Volume 9 Issue 6
Dec.  2023
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Pinghui Liu, Qinsheng Chen, Lianglong Zhang, Chengcheng Ren, Biru Shi, Jingxian Zhang, Shuaiyao Wang, Ziliang Chen, Qi Wang, Hui Xie, Qingxia Huang, Huiru Tang. Rapid quantification of 50 fatty acids in small amounts of biological samples for population molecular phenotyping. Biophysics Reports, 2023, 9(6): 299-308. doi: 10.52601/bpr.2023.230042
Citation: Pinghui Liu, Qinsheng Chen, Lianglong Zhang, Chengcheng Ren, Biru Shi, Jingxian Zhang, Shuaiyao Wang, Ziliang Chen, Qi Wang, Hui Xie, Qingxia Huang, Huiru Tang. Rapid quantification of 50 fatty acids in small amounts of biological samples for population molecular phenotyping. Biophysics Reports, 2023, 9(6): 299-308. doi: 10.52601/bpr.2023.230042

Rapid quantification of 50 fatty acids in small amounts of biological samples for population molecular phenotyping

doi: 10.52601/bpr.2023.230042
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  • Efficient quantification of fatty-acid (FA) composition (fatty-acidome) in biological samples is crucial for understanding physiology and pathophysiology in large population cohorts. Here, we report a rapid GC-FID/MS method for simultaneous quantification of all FAs in numerous biological matrices. Within eight minutes, this method enabled simultaneous quantification of 50 FAs as fatty-acid methyl esters (FAMEs) in femtomole levels following the efficient transformation of FAs in all lipids including FFAs, cholesterol-esters, glycerides, phospholipids and sphingolipids. The method showed satisfactory inter-day and intra-day precision, stability and linearity (R2 > 0.994) within a concentration range of 2–3 orders of magnitude. FAs were then quantified in typical multiple biological matrices including human biofluids (urine, plasma) and cells, animal intestinal content and tissue samples. We also established a quantitative structure-retention relationship (QSRR) for analytes to accurately predict their retention time and aid their reliable identification. We further developed a novel no-additive retention index (NARI) with endogenous FAMEs reducing inter-batch variations to 15 seconds; such NARI performed better than the alkanes-based classical RI, making meta-analysis possible for data obtained from different batches and platforms. Collectively, this provides an inexpensive high-throughput analytical system for quantitative phenotyping of all FAs in 8-minutes multiple biological matrices in large cohort studies of pathophysiological effects.

  • Pinghui Liu, Qinsheng Chen, Lianglong Zhang, Chengcheng Ren, Biru Shi, Jingxian Zhang, Shuaiyao Wang, Ziliang Chen, Qi Wang, Hui Xie, Qingxia Huang, Huiru Tang declar that they have no conflict of interest.
    Research involving human blood plasma and urine samples from Chinese adult volunteers, methods and data have been approved by the Ethic Committee of Fudan University (Approval No.: FE21087). All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000 (5). Informed consent was obtained from all patients for being included in the study. Rabbit liver tissue and cecum contents of C57BL/6 mice was collected according to the procedures approved by Experimental Animal Ethics Committee, School of Pharmacy, Fudan University (Approval No.: 2018-03-YL-GW-01). All institutional and national guidelines for the care and use of laboratory animals were followed.
    Pinghui Liu and Qinsheng Chen contributed equally to this work.

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