超高效液相色谱-高分辨质谱定量检测母乳中14种低聚糖  

Quantification of 14 Human Milk Oligosaccharides by Ultra-High Performance Liquid Chromatography Coupled with High-Resolution Mass Spectrometry

作　　者：郭敏[1] 杨芹 魏小晶 王琳琳[1] 田丰伟[1] 赵建新[1] GUO Min;YANG Qin;WEI Xiaojing;WANG Linlin;TIAN Fengwei;ZHAO Jianxin(School of Food Science and Technology,Jiangnan University,Wuxi 214122,China;Wuxi School of Medicine,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学食品学院,江苏无锡214122 [2]江南大学无锡医学院,江苏无锡214122

出　　处：《食品与生物技术学报》2024年第12期154-163,共10页Journal of Food Science and Biotechnology

基　　金：国家“十四五”重点研发计划项目(2022YFF1100400)。

摘　　要：【目的】建立了超高效液相色谱-高分辨质谱定量检测母乳中14种母乳低聚糖的方法。【方法】样品脱脂后离心取上清液,经Waters ACQUITY UPLC®BEH Amide色谱柱(2.1 mm×100 mm×1.7μm)分离,以含25 mmol/L乙酸铵水溶液和乙腈作为流动相进行梯度洗脱,在正离子模式下,通过平行反应监测模式(parallel reaction monitoring,PRM)采集数据,外标法定量。【结果】14种母乳低聚糖在质量浓度为0.10~100.00μg/mL时线性关系良好,相关系数≥0.9802,检出限为0.05~1.00μg/mL,定量限为0.10~5.00μg/mL,回收率为78.03%~105.23%,相对标准偏差为0.14%~5.86%。【结论】该方法可以快速、有效地检测不同来源母乳样品中母乳低聚糖,为母乳低聚糖的研究提供技术支持。[Objective]To establish a method for quantifying 14 human milk oligosaccharides based on ultra-high performance liquid chromatography coupled with high-resolution mass spectrometry.[Method]The supernatant obtained by centrifugation of defatted human milk was loaded on an Waters ACQUITY UPLC®BEH Amide column(2.1 mm×100 mm×1.7μm)for gradient elution,with 25 mmol/L ammonium acetate solution and acetonitrile as mobile phases.Data of human milk oligosaccharides were collected by parallel reaction monitoring in the positive ion mode.Quantitative analysis was performed with the external standard method.[Result]The calibration curves of 14 human milk oligosaccharides were linear within the mass concentration range of 0.10~100.00μg/mL,with the correlation coefficient≥0.9802.The limits of detection and limits of quantitation were 0.05~1.00μg/mL and 0.10~5.00μg/mL,respectively.The recovery of each oligosaccharide added to human milk ranged from 78.03%to 105.23%,with the relative standard deviations ranging from 0.14%to 5.86%.The established method was then employed to quantify human milk oligosaccharides in the samples from different ethnic groups.[Conclusion]The results showed that this method could rapidly and effectively detect human milk oligosaccharides in the samples from different sources,providing technical support for the research on human milk oligosaccharides.

关 键 词：母乳 母乳低聚糖 平行反应监测 

分 类 号：R28[医药卫生—中药学]