Simultaneous Determination of Vitamin B1 and Vitamin B2 in Compound Feedstuff by HPLC-MS/MS

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.