Abstract:

Objective: To establish a method for the simultaneous determination of 11 B vitamins in infant formula by ultrapressure
liquidchromatography-tandem mass spectrometry (UPLC-MS/MS). Methods: The samples were dissolved in 0.1%
formic acid and then added with xinc acetate to precipitate the proteins. Then the filtrate was separated by an ACQUITY
UPLC HSS T3 column through gradient elution using 0.1% formic acid and acetonitrile as the mobile phase. Mass spectral
acquisition was done in the positive ion mode by applying multiple reaction monitoring and the analysis was completed
in 12 min. Results: High correlation coefficients (R2 > 0.99) for 11 B vitamins in their linear ranges were obtained. The
spiked recovery rates and accuracy (expressed as relative standard deviation, RSD) were determined to be in the ranges of
85%–110% and 1.03%–6.75%, respectively. The limits of quantitation (LOQs) for vitamin B1, vitamin B2, nicotinic acid,
niacinamide, pantothenic acid, pyridoxal, pyridoxine, pyridoxamine, folic acid, vitamin H and vitamin B12 were 40.0, 40.0,
30.0, 30.0, 50.0, 0.9, 1.2, 1.2, 2.0, 2.0, and 0.2 μg/100 g, respectively, an increase by 7.7% to 60% over those from the
national standards of China. Conclusions: The method is characterized by high sensitivity, simple operation, fast analysis and
short time. This method also has good precision and accuracy and can be applied to detect 11 B vitamins in infant formula.

Key words: B vitamins, infant formula milk powder, ultra pressure liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)