超高压液相色谱-串联质谱法同时测定人乳中的硫胺素、核黄素、烟酰胺、泛酸和吡哆醛

Determination of thiamin, riboflavin, nicotinamide, pantothenic acid and pyridoxal in human milk by UPLC-MS/MS

中文关键词:  B族水溶性维生素  人乳  同位素稀释  超高压液质联用

英文关键词:water-soluble vitamins B  human milk  stable isotope dilution  ultra pressure liquid chromatography-mass spectrometry/mass spectrometry

中文摘要:

      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0.2 μg/100 g, 维生素B3(烟酰胺)和维生素B5(泛酸)的LOQ均为0.3 μg/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。

英文摘要:

      Objective To establish a rapid and sensitive quantitative method of several B-vitamins in human milk byultra-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 spectrometry 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%~901.1%. 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.