zone, (ii) further developing the TLC plate by enabling the concentrating solvent to migrate through the chromatographic medium in a dimension orthogonal or oblique to the first dimension by (c)Optionally, tag the precipitate molecules with detectable tags, (i) Moving a fluid equipped with a detectable tag through one or more fluid channels in a fluid chip, and delivering the tagged fluid to a separation zone along the tag-detectable analyte molecule. by, (d) Using a detector, detect a signal from the TLC plate indicating the presence of analyte molecules. Methods that include...

[0233] 80. The method according to Embodiment 80, wherein the liquid sample is an aqueous sample comprising an aqueous solvent, and / or the second liquid is an organic solvent. [Examples]

[0234] Saliva samples are analyzed for the presence of THC using the following method.

[0235] Load the oral fluid and dichloromethane (DCM) onto the column. 1. Insert the cartridge. 2. Engage the manifold five times. 3. Turn on the hot plate and heat it to 80°C. 4. Use syringe pump 1 and valve 4 to aspirate 600 μL of oral fluid. 5. Using syringe pump 3 and valve 3, draw 1 mL of dichloromethane (DCM) from the reservoir bottle. 6. Using valve 2, dispense 1 mL of DCM from syringe pump 3. This moves a 400 mL sample from the fluid channel into the phase transfer assembly.

[0236] Load the DCM sample from the column onto the TLC plate. 7. Use syringe pump 3 and valve 3 to draw 1 mL of DCM from the reservoir bottle. 8. Using valve 2, dispense 800 μL of DCM from syringe pump 3. 9. Using syringe pump 3 and valve 3, draw 800 μL of DCM from the reservoir bottle. 10. Turn on the fan. 11. Using syringe pump 3 and valve 2, dispense 100 μL of DCM. Wait for 5 seconds. Repeat this 4 times. Next, using syringe pump 3 and valve 2, aspirate 300 μL of DCM. Wait for 5 seconds. A layer of cotton, positioned after the silica layer, contains the control CBD. In this step, THC metabolites such as 11-OH-THC and 11-COOH-THC remain on the steady phase, while THC, CBC, and CBD move with the mobile phase into the sample loading area. The sample loading area has a length of approximately 2.5 cm.

[0237] THC and CBD are stacked in a straight line on a TLC plate. 12. Repeat step 11 twice. 13. Turn off the fan. 14. Using syringe pump 3 and valve 3, draw 500 μL of DCM from the reservoir bottle. 15. Using valve 2, dispense 700 μL of DCM from syringe pump 3. Wait for 15 seconds. 16. Turn on the fan. 17. Using syringe pump 3 and valve 2, aspirate 500 μL of DCM. 18. Change the hot plate temperature to 35°C. Wait for 1 minute. 19. Turn off the fan and cover the TLC plate with the glass cover. 20. Using syringe pump 2 and valve 3, draw 700 μL of 5 acetonitrile:1 water (separation solvent) from the reservoir bottle.

[0238] TLC separates CBD, CBC, and THC. Dispense 21,500 μL of 5:1 water:methanol from syringe pump 2 and valve 4. Wait for 40 seconds. Dispense 22,100 μL of 5:1 solution from syringe pump 2 and valve 4. Wait for 75 seconds. 23. Dispense 100 μL of 5:1 from syringe pump 2 and valve 4. Wait for 80 seconds.

[0239] Concentrate THC and CBD onto the spots concentrated from the line 24. Turn on the fan and set the hot plate to 40 °C. Wait for 1 minute. 25. Turn off the fan. 26. Use syringe pump 3 and valve 3 to aspirate 300 μL of DCM from the reservoir bottle. 27. Use valve 1 to dispense 700 μL of DCM from syringe pump 3. 28. Use syringe pump 3 and valve 3 to aspirate 700 μL of DCM from the reservoir bottle. 29. Use valve 1 to dispense 1 mL of DCM from syringe pump 3.

[0240] Perform the second concentration step 30. Use syringe pump 3 and valve 1 to aspirate 200 μL of DCM. 31. Turn on the fan. 32. Use syringe pump 3 and valve 3 to aspirate 800 μL of DCM from the reservoir bottle. 33. Use valve 1 to dispense 1 mL of DCM from syringe pump 3. 34. Use syringe pump 3 and valve 1 to aspirate 200 μL of DCM. 35. Turn on the fan. Wait for 30 seconds. 36. Turn off the fan.

[0241] Application of dye 37. Dispense 300 μL of air from syringe pump 1 and valve 3. 38. Use syringe pump 1 and valve 5 to aspirate 700 μL of fast blue indicator from the reservoir bottle. 39. Use valve 6 to dispense 1 mL of fast blue indicator from syringe pump 1. 40. Turn on peristaltic pump 2 and aspirate / dispense methanol onto the TLC plate. Wait for 30 seconds. 41. Turn off peristaltic pump 2. 42. Turn on the fan to blow away the Fast Blue indicator and methanol from the TLC plate.

[0242] Image acquisition and processing After waiting 43.2 minutes, take a photograph of the TLC plate. 44. Perform background subtraction on the image. 45. Integrate the peaks, compare them to the calibration standard and the unknown sample, and find the unknown concentration.

[0243] When used herein, the following meanings apply unless otherwise specified: The words “can” and “may” are used in a permissive sense (i.e., meaning potential) rather than a mandatory sense (i.e., meaning obligation). The words “include,” “including,” and “includes,” and their equivalents, mean “include,” not limit. The singular forms “a,” “an,” and “the” include plural references. Thus, for example, a reference to “an element” includes a combination of two or more elements, regardless of the use of other terms and phrases relating to one or more elements, such as “one or more.” The phrase “at least one” includes “one,” “one or more,” and “one or a plurality,” and therefore consider the use of the term “a plurality.” The term "or" is non-exclusive unless otherwise indicated, i.e., it encompasses both "and" and "or". The term "any of" between a modifier and a sequence means that the modifier modifies each component of the sequence. For example, the phrase "at least one of 1, 2, or 3" means "at least 1, at least 2, or at least 3". The term "about" refers to a range of ±5% of a number stated within the context of a particular use. The term "consisting essentially of" refers to the inclusion of the enumerated elements with other elements that do not significantly affect the basic and novel properties of the claimed combination.

[0244] It should be understood that the description and drawings are not intended to limit the invention to any particular form disclosed herein, but rather to cover all modifications, equivalents, and substitutions that fall within the spirit and scope of the invention as defined by the appended claims. Further modifications and substitutions of various aspects of the embodiments of the invention will be obvious to those skilled in the art in light of this description. Therefore, this description and drawings should be interpreted as illustrative only and are intended to teach those skilled in the art a general way of practicing the invention. It should be understood that the forms of the invention illustrated and described herein should be taken as examples of embodiments. Elements and materials may be substituted with those illustrated and described herein, parts and processes may be reversed or omitted, and certain features of the invention may be used independently, all of which will be obvious to those skilled in the art after understanding the description of the invention. Modifications may be made to the elements described herein without departing from the spirit and scope of the invention as described in the following claims.

[0245] All publications, patents, and patent applications referenced herein are incorporated herein by reference to the same extent as each individual publication, patent, or patent application is shown to be incorporated by specific and individual reference.

Claims

1. It is a method, (a) Performing thin-layer chromatography ("TLC") on one or more analyte molecules, (i) Moving the liquid sample containing the analyte molecules into the fluid channel of the fluid tip, (ii) Moving the liquid sample through an interphase transfer assembly contained within a fluid channel inside or outside the fluid tip to separate the analyte molecules from contaminants, (iii) Delivering the separated analyte molecules from the port in the fluid chip to the sample loading zone of a TLC plate having a chromatography medium on a solid substrate, (iv) Moving the separation solvent through the fluid channel in the fluid tip, and delivering the separation solvent from the fluid tip to the sample loading zone, (v) Developing the TLC plate by enabling the separation solvent, together with the analyte molecules, to migrate from the sample loading zone into the separation zone extending to the first dimension. by, (b) optionally, to concentrate the analyte, (i) Moving a concentrated solvent through one or more fluid channels in the fluid tip, and delivering the concentrated solvent from the fluid tip to one or more concentrated solvent loading zones on the TLC plate, wherein the one or more concentrated solvent loading zones are positioned laterally to the analyte molecules within the separation zone, (ii) Further developing the TLC plate by enabling the concentrated solvent to migrate through the chromatography medium in an orthogonal or oblique dimension to the first dimension. by, (c) Optionally, tagging the analyte molecules with detectable tags, (i) Moving a fluid containing a detectable tag through one or more fluid channels in the fluid chip, and delivering the fluid containing the tag to a separation zone along the tag-detectable analyte molecule. by, (d) Using a detector, detect a signal from the TLC plate indicating the presence of the analyte molecules. Methods that include...

2. (i) The fluid chip comprises a sample inlet port, a negative pressure port, a positive pressure port, and a liquid delivery port, which are fluidly connected to each other through a network of fluid channels, wherein the channel between the sample inlet port and the positive pressure port is intersected by the channel from the negative pressure port and the channel from the liquid delivery port, thereby defining a channel section. (ii) Using the negative pressure applied to the negative pressure port, draw out a volume of the liquid sample sufficient to fill the channel compartment. (iii) Using the positive pressure applied to the positive pressure port, the volume is pushed through the interphase transfer assembly and the analyte is extracted from the volume. The method according to claim 1.

3. The method according to claim 1, wherein moving the liquid sample into the fluid channel includes engaging a sample container containing the sample liquid with a port in the fluid tip.

4. The method according to claim 1, wherein moving the liquid sample into the fluid channel includes drawing the sample through the sample port in the fluid tip by applying negative pressure to a pressure port in the fluid tip that communicates with the sample port.

5. The method according to claim 1, wherein moving the liquid through the fluid channel includes pumping the liquid using one or more pumps that apply positive pressure to the channel.

6. The method according to claim 1, wherein delivering the analyte molecules to the sample loading zone includes, for example, moving a polar organic solvent through the fluid channel in an amount of about 0.1 mL to about 1 mL.

7. The method according to claim 1, wherein developing the TLC plate includes using a fan to evaporate the solvent from the surface of the TLC plate.

8. The method according to claim 1, wherein unfolding the TLC plate includes heating the TLC plate using a temperature control device.

9. The method according to claim 1, wherein the sample comprises an environmental sample.

10. The method according to claim 1, wherein the sample comprises a biological sample, such as saliva.

11. The method according to claim 1, wherein the analyte comprises a psychotropic drug, for example, THC.

12. The method according to claim 1, wherein the analyte is present in the mixture at a concentration of 1% or less.

13. The method according to claim 1, wherein the analyte comprises THC, and the detectable tag comprises Fastblue.

14. The method according to claim 1, wherein the detection includes detecting an image from the TLC plate using a camera.

15. The method according to claim 1, wherein the detection includes quantifying the amount of the detected analyte.

16. The method according to claim 1, wherein the detection includes detecting images from the TLC plate and the camera, digitizing the images, and using computer software to quantify the amount of the analyte in the digitized images.

17. The method according to claim 1, comprising depositing a trace amount of any of 50 ng, 20 ng, 5 ng, 2 ng, 1 ng, or 0.2 ng of an analyte (e.g., THC) onto the thin-layer chromatography plate and detecting the analyte.

18. The method according to claim 1, comprising passing a 200 to 1,000 microliter sample comprising an analyte at any low concentration of 125 nanograms / milliliter, 50 nanograms / milliliter, 12.5 nanograms / milliliter, 2.5 nanograms / milliliter, or 0.5 nanograms / milliliter through the interphase transfer assembly and detecting the analyte.

19. The method according to claim 1, which is carried out within one hour, 45 minutes, 30 minutes, 15 minutes, or 5 minutes.

20. The method according to claim 1, as implemented by the system described in claim 48.

21. A cartridge for performing thin-layer chromatography (TLC), comprising a fluid tip that engages with a thin-layer chromatography (TLC) plate, (a) The TLC plate comprises a surface having a chromatographic medium, the surface comprising a sample loading zone, a separation zone extending in a first dimension from the sample loading zone, and optionally one or more concentrated solvent loading zones located within the separation zone, (b) The fluid tip is (i) A first fluid circuit comprising one or more internal and optionally externally connected ports, including a sample inlet port, at least one pressure port, and a liquid delivery port, wherein the first fluid circuit is configured to receive a sample from the sample inlet port into the fluid channel by negative pressure delivered through the pressure port, and to deliver at least a portion of the sample from the liquid delivery port out onto the sample loading zone by positive pressure delivered through the pressure port, through a fluid channel comprising a phase-transfer medium, (ii) A second fluid circuit comprising a port connected by one or more fluid channels, the second fluid circuit comprising a separation solvent intake port and a separation solvent delivery port, the separation solvent delivery port being positioned to deliver the separation solvent onto the sample loading zone, and the second fluid circuit optionally sharing ports and / or fluid channels with the first fluid circuit, (iii) Optionally, one or more third fluid circuits, the one or more third fluid circuits comprising ports connected by fluid channels, the one or more concentrated solvent intake ports and the one or more concentrated solvent delivery ports, the one or more concentrated solvent delivery ports being positioned to deliver concentrated solvent to the one or more concentrated solvent loading zones, (iv) Optionally, one or more fourth fluid circuits, each having a port connected by one or more fluid channels, the fourth fluid circuit having a dye intake port and a dye delivery port, the dye delivery port being positioned to deliver the dye onto the separation zone, (v) An opening through the fluid tip that exposes the separation zone A cartridge equipped with these features.

22. The cartridge according to claim 21, wherein the one or more concentrated solvent loading zones are positioned laterally with respect to the sample loading zone.

23. The cartridge according to claim 21, wherein the TLC plate comprises a calibration substance that is dropped onto the chromatography medium.

24. The cartridge according to claim 21, wherein the TLC plate comprises a solid substrate selected from glass, glass, quartz, metal, aluminum, and plastic.

25. The cartridge according to claim 21, wherein the chromatography medium is selected from silica, alumina, cellulose, and polyamide.

26. The cartridge according to claim 21, wherein the first fluid circuit comprises a branched fluid channel connected to the sample inlet port, the first branch being connected to the first pressure port through the branched channel, and the second branch being connected to the liquid delivery port.

27. The cartridge according to claim 21, wherein the fluid channel, which includes the interphase transfer medium, is located outside the fluid tip and is connected to an internal channel through a port.

28. The cartridge according to claim 21, wherein the fluid channel comprising the interphase transfer medium is located inside the fluid tip.

29. The cartridge according to claim 21, wherein the interphase transfer assembly comprises a filter for filtering particulate matter, a hydrophilic resin, and a solid-phase extract.

30. The cartridge according to claim 29, wherein the solid-phase extract medium comprises a reverse-phase resin.

31. The cartridge according to claim 21, wherein the second fluid circuit is not in fluid communication with the first fluid circuit.

32. The cartridge according to claim 21, wherein the fluid chip comprises one or more third fluid circuits.

33. The cartridge according to claim 32, wherein the one or more third fluid circuits comprises first and second concentrated solvent delivery ports.

34. The cartridge according to claim 33, wherein the fluid chip comprises a single third fluid circuit.

35. The cartridge according to claim 21, wherein the fluid chip comprises one or more fourth fluid circuits.

36. The cartridge according to claim 35, wherein the fluid chip comprises a single fourth fluid circuit.

37. The cartridge according to claim 21, comprising one or more loading reservoirs in contact with the surface extending over the sample loading zone, wherein the liquid delivery port and the solvent delivery port are in fluid communication with the one or more loading reservoirs.

38. The cartridge according to claim 37, wherein the liquid delivery port and the solvent delivery port communicate with different loading reservoirs.

39. The cartridge according to claim 21, comprising one or more concentrated reservoirs in contact with the surface extending over one or more concentrated solvent loading zones, wherein one or more concentrated solvent delivery ports are in fluid communication with the one or more loading reservoirs, and optionally the concentrated reservoirs have an elongated shape oriented substantially parallel to the first dimension.

40. The cartridge according to claim 39, wherein the concentrated reservoir has an aspect ratio of at least 5:1 or 10:1 in an elongated dimension.

41. The cartridge according to claim 37 or 39, wherein one or more of the reservoirs are at least partially formed within the fluid tip, and the contact between the fluid tip and the TLC plate surface forms a barrier, thereby allowing the liquid in the reservoir to migrate through the chromatography medium beneath the reservoir.

42. The cartridge according to claim 41, wherein the reservoir has elongated, for example, linear sides that allow the separation medium to proceed as a substantially linear front.

43. The cartridge according to claim 21, comprising a holder configured to support the fluid tip and the TLC plate.

44. The cartridge according to claim 43, wherein the holder is recessed with respect to a portion configured to support the fluid chip and comprises a portion configured to support the TLC chip.

45. The cartridge according to claim 21, further comprising a sample collector that engages with the sample inlet port.

46. The cartridge according to claim 21, wherein the fluid tip comprises interlocked first and second pieces, one or both of the pieces having an open channel on the surface of the piece and / or an opening through the piece, the first and second pieces being required to close the channel and to connect the channel to a plurality of openings.

47. The cartridge according to claim 46, wherein at least one opening through a part communicates with an open channel on the surface of the part, or, when the part is engaged, at least one opening through a part communicates with an open channel on the other part.

48. It is a system, (a) A cartridge interface for engaging with a cartridge, (i) A stage for holding the cartridge, (ii) A fluid manifold having a plurality of fluid ports that, when held by the holder, are adapted to engage with ports in a cartridge. A cartridge interface equipped with, (b) A fluid assembly, (i) Multiple containers for containing fluid, (ii) Multiple fluid lines, (iii) One or more pumps configured to pump liquid from the container through the fluid lines to the fluid ports, and to apply positive or negative pressure to at least one fluid port through at least one fluid line, (iv) Optionally, one or more valves configured to switch the fluid connection between the container and the fluid line connected to the fluid manifold. A fluid assembly comprising, (c) A detector configured to detect a signal from the cartridge, (d) optionally, when the cartridge is engaged with the cartridge interface, a fan is positioned to direct air onto the surface of the thin-layer chromatography plate of the cartridge, (e) A temperature controller, optionally positioned to adjust the temperature of a cartridge engaged with the interface, (f) A computer equipped with operating software, the operating software being for controlling the operation of the cartridge interface, the fluid assembly, and the detector, and, if present, the fan and the temperature controller, (g) A cartridge which can be optionally engaged with the cartridge interface. A system equipped with these features.

49. The system according to claim 48, wherein the stage is slidably mounted to the chassis of the system, and when the stage is extended from the interface, it allows for the installation of the cartridge on the stage, and when the stage is retracted into the system, it allows for the engagement of the fluid manifold.

50. The system according to claim 48, wherein the fluid manifold comprises at least three, four, five, six, seven, or eight fluid ports.

51. The system according to claim 48, wherein the plurality of containers include containers each containing one, two, three, four, or five liquids selected from a separation solvent, an organic polar solvent, a concentration solvent, and a dye in different ways.

52. The system according to claim 51, wherein the separation solvent is selected from dichloromethane, methanol, hexane, water, acetonitrile, dibromomethane, chloroform, toluene, acetone, methyl THF, pentane, cyclopentyl methyl ether, ethyl acetate, and 1-octanol.

53. The system according to claim 51, wherein the organic polar solvent is selected from dichloromethane, methanol, hexane, water, acetonitrile, dibromomethane, chloroform, toluene, acetone, methyl THF, pentane, cyclopentyl methyl ether, ethyl acetate, and 1-octanol.

54. The system according to claim 51, wherein the concentrated solvent is selected from dichloromethane, methanol, hexane, water, acetonitrile, dibromomethane, chloroform, toluene, acetone, methyl THF, pentane, cyclopentyl methyl ether, ethyl acetate, and 1-octanol.

55. The system according to claim 51, wherein the dye is selected from Fast Blue, Fast Blue BB salt, Fast Black K, Fast Red TR salt, and Fast Garnet GBC sulfate.

56. The system according to claim 48, wherein one or more of the pumps are syringe pumps.

57. The system according to claim 48, wherein one of the pumps is a peristaltic pump.

58. The system according to claim 48, wherein the one or more pumps are at least two, three, or four pumps.

59. The system according to claim 48, wherein the fluid assembly comprises one or more valves.

60. The system according to claim 48, wherein the detector comprises a camera.

61. The system according to claim 48, wherein the detector comprises a light source in the range of 500 nm to 600 nm.

62. The system according to claim 48, comprising the aforementioned fan.

63. The system according to claim 48, comprising the temperature controller.

64. The system according to claim 63, wherein the temperature controller comprises a Peltier element.

65. The system according to claim 48, wherein the cartridge is engaged with the cartridge interface.

66. The system according to claim 65, wherein the cartridge comprises the cartridge described in claim 21.

67. The system according to claim 65, wherein the cartridge comprises a fluid tip that meshes with a thin-layer chromatography plate, the tip having an opening that exposes the chromatography surface of the TLC plate, and the system comprises the detector configured to detect a signal from the surface.

68. A method comprising performing thin-layer chromatography ("TLC") by moving a sample from a fluid channel in a fluid tip onto a sample loading zone of a thin-layer chromatography plate; moving a separation solvent from a fluid channel in a fluid tip onto the sample loading zone; and unfolding the TLC plate by allowing the separation solvent to separate the analyte within the separation zone and a first dimension of the TLC plate.

69. The method according to claim 68, further comprising moving a concentrated solvent from one or more fluid channels in the fluid tip onto one or more concentrated solvent loading zones positioned laterally within the separation zone relative to the sample loading zone, and further unfolding the TLC plate by enabling the concentrated solvent to concentrate the analyte in a second dimension within the separation zone.

70. The method according to claim 68 or 69, further comprising moving the detection dye from the channel in the fluid tip onto the separation zone, thereby enabling the dye to tag the analyte.

71. It is a system, (a) A computer, (i) Processor, (II) A memory coupled to the processor, wherein the memory is (1) A module comprising computer executable instructions for implementing the method described in claim 20. It has memory and A system that includes a computer.

72. It is a cartridge, (1) A fluid chip, (a) A first fluid channel that is in fluid communication with the first inlet and the first outlet, (b) A second fluid channel that is in fluid communication with the second inlet and the second outlet, and optionally, (c) A third fluid channel that is in fluid communication with the third inlet and third outlet The first, second, and third fluid channels are provided, and the fluid chip is disposed within a substantially planar substrate positioned between the first and second layers. (2) A chromatography plate attached to at least a portion of the fluid tip, wherein the first, second, and optionally third outlets are in fluid communication with the surface of the chromatography plate. A cartridge equipped with these features.

73. It is a cartridge, (1) A fluid chip, (a) Multiple entrances, (b) Multiple exits, (c) A plurality of channels in the substrate that are in fluid communication with at least one inlet and at least one outlet A fluid chip equipped with, (2) A thin-layer chromatography plate in contact with at least a portion of the fluid chip such that at least two of the plurality of outlets are in fluid communication with the plate. A cartridge equipped with these features.

74. It is a cartridge, (1) A fluid chip, (a) A plurality of reservoirs, each reservoir configured to receive a liquid phase, (b) Multiple channels that are in fluid communication with the multiple reservoirs A fluid chip equipped with, (2) A thin-layer chromatography plate in contact with at least a portion of the fluid chip so that the plurality of reservoirs are in fluid communication with the plate. A cartridge equipped with these features.

75. It is a cartridge, (1) A conduit comprising at least one chromatographic resin, (2) Thin-layer chromatography plate and (3) A fluid chip having at least one channel, wherein the chromatographic resin and the plate are in fluid communication through the channel, and A cartridge equipped with these features.

76. It is a system, (A) A manifold for inserting the cartridge, (B) A cartridge inserted into the manifold, (1) A fluid chip, (a) A first fluid channel that is in fluid communication with the first inlet and the first outlet, (b) A second fluid channel that is in fluid communication with the second inlet and the second outlet, and optionally, (c) A third fluid channel that is in fluid communication with the third inlet and third outlet The first, second, and third fluid channels are provided, and the fluid chip is disposed within a substantially planar substrate positioned between the first and second layers. (2) A chromatography plate attached to at least a portion of the substrate, wherein the first, second, and optionally third outlets are in fluid communication with the surface of the chromatography plate. A cartridge equipped with, (C) A first conduit having fluid communication with the first inlet and the first valve, the second inlet and the second valve, and optionally, a second conduit having fluid communication with the third inlet. A system equipped with these features.

77. It is a method, (A) Delivering a sample comprising at least one analyte to a first inlet of a fluid tip, the fluid tip is (a) A first fluid channel that is in fluid communication with the first inlet and the first outlet, (b) A second fluid channel that is in fluid communication with the second inlet and the second outlet, and optionally, (c) A third fluid channel that is in fluid communication with the third inlet and third outlet The first, second, and third fluid channels are located within a substantially planar substrate positioned between the first and second layers, and further, A chromatography plate is attached to at least a portion of the substrate, and the first, second, and optionally third outlets are in fluid communication with the surface of the chromatography plate. That thing, (B) Dispensing the first solvent through the first channel of the fluid tip such that the solvent transports the sample through the first channel to the first outlet and then onto the chromatography plate, (C) Dispensing a second solvent through the first channel of the fluid tip to the first outlet and onto the chromatography plate, and optionally, so that at least one analyte in the sample is separated along the first length of the chromatography plate, (D) Dispensing the third solvent through the second channel of the fluid tip to the second outlet and onto the chromatography plate so that the at least one analyte is concentrated along the second length of the chromatography plate. Methods that include...

78. A method for automatically performing thin-layer chromatography, (a) Engaging a fluid cartridge containing a sample with the system's cartridge interface, (b) Automatically, using the system, (i) Separating the analyte in the sample from the contaminating material, (ii) Delivering the separated analytes to the surface of a thin-layer chromatography plate, (iii) Unfolding the plate and moving the analyte into the separation zone of the plate, (iv) Optionally, concentrate the analyte with a concentrating solvent, (v) detecting the arbitrarily concentrated analyte on the plate The steps to be taken are to carry out and Methods that include...

79. It is a method, (a) Performing thin-layer chromatography ("TLC") on one or more analyte molecules, (i) Moving the liquid sample containing the analyte molecules into the fluid channel of the fluid tip, (ii) Moving the liquid sample through an interphase transfer assembly contained within a fluid channel inside or outside the fluid tip to exchange and / or transfer the analyte molecules from the liquid sample into the second liquid, (iii) Delivering the second liquid from the port in the fluid chip to the sample loading zone of a TLC plate having a chromatography medium on a solid substrate, (iv) Moving the separation solvent through the fluid channel in the fluid tip, and delivering the separation solvent from the fluid tip to the sample loading zone, (v) Developing the TLC plate by enabling the separation solvent, together with the analyte molecules, to migrate from the sample loading zone into the separation zone extending to the first dimension. by, (b) optionally, to concentrate the analyte, (i) Moving a concentrated solvent through one or more fluid channels in the fluid tip, and delivering the concentrated solvent from the fluid tip to one or more concentrated solvent loading zones on the TLC plate, wherein the one or more concentrated solvent loading zones are positioned laterally to the analyte molecules within the separation zone, (ii) Further developing the TLC plate by enabling the concentrated solvent to migrate through the chromatography medium in an orthogonal or oblique dimension to the first dimension. by, (c) Optionally, tagging the analyte molecules with detectable tags, (i) Moving a fluid containing a detectable tag through one or more fluid channels in the fluid chip, and delivering the fluid containing the tag to a separation zone along the tag-detectable analyte molecule. by, (d) Using a detector, detect a signal from the TLC plate indicating the presence of the analyte molecules. Methods that include...

80. The method according to claim 79, wherein the liquid sample is an aqueous sample, and / or the second liquid is an organic solvent.

Images