UPLC-MS/MS同时测定配合饲料中6种水溶性维生素的含量

Simultaneous determination of the 6 water soluble vitamins in feed by ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS

建立了UPLC-MS/MS同时测定配合饲料中6种水溶性维生素（VB1、VB2、VB3、VB6、VB7、VB12）的方法。饲料样品中水溶性维生素用乙酸铵水溶液（p H=4.0）超声提取,吸取上清液过膜后,供UPLC-MS/MS检测。各待测物用Capcell PAK ADME（2.1 mm I.D×150 mm,3 um）色谱柱进行分离,采用50 mmol乙酸铵-甲醇作流动相,梯度洗脱,串联质谱采用电喷雾离子源正模式,在多反应监测（MRM）模式下对6种水溶性维生素进行测定。结果表明,6种维生素在1~20μg/l范围内线性关系良好,相关系数为0.996~0.999。空白加标浓度为10~1 000μg/kg,这6种水溶性维生素的加标回收率为83.5%~106.4%,相对标准偏差（n=6）为3.5%~9.1%,上述饲料中6种维生素的检出限为0.05~1.97μg/kg,定量下限为0.16~6.5μg/kg。方法快速准确、经济实用,符合法规要求,可满足日常检测的需要。

The research was conducted to establish an analytical method for simultaneous determina-tion of the 6 water soluble vitamins in premix by ultra high performance liquid chromatography-tan-dem mass spectrometry（UPLC- MS/MS）, including thiamine（VB1）, riboflavin（VB2）, niacin（VB3）, pyri-doxine（VB6）, biotin（VB7） and cyanocobalamin（VB12）. The samples were extracted with ammonium ace-tate solution（p H=4.0） and detected by positive electrospray ionization and multiple reaction monitor-ing（MRM）. The six water vitamins were analysed using 50 mmol ammonium acetate-methanol as mo-bile phase, and eluted gradient by tandem mass spectrometry after separating by Capcell PAK ADME（2.1 mm I.D×150 mm,3 μm） chromatographic column. The results showed that： when the concentra-tion of 6 water soluble vitamins ranged from 1 to 20 μg/l, the correlation coefficient is 0.996 to0.999. The average recoveries of the six water vitamins at 10~1 000 μg/kg ranged from 83.5% to106.4% with the relative standard deviations（n=6） from 3.5% to 9.1%. The limits of detection werefrom 0.05 to 1.97 μg/kg, and the limits of quantification were from 0.16 to 6.50 μg/kg. The methodcan meet the needs of daily inspection with rapid accurate, economical and practical, conform to the re-quirements of the regulations.