高效液相色谱法测定奶粉及其他乳制品中的L-羟脯氨酸

Determination of hydroxyproline in milk powder and other dairy products by HPLC

建立了柱前衍生反相高效液相色谱法测定奶粉及其他乳制品中L-羟脯氨酸的方法。采用酸水解样品,经6-氨基喹啉基-N-羟基琥珀酰亚氨基甲酸酯(AQC试剂)衍生水解样品中的L-羟脯氨酸,以磷酸盐缓冲溶液、乙腈、水梯度洗脱,流速1.0ml/min,通过symmetry C18柱将L-羟脯氨酸与其他18种氨基酸分离,紫外检测器在248nm波长下检测。测定结果显示,在0～2.05mmol/L范围内,标准曲线呈良好的线性关系(r=0.9991),加样回收率为95.1%～101%,方法精密度RSD(n=7)为1.3%。该方法具有简单、稳定、灵敏度高、准确等优点。

The method of determining hydroxyproline （Hyp） in milk powder and other dairy products was developed by precolumn derivation RP-HPLC. After acid hydrolysis,Hyp in samples was derived with AQC. A gradient elution was used to separate Hyp from other 18 amino acids by symmetry C18 column. And Hyp was determined with PDA detector at 248.0 nm. The mobile phase was phosphate buffered solution -water-acetonitrile. Good linearity was observed within the range from 0 to 2.05 mmol/L （r=0.99991）. The recoveries were 95.1 - 101%, and the reproducibility（n=7） was 1.3%. The method was simple, steady, sensitive and accurate.