Objective: To astablish a strong popularity, high sensitivity of six human milk oligosaccharides in dairy product by precolumn derivatization high performance liquid chromatography-fluorescence method. Methods: Aquatic product sample was precipitated with glacial acetic acid, filtration by using the filter paper. It was derived by 2-aminobenzamide solution. The extracts were centrifuged and filtered by using the organic filter membrane and analyzed by using a AdvanceBio Glycan Map column to be separated and detected，employing acetonitrile-50 mmol/L ammonium formate solution （pH 4.4） as the mobile phase for gradient elution. Results: Under the experimental conditions, six human milk oligosaccharides in dairy product were linear in the range of 1.00～400.0 mg/L, and the correlation coefficients were all over 0.999. The detection limit of the method was 4.1～10.9 mg/kg, the limits of quantification were 12.3～32.7 mg/kg, the mean recoveries rate was ranged from 71.3% to 90.2% with the intra-day precisions （n=6） in the range of 1.0%～6.3% and inter-day precisions （n=6） less than 10.0%. Conclusion: This method can meet the requirements of the daily detection of six human milk oligosaccharides due to its simplicity and sensitives.