CA2649633A1 - Differential hemolysis of a whole blood sample - Google Patents

Differential hemolysis of a whole blood sample Download PDF

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Publication number
CA2649633A1
CA2649633A1 CA002649633A CA2649633A CA2649633A1 CA 2649633 A1 CA2649633 A1 CA 2649633A1 CA 002649633 A CA002649633 A CA 002649633A CA 2649633 A CA2649633 A CA 2649633A CA 2649633 A1 CA2649633 A1 CA 2649633A1
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CA
Canada
Prior art keywords
sample
analyte
red blood
whole blood
detecting
Prior art date
Legal status (The legal status 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 status listed.)
Granted
Application number
CA002649633A
Other languages
French (fr)
Other versions
CA2649633C (en
Inventor
Uwe Kobold
Thomas Duelffer
Rupert Herrmann
Herbert Von Der Eltz
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
F Hoffmann La Roche AG
Original Assignee
F. Hoffmann-La Roche Ag
Uwe Kobold
Thomas Duelffer
Rupert Herrmann
Herbert Von Der Eltz
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 F. Hoffmann-La Roche Ag, Uwe Kobold, Thomas Duelffer, Rupert Herrmann, Herbert Von Der Eltz filed Critical F. Hoffmann-La Roche Ag
Publication of CA2649633A1 publication Critical patent/CA2649633A1/en
Application granted granted Critical
Publication of CA2649633C publication Critical patent/CA2649633C/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

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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
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5002—Partitioning blood components
    • 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
    • G01N2030/022—Column chromatography characterised by the kind of separation mechanism
    • G01N2030/027—Liquid chromatography
    • 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
    • G01N30/60—Construction of the column
    • G01N30/6004—Construction of the column end pieces
    • G01N30/603—Construction of the column end pieces retaining the stationary phase, e.g. Frits
    • 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
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/72—Mass spectrometers
    • G01N30/7233—Mass spectrometers interfaced to liquid or supercritical fluid chromatograph
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00—Chemistry: analytical and immunological testing
    • Y10T436/24—Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00—Chemistry: analytical and immunological testing
    • Y10T436/25—Chemistry: analytical and immunological testing including sample preparation

Abstract

The invention relates to a method for differentially hemolyzing whole blood. It discloses method for detecting an analyte in a liquid sample known or suspected to comprise red blood cells and suspected or known to comprise eukaryotic cells, the method comprising the steps of processing said liquid sample with a membrane solubilizing agent under conditions appropriate to lyse cell membranes of red blood cells and at the same time not to cause precipitation of sample constituents, subjecting the processed sample to a chromatographic separation, and detecting the analyte. The differential hemolysis of red blood cells is of advantage in a method of detecting an analyte in a liquid sample that may comprise both erythrocytes as well as nucleated cells. The differential solubilization of red blood cells can be easily combined with an online detection methodology, like LC-MS, and is advantageous in the detection of many analytes, e.g. in the detection of folate or of immunosuppressive drugs, like tacrolimus or sirolimus.

Claims (10)

1. A method of detecting an analyte in a liquid sample known or suspected to comprise red blood cells and suspected or known to comprise eukaryotic cells, the method comprising the steps of a) processing said liquid sample with a membrane solubilizing agent under conditions appropriate to lyse cell membranes of red blood cells and at the same time not causing precipitation of sample constituents, b) subjecting the processed sample obtained in step (a) to a chromatographic separation, wherein said chromatographic separation is by high performance liquid chromatography (HPLC) and c) detecting the analyte wherein said conditions are appropriate if the agent mixed with a 1:10 diluted sample of whole blood in a 1:1 ratio and incubated for 30 min leads to a processed whole blood sample of which 50 aliquots of 10 µL can be applied to a filter with a diameter of 2 mm and 0.5 µm pore size and wherein the increase in backpressure for injection 50 is less than 20 bar as compared to the first injection.
2. The method of claim 1, wherein said chromatographic separation is based on column chromatography and performed by use of a column comprising a frit and a bed material or by use of a monolithic column.
3. The method of according to claim 2, wherein said frit has a pore size of 0.2 or 0.5 µm.
4. The method according claims 2 or 3, wherein said bed material is particulate and the particles have a diameter from 1 to 10 µm.
5. The method of claim 1, wherein said analyte is detected by mass spectroscopy.
6. The method of claim 1, wherein said biological sample is selected from cerebrospinal fluid and whole blood.
7. The method according to one of claims 1 to 6, wherein said analyte is at least partially located inside a red blood cell.
8. The method according to one of claims 1 to 7, wherein said analyte is an immunosuppressive drug.
9. Use of a membrane solubilizing agent comprising a chemical selected from KBr, KJ, KSCN or a salt consisting of one or more of the following cations and anions, wherein the cation is selected from wherein m is 0 or 1 and n is 4 or 6 and wherein the anion is selected from chloride, tetrafluoroborate, octylsulfate, iodide and thiocyanate in the processing of a whole blood sample for high performance liquid chromatography (HPLC).
10. Use of a processed blood sample obtained by differential hemolysis with a membrane solubilizing agent according any of the methods of claims 1 to 9 in a liquid chromatography-based analysis.
CA2649633A 2006-06-06 2007-06-04 Differential hemolysis of a whole blood sample Active CA2649633C (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
EP06011604.3 2006-06-06
EP06011604 2006-06-06
PCT/EP2007/004923 WO2007140961A1 (en) 2006-06-06 2007-06-04 Differential hemolysis of a whole blood sample

Publications (2)

Publication Number Publication Date
CA2649633A1 true CA2649633A1 (en) 2007-12-13
CA2649633C CA2649633C (en) 2012-11-27

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
CA2649633A Active CA2649633C (en) 2006-06-06 2007-06-04 Differential hemolysis of a whole blood sample

Country Status (8)

Country Link
US (1) US7951598B2 (en)
EP (1) EP2032979B1 (en)
JP (1) JP5156738B2 (en)
CN (1) CN101467036B (en)
CA (1) CA2649633C (en)
ES (1) ES2690319T3 (en)
HK (1) HK1134135A1 (en)
WO (1) WO2007140961A1 (en)

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JP5819943B2 (en) * 2010-05-07 2015-11-24 ユーティ—バテル エルエルシー System and method for extracting a sample from a surface
JP5452383B2 (en) * 2010-06-15 2014-03-26 株式会社日立ハイテクノロジーズ Biological sample pretreatment method and apparatus
CN103124793B (en) * 2010-08-11 2014-12-31 协和梅迪克斯株式会社 Method for measuring glycated hemoglobin
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US8637813B2 (en) 2010-10-01 2014-01-28 Ut-Battelle, Llc System and method for laser assisted sample transfer to solution for chemical analysis
US8519330B2 (en) 2010-10-01 2013-08-27 Ut-Battelle, Llc Systems and methods for laser assisted sample transfer to solution for chemical analysis
US8409807B2 (en) 2010-10-22 2013-04-02 T2 Biosystems, Inc. NMR systems and methods for the rapid detection of analytes
EP2630517B1 (en) 2010-10-22 2016-03-16 T2 Biosystems, Inc. Nmr systems and methods for the rapid detection of analytes
US8563298B2 (en) 2010-10-22 2013-10-22 T2 Biosystems, Inc. NMR systems and methods for the rapid detection of analytes
US9891149B2 (en) 2011-08-08 2018-02-13 Thermo Fisher Scientific Oy Method and apparatus for automated analysis
FI20115785A0 (en) 2011-08-08 2011-08-08 Thermo Fisher Scientific Oy Method and apparatus for automatic analysis
EP3524692A1 (en) 2012-04-20 2019-08-14 T2 Biosystems, Inc. Compositions and methods for detection of candida species
EP3391041B1 (en) * 2015-12-18 2019-10-16 H. Hoffnabb-La Roche Ag Automated clinical diagnostic system and method
WO2017127731A1 (en) 2016-01-21 2017-07-27 T2 Biosystems, Inc. Nmr methods and systems for the rapid detection of bacteria
CN107561170B (en) * 2016-07-02 2021-07-30 山东新时代药业有限公司 Analysis and detection method of temsirolimus intermediate
CN106770747A (en) * 2016-12-09 2017-05-31 广州迈达康医药科技有限公司 A kind of tacrolimus formulations evaluation method
CN110352062B (en) * 2017-03-07 2024-04-02 豪夫迈·罗氏有限公司 Stable formulations of immunomodulatory macrolides
CN113325171B (en) * 2021-08-02 2021-12-21 天津康博尔生物基因技术有限公司 Kit for detecting human body erythrocyte folic acid content, detection method and application
CN113776903A (en) * 2021-08-27 2021-12-10 深圳市希莱恒医用电子有限公司 Preparation method of pretreatment reagent, pretreatment reagent and pretreatment method

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Also Published As

Publication number Publication date
US20090253210A1 (en) 2009-10-08
CN101467036B (en) 2013-06-19
CA2649633C (en) 2012-11-27
WO2007140961A1 (en) 2007-12-13
US7951598B2 (en) 2011-05-31
EP2032979B1 (en) 2018-07-18
CN101467036A (en) 2009-06-24
HK1134135A1 (en) 2010-04-16
ES2690319T3 (en) 2018-11-20
JP2009540274A (en) 2009-11-19
JP5156738B2 (en) 2013-03-06
WO2007140961A8 (en) 2008-11-06
EP2032979A1 (en) 2009-03-11

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