Survey of several methods deproteinizing human plasma before and within the chloroformate-mediated treatment of amino/carboxylic acids quantitated by gas chromatography

被引：16

作者：

Husek, Petr ^{[1
,2
]}

Svagera, Zdenek ^{[1
,3
]}

Hanzlikova, Dagmar ^{[1
]}

Simek, Petr ^{[2
]}

机构：

[1] Fac Hosp, Inst Clin Biochem, CZ-70852 Ostrava, Czech Republic

[2] Acad Sci Czech Republic, Ctr Biol, CZ-37005 Ceske Budejovice, Czech Republic

[3] Univ Ostrava, Fac Med, CZ-70103 Ostrava 1, Czech Republic

来源：

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS | 2012年 / 67-68卷

关键词：

Plasma (lipo)protein precipitation; Chloroformate-treated supernatant; S-amino acids; GC-MS assay; PROTEIN PRECIPITATION METHODS; ETHYL CHLOROFORMATE; PROTEOMIC ANALYSIS; FATTY-ACIDS; HUMAN SERUM; GC-MS; DERIVATIZATION; REAGENTS; AMINO;

D O I：

10.1016/j.jpba.2012.04.027

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

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. (C) 2012 Elsevier B.V. All rights reserved.

引用

页码：159 / 162

页数：4

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