Determination of Formic Acid and Other Six Volatile Fatty Acids in Rumen Fluid by Gas Chromatography-Mass Spectrometry Method

XU Liwei; GENG Meimei; CHEN Wen; ZHANG Liping; HE Zhen; YUAN Hongzhao; PENG Can; LI Chunyong; HE Zhixiong; WANG Jiurong

doi:10.3969/j.issn.1006-267x.2021.02.055

2021 , Vol. 33 >Issue 2: 1145 - 1152

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.02.055

Determination of Formic Acid and Other Six Volatile Fatty Acids in Rumen Fluid by Gas Chromatography-Mass Spectrometry Method

XU Liwei ,
GENG Meimei ,
CHEN Wen ,
ZHANG Liping ,
HE Zhen ,
YUAN Hongzhao ,
PENG Can ,
LI Chunyong ,
HE Zhixiong ,
WANG Jiurong

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Key Laboratory for Agro-Ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China

Received date: 2020-07-22

Online published: 2021-02-04

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Abstract

A method based on gas chromatography-mass spectrometry was developed in this experiment to determine the formic acid and other six volatile fatty acids in rumen fluid. Rumen fluid samples were collected from Xiangdong black goats and Holstein dairy cows, respectively. Samples were centrifuged at 18 000 r/min for 10 min, and the supernatant was taken. The supernatant was evenly mixed with 25% metaphosphoric acid at the ratio 9:1(v:v). The supernatant was left for 0.5 h and centrifuged at 18 000 r/min for 10 min under the condition of 4 ℃. The supernatant was filtered through a 0.22 μm membrane and then analyzed by gas chromatography-mass spectrometry. Formic acid and other volatile acids including acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid and valeric acid were separated by DB-FFAP capillary column within 9 min. The results showed that the relative standard derivations of retention time and peak area of the seven analytes were less than 0.5% and 5.0%, respectively. Formic acid, acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid and valeric acid showed good linear relationships at a range of the range of 0.8 to 121.0 μg/mL, 0.3 to 3 081.0 μg/mL, 0.4 to 1 165.2 μg/mL, 0.4 to 275.1 μg/mL, 0.2 to 543.0 μg/mL, 0.5 to 129.9 μg/mL and 0.4 to 271.5 μg/mL, respectively, the linear correlation coefficients of them all more than 0.998, and the detection limits (signal to noise ratio=3) ranged from 0.069 to 0.252 μg/mL. The recovery rates of standard addition of rumen fluid samples of goats and dairy cows were 91.8% to 103.8% and 78.7% to 95.2%, respectively. The gas chromatography-mass spectrometry method developed in this experiment to determine the formic acid and other six volatile fatty acids in rumen fluid is selectivity and easy accessibility, using small number of samples, which can provide technical support for ruminant research on rumen metabolism of carbohydrate and methane generation.

Key words： rumen fluid; formic acid; volatile acids; gas chromatography-mass spectrometry

Cite this article

XU Liwei , GENG Meimei , CHEN Wen , ZHANG Liping , HE Zhen , YUAN Hongzhao , PENG Can , LI Chunyong , HE Zhixiong , WANG Jiurong . Determination of Formic Acid and Other Six Volatile Fatty Acids in Rumen Fluid by Gas Chromatography-Mass Spectrometry Method[J]. Chinese Journal of Animal Nutrition, 2021 , 33(2) : 1145 -1152 . DOI: 10.3969/j.issn.1006-267x.2021.02.055

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