| 气相色谱-质谱联用法测定瘤胃液中的甲酸和其他6种挥发性脂肪酸 |
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| 引用本文： | 许丽卫,耿梅梅,陈闻,张丽萍,贺珍,袁红朝,彭灿,李春勇,贺志雄,王久荣.气相色谱-质谱联用法测定瘤胃液中的甲酸和其他6种挥发性脂肪酸[J].动物营养学报,2021,33(2). |
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| 作者姓名： | 许丽卫 耿梅梅 陈闻 张丽萍 贺珍 袁红朝 彭灿 李春勇 贺志雄 王久荣 |
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| 作者单位： | 中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室,长沙 410125;中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室,长沙 410125;中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室,长沙 410125;中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室,长沙 410125;中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室,长沙 410125;中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室,长沙 410125;中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室,长沙 410125;中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室,长沙 410125;中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室,长沙 410125;中国科学院亚热带农业生态研究所,亚热带农业生态过程重点实验室,长沙 410125 |
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| 基金项目： | 中科院STS项目(KFJ-STS-ZDTP-075)。 |
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| 摘 要： | 本试验旨在建立一种测定瘤胃液中甲酸、乙酸、丙酸、异丁酸、丁酸、异戊酸和戊酸的气相色谱-质谱联用方法。样品选用湘东黑山羊和荷斯坦奶牛的瘤胃液,18000 r/min离心10 min,取上层清液。将上层清液与25%的偏磷酸按照9∶1(v∶v)的比例混合均匀,静置0.5 h后,4℃条件下18000 r/min离心10 min,取上清液过0.22μm滤膜,进行气相色谱-质谱分析。以DB-FFAP毛细管柱进行分离,甲酸、乙酸、丙酸、异丁酸、丁酸、异戊酸和戊酸在9 min内达到基线分离。结果表明:这7种组分色谱峰保留时间的相对标准偏差均小于0.5%,峰面积的相对标准偏差均小于5.0%。甲酸、乙酸、丙酸、异丁酸、丁酸、异戊酸和戊酸分别在0.8~121.0μg/mL、0.3~3081.0μg/mL、0.4~1165.2μg/mL、0.4~275.1μg/mL、0.2~543.0μg/mL、0.5~129.9μg/mL和0.4~271.5μg/mL内线性关系良好,线性相关系数均达到0.998以上,检出限(3倍信噪比)在0.069~0.252μg/mL。山羊和奶牛瘤胃液样品的加标回收率分别为91.8%~103.8%和78.7%~95.2%。本试验建立的测定瘤胃液中甲酸以及其他6种挥发性脂肪酸的气相色谱-质谱联用方法选择性强、样品用量少、操作简单,可为反刍动物瘤胃碳水化合物代谢及甲烷生成的相关研究提供技术支撑。
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| 关 键 词： | 瘤胃液 甲酸 挥发性脂肪酸 气相色谱-质谱联用 |
Determination of Formic Acid and Other Six Volatile Fatty Acids in Rumen Fluid by Gas Chromatography-Mass Spectrometry Method |
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| 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].Acta Zoonutrimenta Sinica,2021,33(2). |
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| Authors: | XU Liwei GENG Meimei CHEN Wen ZHANG Liping HE Zhen YUAN Hongzhao PENG Can LI Chunyong HE Zhixiong WANG Jiurong |
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| Institution: | (Key Laboratory for Agro-Ecological Processes in Subtropical Region,Institute of Subtropical Agriculture,Chinese Academy of Sciences,Changsha 410125,China) |
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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 18000 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 18000 r/min for 10 min under the condition of 4℃.The supernatant was filtered through a 0.22μm 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 121.0μg/mL,0.3 to 3081.0μg/mL,0.4 to 1165.2μg/mL,0.4 to 275.1μg/mL,0.2 to 543.0μg/mL,0.5 to 129.9μg/mL and 0.4 to 271.5μg/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 0.252μg/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. |
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| Keywords: | rumen fluid formic acid volatile acids gas chromatography-mass spectrometry |
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