GC-MS with ethyl chloroformate derivatization for comprehensive analysis of metabolites in serum and its application to human uremia

Xiumei Tao, Yumin Liu, Yihuang Wang, Yunping Qiu, Jingchao Lin, Aihua Zhao, Mingming Su, Wei Jia

Research output: Contribution to journalArticle

44 Scopus citations

Abstract

An optimized method based on GC-MS with ethyl chloroformate derivatization has been developed for the comprehensive analysis of endogenous metabolites in serum. Twenty-two reference standards and serum samples were used to validate the proposed method. The correlation coefficient was higher than 0.9900 for each of the standards, and the LOD varied from 125 to 300 pg on-column. The analytical equipment exhibited good repeatability (RSD<10%) for all of the standards. Both the repeatability and the within-48-h stability of the analytical method were satisfactory (RSD<10%) for the 18 metabolites identified in the serum samples. Mean recovery was acceptable for the 18 metabolites, ranging from 70% to 120% with RSDs of less than 10%. Using the optimized protocol and a subsequent multivariate statistical technique, complete differentiation was achieved between the metabolic profile of uremic patients and that of age- and sex-matched normal subjects. Significantly decreased levels of valine, leucine, and isoleucine and increased levels of myristic acid and linoleic acid were observed in the patient group. This work demonstrated that this method is suitable for serum-based metabolic profiling studies.

Original languageEnglish (US)
Pages (from-to)2881-2889
Number of pages9
JournalAnalytical and Bioanalytical Chemistry
Volume391
Issue number8
DOIs
StatePublished - Aug 1 2008
Externally publishedYes

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Keywords

  • Bioanalytical methods
  • Clinical/biomedical analysis
  • Ethyl chloroformate
  • GC
  • GC-MS
  • Metabolic profiling
  • Metabonomics
  • Multivariate statistical analysis
  • Serum

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

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