柱前衍生—高效液相色谱荧光检测法测定乳制品中6种母乳寡糖

目的：建立一种普及性强、灵敏度高，并能同时测定乳制品中6种母乳寡糖的高效液相色谱—荧光检测分析方法。方法：样品经冰醋酸沉淀，滤纸过滤后，经2-氨基苯甲酰胺溶液衍生，离心后经有机滤膜过滤，用AdvanceBio Glycan Map色谱柱分离，以乙腈-50 mmol/L甲酸铵溶液（pH 4.4）为流动相，梯度洗脱分离，并使用荧光检测器检测。结果：6种母乳寡糖在1.00～400.0 mg/L质量浓度范围内线性关系良好，相关系数均＞0.999，方法检出限为4.1～10.9 mg/kg，回收率为71.3%～90.2%，日内相对标准偏差（RSD）为1.0%～6.3%，日间相对标准偏差＜10.0%。结论：该方法简单、快捷，可有效降低衍生峰的干扰，适用于乳制品中3′-岩藻糖基乳糖、2′-岩藻糖基乳糖、乳糖-N-四糖、乳糖-N-新四糖、3′-唾液乳糖和6′-唾液乳糖6种母乳寡糖的日常检测。

Determination of six human milk oligosaccharides in dairy product by precolumn derivatization high performance liquid chromatography with fluorescence detection

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.