Document Detail


A novel colorimetric assay for the determination of lysophosphatidic acid in plasma using an enzymatic cycling method.
MedLine Citation:
PMID:  12809736     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Several methods for measuring concentrations of lysophosphatidic acid (LPA), a lipid mediator, have been reported to date. However, these methods are not routinely used because most of them require specialized instrument and a complicated protocol. METHODS: We developed a novel LPA assay using enzymatic cycling. LPA in a sample is hydrolyzed with lysophospholipase to glycerol-3-phosphate, followed by enzymatic cycling using glycerol-3-phosphate oxidase and glycerol-3-phosphate dehydrogenase. Amplified concentrations of hydrogen peroxides, a product of the enzymatic cycling, are then colorimetrically measured. RESULTS: This method was specific for LPA, being insensitive to the presence of phosphatidic acid or lysophosphatidylcholine. The within-run and between-run CVs were 1.31-1.32% and 0.73-1.03%, respectively. The recoveries of exogenous LPA added to plasma were 100.3-101.6%. In males, LPA concentrations (mean+/-S.D.) of human serum and EDTA-plasma were 0.41+/-0.14 and 0.08+/-0.02 micromol/l, respectively. In females, they were 0.41+/-0.12 and 0.09+/-0.02 micromol/l, respectively. CONCLUSIONS: This novel colorimetric assay for determination of LPA using enzymatic cycling is simple and highly sensitive. It can be used with an automatic analyzer. It may also be useful for further studies of the biological functions of LPA as well as clinical applications in various disorders.
Authors:
Tatsuya Kishimoto; Takeshi Matsuoka; Shigeyuki Imamura; Koji Mizuno
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Clinica chimica acta; international journal of clinical chemistry     Volume:  333     ISSN:  0009-8981     ISO Abbreviation:  Clin. Chim. Acta     Publication Date:  2003 Jul 
Date Detail:
Created Date:  2003-06-17     Completed Date:  2004-02-10     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  1302422     Medline TA:  Clin Chim Acta     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  59-67     Citation Subset:  IM    
Affiliation:
Diagnostic Research and Development Department, R&D Division, Nesco Company, Azwell Inc., 2-24-3 Sho, Osaka 567-0806, Ibaraki, Japan. ta-kishimoto@azwell.co.jp
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MeSH Terms
Descriptor/Qualifier:
Adult
Calibration
Colorimetry / methods
Edetic Acid
Female
Glycerolphosphate Dehydrogenase / metabolism
Glycerophosphates / chemistry,  metabolism
Humans
Lysophospholipase / metabolism
Lysophospholipids / blood*
Male
Middle Aged
Reproducibility of Results
Substrate Cycling
Time Factors
Chemical
Reg. No./Substance:
0/Glycerophosphates; 0/Lysophospholipids; 57-03-4/alpha-glycerophosphoric acid; 60-00-4/Edetic Acid; EC 1.1.-/Glycerolphosphate Dehydrogenase; EC 1.1.3.21/glycerol-3-phosphate oxidase; EC 3.1.1.5/Lysophospholipase

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