EP0974059A1 - Aufbereitung von blutproben für die homocystein- und/oder folatbestimmung - Google Patents
Aufbereitung von blutproben für die homocystein- und/oder folatbestimmungInfo
- Publication number
- EP0974059A1 EP0974059A1 EP98919180A EP98919180A EP0974059A1 EP 0974059 A1 EP0974059 A1 EP 0974059A1 EP 98919180 A EP98919180 A EP 98919180A EP 98919180 A EP98919180 A EP 98919180A EP 0974059 A1 EP0974059 A1 EP 0974059A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- blood
- homocysteine
- collection vessel
- ethylene oxide
- folate
- 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.)
- Withdrawn
Links
Classifications
-
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6806—Determination of free amino acids
- G01N33/6812—Assays for specific amino acids
- G01N33/6815—Assays for specific amino acids containing sulfur, e.g. cysteine, cystine, methionine, homocysteine
-
- 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
- G01N33/82—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving vitamins or their receptors
-
- 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
- Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
- Y10T436/107497—Preparation composition [e.g., lysing or precipitation, etc.]
-
- 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
-
- 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
- Y10T436/2525—Stabilizing or preserving
Definitions
- the present invention relates to a method and a blood collection vessel for the preparation of blood samples for the homocysteine and / or total folate determination.
- Preparation here means in particular the stabilization of the blood samples for the homocysteine determination and the lysis of the erythrocytes to prepare the total 5 folate determination.
- Total folate in the context of this invention is understood to mean the sum of erythrocyte and plasma folate.
- Homocysteine a sulfur-containing amino acid, only occurs in the organism as an intermediate in the methionine-cysteine-glutathione metabolism cycle and is not incorporated into proteins. Due to various hereditary defects
- Tetrahydrofolate reductase MTHFR
- B 12 , B 6 , folate corresponding vitamin cofactors
- homocysteine is not sufficiently broken down and therefore occurs in increased concentrations in the plasma (K. Berg et al.,
- the prevalence of the homozygous MTHFR defect which is associated with an increased homocysteine level, is given as 5% (Goyette et al., Loc.cit), this is of the order of magnitude of the prevalence of diabetes mellitus in the total Caucasian population.
- Homocysteine is currently measured using HPLC (high-pressure liquid chromatography) from EDTA plasma and can be carried out with good accuracy. However, it is necessary to pre-analyze the HPLC (high-pressure liquid chromatography) from EDTA plasma and can be carried out with good accuracy. However, it is necessary to pre-analyze the HPLC (high-pressure liquid chromatography) from EDTA plasma and can be carried out with good accuracy. However, it is necessary to pre-analyze the HPLC (high-pressure liquid chromatography) from EDTA plasma and can be carried out with good accuracy. However, it is necessary to pre-analyze the HPLC (high-pressure liquid chromatography) from EDTA plasma and can be carried out with good accuracy. However, it is necessary to pre-analyze the HPLC (high-pressure liquid chromatography) from EDTA plasma and can be carried out with good accuracy. However, it is necessary to pre-analyze the HPLC (high-pressure liquid chromatography) from EDTA
- the present invention is therefore based in part on the problem of avoiding a significant increase in homocysteine in whole blood, which occurs naturally within one to two hours after venous blood withdrawal (FIG. 1).
- the present invention avoids the previously necessary, time-critical separation of the blood cells from the plasma and stabilizes the homocysteine Concentration in the resulting lysed blood can be reached over 48 hours.
- the homocysteine concentration often correlates inversely with the folate concentration, since folate is a cofactor for the homocysteine-degrading enzyme MTHFR. Folate is therefore often given to bring a pathologically increased homocysteine concentration back to normal. A determination of the basal and current folate level in addition to the homocysteine level is therefore diagnostically useful. Since folate is mainly present (approx. 98%) in the erythrocytes and has an effect, the determination of the erythrocyte folate or the total folate is more meaningful than the usual determination of the plasma or serum folate. The present invention is therefore based on the further problem of preparing a blood sample so that the total folate concentration without can be determined in addition to or instead of the homocysteine concentration.
- the above objects are achieved by a method for the preparation of blood samples for the homocysteine and / or total folate determination, in which the blood sample is taken during or immediately after the blood withdrawal with a) at least one reagent for lysing the blood cells and b) associates at least one inhibitor of the homocysteine-forming and degrading enzymes.
- the blood sample is also particularly preferably associated with c) one or more acids.
- the invention is directed to a blood collection vessel for the preparation of blood samples, in particular for the homocysteine and / or total folate determination, comprising a) at least one reagent for the lysis of the blood cells and b) at least one inhibitor of the homocysteine-forming and - degrading enzymes and preferably also c) one or more acids.
- 1 shows the time course of the homocysteine concentration in blood collection vessels according to the prior art.
- 2 shows the time course of the homocysteine concentration in blood collection vessels according to the present invention.
- Figure 3 shows the correlation of plasma and lysate homocysteine concentrations.
- the reagent for lysing the blood cells a) can in particular be a detergent, but also e.g. Ascorbic acid.
- An anionic detergent e.g. Sodium dodecyl sulfate (SDS), but preferably a nonionic detergent is used, e.g. Nonidet P40 (octylphenol-ethylene oxide condensate with an average of 9 moles of ethylene oxide per mole of phenol) or Triton X.
- Nonidet P40 octylphenol-ethylene oxide condensate with an average of 9 moles of ethylene oxide per mole of phenol
- Triton X Triton X
- Reagents which form chelate complexes with calcium are particularly suitable as inhibitors of the homocysteine-forming and degrading enzymes b).
- Citrates and other chelating agents e.g. Di- or polycarboxylic acids, aminocarboxylic acids or phosphonocarboxylic acids and their salts, especially the alkali salts.
- Preferred is ethylenediaminetetraacetic acid disodium salt (EDTA), which is particularly preferably used in the form of a highly concentrated aqueous solution just below the saturation limit.
- the blood cells are lysed with the help of a highly concentrated detergent while simultaneously
- the same sample can be taken in the blood collection vessel according to the invention using an immunoassay system (e.g. Fa.
- an immunoassay system e.g. Fa.
- the total folate concentration can be determined. From Example 2 it can be seen that the total folate concentrations measured in the lysate stabilized directly during the decrease are comparable to the folate concentrations which were conventionally measured in the laboratory after an aliquot of an EDTA whole blood had been introduced into the lysis reagent presented . The time of lysis of the whole blood does not affect the total folate concentration. According to the prior art, however, two blood samples were usually required to determine both the homocysteine and the total folate concentration because the Homocysteine sample had to be centrifuged immediately. Here, the present invention makes it easier for doctors and patients.
- Blood collection vessel for sample preparation for common HPLC analysis methods without having to modify them, makes it easier for the analyst in the laboratory to interpret the values reliably, since incorrect pre-analytical treatment of the samples (the whole blood is too long) can be excluded. Furthermore, the immediate centrifugation of the sample material, which is usually not possible in clinical wards or with private doctors, can be dispensed with.
- the reagents in commercial blood collection tubes for example, be a Monovette ® (Messrs. Sarstedt) or a Vacutainer ® (Fa. Becton Dickinson) submitted.
- Monovettes ® with EDTA, NaF, heparin and other fillings are already commercially available. It is readily possible to fill a corresponding container with the reagents according to the invention, so that a blood collection vessel according to the invention is available is provided.
- the sample container can be made of polyethylene or another suitable plastic, for example, and can be in the form of a syringe. With the help of a stamp, the blood is drawn into the vein by creating a negative pressure via an injection needle
- Blood collection tube transferred.
- a pre-evacuated vessel with an injection needle can be used.
- "Blood collection vessel” in the sense of the invention also includes a capillary blood collection system, e.g. a glass capillary, understood in combination with a vessel in which the
- Capillary blood is kept until it is processed for measurement.
- Such systems are already on the market.
- the reagent combination according to the invention is usually present in the storage vessel of this capillary blood collection system.
- Nonidet P40, EDTA and citric acid is also advantageous in that it consists of inexpensive components which are already used individually in the clinical-chemical field.
- Example 1 Of a as indicated in Example 1, according to the invention prepared EDTA-Monovette ® "system Assay Access immuno-" Sanofi Diagnostics Pasteur is one of the Fa. The total folate concentration was determined. The resulting values are compared to the total folate concentrations of blood samples which were processed by subsequent lysis of EDTA blood in the laboratory with a submitted lysis reagent from the above-mentioned company. Table 1 below shows the good correlation of folate concentrations.
Abstract
Description
Claims
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19713088 | 1997-03-27 | ||
DE19713088A DE19713088C1 (de) | 1997-03-27 | 1997-03-27 | Verfahren und Blutabnahmegefäß zur Aufbereitung von Blutproben für die Homocystein- und/oder Folatbestimmung |
PCT/EP1998/001830 WO1998044351A1 (de) | 1997-03-27 | 1998-03-27 | Aufbereitung von blutproben für die homocystein- und/oder folatbestimmung |
Publications (1)
Publication Number | Publication Date |
---|---|
EP0974059A1 true EP0974059A1 (de) | 2000-01-26 |
Family
ID=7824922
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP98919180A Withdrawn EP0974059A1 (de) | 1997-03-27 | 1998-03-27 | Aufbereitung von blutproben für die homocystein- und/oder folatbestimmung |
Country Status (7)
Country | Link |
---|---|
US (2) | US6309885B1 (de) |
EP (1) | EP0974059A1 (de) |
AU (1) | AU7212198A (de) |
CA (1) | CA2284674A1 (de) |
DE (1) | DE19713088C1 (de) |
NO (1) | NO994705L (de) |
WO (1) | WO1998044351A1 (de) |
Families Citing this family (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE19836559A1 (de) | 1998-08-12 | 2000-03-23 | Antigen Gmbh | Gefäß zur Entnahme von Blut |
DE10124820A1 (de) | 2001-05-21 | 2002-12-05 | Heinrich Wieland | Zusammensetzung und Verwendung von Substanzen zur Stabilisierung von schwefelhaltigen Aminosäuren im Blut |
JP2005524850A (ja) * | 2002-05-07 | 2005-08-18 | ベクトン・ディキンソン・アンド・カンパニー | 採集装置 |
EP1518118B1 (de) * | 2002-05-08 | 2007-10-03 | Yorktest Laboratories Limited | Behälter zur aufnahme von proben, der eine hydrophile membran zur abtrennung von partikeln enthält |
EP1549434A1 (de) * | 2002-05-13 | 2005-07-06 | Becton Dickinson and Company | Probenahmesystem mit proteasehemmer |
US20050019937A1 (en) * | 2002-11-29 | 2005-01-27 | Industrial Technology Research Institute | Assay and kit for homocysteine |
US7356566B2 (en) * | 2003-10-09 | 2008-04-08 | International Business Machines Corporation | Selective mirrored site accesses from a communication |
US20050124965A1 (en) * | 2003-12-08 | 2005-06-09 | Becton, Dickinson And Company | Phosphatase inhibitor sample collection system |
US8852862B2 (en) * | 2004-05-03 | 2014-10-07 | Handylab, Inc. | Method for processing polynucleotide-containing samples |
US7592139B2 (en) | 2004-09-24 | 2009-09-22 | Sandia National Laboratories | High temperature flow-through device for rapid solubilization and analysis |
US20090136385A1 (en) * | 2007-07-13 | 2009-05-28 | Handylab, Inc. | Reagent Tube |
CN115436540A (zh) * | 2022-09-26 | 2022-12-06 | 汤臣倍健股份有限公司 | 一种同时测定血液中叶酸和同型半胱氨酸含量的方法及试剂盒 |
Family Cites Families (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3972991A (en) * | 1974-03-18 | 1976-08-03 | Case Western Reserve University | Radioisotopic assay and binder therefor |
US4940658A (en) * | 1986-11-20 | 1990-07-10 | University Patents, Inc. | Assay for sulfhydryl amino acids and methods for detecting and distinguishing cobalamin and folic acid deficency |
US5541116A (en) * | 1991-09-30 | 1996-07-30 | B.R.A.H.M.S. Diagnostica Gmbh | Method for the stabilization of endogenous, physiologically active peptides |
CA2128512C (en) * | 1992-01-22 | 1999-12-07 | Erling Sundrehagen | Homocysteine assay |
US5434087A (en) * | 1993-02-24 | 1995-07-18 | Abbott Laboratories | Folate immunoassay utilizing folate binding protein in a multiclonal antibody format |
DE4330213C2 (de) * | 1993-09-07 | 1995-08-10 | Patscheke Heinrich | Mittel zur Stabilisierung von Blut |
US5478729A (en) * | 1994-04-28 | 1995-12-26 | Syntex (U.S.A.) Inc. | Immunoassay for homocysteine |
US5559038A (en) * | 1994-05-04 | 1996-09-24 | The Regents Of The University Of Colorado | Gas chromatography/mass spectrometry determination of oxidized sulfhydryl amino acids |
AU8153098A (en) * | 1997-06-23 | 1999-01-04 | Nexstar Pharmaceuticals, Inc. | Homocysteine assay |
US5985540A (en) * | 1997-07-24 | 1999-11-16 | Anticancer, Inc. | High specificity homocysteine assays for biological samples |
US6107100A (en) * | 1998-04-30 | 2000-08-22 | Metaquant Trust | Compounds and methods for determination of thiols |
-
1997
- 1997-03-27 DE DE19713088A patent/DE19713088C1/de not_active Expired - Fee Related
-
1998
- 1998-03-27 EP EP98919180A patent/EP0974059A1/de not_active Withdrawn
- 1998-03-27 WO PCT/EP1998/001830 patent/WO1998044351A1/de not_active Application Discontinuation
- 1998-03-27 CA CA002284674A patent/CA2284674A1/en not_active Abandoned
- 1998-03-27 AU AU72121/98A patent/AU7212198A/en not_active Abandoned
-
1999
- 1999-09-24 US US09/405,549 patent/US6309885B1/en not_active Expired - Fee Related
- 1999-09-27 NO NO994705A patent/NO994705L/no unknown
-
2001
- 2001-04-24 US US09/841,877 patent/US20020001846A1/en not_active Abandoned
Non-Patent Citations (1)
Title |
---|
See references of WO9844351A1 * |
Also Published As
Publication number | Publication date |
---|---|
NO994705D0 (no) | 1999-09-27 |
WO1998044351A1 (de) | 1998-10-08 |
NO994705L (no) | 1999-11-25 |
DE19713088C1 (de) | 1998-11-26 |
CA2284674A1 (en) | 1998-10-08 |
US6309885B1 (en) | 2001-10-30 |
US20020001846A1 (en) | 2002-01-03 |
AU7212198A (en) | 1998-10-22 |
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