High Throughput and Quantitative Measurement of Microbial Metabolome by Gas Chromatography/Mass Spectrometry Using Automated Alkyl Chloroformate Derivatization.
- Zhao L Shanghai Key Laboratory of Diabetes Mellitus and Center for Translational Medicine, Shanghai Jiao Tong University Affiliated Sixth People's Hospital , Shanghai 200233, China.
- Ni Y Shanghai Key Laboratory of Diabetes Mellitus and Center for Translational Medicine, Shanghai Jiao Tong University Affiliated Sixth People's Hospital , Shanghai 200233, China.
- Su M Shanghai Key Laboratory of Diabetes Mellitus and Center for Translational Medicine, Shanghai Jiao Tong University Affiliated Sixth People's Hospital , Shanghai 200233, China.
- Li H College of Chemistry and Chemical Engineering, Shanghai University of Engineering Science , Shanghai 201620, China.
- Dong F University of Hawaii Cancer Center , Honolulu, Hawaii 96813, United States.
- Chen W University of Hawaii Cancer Center , Honolulu, Hawaii 96813, United States.
- Wei R University of Hawaii Cancer Center , Honolulu, Hawaii 96813, United States.
- Zhang L University of Hawaii Cancer Center , Honolulu, Hawaii 96813, United States.
- Guiraud SP Nestlé Institute of Health Sciences SA , EPFL Innovation Park, 1015 Lausanne, Switzerland.
- Martin FP Nestlé Institute of Health Sciences SA , EPFL Innovation Park, 1015 Lausanne, Switzerland.
- Rajani C University of Hawaii Cancer Center , Honolulu, Hawaii 96813, United States.
- Xie G University of Hawaii Cancer Center , Honolulu, Hawaii 96813, United States.
- Jia W Shanghai Key Laboratory of Diabetes Mellitus and Center for Translational Medicine, Shanghai Jiao Tong University Affiliated Sixth People's Hospital , Shanghai 200233, China.
- 2017-04-25
Published in:
- Analytical chemistry. - 2017
Automation
Escherichia coli
Feces
Formates
Formic Acid Esters
Gas Chromatography-Mass Spectrometry
Humans
Metabolome
Principal Component Analysis
Reproducibility of Results
Serum
Urine
English
The ability to identify and quantify small molecule metabolites derived from gut microbial-mammalian cometabolism is essential for the understanding of the distinct metabolic functions of the microbiome. To date, analytical protocols that quantitatively measure a complete panel of microbial metabolites in biological samples have not been established but are urgently needed by the microbiome research community. Here, we report an automated high-throughput quantitative method using a gas chromatography/time-of-flight mass spectrometry (GC/TOFMS) platform to simultaneously measure over one hundred microbial metabolites in human serum, urine, feces, and Escherichia coli cell samples within 15 min per sample. A reference library was developed consisting of 145 methyl and ethyl chloroformate (MCF and ECF) derivatized compounds with their mass spectral and retention index information for metabolite identification. These compounds encompass different chemical classes including fatty acids, amino acids, carboxylic acids, hydroxylic acids, and phenolic acids as well as benzoyl and phenyl derivatives, indoles, etc., that are involved in a number of important metabolic pathways. Within an optimized range of concentrations and sample volumes, most derivatives of both reference standards and endogenous metabolites in biological samples exhibited satisfactory linearity (R2 > 0.99), good intrabatch reproducibility, and acceptable stability within 6 days (RSD < 20%). This method was further validated by examination of the analytical variability of 76 paired human serum, urine, and fecal samples as well as quality control samples. Our method involved using high-throughput sample preparation, measurement with automated derivatization, and rapid GC/TOFMS analysis. Both techniques are well suited for microbiome metabolomics studies.
- Language
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- English
- Open access status
- green
- Identifiers
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- DOI 10.1021/acs.analchem.7b00660
- PMID 28437060
- Persistent URL
- https://folia.unifr.ch/global/documents/80341
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