Objective:To establish an HPLC method for the determination of content and content uniformity of vitamin B12 tablets. Method:The HPLC method was conducted on a C18 column. The mobile phase was 0. 02 mol·L-1 disodium hydrogen phosphate solu-tion-methanol(74∶26). The flow rate was 1. 0 ml·min-1 and the detection wavelength was 225nm. The column temperature was 30℃and the injection volume was 20 μl. Result: The linearity of vitamin B12 was good within the range of 1. 67-41. 72 μg·ml-1 ( r=1. 000 0). The average recovery was 99. 9%(RSD=0. 40%,n=9). Conclusion:The method is accurate,reliable and reproducible, and can be used in the determination of content and content uniformity of vitamin B12 tablets.