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Article: A targeted metabolomic protocol for short-chain fatty acids and branched-chain amino acids

TitleA targeted metabolomic protocol for short-chain fatty acids and branched-chain amino acids
Authors
KeywordsBiological samples
Branched-chain amino acids
Gas chromatography-mass spectrometry
Propyl chloroformate
Short-chain fatty acids
Targeted metabolomics
Issue Date2013
Citation
Metabolomics, 2013, v. 9, n. 4, p. 818-827 How to Cite?
AbstractResearch in obesity and metabolic disorders that involve intestinal microbiota demands reliable methods for the precise measurement of the short-chain fatty acids (SCFAs) and branched-chain amino acids (BCAAs) concentration. Here, we report a rapid method of simultaneously determining SCFAs and BCAAs in biological samples using propyl chloroformate (PCF) derivatization followed by gas chromatography-mass spectrometry (GC-MS) analysis. A one-step derivatization using 100 μL of PCF in a reaction system of water, propanol, and pyridine (v/v/v = 8:3:2) at pH 8 provided the optimal derivatization efficiency. The best extraction efficiency of the derivatized products was achieved by a two-step extraction with hexane. The method exhibited good derivatization efficiency and recovery for a wide range of concentrations with a low limit of detection for each compound. The relative standard deviations of all targeted compounds showed good intra- and inter-day (within 7 days) precision (<10 %), and good stability (<20 %) within 4 days at room temperature (23-25 °C), or 7 days when stored at -20 °C. We applied our method to measure SCFA and BCAA levels in fecal samples from rats administrated with different diet. Both univariate and multivariate statistical analysis of the concentrations of these targeted metabolites could differentiate three groups with ethanol intervention and different oils in diet. This method was also successfully employed to determine SCFA and BCAA in the feces, plasma and urine from normal humans, providing important baseline information of the concentrations of these metabolites. This novel metabolic profile study has great potential for translational research. © 2013 Springer Science+Business Media New York.
Persistent Identifierhttp://hdl.handle.net/10722/342450
ISSN
2021 Impact Factor: 4.747
2020 SCImago Journal Rankings: 0.919
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorZheng, Xiaojiao-
dc.contributor.authorQiu, Yunping-
dc.contributor.authorZhong, Wei-
dc.contributor.authorBaxter, Sarah-
dc.contributor.authorSu, Mingming-
dc.contributor.authorLi, Qiong-
dc.contributor.authorXie, Guoxiang-
dc.contributor.authorOre, Brandon M.-
dc.contributor.authorQiao, Shanlei-
dc.contributor.authorSpencer, Melanie D.-
dc.contributor.authorZeisel, Steven H.-
dc.contributor.authorZhou, Zhanxiang-
dc.contributor.authorZhao, Aihua-
dc.contributor.authorJia, Wei-
dc.date.accessioned2024-04-17T07:03:54Z-
dc.date.available2024-04-17T07:03:54Z-
dc.date.issued2013-
dc.identifier.citationMetabolomics, 2013, v. 9, n. 4, p. 818-827-
dc.identifier.issn1573-3882-
dc.identifier.urihttp://hdl.handle.net/10722/342450-
dc.description.abstractResearch in obesity and metabolic disorders that involve intestinal microbiota demands reliable methods for the precise measurement of the short-chain fatty acids (SCFAs) and branched-chain amino acids (BCAAs) concentration. Here, we report a rapid method of simultaneously determining SCFAs and BCAAs in biological samples using propyl chloroformate (PCF) derivatization followed by gas chromatography-mass spectrometry (GC-MS) analysis. A one-step derivatization using 100 μL of PCF in a reaction system of water, propanol, and pyridine (v/v/v = 8:3:2) at pH 8 provided the optimal derivatization efficiency. The best extraction efficiency of the derivatized products was achieved by a two-step extraction with hexane. The method exhibited good derivatization efficiency and recovery for a wide range of concentrations with a low limit of detection for each compound. The relative standard deviations of all targeted compounds showed good intra- and inter-day (within 7 days) precision (<10 %), and good stability (<20 %) within 4 days at room temperature (23-25 °C), or 7 days when stored at -20 °C. We applied our method to measure SCFA and BCAA levels in fecal samples from rats administrated with different diet. Both univariate and multivariate statistical analysis of the concentrations of these targeted metabolites could differentiate three groups with ethanol intervention and different oils in diet. This method was also successfully employed to determine SCFA and BCAA in the feces, plasma and urine from normal humans, providing important baseline information of the concentrations of these metabolites. This novel metabolic profile study has great potential for translational research. © 2013 Springer Science+Business Media New York.-
dc.languageeng-
dc.relation.ispartofMetabolomics-
dc.subjectBiological samples-
dc.subjectBranched-chain amino acids-
dc.subjectGas chromatography-mass spectrometry-
dc.subjectPropyl chloroformate-
dc.subjectShort-chain fatty acids-
dc.subjectTargeted metabolomics-
dc.titleA targeted metabolomic protocol for short-chain fatty acids and branched-chain amino acids-
dc.typeArticle-
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1007/s11306-013-0500-6-
dc.identifier.scopuseid_2-s2.0-84880627110-
dc.identifier.volume9-
dc.identifier.issue4-
dc.identifier.spage818-
dc.identifier.epage827-
dc.identifier.eissn1573-3890-
dc.identifier.isiWOS:000322019100008-

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