柱前衍生RP-HPLC法测定阿胶中13种氨基酸  

Determination of 13 amino acids in Asini Corii Colla by RP-HPLC with precolumn derivatization

作　　者：陈萍红[1] 王书芳[1] 田守生 龚行楚[1] 张淹 

机构地区：[1]浙江大学药物信息学研究所,浙江杭州310058 [2]国家胶类中药工程技术研究中心,山东聊城252201

出　　处：《中草药》2013年第14期1995-1999,共5页Chinese Traditional and Herbal Drugs

基　　金：国家“重大新药创制”科技重大专项(2011ZX09201-201-10)

摘　　要：目的采用RP-HPLC法同时测定阿胶中13种氨基酸的量。方法阿胶以6 mol/L盐酸于120℃水解3 h,以异硫氰酸苯酯柱前衍生,用反相高效液相色谱法测定氨基酸的量。采用Agilent EclipseAAA色谱柱(75 mm×4.6 mm,3.5μm),以乙腈-110 mmol/L醋酸胺溶液(1∶99)为流动相A、醋酸-乙腈-水(0.05∶80∶20)为流动相B进行梯度洗脱,体积流量0.6 mL/min,柱温42℃,检测波长为254 nm。结果在36 min内可完成对阿胶中13种氨基酸的分离测定,各氨基酸的峰面积与浓度的线性关系良好(r>0.997 7),加样回收率为95.2%～104.4%,RSD<5.0%。结论方法简便、准确,重复性好,可用于阿胶的质量控制,亦可为其他药物中氨基酸的分析提供参考。Objective To establish a RP-HPLC method for the simultaneous determination of 13 amino acids in Asini Corii Colla.Methods Asini Corii Colla was hydrolyzed for 3 h at 120 ℃ using 6 mol/L hydrochloric acid,and the amino acids were determined by RP-HPLC after being derived with phenyl isothiocyanate.HPLC was performed on an Eclipse AAA column（75 mm × 4.6 mm,3.5 μm） with acetonitrile-110 mmol/L ammonium acetate solution（1 ： 99） as mobile phase Aand acetic acid-acetonitrile-water（0.05 ： 80 ： 20） as mobile phase B in the gradient mode at a flow rate of 0.6 mL/min.The column temperature was 42 ℃,and the detection wavelength was set at 254 nm.Results The 13 amino acids were separated within 36 min with a good linearity（r 0.9977） in the range of the test concentration.The average recoveries（n = 3） were 95.2%—104.4% and the RSD values were less than 5.0%.Conclusion The method is simple,accurate,and repeatable,which could be available for the quality control of Asini Corii Colla.It may also serve as a good reference for the determination of amino acids in other pharmaceuticals.

关 键 词：阿胶 衍生化 氨基酸 异硫氰酸苯酯 高效液相色谱 

分 类 号：R286.022[医药卫生—中药学]