采用高效液相色谱-串联质谱法同时测定配合饲料中维生素B1和维生素B2。样品经20 mmol·L-1甲酸铵溶液提取后,采用Tigerkin C8色谱柱分离,以甲醇-20 mmol·L-1甲酸溶液为流动相进行梯度洗脱,采用电喷雾正离子源-选择多反应监测模式检测,外标法定量。维生素B1和维生素B2的质量分数在20.0~500 μg·kg-1范围内与其峰面积呈线性关系,检出限(3S/N)均为50 μg·kg-1。对虾饲料进行加标回收试验,回收率在88.0%~120%之间,测定值的相对标准偏差(n=6)为0.90%~2.7%。方法已应用于市售配合饲料中维生素B1和维生素B2的测定。

A method of HPLC-MS/MS was applied for simultaneous determination of vitamin B1 and vitamin B2 in compound feedstuff. The samples were extracted with 20 mmol·L-1 ammonium formate solution. The extract was then separated on a Tigerkin C8 column, using a mixture of methanol and 20 mmol·L-1 formic acid as mobile phase for gradient elution. ESI+ and MRM were adopted in MS/MS. External standard method was adopted for quantification. The linear relationship between the peak area and the mass ratio of vitamin B1 and vitamin B2 was in the range of 20.0-500 μg·kg-1, with detection limit (3S/N) of 50 μg·kg-1. Recovery rate measured by standard addition method was in the range of 88.0%-120%, with RSD′s (n=6) in the range of 0.90%-2.7%. The method was applied to the analysis of vitamin B1 and vitamin B2 in commercial compound feedstuff.