Determination Method of Eight Acidic Oligosaccharides in Animals' Intestinal Contents by Liquid Chromatography-Mass Spectrometry

XU Liwei; SUN Dehui; GENG Meimei; CHEN Wen; HE Zhixiong; WANG Jiurong

doi:10.3969/j.issn.1006-267x.2022.10.067

2022 , Vol. 34 >Issue 10: 6748 - 6757

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.10.067

RESEARCH PAPER

Determination Method of Eight Acidic Oligosaccharides in Animals' Intestinal Contents by Liquid Chromatography-Mass Spectrometry

XU Liwei ,
SUN Dehui ,
GENG Meimei ,
CHEN Wen ,
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: 2022-03-24

Online published: 2022-10-17

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Abstract

The purpose of this experiment was to establish a method based on liquid chromatography-mass spectrometry to determine the eight acidic oligosaccharides of 3'-salivary lactose (3'-SL), 6'-salivary lactose (6'-SL), 3'-sialyl-N-acetylated lactose (3'-SLN), 6'-sialyl-N-acetylated lactose (6'-SLN), 3'-sialyl-3-fucosyl lactose (3'-S, 3-FL), sialyl lactose-N-tetrasaccharide a (LSTa), sialyl lactose-N-tetrasaccharide c (LSTc) and disasialyl lactose (DSL) in animals' intestinal contents. First, the intestinal contents were freeze-dried and pulverized into powder, then ultrasonic extraction with water, centrifuged, and the supernatant was taken. The procedures repeated once and supernatant was merged. Supernatant was treated with acetonitrile, centrifuged. The supernatant was purified by solid phase extraction column, and the purified liquid was filtered by 0.22 μm membrane for liquid chromatography-mass spectrometry analysis. The eight acidic oligosaccharides were separated by Acquity UPLC^? BEH Amide column within 7 minutes of eight acidic oligosaccharides. The results showed that the relative standard deviations of retention time and peak area of eight target analytes were less than 0.5% and 11.0%. The linear range of 3'-SL, 6'-SLN, LSTa, 3'-S, 3-FL, LSTc, 3'-SLN, DSL and 6'-SL were 5.00 to 2 000.00 ng/mL, 2.50 to 100.00 ng/mL, 9.59 to 94.00 ng/mL, 10.00 to 100.00 ng/mL, 27.80 to 200.00 ng/mL, 4.00 to 80.00 ng/mL, 6.00 to 120.00 ng/mL, 16.10 to 642.00 ng/mL, respectively. The linearity of the eight target analytes was good in the linear range, the correlation coefficients were all above 0.991, the detection limits were ranged from 0.335 to 8.330 ng/mL, and the average recoveries were 77.3% to 122.9%. In conclusion, the method has high selectivity, low sample consumption and simple operation, which can be used for the determination of eight acidic oligosaccharides in contents of different intestinal segments. The established method can provide technical reference for the healthy mechanism study of acidic oligosaccharides in animals.

Key words： intestinal contents; acidic oligosaccharides; liquid chromatography-mass spectrometry

Cite this article

XU Liwei , SUN Dehui , GENG Meimei , CHEN Wen , HE Zhixiong , WANG Jiurong . Determination Method of Eight Acidic Oligosaccharides in Animals' Intestinal Contents by Liquid Chromatography-Mass Spectrometry[J]. Chinese Journal of Animal Nutrition, 2022 , 34(10) : 6748 -6757 . DOI: 10.3969/j.issn.1006-267x.2022.10.067

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