| 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 |
Related Documents
:
|
10709616 - Improved hplc determination of acidic opines by phenylisothiocyanate derivatization and... 18963876 - Extraction-spectrophotometric determination of boron and antimony in carbon and low-all... 7710076 - Quantitative lipoxygenase product profiling by gas chromatography negative-ion chemical... 6469896 - Rapid differential pulse polarographic determination of tiaprofenic acid. 23670636 - D-lactic acid biosynthesis from biomass-derived sugars via lactobacillus delbrueckii fe... 17434686 - Induction of neurite outgrowth in pc12 cells by the medium-chain fatty acid octanoic acid. |
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 |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| 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 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Performance characteristics of the Dimension RxL iron and total iron-binding capacity methods.
Next Document: Chronic cold exposure affects the antioxidant defense system in various rat tissues.