液相色谱-串联质谱法测定保健品中 L-羟脯氨酸含量

Determination of L-hydroxyproline content in health products by high performance liquid chromatography-tandem mass spectrometry

中文关键词:  LL-羟脯氨酸  保健品  液相色谱-串联质谱法  含量测定

英文关键词:L-hydroxyproline  health care products  liquid chromatography-tandem mass spectrometry  content determination

中文摘要:

      目的 通过优化保健品中L-羟脯氨酸的测定条件, 建立保健品中L-羟脯氨酸的液相色谱-串联质谱(LC-MS/MS)定量检测的分析方法。方法 采用酸水解的方法处理L-羟脯氨酸, 色谱柱(EC-C18, 4.6 mm×50 mm, 2.7 μm), 流动相为水相和乙腈按一定的梯度进行洗脱, 流速0.5 mL/min, 采用LC-MS/MS在正离子模式下检测, 外标法定量。结果 LC-MS/MS法在0.00284 mg/mL~0.0284 mg/mL范围内, L-羟脯氨酸的浓度和峰面积的线性良好, 相关系数R2＞0.99, 方法的检出限和定量限分别为1.42 μg/g和4.7 μg/g, 放置24 h内L-羟脯氨酸的稳定性良好, 在不同添加水平下, 方法的回收率范围为95.0%~98.8%, 相对标准偏差为1.7%(n=9)。结论 高效液相色谱-串联质谱法灵敏度高、准确、重现性好, 适用于保健品中L-羟脯氨酸的含量测定。

英文摘要:

      Objective By optimizing the determination conditions of L-hydroxyproline in health foods, to establish a method for quantitative determination of L-hydroxyproline by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Methods L-hydroxyproline was of acid hydrolysis and chromatographic analysis was performed on column (EC-C18, 4.6 mm×50 mm, 2.7 μm) eluted with an acetonitrile-water flow rate of 0.5 mL/min, which was detected by LC-MS/MSin the positive ion mode and quantified by external standard method. Results The linear relationship between L-hydroxyproline concentration and peak area were good within 0.00284~0.0284 mg/mL range, R2>0.99, the detection limit and the limits of quantification were 1.42 μg/g and 4.7 μg/g, with a good stability of L-hydroxyproline within 24 h, and the recovery of different adding levels was 95.0%~98.8%, and the relative standard deviation was 1.7% (n=9). Conclusion High performance liquid chromatography-tandem mass spectrometry method is sensitive, accurate, reproducible, and applicable to the determination of L-hydroxyproline content in health foods.