WO1993013219A1 - Composition de determination d'ions calcium - Google Patents

Composition de determination d'ions calcium Download PDF

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Publication number
WO1993013219A1
WO1993013219A1 PCT/JP1992/001680 JP9201680W WO9313219A1 WO 1993013219 A1 WO1993013219 A1 WO 1993013219A1 JP 9201680 W JP9201680 W JP 9201680W WO 9313219 A1 WO9313219 A1 WO 9313219A1
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WO
WIPO (PCT)
Prior art keywords
reagent
measuring
composition
calcium
lysolecithin
Prior art date
Application number
PCT/JP1992/001680
Other languages
English (en)
Japanese (ja)
Inventor
Shigeyuki Imamura
Original Assignee
Asahi Kasei Kogyo Kabushiki Kaisha
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 Asahi Kasei Kogyo Kabushiki Kaisha filed Critical Asahi Kasei Kogyo Kabushiki Kaisha
Publication of WO1993013219A1 publication Critical patent/WO1993013219A1/fr

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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
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • 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
    • G01N33/84—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving inorganic compounds or pH
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/916—Hydrolases (3) acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
    • G01N2333/918—Carboxylic ester hydrolases (3.1.1)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2405/00—Assays, e.g. immunoassays or enzyme assays, involving lipids
    • G01N2405/04—Phospholipids, i.e. phosphoglycerides

Definitions

  • the present invention relates to a composition for measuring calcium ions. More particularly calcium I O emissions measured composition having Complex Li phospholipids, phospholipase A 2 and lysolecithin measuring reagent.
  • Conventional technology having Complex Li phospholipids, phospholipase A 2 and lysolecithin measuring reagent.
  • Measurement of calcium in body fluids provides useful information on various physiological functions such as nerve and muscle excitability, blood coagulation reaction, cell membrane permeability, and parathyroid function, and has high clinical significance.
  • methods such as an electrode method using an ion-selective electrode have been put to practical use and generally used for measuring calcium ions in body fluids.
  • this method requires a dedicated device, and therefore requires attention to the maintenance and management of the device, and has problems such as poor sample analysis processing efficiency.
  • An object of the present invention is to provide a simple calcium ion measuring reagent which does not require maintenance and management of a special measuring device, has excellent measurement accuracy, and gives accurate analysis results even with a high-value calcium sample.
  • the present invention is a phospholipid, calcium I O emissions measured composition characterized by containing the phospholipase A 2 and 1 / Zoreshichin measuring reagent.
  • the phospholipid used in the present invention is represented by the following general formula.
  • Rz represents an acyl group having 6 to 22 carbon atoms
  • X represents collin or ethanolamine.
  • this phospholipid examples include natural lecithins such as egg yolk lecithin and soy lecithin, giorail lecithin, dipalmitoyl lecithin, distearoyl lecithin, zimiris toy / lelecithin, zirino leis relecithin, zirino leioinolecithin, I—Palmi Toinole 1—Oleyl Lecithin, 1— ⁇ Lumi Toilet 1—Lino Rail Lecithin, 1—Palmi Toileu 2—Lino Leisleine, 1—Stear Mouth 2—Oleino Lecithin, 1—Stearoyl 2—Lino Lele Lecithin And synthetic lecithin.
  • natural lecithins such as egg yolk lecithin and soy lecithin, giorail lecithin, dipalmitoyl lecithin, distearoyl lecithin, zimiris toy /
  • the reagent for measuring lysolecithin is composed of a combination of enzymes and reagents for detecting lysolecithin, which is a reaction product of phospholipase and the phospholipid represented by the above general formula. That is, at least lysophospholipase, glycerylphosphorylcholine phosphogen
  • the amount of hydrogen peroxide produced by the reaction using a lysolecithin measurement reagent consisting of a sterase and glycerophosphate oxidase or choline oxidase'- or ethanolamine oxidase can be determined.
  • glycerylphosphorylcholine phosphodiesterase is used as a substrate when X is ethanolamine in the above general formula, so that the reaction proceeds even when glyceryl phosphorylethanolamine is used.
  • the bi venom, bee venom forces the enzyme of origin such as the present the animal ⁇ it is known maximum activation with calcium I O emissions lightly-doped region It is known to happen.
  • phosphorylase Horipaze eight 2 from scan Torebutomaisesu is activated in proportion to the concentration to a relatively high concentration of calcium I-one as shown in FIG. 1 can be seen.
  • Reputomaisesu derived phospholipase eight 2 (Toyo Jozo Co., catalog T one 3 1) is advantageous, also Re Yu calcium I O emissions measured composition of the present invention this point It is a point.
  • the source of lysophospholipase as a reagent for measuring lysolecithin is not particularly limited, but an enzyme obtained from animals, plants, and microorganisms and which does not act on lecithin is used.
  • Suitable enzymes derived from microorganisms include enzymes derived from the genus Vibrio (Catalog T-32, manufactured by Toyo Brewing Co., Ltd.). Glyceryl phosphorylco
  • the origin of the phosphodiesterase is not particularly limited, and enzymes derived from, animals and microorganisms can be used. Similarly, glycerophosphate
  • the origin of the enzyme, choline oxidase and ethanolamine oxidase is not limited at all, and enzymes derived from microorganisms can be preferably used.For these various enzymes, commercially available enzymes are used. That It is simple.
  • any buffer solution such as a phosphate buffer, an ethanol-lamine buffer, a borate buffer, and various good buffers can be used in the composition for measuring calcium ion of the present invention. What is the pH of the buffer? ⁇ 8 is preferred.
  • composition for measuring calcium ion of the present invention phospholipids 2 to 5 mM, phospholipase 21 to; L0 UZml, lysophospholipase 0.5 to 5 UZml, glycerylphosphorylcholine phosphodiesterase 0.05 ⁇ 1 U / m 1, glycemic acid phosphate 2 ⁇ 201 1 111, choline oxide 2 ⁇ 20 U / m 1, ethano-lamine oxidase 2 ⁇ 20 U / m ⁇ , Peroxidase 1 to 10 U / m 1, 4-aminoantipyrine 0.02% to 0.2%, phenol or aniline derivative 0.02% to 0.2% .
  • a surfactant for example, a nonionic surfactant such as 0.1 to 0.5% of Triton X—100, P It is preferable to have a preservative, a stabilizing agent, etc., and to easily disperse the phospholipid in the composition for measurement by ultrasonic treatment, or to disperse easily by using a surfactant in combination. I just need.
  • body fluids such as blood, serum, and urine.
  • the measurement of hydrogen peroxide is carried out by a conventional method, which is described in, for example, Japanese Patent Publication No. 2-319600.
  • Reagents may be divided into two or more and appropriately combined. Specifically, calcium ion measurement consisting of the above composition Add a serum sample of 100 to 201 to the reaction solution containing the composition for use, and then react for 5 to 10 minutes at 37 ° C. In the case of a color reaction, the absorbance at 500 to 550 nm may be measured.
  • composition for measuring calcium ion of the present invention has good fractionation efficiency, excellent measurement accuracy, good linearity, and can measure accurately and easily up to a high concentration region.
  • this measurement reagent is applicable to continuous measurement using an automatic analyzer often used in clinical laboratories.
  • Figure 1 shows the results of measurement of enzymes derived from Streptomyces and bee venom.
  • FIG. 2 shows the measurement results using the reagent in Example 1 of the present invention.
  • FIG. 3 shows a measurement result using a reagent in Example 3 of the present invention.
  • FIG. 4 shows the measurement results using the reagent in Example 4 of the present invention.
  • the calcium ion concentration was measured by the following measurement method using the following reagents.
  • Human serum was used as a test solution.
  • the reagents and the measurement method were measured under the same conditions as in Example 1. Incidentally, it issued Ki serum calcium ⁇ beam concentration 1 mMC a CI Z as a standard solution. The results are shown in Table 1.
  • test solution 0.5 mM CaCl 2
  • Reagent 1 having the above composition
  • the increase in absorbance at 0 nm was measured continuously with a spectrophotometer. The results are shown in FIG. Example 4
  • the calcium ion concentration was measured by the following measurement method using the following reagent.
  • composition for measuring calcium ion having the following composition
  • composition for measuring calcium ion having the following composition
  • a composition for measuring calcium ion having the following composition.

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  • 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)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Microbiology (AREA)
  • Urology & Nephrology (AREA)
  • Analytical Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Inorganic Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biophysics (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

Composition de détermination d'ions calcium, contenant un phospholipide, une phospholipase A2, et un réactif servant à déterminer une lysolécithine en tant que composants pour déterminer la teneur en ions calcium d'un échantillon biologique. La composition facilite la détermination et donne des résultats analytiques précis même avec un échantillon présentant une teneur élevée en calcium.
PCT/JP1992/001680 1991-12-24 1992-12-22 Composition de determination d'ions calcium WO1993013219A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP3/340702 1991-12-24
JP34070291A JPH05168498A (ja) 1991-12-24 1991-12-24 カルシウムイオン測定用組成物

Publications (1)

Publication Number Publication Date
WO1993013219A1 true WO1993013219A1 (fr) 1993-07-08

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Application Number Title Priority Date Filing Date
PCT/JP1992/001680 WO1993013219A1 (fr) 1991-12-24 1992-12-22 Composition de determination d'ions calcium

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JP (1) JPH05168498A (fr)
WO (1) WO1993013219A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997028270A1 (fr) * 1996-02-02 1997-08-07 Biomolecular Products, Inc. Procedes de preparation de lysophosphatidylcholine
US5891466A (en) * 1990-08-13 1999-04-06 Yesair; David W. Mixed Liped-Bicarbonate colloidal particles for delivering drugs or calories

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4969920B2 (ja) * 2006-06-02 2012-07-04 三菱化学メディエンス株式会社 カルシウム測定試薬及び測定方法
JP6063207B2 (ja) * 2012-10-24 2017-01-18 国立大学法人福島大学 グリセロール−３−ホスホコリン（ｇｐｃ）などのグリセロール−３−ホスホジエステルに対して加水分解作用を有する酵素及びその製造方法
JP6080254B2 (ja) * 2012-12-07 2017-02-15 国立大学法人福島大学 グリセロール−３−ホスホエタノールアミン（ｇｐｅ）に対して加水分解作用を有する酵素及びその製造方法
DE112017003142T8 (de) 2016-06-22 2019-08-08 Asahi Kasei Pharma Corporation Messung der Aktivität der LP-PLA2

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ANAL BIOCHEM, Vol. 164, No. 1, (1987), KASURINEN J. et al., "An Enzymatic Colorimetric Assay of Calciumdependent Phospholipases A", p. 96-101. *
TEST AND TECHNOLOGY, Vol. 19, No. 6, (1991), YUZO KAYAMORI et al., "Measurement of Electrolyte by Enzymic Method Focused on Cholor, Magnesium, Calcium". *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5891466A (en) * 1990-08-13 1999-04-06 Yesair; David W. Mixed Liped-Bicarbonate colloidal particles for delivering drugs or calories
WO1997028270A1 (fr) * 1996-02-02 1997-08-07 Biomolecular Products, Inc. Procedes de preparation de lysophosphatidylcholine

Also Published As

Publication number Publication date
JPH05168498A (ja) 1993-07-02

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