Keywords: 
• Compound Trivitamin and Calcium Pantothenate Syrup /
• ultra hydrophilic interaction chromatography-mass spectrometry/mass spectrometry /
• thiamine /
• riboflavin /
• pyridoxine /
• nicotinamide /
• calcium pantothenate /
• content determination

Abstract: Objective To establish a ultra hydrophilic interaction chromatography-tandem mass spectrometry (UHILIC-MS/MS) for the simultaneous determination of 5 B-vitamins including thiamine, riboflavin, pyridoxine, nicotinamide and calcium pantothenate in Compound Trivitamin and Calcium Pantothenate Syrup. Methods The chromatographic separation was performed on a Waters ACQUITY UHILIC BEH Amide (2.1 mm×100 mm,1.7 μm) column with the mobile phase consisting of 10 mmol/L ammonium formate water and 90% acetonitrile both containing 0.5% formic acid water in a gradient elution at a flow rate of 0.30 ml/min. An electrospray positive ion multiple reaction monitoring(MRM) was used as a detector. Results Five B-vitamins in the sample achieved excellent linearity calibration curves over the concentration range tested (r>0.998 4) within 5 min. The overall average recovery rates were in the range of 93.27%-100.39% with RSD of 1.41%-4.96%. The contents of thiamine, riboflavin, pyridoxine, nicotinamide and calcium pantothenate in ten samples were 32.40-38.91, 7.002-8.462, 9.677-11.17, 33.64-39.58, 3.276-3.771 mg/250 g, respectively. Conclusion The UHILIC-MS/MS method established in this study could quickly achieve qualitative identification or quantitative detection of five B-vitamins components in Compound Trivitamin and Calcium Pantothenate Syrup, which provided a reliable technical detection method for the development, utilization and quality evaluation of Compound Trivitamin and Calcium Pantothenate Syrup.