Development and Single-Lab Validation of an UHPLC-APCI-MS/MS Method for Vitamin K1 in Infant Formulas and Other Nutritional Formulas

Accurate analysis of vitamins is essential to help the public maintain adequate intakes of vitamins. Currently in Atlanta Center for Nutrient Analysis (ACNA), AOAC Method 999.15 with fluorescence detection is utilized for the analysis of vitamin K in infant formulas, dietary supplements and other medical foods. An UHPLC-(+)-APCI-MS/MS method for vitamin K1 (phylloquinone) was developed to improve the accuracy, selectivity and eciency of the analysis. SRM1849a and infant formula samples were used to demonstrate that vitamin K1 data by LC-MS/MS analysis matched well with those from the AOAC Method 999.15. A single-laboratory validation of an UHPLC-MS/MS method for vitamin K1 analysis in SRM1849a showed good accuracy with a mean value of 99.6% of the certified value (n = 8). Recoveries of vitamin K1 at two dierent spike levels were 99.6 and 103.7% from SRM1849a. Mean recovery of vitamin K1 from four dierent infant formula samples was 102.4% ranging from 95.6 to 115.5% with %RSD of 7.8 15.6. Precision, measured as repeatability (%RSDr), was 8.7 for SRM1849a and ranged from 3.7 to 13.4 for four infant formula samples. Application of this method will help ACNA facilitate the accurate analysis of vitamin K in infant formulas and other samples.