High-Throughput Determination of Vitamins (B1, B2, B3, B5, B6, B7, and B9) using UPLC-ESI-MS

Background: To prevent vitamin deficiencies or to restore their levels, multi-vitamin formulations are indicated for the elderly, adults, children, and infants. Objective: To provide a valid high-throughput method for simultaneous determination of vitamins in multi-vitamin formulations. Methods: A high-throughput UPLC-ESI-MS method was developed and validated for simultaneous determination of vitamins B1, B2, B3, B5, B6, B7, and B9. Analytes were eluted on an Acquity UPLC® BEH C18 1.7 μm, 2.1 x 50 mm column at 40 ± 5°C. Mobile phase containing acetonitrile (0.1% formic acid) and water (0.1% formic acid) in 30:70% ratio was pumped at 300 μL/min under isocratic control. Protonated ions of vitamin B1, B2, B3, B5, B6, B7, and B9 were monitored in single ion recording mode, using an electrospray ionization probe in positive mode. Results: The m/z ratios of positive ions of vitamin B1, B2, B3, B5, B6, B7, and B9 were 265.1, 377.2, 122.95, 220, 169.98, 244.9, and 442.1, respectively. The calibration curve of different linearity range (ng/ml) was prepared for each vitamin. Linearity range for vitamin B1, B2, B3, B5, B6, B7, and B9 were 60-1000, 25-1000, 75-5000, 30-1000, 30/1000, 25/1000 and 30-900 25-1000 ng/mL, respectively. Coefficient of variation for intra-day and inter-day precision for vitamin B1, B2, B3, B5, B6, B7, and B9 at the middle and higher limit of quantitation were less than 15%. Conclusion: The method was successfully developed and validated, and three different brands of multi-vitamin tablets were assayed for water-soluble vitamins.