亲水色谱-超高效液相-串联质谱法测定滇橄榄中水解氨基酸研究

Determination of Hydrolyzed Amino Acids by Hydrophilic Chromatography-Ultra high performance liquid-tandem mass spectrometry in Phyllanthus emblica Linn.

目的:建立滇橄榄中的水解氨基酸的亲水色谱-超高效液相-串联质谱法测定方法。方法:采用盐酸水解滇橄榄中的17种氨基酸,DE-52纤维素除氯离子后,利用亲水色谱柱,结合超高效液相色谱-串联质谱仪,采用正离子模式,多反应监测(MRM),外标法定量,测定滇橄榄中17种水解氨基酸。结果:DE-52纤维素能够有效去除水解溶液中的氯离子,亲水色谱柱对于亮氨酸和异亮氨酸等具有较好的分离效果,在0.0003~2.0μg·mL^-1质量浓度范围内,17种氨基酸的线性关系较好,检出限为0.015~13.0 mg·kg^-1,回收率为73.8%~98.8%,RSD为3.25%~7.79%。结论:该方法重复性好、灵敏度高,适合滇橄榄中17种水解氨基酸的测定。

Objective:To establis a method for the determination of hydrolyzed amino acids in the Phyllanthus emblica Linn.by hydrophilic chromatography-ultra-high performance liquid chromatography-tandem mass spectrometry.Methods:Hydrochloric acid was used to hydrolyze 17 amino acids in the P.emblica Linn.,DE-52 cellulose was dechlorinated,and then a hydrophilic chromatography column was used in combination with ultra performance liquid chromatography-tandem mass spectrometry.Positive ion mode,multiple reactions monitoring(MRM),and external standard method were used for quantify 17 kinds of hydrolyzed amino acids in in the Phyllanthus emblica Linn..Results:The results show that DE-52 cellulose can effectively remove the chloride ions in the hydrolysis solution,and the hydrophilic column has good separation effect for leucine and isoleucine.In the concentration range of 0.0003-2.0μg·mL^-1,the linear relationship of 17 amino acids was good,the detection limit ranged from 0.015 to 13.0 mg·kg^-1,the recovery ranged from 73.8%to 98.8%,and the relative standard deviation(RSD)range was 3.25%-7.79%.Conclusion:This method has good reproducibility and high sensitivity,and is suitable for the determination of 17 kinds of hydrolyzed amino acids in the P.emblica Linn..