OBJECTIVE To establish a HPLC method for the simultaneous determination of Vitamin B₁， Vitamin C， Vitamin B₆ and niacinamide in Vitamins with Minerals Tablets（29）. METHODS An Alltima C₁₈ column （4.6 mm×250 mm， 5 μm） was used for the separation with 50 mmol·L-1 ammonium dihydrogen phosphate （adjusted pH to 3.0 with H₃PO₄） and acetonitrile （95∶5） as a mobile phase at a flow rate of 0.5 mL·min-1. The detection wavelength was 275 nm. RESULTS The method showed good linearity for Vitamin B₁， C， B₆ and niacinamide with all correlation coefficients （r） greater than 0.999 7. The specificity study showed satisfactory resolutions between Vitamin B₁， C， B₆， niacinamide， other ingredients and forced degradation products. The precisions were satisfactory with all relative standard deviations （RSD） lower than 2.1%. The spiked recovery for four compounds ranged from 98.82% to 100.3%. The stability were acceptable with a 4 h stable duration for Vitamin C and a method comparison indicated coincident RESULTS for Vitamin C between this method and standard method. CONCLUSION This method is an accurate， fast and simple for the simultaneous determination of Vitamin B₁， C， B₆ and niacinamide in Vitamins with Minerals Tablets（29）.