Simultaneous determination of nine water-soluble vitamins in pharmaceutical preparations by high-performance liquid chromatography with diode array detection

Abstract

A simple and rapid method for the simultaneous determination of nine water-soluble vitamins was developed by high performance liquid chromatography with diode array detection. The water-soluble vitamins were analyzed by HPLC on a μBondapak C18 column (300 × 3.9 mm, 10 μm) with methanol-0.1 mol/L KH2PO4 buffer (pH 7.0) as mobile phase (1.5 mL/min) in gradient mode as follows: in the range 0-7.0 min, 10:90; and in the range 8.0-15.0 min, 40:60 (v/v). All nine water-soluble vitamins were separated in a single HPLC run within less than 15 min. The linearity of the calibration graphs was compound-dependent with correlation coefficients of 0.9986 to 0.9999 (n = 6). The detection limits ranged from 0.02 μg/mL to 0.5 μg/mL. The method was successfully applied to the determination of vitamins in pharmaceutical preparations. The recoveries were from 95% to 102% and the relative standard deviations were in the range of 0.8% to 4.2%.