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WO2001011355A1 - Procede et dispositif analytique - Google Patents

Procede et dispositif analytique Download PDF

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Publication number
WO2001011355A1
WO2001011355A1 PCT/SE2000/001510 SE0001510W WO0111355A1 WO 2001011355 A1 WO2001011355 A1 WO 2001011355A1 SE 0001510 W SE0001510 W SE 0001510W WO 0111355 A1 WO0111355 A1 WO 0111355A1
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WO
WIPO (PCT)
Prior art keywords
flow
membrane
components
matrix
flow matrix
Prior art date
Application number
PCT/SE2000/001510
Other languages
English (en)
Inventor
Jan Carlsson
Maria LÖNNBERG
Original Assignee
Pharmacia Diagnostics Ab
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
Priority claimed from SE9902855A external-priority patent/SE9902855D0/xx
Priority claimed from SE9903970A external-priority patent/SE9903970D0/xx
Application filed by Pharmacia Diagnostics Ab filed Critical Pharmacia Diagnostics Ab
Priority to AU63313/00A priority Critical patent/AU770715B2/en
Priority to CA002381137A priority patent/CA2381137A1/fr
Priority to JP2001515960A priority patent/JP2003525426A/ja
Priority to EP00950177A priority patent/EP1210594A1/fr
Publication of WO2001011355A1 publication Critical patent/WO2001011355A1/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
    • G01N30/96—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 using ion-exchange
    • 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
    • G01N30/94—Development
    • 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
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/38—Flow patterns
    • G01N2030/388—Elution in two different directions on one stationary phase
    • 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

Definitions

  • the present invention relates to a method of determining one or more components in a sample, wherein the sample components are separated by chromatography prior to the determination.
  • the invention also relates to a device therefor.
  • PhastSystemTM Anamersham Pharmacia Biotech AB, Sweden
  • PhastSystemTM Anamersham Pharmacia Biotech AB, Sweden
  • electrophoresis on pre-prepared gels. While this system heavily facilitates the labour and time for the analytical operator, the system is still rather laborious and expensive.
  • US-A-4,469,601 discloses a method and system for multi-dimensional chromatography in a thin-layer chromatographic plate wherein a sample is separated into an array of constituents. These constituents are then separated into a second array of sub-constituents by pumping a fluid through the plate in a direction crossing the array, and the sub-constituents are detected as they flow past fixed positions in this second direction.
  • Thin layer chromatography is, however, restricted to the separation of small (i.e. low molecular weight) molecules, and does not permit the separation of biomolecules, such as proteins, for example.
  • Pristoupil, T.I., Chromatog. Rev., 12 (1970) 109-125 describes the use of nitrocellulose filters in chromatography and electrophoresis. Chromatography in aqueous solution was performed with a nitrocellulose membrane in a horizontal position in a plexiglass chamber. Proteins were detected by immersing the membrane in a staining solution, and other substances were detected by usual spray or sandwich techniques. On the intact membrane, proteins having a molecular weight of the order of
  • the present invention provides a chromatographic assay method for qualitative, semi-quantitative or quantitative analysis, which method comprises the steps of: a) providing a membrane type flow matrix attached to a liquid-impervious backing, which flow matrix permits a capillary force assisted lateral fluid flow therethrough, and at least a part of which flow matrix contains ion-exchange functions; b) treating the flow matrix to reduce or eliminate unspecific adsorption properties of the flow matrix; c) applying to the flow matrix a sample containing at least two components; d) initiating a first lateral flow of aqueous fluid to transport the sample through the flow matrix and separate said components therein; e) interrupting said lateral flow; and either fl) detecting at least one of said separated components on the flow matrix in the position reached by the respective component when the flow was interrupted; or f2a) initiating a second flow of aqueous fluid to transport the components in a direction substantially transverse to the direction of the first lateral flow; f2b)
  • the present invention therefore provides a chromatographic device comprising a membrane type matrix attached to a liquid-impervious backing, which membrane permits a capillary force assisted lateral fluid flow therethrough and at least a part of which is modified to support ion-exchange functions.
  • the present invention provides an apparatus for determining components in a sample, which apparatus comprises a chromatographic device as above, and means for initiating and maintaining a liquid flow through the membrane.
  • the present invention is based on ion-exchange chromatograhpy of a sample on thin membranes with a liquid tight backing and detection of one or more of the separated components of the sample directly on the membrane, or optionally after transverse liquid transport of the components to another part of the membrane or to another membrane joined thereto without ion-exchange functions. While the sample may contain several (two or more) components, it may sometimes be of interest only to detect and determine one or a few of the separated components.
  • the membrane should have a fine foam-like structure and a standardized narrow distribution of pore sizes, typically in the range of 0.01 to 100 ⁇ m, preferably 0.01 to 20 ⁇ m.
  • the inner surface of the flow channels or pores in the membrane should, of course, be sufficiently hydrophilic to permit aqueous media, such as buffer, serum, plasma, blood, saliva, etc to be transported through the matrix. This transport may be achieved by capillary forces in the matrix itself, but is usually achieved by auxiliary means, such as an absorbent pad of cellulose or the like.
  • the membrane material is usually polymeric, and exemplary polymers are nitrocellulose, polyester, polyethersulphone, nylon, cellulose nitrate, and regenerated cellulose.
  • the membrane thickness is usually less than about 500 ⁇ m, e.g. in the range of from about 25 to about 500 ⁇ m, and preferably less than about 150 ⁇ m, e.g. in the range of from about 75 to about 150 ⁇ m.
  • the homogeneity of a chromatographic material affects its chromatographic quality and may therefore be reflected in terms of theoretical plate height.
  • the lower height of the theoretical plate the better the material.
  • the membrane for use in the present invention should thus have a height of theoretical plate (HETP) of less than about 500 ⁇ m, preferably less than about 100 ⁇ m.
  • the ion-exchange ligands that make the chromatograhic separation in the membrane possible may be anionic, cationic or amphoteric, and may be physically introduced into the matrix in the manufacturing process, or may be anchored to the membrane, either by covalent binding to the membrane, or via physical adsorption.
  • the anchorage of the ion-exchange ligands to the membrane may take place via a polymer or other substituent which in turn carries covalently, physically adsorptively, or biospecifically bound ligands.
  • Another possibility is deposition of polymer particles which exhibit a desired type of ion-exchange ligand.
  • the particles may be of hydrophilic or hydrophobic character, and the ligand structure may be exhibited by a compound adsorbed or covalently bound to the particles.
  • the technique for binding an ion-exchange ligand to the matrix it may, for example, be referred to our previously filed International (PCT) applications WO 99/36780, WO 99/36776 and WO 99/36777 (the disclosures of which are hereby incorporated by reference herein).
  • the ligand density (substitution degree) is selected to obtain the ⁇ desired isocratic separation.
  • the membrane may have different ligand densities or a gradient of ligand densities along the separation direction.
  • the use of a polybuffer with the ion- exchange membrane permits separation by chromatofocusing, in which a pH gradient is formed in the membrane.
  • ion-exchange functional groups include anion exchangers, such as diethyl aminoethyl (DEAE), trimethyl hydroxypropyl (QA), quaternary aminoethyl (QAE), quaternary aminomethyl (Q), diethyl-(2-hydroxypropyl)-aminoethyl, triethyl aminomethyl (TEAE), triethylaminopropyl (TEAP), polyethyleneimine (PEI), and cation-exchangers, such as methacrylate, carboxymethyl (CM), orthophosphate (P), sulfonate (S), sulfoethyl (SE), sulfopropyl (SP).
  • anion exchangers such as diethyl aminoethyl (DEAE), trimethyl hydroxypropyl (QA), quaternary aminoethyl (QAE), quaternary aminomethyl (Q), diethyl-(2-hydroxypropyl)-aminoethyl, trie
  • the membrane is treated with a detergent or other suitable agent to substantially reduce or eliminate undesired background or unspecific adsorption effects of the membrane matrix as is per se known in the art.
  • the sample containing the analyte or analytes to be determined may be added directly on the membrane surface, but usually it is added to a separate sample application membrane or pad in liquid contact with the membrane, either in edge to edge contact therewith or, preferably, mounted on top of the membrane.
  • the conditions for the separation of the components in the membrane are generally isocratic or with stepwise or continuously changed ion-strength.
  • Detection and quantification of separated components in the detection zone may take place in various ways. If the separated component or components to be determined are enzymatically active, they may be detected directly by their action on a suitable substrate, e.g. a colour change. Usually, however, detection is performed by protein staining, lipid staining, carbohydrate staining or DNA-staining, or by a biospecific detectable reagent. Such a substrate or reagent may be added via a fluid flow in the matrix, either (i) from one of the membrane sides transverse to the separation direction of the membrane, or (ii) from one of the sides extending in the separation direction of the membrane, or (iii) on top of the membrane.
  • the substrate or reagent may be added by incubating the membrane in a solution or suspension thereof wherupon the excess is washed away.
  • Another alternative is spraying the substrate or reagent onto the membrane.
  • the separated components are immobilized in the membrane prior to detection as is per se well known in the art. e.g. chemically by a cross-linking agent, such as glutaraldehyde.
  • Other means of immobilization are e.g. denaturation by heating or exposure to an organic solvent.
  • Staining for lipids may be performed by conventional methods, for example with Oil Red O, Sudan Black B or Fat Red 7b as described in G. Bittolo-Bon and G. Cazzalato, J. Lipid Res. 40, 170-176 (1999); and in O. Gaal et al., Electrophoresis in the Separation of Biological Macromolecules, John Wiley & Sons, 1980, pp. 327-335.
  • Staining for carbohydrates may be performed by conventional methods, for example as described in A.H. Wardi and G.A. Michos, Anal. Biochem. 49, 607-609 (1972); and G. Dubray and G. Bezard, Anal. Biochem. 119, 325-329 (1982).
  • the biospecific detectable reagent may be a biospecific affinity reactant which is labelled with an analytically detectable group, such as an enzymatically active group (e.g. colour formation upon action on substrate), fluorescent group, chromogenic group, hapten, biotin, radiolabel (autoradiography), particles, etc.
  • an analytically detectable group such as an enzymatically active group (e.g. colour formation upon action on substrate), fluorescent group, chromogenic group, hapten, biotin, radiolabel (autoradiography), particles, etc.
  • a usual form of analytically labelled reactants is labelled antibody.
  • a particularly useful labelling group is particles, for example black-coloured carbon particles which may be measured directly, e.g. with a conventional type scanner.
  • the particles contain one of the above mentioned detectable groups, such as fluorophoric group or chromogenic group (fluorescent and coloured particles, respectively).
  • Useful particles often have a size in the range 0.001 to 5 ⁇ m, with preference for the range 0.05 to 5 ⁇ m.
  • the particles may be of colloidal dimensions, so- called sol (i.e. usually spherical and monodisperse having a size in the range 0.001 to 1 ⁇ m).
  • sol i.e. usually spherical and monodisperse having a size in the range 0.001 to 1 ⁇ m.
  • metal particles for example, gold sol
  • non-metal particles for example Si ⁇ 2, carbon, latex and killed erythrocytes and bacteria.
  • particles of non-colloidal dimensions have been used. These have been more or less irregular and more or less polydisperse (for example, carbon particles ⁇ 1 ⁇ m; see e.g. our WO 96/22532).
  • the complexes formed in the detection zone may often be detected visually or by optical measuring equipment (e.g. a CCD camera coupled to a computer with special software for image analysis or laser scanner).
  • optical measuring equipment e.g. a CCD camera coupled to a computer with special software for image analysis or laser scanner.
  • WO 88/08534 Unilever
  • US-A-5, 120,643 Abbott Labs.
  • EP-A-284,232 Becton Dickinson
  • the ion-exchange membrane may not permit detection directly thereon (e.g. due to a particular ion-exchange group). In such a case, it may be necessary to provide a parallel differently or unmodified membrane section, or another membrane joined thereto, to which the separated components may be forced to migrate by a transverse liquid flow initiated after completed separation in the ion-exchange membrane.
  • a detection procedure is described in the illustrative embodiment and the specific Example below.
  • the analytical method of invention is well suited for the analysis of biological samples, for example, blood (serum, plasma, whole blood), saliva, tear fluid, urine, cerebrospinal fluid, sweat, etc.
  • biological samples for example, blood (serum, plasma, whole blood), saliva, tear fluid, urine, cerebrospinal fluid, sweat, etc.
  • the invention is also applicable to other types of samples, such as fermentation solutions, reaction mixtures, etc.
  • Sample components that it may be desired to determine are usually high-molecular components, e.g. proteins, peptides, nucleic acids, or polynucleotides.
  • FIG. 1 illustrates schematically a membrane that may be used for the analysis of e.g. proteins in accordance with the method of the invention.
  • the membrane consists in the illustrated case of two combined parts of different materials, a separation part 1 and a detection part 2, joined by a piece of adhesive tape (not shown) on the backside of the combined membrane and in liquid receiving contact with each other by a thin membrane band 3 as an overlap.
  • This membrane band 3 is secured to the separation/detection membrane by a piece of adhesive tape 4.
  • the separation part defines a separation zone on the combined membrane.
  • the detection part defines a detection zone on the combined membrane.
  • the short-sides of the membrane are indicated in Fig. 1 by a and c and the long-sides by b and d, respectively.
  • the membrane may be used as follows with reference to Figs. 2A to 2C.
  • a sample containing two components to be analysed (referred to as analytes 1 and 2 below) is applied at 5 on the separation zone 1 (Fig. 2A).
  • a pad 6 containing separation buffer is applied at short-side a of the membrane and a sucking pad 7 at the opposite short-side c. This will cause a buffer flow of in the direction of the arrow in Fig. 2 A, separating the two analytes as indicted by the dots at 8 (analyte 1) and 9 (analyte 2) in Fig. 2A.
  • pads 6 and 7 in Fig. 2A are then removed and an eluent-containing pad 10 is mounted to the long-side d, and a sucking pad 11 is mounted to long-side b.
  • This causes a flow of eluent in the direction of the arrow in Fig. 2B, transporting the separated analytes to positions 12 and 13 in the detection zone where the analytes optionally are immobilized by chemical crosslinking, for example.
  • the pads 10 and 11 are removed and replaced by a sucking pad 14 at the long-side d, and a container 15 with a solution or suspension of labelled reactant at the long-side b.
  • labelled reactant will migrate in the direction of the arrow and bind to the immobilized analytes at 16 and 17 in Fig. 2C.
  • the labelled complexes, and thereby the corresponding analytes may then be detected and quantified by reading the intensity of the signals from the label in the detection area and calculating the respective amounts.
  • the measurements may advantageously be performed with a scanner.
  • nitrocellulose membrane (3 ⁇ m, nitrocellulose on polyester backing, Whatman international Ltd, England) was placed in a solution of 0.1 % polyethyleneimine (PEI, Sigma, St Louise, MO, USA). The mixture was shaken for three hours and the membrane was then placed in 0.1 % Tween 20 for 30 minutes, dried in air and then stored in a plastic bag at +4°C.
  • PEI polyethyleneimine
  • the separation membrane was cut to 1.5 x 5 cm and a plain nitrocellulose membrane was cut to 3.5 x 5 cm.
  • the two membranes were put tightly together along the long sides and joined by means of adhesive tape on the underside.
  • a piece of nitrocellulose membrane (0.3 cm x 5 cm, AE99, Schleicher and Schuell, Dassel, Germany) was placed on the top side of the two membranes as an overlap.
  • This membrane was anchored by means of a 1 x 4 cm self-adhesive polyester film ( Gelman adhesive polyesterfilm, 3 mil) placed such that 0.5 cm at the short side on the formed combined separation/plain nitrocellulose membrane remained uncovered.
  • the short sides of the combination membrane are referred to as a and c, respectively, and the two long sides as b and d, respectively ( see Fig.l).
  • Carbon black stock solution 1.5 g of carbon black particles ( sp 5, Degussa, Germany) were suspended in 150 ml 5 mM borate buffer, pH 8.4 and sonicated (VibraCell 600 W, 1.5 cm probe) in a plastic beaker for 5 minutes at 100 % amplitude and 5+5 seconds pulse. 1.5 ml Tween 20 (Sigma, St Louise, MO, USA) were added and the solution was sonicated for 5 minutes at 100 % amplitude and 5+5 seconds pulse. Carbon black work solution: 4 ml of 10 mg/ml of carbon black stock solution were diluted in 35 ml 5 mM borate buffer. pH 8.4 and 1.2 ml Tween 20 were added. The solution was sonicated (VibraCell 600 W, micro probe) for 5 minutes at 30 % amplitude and 5 + 5 seconds pulse.
  • Bovine albumin Bovine albumin (Intergen company, Purchase, NY, USA ) was purified by ion-exchange chromatography on Mono Q (Amersham Pharmacia Biotech AB, Sweden). The most negatively charged part of the material was isolated.
  • Step 1 Wetting of membrane from short side a to short side c
  • the combination membrane is wetted by adding elution buffer to a 1 x 5 x 0.5 cm PVA sponge (PVA D, 60 ⁇ m, Kanebo Ltd, Japan) and then placing the sponge along short side a of the membrane.
  • a 2 x 5 cm sucking cellulose membrane G 004, Schlecher and Schuell.
  • the elution buffer is 20 mM bis-Tris, 0.1 % Tween 20, 10 mM NaCl, pH 6.31.
  • Step 2 Sample application and elution from short side a to short side c
  • Step 3 Elution from long side d (separation membrane) to long side b (detection membrane)
  • the combination membrane is dried by a hairdryer for about 1 -2 minutes. Then the membrane is sprayed with 12.5 % glutaraldhyde (Merck), the excess is wiped off and the reaction is continued for 4 minutes.
  • glutaraldhyde Merck
  • a 2 x 5 cm sucking cellulose membrane (GB 004, Schlecher and Schuell) is mounted along long side d (separation membrane part), and along long side b is placed a 1 x 5 x 0.5 cm PVA sponge (PVA D, 60 ⁇ m, Kanebo Ltd, Japan) wetted by 1 mg/ml of carbon black in 3 % Tween 20.
  • the carbon black particles are allowed to pass the immobilised proteins for 7 minutes and then the PVA sponge is replaced with an identical PVA sponge filled with elution buffer (see step 1). This washing is continued for 10 minutes and then the PVA sponge and the sucking membrane are removed and the combination membrane is dried.
  • the membrane is placed in a scanner (Agfa Acus II Scanner) for measuring the grey scale in a 1 cm broad line along the plain nitrocellulose membrane where the proteins are immobilised and coloured with carbon black.
  • the grey scale is read with a 12 bits grey scale resolution (4096 levels) and 600 points per inch (ppi) optical resolution.
  • the image obtained is digitalised and the intensity values are processed by means of Microsoft Excel.
  • the method of the invention permits excellent separation between proteins of different isoelectric points.
  • the quality of a chromatographic material can be evaluated by the concept of the theoretical plate. The lower the height of the theoretical plate (HETP), the better the material.
  • the nitrocellulose membrane used above (3 ⁇ m nitrocellulose on polyester backing, Whatman international Ltd, England) was tested by using thin lines of bromophenol blue placed on the membrane, and after the elution had started, pictures were taken by a digital camera (Agfa 1280). Pictures were taken between 2.8 - 47 mm of migration. The pictures were scanned (Agfa Arcus II scanner), digitilised and processed (Microsoft Excel).
  • the theoretical plate height (HETP) was calculated by Formula 2 where L is the migration distance.
  • N The theoretical plate height depends on the migration distance, as shown in the table below.
  • the HETP is about 25 ⁇ m.
  • the migration speed was 0.65 cm/minute.

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  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

L'invention concerne un procédé analytique chromatographique dont les étapes sont les suivantes: a)mettre en place une matrice d'écoulement pourvue d'une membrane attachée à un élément de protection imperméable aux liquides aqueux, cette matrice d'écoulement permettant au fluide latéral aidé par une force capillaire de s'écouler, au moins une partie de la matrice d'écoulement étant modifiée afin de favoriser des fonctions échangeuses d'ions. b) traiter la matrice d'écoulement afin de réduire ou d'éliminer les propriétés d'absorption non-spécifiques de la matrice d'écoulement; c) placer dans la matrice d'écoulement un échantillon contenant au moins deux composants; d) provoquer un premier écoulement latéral de fluide aqueux afin que l'échantillon traverse la matrice d'écoulement et sépare les composants qui s'y trouvent; e) interrompre l'écoulement latéral; f1) détecter au moins un des composants séparés sur la matrice d'écoulement dans la position atteinte par le composant respectif quand l'écoulement a été interrompu; ou f2a) provoquer un second écoulement latéral de fluide aqueux afin que les composants soient transportés dans une direction sensiblement transversale par rapport à la direction du premier écoulement latéral. f2b) interrompre le second écoulement latéral; et f2c) détecter au moins un des composants séparés sur la matrice d'écoulement dans la position atteinte par le composant respectif quand l'écoulement a été interrompu. L'invention concerne également un dispositif chromatographique comprenant une matrice d'écoulement pourvue d'une membrane attachée à un élément de protection imperméable aux liquides aqueux, cette matrice d'écoulement permettant au fluide latéral aidé par une force capillaire de s'écouler, au moins une partie de la matrice d'écoulement étant modifiée afin de favoriser des fonctions échangeuses d'ions.
PCT/SE2000/001510 1999-08-06 2000-07-21 Procede et dispositif analytique WO2001011355A1 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
AU63313/00A AU770715B2 (en) 1999-08-06 2000-07-21 Analytical method and device
CA002381137A CA2381137A1 (fr) 1999-08-06 2000-07-21 Procede et dispositif analytique
JP2001515960A JP2003525426A (ja) 1999-08-06 2000-07-21 分析方法及びデバイス
EP00950177A EP1210594A1 (fr) 1999-08-06 2000-07-21 Procede et dispositif analytique

Applications Claiming Priority (8)

Application Number Priority Date Filing Date Title
SE9902855-7 1999-08-06
SE9902855A SE9902855D0 (sv) 1999-08-06 1999-08-06 Analytical method and apparatus
US14856699P 1999-08-13 1999-08-13
US60/148,566 1999-08-13
SE9903970-3 1999-11-01
SE9903970A SE9903970D0 (sv) 1999-08-06 1999-11-01 Analytical method and device
US16414799P 1999-11-08 1999-11-08
US60/164,147 1999-11-08

Publications (1)

Publication Number Publication Date
WO2001011355A1 true WO2001011355A1 (fr) 2001-02-15

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EP (1) EP1210594A1 (fr)
JP (1) JP2003525426A (fr)
AU (1) AU770715B2 (fr)
CA (1) CA2381137A1 (fr)
WO (1) WO2001011355A1 (fr)

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WO2004047947A1 (fr) * 2002-11-26 2004-06-10 Prime Separations Incorporated Procede de separation chromatographique
WO2007120272A2 (fr) * 2005-12-06 2007-10-25 Pall Corporation Systeme et procede pour tester des milieux et dispositifs de chromatographie
US8043811B2 (en) 2006-03-11 2011-10-25 The Food & Environment Research Agency (FERA) representing the Secretary of State for Environment, Food and Rural Affairs Purification method and kits
EP2579034A1 (fr) * 2010-05-27 2013-04-10 Daicel Corporation Procédé de détection dans un échantillon par chromatographie sur couche mince, plaque de chromatographie sur couche mince, et procédé de production associé

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JP6999110B2 (ja) * 2017-10-10 2022-01-18 国立大学法人 東京大学 浸透圧調整物質定着用アニオン性イオン交換膜、浸透圧調整物質検出方法、および浸透圧調整物質検出キット

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WO2004047947A1 (fr) * 2002-11-26 2004-06-10 Prime Separations Incorporated Procede de separation chromatographique
WO2007120272A2 (fr) * 2005-12-06 2007-10-25 Pall Corporation Systeme et procede pour tester des milieux et dispositifs de chromatographie
WO2007120272A3 (fr) * 2005-12-06 2008-01-10 Pall Corp Systeme et procede pour tester des milieux et dispositifs de chromatographie
US7732216B2 (en) 2005-12-06 2010-06-08 Pall Corporation System and method for testing chromatography media and devices
US8043811B2 (en) 2006-03-11 2011-10-25 The Food & Environment Research Agency (FERA) representing the Secretary of State for Environment, Food and Rural Affairs Purification method and kits
EP2579034A1 (fr) * 2010-05-27 2013-04-10 Daicel Corporation Procédé de détection dans un échantillon par chromatographie sur couche mince, plaque de chromatographie sur couche mince, et procédé de production associé
EP2579034A4 (fr) * 2010-05-27 2014-07-16 Daicel Corp Procédé de détection dans un échantillon par chromatographie sur couche mince, plaque de chromatographie sur couche mince, et procédé de production associé
US9234881B2 (en) 2010-05-27 2016-01-12 Daicel Corporation Sample detection method by thin-layer chromatography, thin-layer chromatography plate, and method for producing same

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CA2381137A1 (fr) 2001-02-15

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