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目的建立柱前衍生反相高效液相色谱测定鲜乳及乳粉中L-羟脯氨酸方法。方法样品经酸水解,采用异硫氰酸苯酯(PITC)与水解液中的L-羟脯氨酸衍生,以C18柱分离,紫外检测器254 nm波长下检测。结果在2～10 mg/L范围内,L-羟脯氨酸质量浓度与峰面积具有良好的线性关系,样品加标平均回收率为95.0%～100.3%,方法精密度为0.4%～4.0%。鲜乳的检出限为2.0 mg/kg,乳粉的检出限为14 mg/kg。结论该方法易于操作、稳定、准确,适用于鲜乳及乳粉中L-羟脯氨酸检测。
OBJECTIVE To establish a method for the determination of L-hydroxyproline in fresh milk and milk powder by precolumn derivatization reversed-phase high performance liquid chromatography. Methods The samples were acid-hydrolyzed and derivatized with phenyl isothiocyanate (PITC) and L-hydroxyproline in the hydrolyzate. The samples were separated on a C18 column and detected by UV detector at 254 nm. Results There was a good linear relationship between the concentration of L-hydroxyproline and the peak area in the range of 2 ~ 10 mg / L. The average recoveries of spiked samples ranged from 95.0% to 100.3% with the precision of 0.4% -4.0% . The detection limit of fresh milk is 2.0 mg / kg and the detection limit of milk powder is 14 mg / kg. Conclusion The method is easy to operate, stable and accurate. It is suitable for the detection of L-hydroxyproline in fresh milk and milk powder.