高效液相色谱法测定混菌发酵乳中乳糖、葡萄糖及半乳糖含量的变化

Changes of lactose,glucose and galactose contents in mixed fermented milk detected by HPLC method

利用高效液相色谱法测定混菌发酵乳中乳糖、葡萄糖和半乳糖含量及其变化。样品经TCA沉淀蛋白,采用AminexHPX-87C色谱柱,超纯水为流动相,流速0.3mL/min,使用示差折光检测器进行检测。结果表明:利用该方法进行色谱分析,各组分能够达到很好的分离,各糖组分在0.5～10mg/mL范围内呈现良好的线性关系,相关系数均达到0.9999以上,检出限为0.012～0.016mg/mL,样品加标平均回收率为91.30%～103.28%,精密度实验表明,相对标准偏差在0.49%～3.82%之间(n=6),该法用于实际样品的测定可靠、准确。

A high performance liquid chromatograph （HPLC） with refractive index detector for the determination of lactose,glucose and galactose and its changes in mixed fermented milk was established.Proteins from samples were precipitated with TCA reagent. The chromatographic separation was achieved on an Aminex HPX-87C column using ultrapure water as mobile phase at a flow rate of 0.3mL/min.The results demonstrated that good separation was obtained for the analytes. The calibration curves established showed good linearity over the concentration range of 0.5- 10mg/mL.The correlation coefficient all reached more than 0.9999 and the detection limit varied from 0.012mg/mL to 0.016 mg/mL.The average recoveries for all three analytes ranged from 9].30% to 103.28% with a RSD between 0.49% and 3.82% （ n = 6） .The method was applied to determine the sugars in fermented milk was reliable and accurate.