Evaluating different extraction solvents for GC-MS based metabolomic analysis of the fecal metabolome of adult and baby giant pandas.pdf (2.33 MB)
Evaluating different extraction solvents for GC-MS based metabolomic analysis of the fecal metabolome of adult and baby giant pandas.
journal contributionposted on 2019-09-04, 11:41 authored by Y Yang, Y Yin, X Chen, C Chen, Y Xia, H Qi, PN Baker, H Zhang, T-L Han
The gut microbiome plays a fundamental role in host health and the fecal metabolome can be analysed to assess microbial activity and can be used as an intermediate phenotype monitoring the host-microbiome relationship. However, there is no established extraction protocol to study the fecal metabolome of giant pandas. The aim of this research is to optimize extraction of the fecal metabolome from adult and baby pandas for high throughput metabolomics analysis using gas chromatography-mass spectrometry (GC-MS). Fecal samples were collected from eight adult pandas and a pair of twin baby pandas. Six different extraction solvents were investigated and evaluated for their reproducibility, metabolite coverage, and extraction efficiency, particularly in relation to the biochemical compound classes such as amino acids, tricarboxylic acid (TCA) cycle intermediates, fatty acids, secondary metabolites, and vitamin and cofactors. Our GC-MS results demonstrated that the extraction solvents with isopropanol: acetonitrile: water (3:2:2 ratio) and 80% methanol were the most appropriate for studying the fecal metabolome of adult and baby giant pandas respectively. These extraction solvents can be used in future study protocols for the analysis of the fecal metabolome in giant pandas.
The authors would like to acknowledge the support of the mass spectrometry center of the Maternal and Fetal Medicine of Chongqing Medical University and the Chongqing Zoo. This research was supported by the National Natural Science Foundation of China (No. 81871185, 81571453, 81771607, 81701477), The 111 Project (Yuwaizhuan (2016)32 The National Key Research and Development Program of Reproductive Health & Major Birth Defects Control and Prevention (2016YFC1000407), Health Commission (2018ZDXM024, 2017ZDXM008), and Chongqing Science & Technology Commission (cstc2017jcyjBX0062).
CitationScientific Reports, 2019, volume 9, Article number: 12017
Author affiliation/Organisation/COLLEGE OF LIFE SCIENCES
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