动物营养学报 >

2020 , Vol. 32 >Issue 10: 4972 - 4980

DOI: https://doi.org/10.3969/j.issn.1006-267x.2020.10.048

实验方法与实验动物 EXPERIMENTAL METHOD AND ANIMAL

高效液相色谱-串联质谱-多反应监测技术检测猪肠道内容物和粪便中B族维生素含量

  • 陈一波 ,
  • 易宏波 ,
  • 王丽 ,
  • 蒋宗勇
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  • 1. 广东省农业科学院动物科学研究所, 畜禽育种国家重点实验室, 农业部华南动物营养与饲料重点实验室, 广东省畜禽育种与营养研究重点实验室, 岭南现代农业科学与技术广东省实验室茂名分中心, 广州 510640;
    2. 华南农业大学兽医学院, 广州 510642
陈一波(1997-),女,湖南邵阳人,硕士研究生,研究方向为猪营养与肠道健康。E-mail:544055245@qq.com

收稿日期: 2020-04-30

  网络出版日期: 2020-10-22

基金资助

国家重点研发计划项目（2018YFD0501101）；国家自然科学基金项目（31902199）；科技创新战略专项资金（高水平农科院建设）；现代农业产业技术体系建设专项资金（CARS-35）；广东省农业科学院科技计划项目（201601TD）

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Determination of B Vitamins by Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry-Multi-Reaction Monitoring in Intestinal Contents and Feces of Pigs

  • CHEN Yibo ,
  • YI Hongbo ,
  • WANG Li ,
  • JIANG Zongyong
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  • 1. State Key Laboratory of Livestock and Poultry Breeding, Key Laboratory of Animal Nutrition and Feed Science in South China, Ministry of Agriculture and Rural Affairs, Guangdong Key Laboratory of Animal Breeding and Nutrition, Maoming Branch of Guangdong Laboratory for Lingnan Modern Agriculture, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China;
    2. College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China

Received date: 2020-04-30

  Online published: 2020-10-22

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摘要

B族维生素是肠道微生物的重要代谢产物，在肠道发育、免疫功能及营养代谢等方面有重要作用，被作为饲料添加剂广泛应用于畜禽养殖。由于肠道内成分复杂且B族维生素含量极低，现有方法较难准确监测肠道内B族维生素含量的动态变化。本研究利用超高效液相色谱-串联质谱-多反应监测（UPLC-MS/MS-MRM）的靶向代谢组学技术，建立了一种高效快速检测猪肠道内容物和粪便中B族维生素含量的方法，该方法7 min内完成8种B族维生素含量检测，标准曲线线性关系好（r≥0.995），检出下限低（0.01~0.10 ng/mL）。在3个水平添加回收率试验中，每个水平重复8次，其平均回收率在89%~110%，质控样本相对偏差<18%。利用该方法成功检测了仔猪肠道内容物和粪便样品中8种B族维生素含量，维生素B1、维生素B2、烟酸、烟酰胺、吡哆醛、吡哆醇、维生素B9、维生素B12含量在粪便样品中的平均含量分别为806、11 616、19 620、448、576、206、90、18 ng/g，在盲肠内容物中的平均含量分别为246、6 202、15 058、468、238、14、80、13 ng/g。结果显示，粪便中各种B族维生素含量普遍高于盲肠内容物，且烟酸含量远高于其他B族维生素。综上所述，本研究利用ULPC-MS/MS-MRM技术开展靶向代谢组学分析，高效、定量地检测了猪肠道内容物和粪便样品中8种B族维生素的含量，以期为后续畜禽肠道B族维生素功能研究提供技术基础。

关键词： B族维生素; 猪; 肠道内容物; 粪便; 靶向代谢组学

本文引用格式

陈一波 , 易宏波 , 王丽 , 蒋宗勇 . 高效液相色谱-串联质谱-多反应监测技术检测猪肠道内容物和粪便中B族维生素含量[J]. 动物营养学报, 2020 , 32(10) : 4972 -4980 . DOI: 10.3969/j.issn.1006-267x.2020.10.048

Abstract

B vitamin are important metabolites of intestinal microorganisms, and play important roles in the development, immune function and nutritional metabolism of the intestine. It is widely used as a feed additive in livestock and poultry. Due to the complex components and low B vitamin content in the intestinal tract, it is difficult to accurately monitor the dynamic changes of B vitamin in the intestinal tract with the existing methods. In this study, a highly efficient and rapid method for the detection of B vitamin in intestinal contents and feces of pig using the targeted metabolomics technique of ultra performance liquid chromatography-tandem mass spectrometry-multi-reaction monitoring (UPLC-MS/MS-MRM) was established. Eight kinds of B vitamin could be detected in 7 min by this method as well as exhibiting a good linear relationship (r≥0.995) with low detection limit (0.01 to 0.10 ng/mL). The recovery rate was tested at three levels with eight repeats at each level. The results showed that the average recovery rate ranged from 88.0% to 110.0%, with relative standard deviation of <18% in quality control samples. Eight kinds of B vitamin in intestinal contents and feces of pigs were successfully detected by this method. The average contents of vitamin B1, vitamin B2, nicotinic acid, nicotinamide, pyridoxal, pyridoxine, vitamin B9 and vitamin B12 in fecal samples were 806, 11 616, 19 620, 448, 576, 206, 90 and 18 ng/g, respectively. Their average concentrations in cecal contents were 246, 6 202, 15 058, 468, 238, 14, 80 and 13 ng/g, respectively. The results showed that the content of B vitamin in feces was higher than that in cecal contents, and the content of nicotinic acid was much higher than that of other B vitamin. In conclusion, 8 kinds of B vitamin in intestinal contents and feces of pigs are detected efficiently and quantitatively by the targeted metabolomics with ULPC-MS/MS-MRM technology in this study, providing technical basis for the following studies on functions of B vitamin in intestines of livestock and poultry.

Key words： B vitamin; pig; intestinal contents; feces; targeted metabolomics

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