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A novel ultra-performance liquid chromatography electrospray ionization tandem triple quadrupole mass spectrometry method for the simultaneous determination of four water-soluble vitamins, including vitamin B5 (VB5), vitamin B8 (VB8), vitamin B9 (VB9), and vitamin B12 (VB12) in fortified infant foods is developed and validated. A reverse phase UPLC separation system consisting of a Waters ACQUITY UPLC BEH C-18 column (2.1 mm × 100 mm i.d., 1.7 æm) and a binary gradient acetonitrile-water mobile phase is applied for the separation of the four water-soluble vitamins. Formic acid is spiked into the mobile phase to enhance the ionization efficiency. Tandem MS-MS analysis is performed in multi-reaction monitoring mode (MRM). Product-ion traces at  m/z  220.1 → 89.9 for VB5, 245.1 → 227.1 for VB8, 442.3 → 295.2 for VB9, and 678.9 → 147.0 for VB12 are used for quantitation of the corresponding vitamins, and traces at  m/z  455.5 → 308.0 are used for methotrexate (internal standard). Limits of quantitation (LOQs) are 0.016, 0.090, 0.020, and 0.019 µg/L for VB5, VB8, VB9, and VB12, respectively. Intra- and inter-day precisions for the determination of the four vitamins are better than 6.84% and 12.26% in relative standard deviations, and recoveries for the four vitamins are in the range of 86.0∼101.5%. The developed approach is applied for the determination of the trace amounts of the vitamins in fortified milk powers and fortified rice powers.

journal of chromatographic science

Simultaneous Determination of Four Water-Soluble Vitamins in Fortified Infant Foods by Ultra-Performance Liquid Chromatography Coupled with Triple Quadrupole Mass Spectrometry