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Survey of several methods deproteinizing human plasma before and within the chloroformate-mediated treatment of amino/carboxylic acids quantitated by gas chromatography.
MedLine Citation:
PMID:  22633606     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Trichloroacetic acid, perchloric acid, phosphotungstic acid, acetonitrile, methanol and some other organic solvents were evaluated for their ability to provide protein and lipid-free plasma supernatants. The residual proteins, total cholesterol and triacylglycerols were assayed in the supernatant on a Beckman Analyzer instrument. The free cholesterol and the neutral lipids were further analyzed by means of high-temperature GC analysis. The conditions for the deproteinizing step were optimized for minimal lipoprotein disruption. A substantial difference regarding contamination by the lipids was found if the plasma supernatant or the whole serum were treated with an alkyl chloroformate reagent. Three plasma sulfur amino acids and the aromatic ones were chosen as model compounds to evaluate compatibility of the precipitation methods with a subsequent methyl chloroformate-mediated derivatization and GC-MS analysis. The results of the total homocysteine assay matched well with that obtained using a commercial immunoassay. Precipitation with trichloroacetic acid has proven to be a method of choice for the analysis of the acido-basic analytes by GC-MS via chloroformates.
Authors:
Petr Hušek; Zdeněk Svagera; Dagmar Hanzlíková; Petr Simek
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-5-7
Journal Detail:
Title:  Journal of pharmaceutical and biomedical analysis     Volume:  -     ISSN:  1873-264X     ISO Abbreviation:  -     Publication Date:  2012 May 
Date Detail:
Created Date:  2012-5-28     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8309336     Medline TA:  J Pharm Biomed Anal     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2012 Elsevier B.V. All rights reserved.
Affiliation:
Institute of Clinical Biochemistry, Faculty Hospital, 17. listopadu 1790, CZ-70852 Ostrava, Czech Republic; Biology Centre, Czech Academy of Sciences, Branišovská 31, CZ-370 05 České Budějovice, Czech Republic.
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