北京中医药大学学报2017，Vol.40Issue(6)：503-508,6.DOI:10.3969/j.issn.1006-2157.2017.06.010
柱前衍生HPLC-UV同时测定人工牛黄中胆酸及猪去氧胆酸的含量
Simultaneous testing of bile acid and hyodeoxycholic acid in artificial calculus bovis using HPLC-UV with pre-column derivatization
摘要
Abstract
Objective To establish a pre-column derivation of HPLC-UV and determine bile acid and hyodeoxycholic acid in artificial calculus bovis.Methods Bile acid and hyodeoxycholic acid were derived by 2-bromo-2'-acetonaphthoneat using triethylamine as the catalyst in 60 ℃ water bath.The effects of derivatizing agent usage, catalyst and derivatizing time were investigated to identify the optimal parameters.HPLC-UV parameters were established to determine the content of hyodeoxycholic acid in artificial calculus bovis.Results When the ratio of molar amount of derivatizing agents to that of cholic acid and hyodeoxycholic was over 30∶1, and the ratio of molar amount of catalyst to that of cholic acid and hyodeoxycholic ranged from 5∶1 to 60∶1, the reaction was completed in 60℃ water bath for 50 minutes.The linear relationship was found with agent concentration (0.4 ~5.0 μg) in cholic acid (r=0.999 5) and the linear relationship was also present in hyodeoxycholic (0.12~1.5 μg, r=0.999 8).The average recovery rate of bile acid (n=6) was 100% and that of the RSD was 2.40%.The average recovery rate of hyodeoxycholic acid was 101% and that of the RSD was 1.33%.In this sample, the content of cholic acid was 5.55% and the content of hyodeoxycholic was 2.55%.Conclusion The method is replicable, reliable and accurate to determine bile acid and hyodeoxycholic acid in artificial calculus bovis.关键词
人工牛黄/胆酸/猪去氧胆酸/柱前衍生/HPLC-UVKey words
artificial calculus bovis/cholic acid/hyodeoxycholic acid/pre-column derivatization/HPLC-UV分类
医药卫生引用本文复制引用
杨欢,邱仁杰,丁月珠,王雄飞,袁瑞娟..柱前衍生HPLC-UV同时测定人工牛黄中胆酸及猪去氧胆酸的含量[J].北京中医药大学学报,2017,40(6):503-508,6.基金项目
国家自然科学基金项目(No.81403071),北京市自然科学基金(No.7172130),国家级大学生创新训练计划项目(No.201410026033)National natural science foundation of China(No.81403071),Beijing natural science foundation(No.7172130) （No.81403071）
