| Determination of lipid hydroperoxides in native low-density lipoprotein by a chemiluminescent flow-injection assay. | |
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MedLine Citation:
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PMID: 8418885 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Lipid hydroperoxides have been implicated in the pathogenesis of atherosclerosis. This work was therefore set up to obtain a fast and specific chemiluminescent assay for measuring hydroperoxides in native low-density lipoprotein (LDL). The apparatus was a complete HPLC system including two pumps, an autosampler, a computer and a chemiluminescent detector with a T-mixing coil in the place of the column. Samples were injected from the autosampler and mixed with luminescent reagent (3 microM luminol and 1 microM microperoxidase in 0.1 M carbonate buffer (pH 10)) in the T-piece. To generate a calibration curve, linoleic acid hydroperoxide was obtained by incubating soybean lipoxygenase with linoleic acid. The calculated conjugated diene concentration was in good agreement with the nominal linoleic acid hydroperoxide concentration. The chemiluminescence was linear with the amount of linoleic acid hydroperoxide injected and the detection limit was about 3 pmol linoleic acid hydroperoxide. The chemiluminescence induced by copper-oxidized LDL was linear with concentration; the detection limit, when compared with linoleic acid hydroperoxide, was similar. The reproducibility of the linoleic acid hydroperoxide and of oxidized LDL hydroperoxide was examined in single pools. The coefficient of variation on the triplicates of each pool was about 3%. The titre of the linoleic acid hydroperoxide and oxidized LDL peroxides was quite stable for at least 10 days when stored under argon at 4 degrees C in the presence of EDTA. The mean value of the LDL hydroperoxides in 16 control subjects was 145.20 +/- 98.81 pmol/mg LDL protein. In conclusion, the microperoxidase-luminol-dependent chemiluminescence flow-injection assay is a rapid, sensitive and selective method for measuring lipid hydroperoxides in native LDL. |
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Authors:
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L Cominacini; A M Pastorino; A McCarthy; M Campagnola; U Garbin; A Davoli; A De Santis; V Lo Cascio |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Biochimica et biophysica acta Volume: 1165 ISSN: 0006-3002 ISO Abbreviation: Biochim. Biophys. Acta Publication Date: 1993 Jan |
Date Detail:
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Created Date: 1993-02-10 Completed Date: 1993-02-10 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0217513 Medline TA: Biochim Biophys Acta Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 279-87 Citation Subset: IM |
Affiliation:
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Istituto di Semeiotica e Nefrologia Medica, Università di Verona, Italy. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Chemiluminescent Measurements* Chromatography, High Pressure Liquid / methods Humans Kinetics Linoleic Acid Linoleic Acids / chemistry Lipid Peroxidation Lipid Peroxides / analysis* Lipoproteins, LDL / blood*, chemistry Lipoxygenase / chemistry Luminol Peroxidases |
| Chemical | |
Reg. No./Substance:
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0/Linoleic Acids; 0/Lipid Peroxides; 0/Lipoproteins, LDL; 2197-37-7/Linoleic Acid; 521-31-3/Luminol; EC 1.11.1.-/Peroxidases; EC 1.11.1.-/microperoxidase; EC 1.13.11.12/Lipoxygenase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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