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Article: High Throughput and Quantitative Measurement of Microbial Metabolome by Gas Chromatography/Mass Spectrometry Using Automated Alkyl Chloroformate Derivatization

TitleHigh Throughput and Quantitative Measurement of Microbial Metabolome by Gas Chromatography/Mass Spectrometry Using Automated Alkyl Chloroformate Derivatization
Authors
Issue Date2017
Citation
Analytical Chemistry, 2017, v. 89, n. 10, p. 5565-5577 How to Cite?
AbstractThe 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.
Persistent Identifierhttp://hdl.handle.net/10722/342544
ISSN
2023 Impact Factor: 6.7
2023 SCImago Journal Rankings: 1.621
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorZhao, Linjing-
dc.contributor.authorNi, Yan-
dc.contributor.authorSu, Mingming-
dc.contributor.authorLi, Hongsen-
dc.contributor.authorDong, Fangcong-
dc.contributor.authorChen, Wenlian-
dc.contributor.authorWei, Runmin-
dc.contributor.authorZhang, Lulu-
dc.contributor.authorGuiraud, Seu Ping-
dc.contributor.authorMartin, Francois Pierre-
dc.contributor.authorRajani, Cynthia-
dc.contributor.authorXie, Guoxiang-
dc.contributor.authorJia, Wei-
dc.date.accessioned2024-04-17T07:04:34Z-
dc.date.available2024-04-17T07:04:34Z-
dc.date.issued2017-
dc.identifier.citationAnalytical Chemistry, 2017, v. 89, n. 10, p. 5565-5577-
dc.identifier.issn0003-2700-
dc.identifier.urihttp://hdl.handle.net/10722/342544-
dc.description.abstractThe 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.-
dc.languageeng-
dc.relation.ispartofAnalytical Chemistry-
dc.titleHigh Throughput and Quantitative Measurement of Microbial Metabolome by Gas Chromatography/Mass Spectrometry Using Automated Alkyl Chloroformate Derivatization-
dc.typeArticle-
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1021/acs.analchem.7b00660-
dc.identifier.pmid28437060-
dc.identifier.scopuseid_2-s2.0-85020644069-
dc.identifier.volume89-
dc.identifier.issue10-
dc.identifier.spage5565-
dc.identifier.epage5577-
dc.identifier.eissn1520-6882-
dc.identifier.isiWOS:000401674800051-

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