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

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

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

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

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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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

Cite this article

CHEN Yibo , YI Hongbo , WANG Li , JIANG Zongyong . Determination of B Vitamins by Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry-Multi-Reaction Monitoring in Intestinal Contents and Feces of Pigs[J]. Chinese Journal of Animal Nutrition, 2020 , 32(10) : 4972 -4980 . DOI: 10.3969/j.issn.1006-267x.2020.10.048

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