Abstract: 【Objective】 The content of vitamin B₁ in mosquito repellent cream comparatively determine by HPLC and UV spectrophotometry. 【Methods】 The detection wavelength of UV spectrophotometry 246 nm, and the content of vitamin B₁ calculated by the method of percentile absorption coefficient; the detection wavelength of HPLC 267 nm, chromatographic column C18. 【Results】 The results showed that the UV method good linear relationship in the 7.47~14.94 μg/ml range (r=0.999,3), RSD of precision experiment 0.30%, RSD of repetitive experiment 1.04%, the average recovery rate was 105.13%, RSD1.72%, and the sample was stabilized within 30 minutes after treatment (RSD=0.91%); the HPLC method good linear relationship in the 4.85~11.31 μg/ml range (r=0.999,9), RSD of precision experiment 0.28%, RSD of repetitive experiment 0.90%, the average recovery rate was 101.77%, RSD1.93%, and the sample was stabilized within 12 hours after treatment (RSD=0.39%). 【Conclusion】 The average contents of three batches of samples determined by HPLC method were 8.39, 8.39 and 8.45 mg/g, respectively. The average contents of the three batches of samples were 8.73, 8.70 and 8.77 mg/g. The precision and repeatability of UV and HPLC method both good, but the content and recovery of UV method were higher than of labeled amount, the accuracy of determination of UV was not as good as that of HPLC method. Furthmore, the stability of UV method was poor. Therefore, HPLC method more suitable for determining the content of vitamin B₁ in mosquito repellent cream.