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WO1993001494A1 - Methode et appareil pour extraction en phase solide amelioree - Google Patents

Methode et appareil pour extraction en phase solide amelioree Download PDF

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Publication number
WO1993001494A1
WO1993001494A1 PCT/US1992/005768 US9205768W WO9301494A1 WO 1993001494 A1 WO1993001494 A1 WO 1993001494A1 US 9205768 W US9205768 W US 9205768W WO 9301494 A1 WO9301494 A1 WO 9301494A1
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WO
WIPO (PCT)
Prior art keywords
disc
sample
analyte
fluid
inlet
Prior art date
Application number
PCT/US1992/005768
Other languages
English (en)
Inventor
C. Michael O'donnell
Stephen K. Schultheis
Original Assignee
Toxi Lab, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Toxi Lab, Inc. filed Critical Toxi Lab, Inc.
Publication of WO1993001494A1 publication Critical patent/WO1993001494A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
    • G01N30/92—Construction of the plate
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
    • B01D15/22—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the construction of the column
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
    • B01D15/36—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving ionic interaction, e.g. ion-exchange, ion-pair, ion-suppression or ion-exclusion
    • B01D15/361—Ion-exchange
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/405—Concentrating samples by adsorption or absorption
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N2030/009—Extraction
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N2030/062—Preparation extracting sample from raw material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/50—Conditioning of the sorbent material or stationary liquid
    • G01N30/52—Physical parameters
    • G01N2030/524—Physical parameters structural properties
    • G01N2030/527—Physical parameters structural properties sorbent material in form of a membrane
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
    • G01N30/91—Application of the sample

Definitions

  • the field of the present invention relates to the process of solid phase extraction. More specifically, the present invention relates to the use of a process of solid phase extraction to isolate a specific chemical from a prepared liquid sample.
  • One common application of this process is in the drug screening field.
  • the sample to be screened must first be prepared in a liquid form, e.g., by dissolving the sample in a solvent.
  • the liquid is run through a column with loose silica material packed in the bottom.
  • loose silica is suspended in between two porous stops called frits at the base of the column.
  • the silica In order to retain the analyte of interest, the silica must be treated with a phase such as an ion or cation exchange agent to attract specific drugs.
  • a vacuum, positive pres- sure or a centrifuge is used to aid in passage of the liquid through the bed of silica.
  • the silica is washed several times with an extracting solvent to remove the analyte.
  • the resulting solution containing the drug is dried out and concentrate. This concentrate is then screened for the presence of
  • the concentrate can be used in its solid form or reconstituted depending on the type of screen that is to be performed.
  • Some common screening processes include gas, liquid, or thin layer chromatography. While gas and liquid chromatography are used for quantitative results, they are time intensive and expensive. If only qualitative results are needed, thin film or thin layer chromatography is sued because of the simplicity and low cost.
  • the process of thin layer chromato ⁇ graphy as is well known in the art, characterizes analytes based on their ability to migrate in a known medium. An example of this process is disclosed in U.S. Patent No. 3,714,035.
  • the foregoing conventional column-based solid phase extraction process uses chemical agents to attract the desired chemical to the silica by forcing the sample to pass through a bed of the treated silica particles.
  • the loose silica must be carefully packed between the frits to insure that the sample, when poured into the column, will pass through the treated particles. If the silica is improperly packed, there is the possibility that small channels will form in between the loose silica fibers. These channels act as passageways of low resistance which allow the liquid to flow through the silica bed without coming into contact with enough of the reagents to pick up sufficient amounts of the analyte. This phenomena is commonly known as channeling and is the main cause of inaccuracy in the solid phase extraction process.
  • a solid phase extraction device for extracting a desired analyte from a sample, comprising an upper cartridge having a fluid inlet, a lower cartridge having a fluid outlet, and at least one concentrator disc positioned intermediate the inlet and the outlet.
  • the fluid outlet is in fluid communication with the fluid inlet.
  • the inlet and the outlet can be provided with luer fittings.
  • the disc is comprised of silica embedded in a rigid matrix, and has associated therewith a phase capable of binding the analyte.
  • the rigid matrix comprises a material selected from the group consisting of glass, a crystalline polymer and a non-crystalline polymer.
  • the phase is an ion exchange resin, including cation exchange resins such as activated silica, sulfonyl and cyanopropyl, and anion exchange resins, such as trimethyl amino propyl.
  • the upper and lower cartridges are formed from a single unitary piece.
  • the upper and lower cartridges can be press fit together to substantially prevent removal of the disc by hand.
  • the upper and lower cartridges are separable so as to allow removal of the disc for further processing.
  • the upper and lower cartridges preferably each comprise a handle to facilitate
  • One preferred phase is an alkyl silane, including C2, C8 and C 18 alkyl silanes.
  • Other preferred phases include a cation exchange resin, such as sulfonyl and cyanopropyl.
  • the device is preferably provided with a visually discernable code associated with the type of the phase.
  • the device includes a reservoir in fluid communication with the inlet.
  • This reservoir preferably comprises a filter to substantially prevent undissolved solids from entering the inlet.
  • a vacuum source in connection with the outlet is preferably used with the device.
  • the device can in ⁇ clude more than one disc and discs can be treated with different phases.
  • the disc has a mass under 15 mg.
  • the disc is comprised of a material sufficiently rigid to be used for thin layer chromatography, such as glass matrix produced from an inert glass membrane dipped in a silica gel.
  • the inlet is preferably sized to allow insertion of a conventional piston apparatus.
  • the upper cartridge is sized for use with a centrifuge tube, and the lower cartridge is sized to hold the disc.
  • the present invention also provides a method of extracting a desired analyte from a fluid sample
  • SUB comprising the steps of inputting the fluid sample into a solid phase extraction device, contacting the fluid sample with a concentrator disc iocated inside the extraction device, and binding the analyte to the disc, allowing the substantial part of the fluid com ⁇ ponent of the sample to pass through the disc.
  • the disc used in this method is comprised of silica em ⁇ bedded in a rigid matrix, and has associated therewith a phase capable of binding the analyte.
  • the contacting step comprises as ⁇ piration of the sample using a vacuum.
  • the contacting step comprises centrifuging the solid phase extraction concentrator.
  • the method preferably also comprises washing the disc with wash reagents.
  • the inputting step preferably additionally comprises using a piston to input the fluid sample.
  • the method includes eluting the analyte from the disk.
  • the elution step preferably comprises selecting an elution solvent that will remove the analyte from the disc, and binding the analyte with the elution solvent.
  • the disc resulting from the disc can be dried.
  • the present inven ⁇ tion also provides a method of storing specimens comprising using a preferred method and storing the dried disc produced therefrom.
  • the method is performed using a sample, such as urine, whole blood, serum, saliva, or other body fluid.
  • the method preferably additionally comprises identifying the disc as related to a particular sample, and transferring the disc while ensuring the chain of custody when the disc is transferred.
  • a chromatographic detection kit for detecting a desired analyte from a fluid sample comprises a solid phase extraction device
  • SUBSTITUTE SHEET having a fluid inlet and a fluid outlet, at least one concentrator disc positioned intermediate the inlet and the outlet, and a chromatographic development sheet of the same material as the disc.
  • the sheet has a sample space thereon sized and shaped to accept the disc.
  • the disc is of a material comprised of silica embedded in a rigid matrix.
  • the disc comprises a phase for binding the analyte.
  • a method of performing thin layer chroma ⁇ tography to detect the presence of a desired analyte in a fluid sample comprising extracting the analyte from the sample onto an extraction concentrator disc by inputting the sample into a fluid inlet of a solid phase extraction device having the disc positioned therein, the disc being of a material comprised of silica embedded in a glass matrix, inoculating the disc having the extracted sample thereon into a sample space on a thin layer chromatography sheet of the same material as the disc, developing the thin layer chro ⁇ matography sheet using a developing solution which will migrate along the sheet, so as to differentiate the analyte from other components in the sample based on the analyte*s ability to migrate through the sheet as the developing solution migrates along the sheet, and detecting the analyte as a spot on the sheet.
  • the detecting step preferably additionally comprises iden ⁇ tifying the spot by a color reaction and/or identify- ing the spot under UV light.
  • the extracting step preferably comprises extracting the sample onto a disc treated with a phase capable of binding the analyte.
  • Figure 1 is a perspective view of an assembled apparatus of the preferred embodiment of the present invention.
  • Figure 2 is an exploded view of the apparatus of the preferred embodiment of the present invention.
  • Figure 3 is a cross sectional view of the assembled apparatus of the preferred embodiment of the present invention.
  • Figure 4 is a perspective view of an assembled apparatus of an alternate embodiment of the present invention.
  • Figure 5 is an exploded view of the apparatus of the alternate embodiment of the present invention.
  • Figure 6 is a cross sectional view of the assembled apparatus of the alternate embodiment of the present invention.
  • Figure 7 is an exploded perspective view of the thin film chromatographic developing sheet with a concentrator disc aligned with its respective sample space.
  • the present invention advantageously provides an apparatus that simplifies the process of solid phase extraction.
  • the present invention also eliminates the channeling effect common in loose particle based extraction methods.
  • the present invention also allows a user thereof to quickly adapt the results of the solid phase extraction to thin layer chromatography in order to identify the analyte while providing means to use the analyte in any other type of identification process.
  • the present invention is an apparatus for performing solid phase extraction that uses a concentrator disc made from silica embedded in a rigid matrix instead of the loose silica used in the prior art.
  • the concentrator disc of the present invention has several advantages over the loose silica of the prior art.
  • One advantage is that significantly less silica material is re ⁇ quired. Normally between 50 and 100 mg of loose silica is required but each disc of the present inven ⁇ tion requires less than 15 mg of silica.
  • Another ad ⁇ vantage is that the concentrator disc does not suffer from channeling that may occur in loose silica and therefore provides more accurate test results.
  • Still another advantage is that the disc may immediately be placed in a thin layer chromatographic plate which eliminates the need for several steps of the conven ⁇ tional solid phase extraction processes.
  • the disc is made from a medium that is rigid enough to be placed in the apparatus used for the standard thin layer chromatographic process.
  • the elution process involves several steps: washing the silica several times with a specific reagent to remove the analyte; drying down the eluted solution; concen ⁇ trating the resultant; and then reconstituting the solid so that it can be spotted on the thin layer chromatographic medium.
  • the method and apparatus of the present invention remove the need for several of these steps, as the disc can be removed from the apparatus of the present invention, dried and then placed directly into the thin layer chroma ⁇ tography apparatus.
  • the discs are designed to facilitate the thin layer chromatographic process, they may be treated like loose silica in the conventional solid phase extraction process.
  • the analyte can be eluted as well known in the art, for processing by liquid or gas chromatography.
  • Another advantage of the present invention is that multiple discs may be placed in the apparatus. Each collects a portion of the sample and can be used individually to perform a different screening process. For example, one disc can be used in a thin layer screen while the others can be prepared for liquid or gas chromatography screens. Alternatively, depending on the sample that is being screened, each disc can be treated with a different phase to try to detect the presences of multiple types of drugs while only requiring one decanting. Further, while some drugs are stable, the samples that they come from are often unstable, e.g., urine. Such analytes can be preserved by extracting them from the sample onto the concen- trator discs and then drying and storing the discs for later testing.
  • FIG. 10 one preferred embodiment of the solid phase extraction device of the present invention.
  • This embodiment comprises a two-piece unit that when
  • SUBST assembled forms a hollow drum shaped unit with a centrally located cylindrical passage 15.
  • the upper cartridge 20 and lower cartridge 25 are sep- arable to allow for insertion and removal of one or more concentrator discs 30. These discs 30 are described in greater detail hereinbelow.
  • the upper cartridge 20 preferably comprises a fluid inlet piece 35, a central passage 15, and an annular portion 45.
  • the annular portion serves as a handle 50 for manipulation of the device as a whole and for removing the upper cartridge 20 from the lower cartridge 25, as will be described below.
  • the fluid inlet piece 35 defines a fluid inlet 40 through which the sample can be introduced into the device 10.
  • the central passage 15 through the upper cartridge 20 preferably comprises two concentrically aligned passages: the fluid inlet piece 35 and an interior passage tube 55.
  • the fluid inlet piece 35 and the upper interior passage tube 55 are concen ⁇ trically aligned with each other and with the annular portion 45.
  • the fluid inlet piece 35 extends perpen ⁇ dicularly from an upper surface 47 of the annular portion 45, and is preferably sized to allow insertion of a conventional piston nozzle of a syringe.
  • the fluid inlet piece 35 can be provided with conventional luer fittings to provide for a solid connection with various types of input apparatuses.
  • the lower cartridge 25 preferably comprises a fluid outlet piece 60, a central passage 15, and a lower annular portion 65, also defining a handle 50, as described above.
  • the fluid outlet piece 60 defines a fluid outlet 75 through which the extracted sample can exit the device 10.
  • the central passage 15 is concentrically aligned with the annular portion 65 and is preferably comprised of two concentrically aligned passages: the lower interior passage tube 80 and the fluid outlet piece 60.
  • the fluid outlet 75 is in fluid communication with the fluid inlet 40 of the upper cartridge 20 through the central passage 15.
  • the lower interior passage tube 80 extends perpendicu ⁇ larly from the top surface 67 of the annular portion 65, and contains an inner shoulder 85 located approxi ⁇ mately a third of the way from a top surface 90 of the lower interior passage 80.
  • the fluid outlet piece 60 extends perpendicularly from a lower surface 68 of the annular portion 65.
  • the fluid outlet piece 60 has a significantly smaller diameter than the lower interior passage 80 and tapers slightly to enable the use of a conventional vacuum apparatus.
  • the outlet piece 60 is provided with standard luer fittings to provide for a solid connec- tion with various types of receiving apparatuses.
  • each handle 50 is provided with a plurality of traction ridges 52 that are oriented in parallel with the central passageway 15 and extend from the edge of the annular portions 45, 65 to the rim of the handle 50 itself.
  • the traction ridges 52 provide for grippability when the upper cartridge 20 and lower cartridge 25 are separated or assembled.
  • the handles 50 extend radially symmet ⁇ rically from the fluid inlet 40 and the fluid outlet 75. In the upper cartridge 20, the handle 50 extends
  • both the upper cartridge 20 and lower 25 cartridges are formed from a single unitary piece, but in the pre- ferred embodiment, as illustrated in Figures 1-3, the upper and lower cartridges are two separate pieces.
  • one or more concentrator discs 30 are placed within the lower interior passage tube 80 and are oriented parallel to the top surface 67 of the annular portion 65. The first concentrator disc 30 is inserted until it abuts the interior shoulder 85 of the lower in ⁇ terior passage tube 80. Additional discs 30 can be inserted with the same orientation as described above and are placed adjacent to each other. In the pre ⁇ ferred embodiment, sufficient space is provided to allow for the placement of three discs 30 and/or filter units 95.
  • a filter unit 95 may be placed above the concentrator discs 30 to prevent undesired large particles from coming into contact with the concen ⁇ trator discs 30.
  • the upper cartridge 20 is mated with the lower cartridge 25 about the concentrically aligned central passage 15.
  • the upper interior passage tube 55 fits within the lower interior passage tube 80 and the upper cartridge 20 and the lower cartridge 25 are squeezed together with the two handles 50 moving towards each other.
  • the two car- tridges 20 and 25 are preferably assembled tightly so that a bottom surface 97 of the upper interior passage tube 55 is pressing the concentrator discs 30 and optional filter 95 against the inner shoulder 85 of the lower interior passage tube 80.
  • the central passage 15 should be prop ⁇ erly sealed as described above, the handles 50 are located proximate to each other, and the hollow drum ⁇ like shape of the unit has been achieved.
  • the upper interior passage tube 55 and the lower interior passage tube 55 are provided with a series of concentric receiving and engaging ridges.
  • the ridges are aligned so that the upper cartridge 20 and lower cartridge 25 can be press fit together, to prevent the removal of the concen ⁇ trator discs 30 from the unit.
  • This embodiment does not have the traction ridges 52 all around the handle 50 of the cartridges as the upper cartridge 20 and lower cartridge 25 need not be separated. Instead, this embodiment has these traction ridges 52 in enough locations on the handle 50 to provide for improved traction, so that the cartridge will not slip out of the hands of the user.
  • an alternative embodiment 100 is provided to allow for use with multiple sizes of centrifuge tubes.
  • This embodiment comprises an inner passageway 110 through an upper cartridge 120 and a lower cartridge 125 with a disc 30 held therebetween.
  • this embodiment also serves as a disc 30 held therebetween.
  • SUBSTITUTE SHEET includes a hollow cylindrical column 130 with two ends, with one end being covered with a filter 135 for keeping undesired large particles suspended in the sample as they pass through the passageway 110.
  • the column 130 extends to a larger diameter at its upper portion 140 and has an outer extending shoulder 145 at its edge.
  • the upper portion 140 is open at its end, thereby providing a fluid inlet 150.
  • the upper portion 140 is sized to provide an interference fit with the rim of a standard 50 ml centrifuge tube.
  • the shoulder 145 preferably extends beyond the rim of the centrifuge tube in order to keep the device 100 in the upper portion of a standard centrifuge tube.
  • the column 130 provides a reservoir for approximately 5 ml of the unfiltered sample used in the solid phase extraction method of the present invention.
  • the upper cartridge 120 comprises a hollow cylinder 157 having a diameter sized to allow inser- tion of the column 130.
  • the hollow cylinder 157 serves as a fluid inlet for the upper cartridge 120.
  • This hollow cylinder 157 tapers to form a smaller cylinder 158 sized to fit within an upper cylinder 159 of the lower cartridge 125.
  • the upper cylinder 159 of the lower cartridge 125 is sized to hold the concen ⁇ trator discs 30.
  • the discs 30 are placed in parallel alignment with an inner shoulder 162 of the lower cartridge 125 as it tapers to form a fluid outlet 165.
  • the smaller cylinder 158 of the upper cartridge 120 is sized to provide for an interference fit with a standard small centrifuge tube, such that the cylinder 158 can hold a substantial portion of the upper cartridge 120 above the centrifuge tube while the lower cartridge 125 is suspended inside the tube.
  • the alternate embodiment 100 can be assembled as follows: the concentrator discs 30 are placed within the lower cartridge 125, the upper cartridge 20 is placed within the lower cartridge 125 such that smaller cylinder 158 of the upper cartridge 120 squeezes the concentrator discs 30 against the inner shoulder 162 of the lower cartridge 125 and thereby seals the inner passageway 110 with the lower cartridge 125; finally, the column 130 complete with filter 135 is placed within the upper cartridge 120 to complete the inner passageway 110 and maintain a proper seal.
  • the device 10 comprises one or more concentrator discs 30.
  • the discs 30 are com- prised of silica embedded in a rigid matrix.
  • the matrix be sufficiently rigid that the sheets made of the same material as the discs can be used in conventional thin layer chromatography.
  • preferred matrix materials include glass, crystalline polymers, such as acrylic, and non-crys ⁇ talline polymers.
  • silica is embedded in glass by dipping raw glass fiber paper into a concentrated sodium silicate solution and it is immersed in an ammonium chloride solution to gel. The paper is then dried, water rinsed and dried again.
  • the silica embedded paper is shaped into a specif ⁇ ically sized disc depending on the required absorbency of the disc. The amount of silica provided in each disc will depend on the analyte being tested, the
  • the disc will have an amount of silica in the range of 1.5 to 15 mg. In one embodiment, each disc 30 has approximately 1.5 to 2.0 mg of silica embedded in a glass matrix.
  • the concentrator discs 30 of the present inven ⁇ tion that are made from silica embedded in a glass matrix advantageously provide silica in a form that is not loose as is common in the prior art.
  • the silica on the concentrator discs 30 is held in place in the glass matrix, so there are no areas where channels can form, as is common in loose silica.
  • the concentrator discs 30 of the present invention there is always a resistive layer embedded with silica for the analyte to bind.
  • the concentrator disc 30 itself provides for the more accurate extraction process of the present invention.
  • each disc 30 can then be used for a different screening process, for storing the analyte or for repeat testing.
  • each disc 30 can be doped with a different phase in order to extract different analytes onto different discs from the same sample.
  • the same disc 30 can be doped with more than one phase.
  • the concentrator discs 30 of the present invention are preferably doped with a phase that will attract a desired analyte.
  • a phase that will attract a desired analyte There are many different types of phases available, as will be known to those with ordinary skill in the art.
  • an alkyl silane phase may be used, such as a C2, C8, or C18 silane.
  • shorter chain silanes are preferable in order to allow for elution of the analyte from the phase.
  • a cyclohexyl (CH) phase can be chosen.
  • a phenyl phase can be used which provides a different selectivity due to its aromaticity.
  • anionic and cationic phases that rely on the properties of ionic bonding to attract the analyte.
  • An example of a strong anionic phase is trimethyl amino propyl.
  • An example of a strong cationic phase, or cationic resin, is sulfonyl.
  • weaker cation exchange phases such as cyanopropyl (CN) and activated silica. Use of the weaker ion exchange phases is appropriate for strongly ionic analytes, which would be difficult to elute from the strong ion exchange phases.
  • silica embedded paper is doped with C18 as follows: The silica paper is dipped in a 5% solution of octadecyl (C18) silane, rinsed in toluene and then dried. In order to dope the silica
  • SUBSTITUTE SHEET paper with a cationic phase such as sulfonyl the silica paper is dipped in a 5% solution of 2(4- cholorosulfonylphenyl) ethyl trichlorosilane/toluene solution, rinsed in methanol, dipped in a sodium bicarbonate solution and then dried. Doping the silica paper with other solutions may require slightly different processes, but the two illustrated examples are considered the best mode solutions.
  • the devices 10 are coded with a visually discernable marking, preferably using a color code, to indicate what phases are contained within each apparatus.
  • devices employing discs treated with a cationic exchange resin can be constructed from blue plastic.
  • the sample to be tested is first poured into the fluid inlet 40. If desired, the sample can be forced through the inlet 40 by a piston, such as that of a syringe.
  • the fluid sample will pass through the passage 15 of the upper cartridge 20, through a filter 95, if present, and then come into contact with at least one concentrator disc 30.
  • the disc 30 will be comprised of silica embedded in a rigid matrix, such as a glass matrix, and will be doped with a phase capable of binding the desired analyte.
  • the desired analyte will be bound to the concentrator disc 30 and the remainder of the sample will pass through the disc 30 and out the fluid outlet 75.
  • the fluid may be aspirated through the unit by attaching a conventional vacuum apparatus to the fluid outlet piece 60. Additionally, the fluid may be forced through the concentrator discs 30 in the apparatus of the alterna ⁇ tive embodiment 100 illustrated in Figures 4-6 by using the forces of a centrifuge.
  • wash reagent is a solvent which will not elute the analyte off the phase yet will still remove unbound foreign material.
  • distilled water can be used as a wash reagent.
  • the discs 30 may be removed from the solid phase extraction device for further screening, or the analyte can be eluted from the disc 30 using an elution solvent which will remove the analyte from the disc 30.
  • the elution process is used to remove the analyte from the concentrator discs 30, no direct access to the discs 30 is required. Therefore, if embodiment 10 is used, the upper and lower cartridges 20, 25 need not be opened as the discs 30 can remain in the apparatus 10. The elution process requires that the discs 30 be washed with a solvent to remove
  • SUBSTITUTE SHEET the analyte from the phase that bound it to the concentrator disc.
  • the solvent is poured into the fluid inlet 40 and the solution that exits the fluid outlet 75 will contain the analyte that has been removed from the concentrator discs 30.
  • the solvent is chosen based on the analyte of interest and the phase that the disc 30 was doped with.
  • the preferred use of this embodiment 100 requires some variations in the method described above in connection with the embodiment of Figures 1-3.
  • the column 130 is removed which automati- cally removes the filter 135.
  • the analyte can be eluted from the discs 30 or the disc 30 removed for further processing.
  • the remaining upper cartridge 120 and lower cartridge 125 can be balanced on a smaller centrifuge tube while the elution solvent is poured into the disc retainer portion 155 of the upper cartridge 120.
  • the elution solution is pulled through the concentrator discs 30 and then through the fluid outlet 165 in the lower cartridge 125.
  • this process is aided by the forces of the centrifuge.
  • the solution containing the analyte is captured in the centrifuge tube, and can then be dried.
  • the dried analyte can then be processed in several different ways depending on the type of screen required.
  • the analyte is removed from the disk using an elution solvent. Whether this elution solvent is used for removal of the analyte from the disc for further screening or whether the solvent is used to chromatograph the analyte directly from the disc, such as through thin layer chroma-
  • elution solvent and phase on the disc can be used to select for desired analytes, in a manner which will be known to those of ordinary skill in the art.
  • the analyte In order to elute a desired analyte, the analyte should have a higher affinity for the elution solvent than for the phase.
  • highly non-polar molecules will bind very tightly to long chain alkyl silane, such as C18 phases.
  • a C18 phase can be used with a methanol/acetone solvent.
  • the concentrator disc 30 can be removed from the apparatus and dried.
  • the dried disc may be stored or used for a variety of screening processes. Many samples, particularly urine, can be unstable and will break down over time. Thus, it is advantageous to extract the analyte of interest from the sample as the analytes are often more stable than the sample.
  • the apparatus 10 can be used in the improved method of solid phase extraction described above to capture the analyte of interest onto one or more concentrator discs 30. The apparatus is then opened, by separating the upper cartridge 20 and the lower cartridge 25, and one or more of the discs 30 can be removed and dried.
  • the discs 30 containing the desired analyte can then be identified by relating the disc to the sample from which it was extracted.
  • the disc 30 can be marked directly or can be placed within a marked container to associate the analyte with the sample from which it was extracted.
  • the concentrator disc 30 can be stored as evidence or it can be stored for further testing at a later date. As the disc 30 can be transferred or stored in other locations, a chain of custody can be maintained to ensure the origin of the disc 30 for use as evidence of the origin of the sample stored on the disc 30.
  • a chromatographic detection kit of the present invention can be used.
  • the chromatographic detection kit includes a solid phase extraction device, such as that illustrated in Figures 1-3 or in Figures 4-6.
  • the device of the kit includes a fluid inlet 40 and a fluid outlet 75 and at least one concentrator disc 30 of a material comprised of silica embedded in a rigid matrix.
  • the kit includes a chromatographic sheet 200, as illustrated in Figure 7.
  • the sheet 200 is comprised of the same material as the concentrator disc 30, and has a sample space 210 sized and shaped to receive the disc 30 from the device.
  • the disc 30 is preferably comprised of a glass matrix that is rigid enough to be used for thin layer chromatography, although other rigid materials can also be used.
  • the disc is doped with a phase to bind a desired analyte.
  • the desired analyte is extracted from the sample by pouring the sample into the fluid inlet 40 of the solid phase extraction device, allowing it to come into contact with the concentrator disc 30.
  • the concentrator disc 30 is treated with a phase that will bind with the analyte of interest and allow other materials, including most of the fluid component of the sample, to pass through the fluid outlet 75.
  • the disc is removed, such as by separating the upper 20 and lower cartridges 25.
  • the disc can then be dried or can be immediately placed into the specifically sized sample space 210 of thin layer chromatography development sheet 200 in order to identify the analyte of interest.
  • SUBSTITUTE SHEET comprises developing the thin layer chromatographic sheet 200 with a developing solution that will migrate along the sheet and the analyte of interest will migrate along with the solution.
  • the developing solution must be selected as discussed above, to ensure that the analyte is eluted from the phase.
  • the characteristics of the analyte can be determined by its ability to migrate along the thin layer medium in the developing solution.
  • the desired analyte can be detected as a spot on the developing sheet 200 having the appropriate characteristics.
  • the thin layer chromatographic developing sheet 200 can be placed under ultraviolet (UV) light and the analyte can be determined by its color change under the light.
  • UV ultraviolet
  • the thin film chromatographic developing sheet 200 can be dipped in water or a staining solution and based on the color change of the developing paper 200, the analyte can be identified.
  • any combination of methods can be used to determine the identity of a given analyte, with more testing providing more accurate results.

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  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

Dispositif (10, 100) d'extraction en phase solide permettant d'extraire d'un échantillon un analyte souhaité. Le dispositif comprend une cartouche supérieure (20, 120) munie d'une entrée de fluide (40, 157), une cartouche inférieure (25-125) munie d'une sortie de fluide (75, 165) communiquant avec l'entrée de fluide (40, 157), et au moins un disque concentrateur (30) placé entre l'entrée (40, 157) et la sortie (75, 165). Le disque ((30) est constitué de silice noyée dans une matrice rigide, et possède une phase capable de lier l'analyte qui lui est associé. Est également décrit un ensemble comprenant un dispositif d'extraction (10, 100) et une feuille (200) de développement chromatographique. Sont également décrites des méthodes d'extraction et de stockage d'un analyte souhaité.
PCT/US1992/005768 1991-07-12 1992-07-08 Methode et appareil pour extraction en phase solide amelioree WO1993001494A1 (fr)

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US72935691A 1991-07-12 1991-07-12
US729,356 1991-07-12

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Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994019687A1 (fr) * 1993-02-26 1994-09-01 Merck Patent Gmbh Separateur
WO1995017247A1 (fr) * 1993-12-22 1995-06-29 Minnesota Mining And Manufacturing Company Matieres en feuille pour extractions en phase solide ou reactions en phase solide
WO1996014931A1 (fr) * 1994-11-10 1996-05-23 Minnesota Mining And Manufacturing Company Extraction en phase solide avec une feuille composite permettant la mesure directe de la radioactivite
WO1996028248A1 (fr) * 1995-03-15 1996-09-19 City Of Hope Cartouche jetable pour le stockage et l'administration de reactif
WO1997049988A1 (fr) * 1996-06-21 1997-12-31 Kuehn Michael Procede et melange pour fabriquer un materiau autoportant, et materiau
US5728301A (en) * 1994-09-29 1998-03-17 Truett; William L. Apparatus and method for thin layer chromatography
WO1999016526A1 (fr) * 1997-09-30 1999-04-08 Hanns-Knöll-Institut Für Naturstoff-Forschung E.V. Procede permettant de separer des melanges complexes par extraction de phases solides aux fins de recherche de principes actifs et d'analyse
WO2000075623A1 (fr) * 1999-06-08 2000-12-14 Dna Research Innovations Limited Dispositif de traitement d'echantillons
US6177282B1 (en) * 1997-08-12 2001-01-23 Mcintyre John A. Antigens embedded in thermoplastic
WO2001014852A1 (fr) * 1999-08-25 2001-03-01 University Of Portsmouth Dispositif passif d'echantillonnage aquatique et ses procedes d'utilisation
FR2839458A1 (fr) * 2002-05-07 2003-11-14 Univ Lille Sciences Tech Procede d'extraction sur phase solide, notamment pour l'analyse d'echantillons biologiques et dispositif pour la mise en oeuvre du procede
WO2010101865A1 (fr) * 2009-03-02 2010-09-10 Zymo Research Corporation Colonne universelle
WO2011010265A1 (fr) * 2009-07-24 2011-01-27 Camag Procédé et dispositif permettant de préparer des substances pour des analyses quantitatives et qualitatives
EP2649194A4 (fr) * 2010-12-06 2014-05-28 3M Innovative Properties Co Procédé et dispositif de concentration de microorganismes
EP2649193A4 (fr) * 2010-12-06 2014-05-28 3M Innovative Properties Co Procédé et dispositif de concentration de microorganismes
JP2018503069A (ja) * 2014-11-21 2018-02-01 オッカム バイオラブス，インコーポレイティド 核酸サンプルを収集するためのシステム及び方法
EP1399727B1 (fr) * 2001-06-15 2018-12-26 Life Technologies Corporation Dispositifs de colonnes de filtration et procedes de filtration associes
EP4244597A4 (fr) * 2020-11-16 2024-12-18 Ankom Technology Corporation Colonne d'extraction en phase solide (spe)

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US4787971A (en) * 1987-01-23 1988-11-29 Alan Donald Miniaturized column chromatography separation apparatus and method of assaying biomolecules employing the same
US4961916A (en) * 1988-06-02 1990-10-09 Irsst-Institut De Recherche En Sante Et En Securite Du Travail Du Quebec Sampling device
US5124041A (en) * 1989-07-28 1992-06-23 Applied Biosystems, Inc. Biomolecule sample immobilization

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US3714035A (en) * 1971-05-19 1973-01-30 D Jones Method and apparatus for thin layer chromatography
US3963421A (en) * 1974-07-12 1976-06-15 Sierra Laboratories, Inc. TLC method for drug detection
US4787971A (en) * 1987-01-23 1988-11-29 Alan Donald Miniaturized column chromatography separation apparatus and method of assaying biomolecules employing the same
US4961916A (en) * 1988-06-02 1990-10-09 Irsst-Institut De Recherche En Sante Et En Securite Du Travail Du Quebec Sampling device
US5124041A (en) * 1989-07-28 1992-06-23 Applied Biosystems, Inc. Biomolecule sample immobilization

Cited By (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994019687A1 (fr) * 1993-02-26 1994-09-01 Merck Patent Gmbh Separateur
WO1995017247A1 (fr) * 1993-12-22 1995-06-29 Minnesota Mining And Manufacturing Company Matieres en feuille pour extractions en phase solide ou reactions en phase solide
US5688370A (en) * 1993-12-22 1997-11-18 Minnesota Mining And Manufacturing Company Sheet materials for solid phase extractions and solid phase reactions
US5702610A (en) * 1993-12-22 1997-12-30 Minnesota Mining And Manufacturing Company Sheet materials for solid phase extractions and solid phase reactions
US5728301A (en) * 1994-09-29 1998-03-17 Truett; William L. Apparatus and method for thin layer chromatography
WO1996014931A1 (fr) * 1994-11-10 1996-05-23 Minnesota Mining And Manufacturing Company Extraction en phase solide avec une feuille composite permettant la mesure directe de la radioactivite
US5637506A (en) * 1994-11-10 1997-06-10 Minnesota Mining And Manufacturing Company Solid phase extraction using composite sheet for direct measurement of radioactivity
US5891559A (en) * 1994-11-10 1999-04-06 Minnesota Mining And Manufacturing Co. Solid phase extraction using composite sheet for direct measurement of radioactivity
WO1996028248A1 (fr) * 1995-03-15 1996-09-19 City Of Hope Cartouche jetable pour le stockage et l'administration de reactif
US5766550A (en) * 1995-03-15 1998-06-16 City Of Hope Disposable reagent storage and delivery cartridge
WO1997049988A1 (fr) * 1996-06-21 1997-12-31 Kuehn Michael Procede et melange pour fabriquer un materiau autoportant, et materiau
US6872576B1 (en) * 1997-08-12 2005-03-29 Embedded Concepts, Llc Antigens embedded in thermoplastic
US6177282B1 (en) * 1997-08-12 2001-01-23 Mcintyre John A. Antigens embedded in thermoplastic
WO1999016526A1 (fr) * 1997-09-30 1999-04-08 Hanns-Knöll-Institut Für Naturstoff-Forschung E.V. Procede permettant de separer des melanges complexes par extraction de phases solides aux fins de recherche de principes actifs et d'analyse
WO2000075623A1 (fr) * 1999-06-08 2000-12-14 Dna Research Innovations Limited Dispositif de traitement d'echantillons
WO2001014852A1 (fr) * 1999-08-25 2001-03-01 University Of Portsmouth Dispositif passif d'echantillonnage aquatique et ses procedes d'utilisation
US7059206B1 (en) 1999-08-25 2006-06-13 University Of Portsmouth Aquatic passive sampling device and methods for its use
EP1399727B1 (fr) * 2001-06-15 2018-12-26 Life Technologies Corporation Dispositifs de colonnes de filtration et procedes de filtration associes
FR2839458A1 (fr) * 2002-05-07 2003-11-14 Univ Lille Sciences Tech Procede d'extraction sur phase solide, notamment pour l'analyse d'echantillons biologiques et dispositif pour la mise en oeuvre du procede
WO2010101865A1 (fr) * 2009-03-02 2010-09-10 Zymo Research Corporation Colonne universelle
WO2011010265A1 (fr) * 2009-07-24 2011-01-27 Camag Procédé et dispositif permettant de préparer des substances pour des analyses quantitatives et qualitatives
EP2649194A4 (fr) * 2010-12-06 2014-05-28 3M Innovative Properties Co Procédé et dispositif de concentration de microorganismes
EP2649193A4 (fr) * 2010-12-06 2014-05-28 3M Innovative Properties Co Procédé et dispositif de concentration de microorganismes
JP2018503069A (ja) * 2014-11-21 2018-02-01 オッカム バイオラブス，インコーポレイティド 核酸サンプルを収集するためのシステム及び方法
JP2020190560A (ja) * 2014-11-21 2020-11-26 オッカム バイオラブス，インコーポレイティド 核酸サンプルを収集するためのシステム及び方法
EP4244597A4 (fr) * 2020-11-16 2024-12-18 Ankom Technology Corporation Colonne d'extraction en phase solide (spe)

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