Objective To establish a rapid and sensitive quantitative method of several B-vitamins in hu-man milk by ultra-pressure liquid-chromatography tandem mass spectrometry (UPLC-MS/MS). Methods The non-polar substances and protein were removed from samples by ether and methanol respectively. The sample extracts were separated by a UPLC HSS T3 column. The detection was performed with mass spectro-metry under multiple reaction monitoring (MRM) mode with stable isotopedilution method. Results LOQs of thiamin, riboflavin, pyridoxalnicotinamide and pantothenic acid were between 0.2 and 0.3 µg/100 g. The spiked recoveries were 93.1%~101.1%. The precision RSD were below 6.2%.The lineal range had got high correlation coefficients (R2>0.99). Conclusion A simultaneous quantification of water-soluble vitamin B in human milk was developed by using a UHPLC-MS/MS method. The preparation of sample was simple, rapid with small volume of samples. Five stable isotope standards for each vitamin B were used which showed high precisive and accurate. The developed method is suitable for the study of vitamin B concentration in human milk and can provide the technical support for infant formula.%目的：建立一种快速、灵敏的同时测定人乳中多种B族维生素的超高压液相色谱-串联质谱法。方法样品经甲醇沉淀蛋白、乙醚去除非极性杂质, UPLC HSS T3色谱柱分离后采用保留时间和多反应离子检测(MRM)定性、定量,同位素内标校正。结果维生素B1(硫胺素)、维生素B2(核黄素)和维生素B6(吡哆醛)的定量下限(LOQ)均为0.2µg/100 g,维生素B3(烟酰胺)和维生素B5(泛酸)的LOQ均为0.3µg/100 g。标准加入法测定所有被测物的高、中、低三个浓度加标水平的回收率在93.1%~101.1%之间,相对标准偏差(RSD)在2.8%~6.2%之间。5种水溶性维生素在各自浓度范围内线性良好, R2均大于0.998。结论本文建立了同时测定人乳中5种 B 族水溶性维生素的超高压液相色谱-串联质谱法。方法前处理简单、快速,样品需要量少；采用质谱检测器同位素内标一对一校正定量,检测结果准确、灵敏。方法可以为人乳中维生素含量的检测、调查、研究提供技术支持。
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