GC-MS with ethyl chloroformate derivatization for comprehensive analysis of metabolites in serum and its application to human uremia |
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Authors: | Xiumei Tao Yumin Liu Yihuang Wang Yunping Qiu Jingchao Lin Aihua Zhao Mingming Su Wei Jia |
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Institution: | (1) School of Pharmacy, Shanghai Jiao Tong University, Shanghai, 200240, China;(2) Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China;(3) Shanghai Institute for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, 200240, China |
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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. |
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Keywords: | Metabolic profiling GC-MS Ethyl chloroformate Serum Multivariate statistical analysis Metabonomics GC Bioanalytical methods Clinical/biomedical analysis |
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