Abstract
A novel liquid chromatography tandem mass spectrometry (LC–MS/MS) method was developed and validated for the determination of four water-soluble vitamins (B1, B2, B6 and B9) in human serum and applied to compare with the electrochemical (EC) method clinically used for routine vitamin determination in patients. Vitamin separation were achieved at 30 °C on a Venusil XBP-C18 column (150 × 4.6 mm, 3 μm, 100A), and eluted with the mobile phase of 5 mM ammonium acetate: methanol (40:60, v/v) at a flow rate of 0.5 mL min−1, and with a run time of 6 min. The established LC–MS/MS method linearity was in the ranges 1–200, 5–40, 1–80, 5–40 ng mL−1 for vitamin B1, B2, B6 and B9, respectively. LC–MS/MS method met the precision and accuracy criteria (RSD% <15%) and the limits of quantitation were 1, 5, 1, 5 ng mL−1 for vitamin B1, B2, B6 and B9, respectively. LC–MS/MS was successfully used for determination of vitamins in human serum, and the method was found to be closely related to the EC method, with coefficients of determination (r 2) of 0.963, 0.953, 0.901 and 0.978 or 95% confidence intervals (CI) of −2.1 to 2.6, −0.29 to 1.00, −1.7 to 2.6, −2.12 to −1.15 ng mL−1 for vitamin B1, B2, B6 and B9, respectively. The clinical application of this automated EC method for determination of vitamin B1, B2, B6 and B9 in human serum may serve as a validated tool for vitamin therapeutic monitoring and for vitamin-related disease diagnosis.
Published Version
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