利用柱前在线衍生-高效液相色谱法测定培植牛黄及天然牛黄中18种氨基酸的含量  

Determination of 18 Kinds of Amino Acids in Cultured Bezoar and Bovisc Alculus Using Pre-column Derivatization HPLC

作　　者：温浩然 冀艳华 韩星 吕宝俊 汪国鹏 刘洋[1] 唐双喜 WEN Hao-ran;JI Yan-hua;HAN Xing;LYU Bao-jun;WANG Guo-peng;LIU Yang;TANG Shuang-xi(Beijing University of Chinese Medicine,Beijing 102488,China;ShangYaoJu Pharmaceutical Co.,Ltd.,Yinchuan 750004,China;SHIMADZU Corporation,Beijing 100020,China;ZhongCaiHanXi (Beijing) Biotechnology Development Co., Ltd.,Beijing 101503,China)

机构地区：[1]北京中医药大学,北京102488 [2]尚药局制药有限公司,宁夏银川750004 [3]岛津公司,北京100020 [4]中财瀚熙(北京)生物科技发展有限公司,北京101503

出　　处：《中国现代中药》2020年第2期231-236,共6页Modern Chinese Medicine

摘　　要：目的:建立1种利用高效液相色谱同时测定培植牛黄药材中18种氨基酸含量的方法。方法:以邻苯二甲醛(OPA)和9-芴甲基氯甲酸酯(FMOC)为柱前衍生化试剂。色谱柱为AJS-02氨基酸分析专用柱C18(150 mm×4.6 mm,3μm);流动相A为磷酸氢二钠-四硼酸钠缓冲液(pH 8.2),流动相B为甲醇-乙腈-水(9∶9∶2),梯度洗脱;流速为1.6 mL·min^-1;柱温为50℃;检测波长为338 nm(一级氨基酸)和262 nm(二级氨基酸)。结果:各个氨基酸在线性浓度范围内,峰面积与内标比值与浓度呈良好的线性关系(除Lys外,r>0.9990),平均回收率为90%~110%。结论:本方法准确、灵敏、简便,适用于培植牛黄药材中门冬氨酸等18种氨基酸的含量测定。Objective:To establish an HPLC method for the determination of 18 kinds of amino acids in cultured bezoar.Methods:The OPA and FMOC were used as pre-column derivatization reagent.Analysis was performed on a AJS-02 amino acids analysis column(C18,150 mm×4.6 mm,3μm);Using Na2HPO4 and Na2B4O7 buffer solution(pH 8.2)as mobile phase A and methanol-acetonitrile-water(9∶9∶2)as mobile phase B in gradient mode,the flow rate was 1.6 mL·min^-1.The column temperature was maintained at 50℃and the detection wavelength was set at 338 nm(primary amino acid)and 262 nm(secondary amino acid).Results:All calibration curves of eighteen amino acids showed a good linear relationship(except Lys,r>0.9990).the average recoveries were 90%-110%.Conclusion:The method is accurate,sensitive and simple,and proved to be suitable for the quantification of each amino acid in cultured bezoar.

关 键 词：培植牛黄 氨基酸 高效液相色谱法 柱前衍生化 

分 类 号：R284.1[医药卫生—中药学] O657.72[医药卫生—中医学]