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WO1991015597A1 - Procede permettant de determiner les metabolites d'importance clinique - Google Patents

Procede permettant de determiner les metabolites d'importance clinique Download PDF

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Publication number
WO1991015597A1
WO1991015597A1 PCT/FI1991/000094 FI9100094W WO9115597A1 WO 1991015597 A1 WO1991015597 A1 WO 1991015597A1 FI 9100094 W FI9100094 W FI 9100094W WO 9115597 A1 WO9115597 A1 WO 9115597A1
Authority
WO
WIPO (PCT)
Prior art keywords
peroxide
hppa
transformed
procedure according
cholesterol
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.)
Ceased
Application number
PCT/FI1991/000094
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English (en)
Inventor
Osmo Suovaniemi
Matti HÄRKONEN
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.)
ELOMIT Oy
Original Assignee
ELOMIT Oy
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 ELOMIT Oy filed Critical ELOMIT Oy
Publication of WO1991015597A1 publication Critical patent/WO1991015597A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
    • C12Q1/28—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase involving peroxidase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/60—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving cholesterol
    • 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
    • G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2326/00—Chromogens for determinations of oxidoreductase enzymes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/04—Endocrine or metabolic disorders
    • G01N2800/044—Hyperlipemia or hypolipemia, e.g. dyslipidaemia, obesity

Definitions

  • the present invention concerns a procedure-for determining clinically significant metabolites, such as cholesterol, HDL cholesterol, triglycerides, creatinine and uric acid, in body fluids.
  • the object of the present invention is to dis ⁇ close a novel procedure for determining, in a simple way, rapidly and economically, significant metabolites, such as cholesterol, HDL cholesterol, triglycerides, creatinine and uric acid. It is a particular object of the invention, to disclose a novel procedure for simul ⁇ taneous determination of total cholesterol and HDL cho ⁇ lesterol, e.g. in plasma or serum.
  • the invention is based on the fundamental idea that the respective metabolite is made to produce per ⁇ oxide.
  • the peroxide which has been formed is determin ⁇ ed, based on the dimer constituted by 3-(4-hydroxy- phenyl)propionic acid (HPPA) with peroxide (dihydroxy- diphenyl-dipropionic acid) .
  • HPPA 3-(4-hydroxy- phenyl)propionic acid
  • peroxide dihydroxy- diphenyl-dipropionic acid
  • cholesterolester for instance, may be hydrolyzed to cholesterol and fatty acid: cholesterolester + H 2 0 —> cholesterol + fatty acids
  • the cholesterol is oxidized e.g. enzymatically with the aid of cholesteroloxidase:
  • the peroxide and HPPA are constituted to form the powerfully fluorescent HPPA dimer, e.g. with the aid of peroxidase.
  • the strongly fluorescent HPPA dimer (HPPA-HPPA) can be quantitatively determined e.g. at excitation wavelength 320 nm and emission wavelength 405 nm.
  • the glycerol is transformed into glycerol-3- phosphate, e.g. enzymatically with the aid of glycero- kinase:
  • the glycerol-3-phosphate is oxidized to di- hydroxyacetonephosphate and peroxide, e.g. enzymatical ⁇ ly with the aid of glycerol-3-P-oxidase:
  • creatinine For determination of creatinine, it is hydro- lyzed to creatine, e.g. enzymatically with the aid of creatinase:
  • the creatine is further hydrolyzed to sarco- sine, e.g. enzymatically with the aid of creatinase:
  • the sarcosine is oxidized to glycine, form- aldehyde and peroxide, e.g. enzymatically with the aid of sarcosinoxidase:
  • uric acid For determining uric acid, it is oxidized to allantoin, carbon dioxide and peroxide, e.g. enzymati ⁇ cally with the aid of uricase:
  • the peroxide is determined as HPPA dimer in the manner described.
  • the procedure is applicable in principle in determination of all clinically significant metabo ⁇ lites, hormones and proteins which can be made to pro ⁇ Jerusalem peroxide (H 2 0 2 ) , e.g. with the aid of various purified enzymes.
  • the metabolite may be reacted with peroxidase-labelled antibodies which pro ⁇ Jerusalem peroxide quantitatively.
  • the peroxide determina ⁇ tion is ultimately made as described, as HPPA dimers.
  • the procedure of the invention can be imple ⁇ mented most rapidly, e.g. within 6 to 7 min. after drawing a blood sample. As a whole, the procedure is readily implementable and applicable in automated sample handling and analysis.
  • the proce ⁇ dure is appropriate for determining total cholesterol and HDL cholesterol.
  • a blood sample was taken into a heparinized tube, plasma and cells were separated in the centri ⁇ fuge, 30 sec, 8000 g.
  • dextran sulphate-Mg ++ precipitation was carried out, alternatively phosphotungstic acid-Mg ++ precipitation, the precipitate was separated in the centrifuge, 2 min., 8000 g.
  • Cholesterolesterase 1.0 U/ml Cholesteroloxidase 1.0 U/l
  • Serum or plasma can be used for sample.
  • the sample can be stored one week at +4°C, and for instance several months at -20°C.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Zoology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Wood Science & Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Urology & Nephrology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • General Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • Cell Biology (AREA)
  • Endocrinology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Il s'agit d'un procédé à l'aide duquel on peut déterminer les métabolites d'importance clinique, comme c'est le cas pour le cholestérol, le cholestérol HDL, les triglycérides, la créatinine et l'acique urique présents dans les fluides organiques. On fait produire du peroxyde au composé, et le peroxyde ainsi produit est déterminé à l'aide du dimère de peroxydase et de l'acide 3-(4-hydroxyphényle) propionique (HPPA), ce qui donne un dimère HPPA fortement fluorescent qui est analysé de manière fluorométrique.
PCT/FI1991/000094 1990-04-04 1991-04-02 Procede permettant de determiner les metabolites d'importance clinique Ceased WO1991015597A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FI901703A FI901703L (fi) 1990-04-04 1990-04-04 Foerfarande foer bestaemmande av kliniskt relevanta metaboliter.
FI901703 1990-04-04

Publications (1)

Publication Number Publication Date
WO1991015597A1 true WO1991015597A1 (fr) 1991-10-17

Family

ID=8530207

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/FI1991/000094 Ceased WO1991015597A1 (fr) 1990-04-04 1991-04-02 Procede permettant de determiner les metabolites d'importance clinique

Country Status (2)

Country Link
FI (1) FI901703L (fr)
WO (1) WO1991015597A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006061646A1 (fr) * 2004-12-11 2006-06-15 The Science And Technology Facilities Council Dosage pour produire un profil lipidique au moyen d'une mesure de fluorescence

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0218127A1 (fr) * 1985-09-18 1987-04-15 Roche Diagnostics GmbH Procédé et réactif pour la détermination spécifique de HDL-cholestérol dans le sérum

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0218127A1 (fr) * 1985-09-18 1987-04-15 Roche Diagnostics GmbH Procédé et réactif pour la détermination spécifique de HDL-cholestérol dans le sérum

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
CHEMCIAL ABSTRACTS, Volume 94, No. 21, 25 May 1981, (Columbus, Ohio, US), see page 354, Abstract 170650v; & JP,A,56 014 157, (OKURA, OYSUKE), 1981. *
CHEMICAL ABSTACTS, Volume 102, No. 23, 10 June 1985, (Columbus, Ohio, US), HAMADA, CHIZUKO et al.: "Enzymic Microassay of Glucose, Uric Acid and Cholesterol using 3-(P-Hydroxyphenyl), Propionic Acid as Fluorogenic Substrate for Horseradish Peroxidase", see page 306, Abstract 200512e; & RINSHO KAGAKU, 1984, 13(5), *
CHEMICAL ABSTRACTS, Volume 104, No. 16, 21 April 1986, (Columbus, Ohio, US), HAYASHI, YOHJI et al.: "Flow-Injection Analysis for Hydrogen Peroxide by using Immobilized Horseradish Peroxidase and its Fluorogenic Substrate 3-(P-Hydroxyphenyl) Propionic Acid", see page 757, Abstract 141252m; & ANAL. SCI., 1985, 1(1), 65-68. *
CHEMICAL ABSTRACTS, Volume 109, No. 9, 29 August 1988, (Columbus, Ohio, US), GU, YANSONG et al.: "A Flurogenic Substrate for Horseradish Peroxidase and its use in Elisa", see page 361, Abstract 69853s; & ZHONGHUA YIXUE JIANYAN ZAZHI, 1987, 10(6), 343-345. *
CHEMICAL ABSTRACTS, Volume 94, No. 3, 19 January 1981, (Columbus, Ohio, US), ZAITSU, KIYOSHI et al.: "New Fluorogenic Substrates for Horseradish Peroxidase: Rapid and Sensitive Assays for Hydrogen Peroxide and the Peroxidase", see page 125, Abstract 12014m; & ANAL. BIOCHEM., 1980, 109(1), 109-113. *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006061646A1 (fr) * 2004-12-11 2006-06-15 The Science And Technology Facilities Council Dosage pour produire un profil lipidique au moyen d'une mesure de fluorescence

Also Published As

Publication number Publication date
FI901703A0 (fi) 1990-04-04
FI901703A7 (fi) 1991-10-05
FI901703L (fi) 1991-10-05

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