护肝片中两种主要结合型胆酸的HPLC-ELSD含量测定
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摘要
目的:建立HPLC-ELSD同时测定护肝片中两种主要结合型胆酸甘氨猪去氧胆酸、甘氨鹅去氧胆酸的含量测定方法。方法:采用Diamonsil C_(18)色谱柱(250 mm×4. 6 mm,5μm),流动相为乙腈-0. 2%甲酸水溶液,梯度洗脱;流速为1. 0 ml·min~(-1); ELSD漂移管温度为60℃,氮气流量为1. 5 L·min~(-1),增益为1。结果:甘氨猪去氧胆酸在26. 52～265. 20μg·ml~(-1)范围内线性关系良好(r=0. 999 8),平均加样回收率为97. 19%,RSD为3. 23%(n=6)。甘氨鹅去氧胆酸在19. 84～198. 40μg·ml~(-1)线性关系良好(r=0. 999 1),平均加样回收率为103. 33%,RSD为4. 00%(n=6)。所检测的16个厂家共30批护肝片均检出两种结合型胆酸。其中,每片护肝片中甘氨猪去氧胆酸的含量在3. 03～6. 10 mg之间,甘氨鹅去氧胆酸的含量在1. 54～3. 39 mg之间,含量变化较大。结论:该法专属性强、准确度高,适合对护肝片中结合型胆酸进行定量分析,对护肝片质量标准提升具有参考价值。
Objective: To establish an HPLC-ELSD method for the simultaneous determination of two major conjugated bile acids glycohyodeoxycholic acid and glycochenodeoxycholic acid in Hugan tablets. Methods: The analysis was performed on a Diamonsil C_(18)column (250 mm × 4. 6 mm,5 μm) with the mobile phase of acetonitrile-0. 2% formic acid with gradient elution. The flow rate was1. 0 ml·min~(-1). The temperature of the ELSD drift tube was 60℃,the flow rate of nitrogen was 1. 5 L·min~(-1),and the gain was 1.Results: The calibration curves of glycohyodeoxycholic acid and glycochenodeoxycholic acid were linear within the range of 26. 52-265. 20 μg·ml~(-1) (r = 0. 999 8) and 19. 84~(-1)98. 40 μg·ml~(-1) (r = 0. 999 1),respectively. The average recoveries were 97. 19% and 103. 33%,and the RSDs were 3. 23% and 4. 00% (n = 6),respectively. The two kinds of conjugated bile acids were detected in 30 batches of Hugan tablets from 16 manufacturers. Among them,the content of glycohyodeoxycholic acid in each tablet was between 3. 03 mg and 6. 10 mg,and the content of glycochenodeoxycholic acid was between 1. 54 mg and 3. 39 mg. Conclusion: The method is highly specific and accurate,and suitable for the quantitative analysis of bile acids in Hugan tablets,which provides reference for improving the quality standard of Hugan tablets.
Objective: To establish an HPLC-ELSD method for the simultaneous determination of two major conjugated bile acids glycohyodeoxycholic acid and glycochenodeoxycholic acid in Hugan tablets. Methods: The analysis was performed on a Diamonsil C_(18)column (250 mm × 4. 6 mm,5 μm) with the mobile phase of acetonitrile-0. 2% formic acid with gradient elution. The flow rate was1. 0 ml·min~(-1). The temperature of the ELSD drift tube was 60℃,the flow rate of nitrogen was 1. 5 L·min~(-1),and the gain was 1.Results: The calibration curves of glycohyodeoxycholic acid and glycochenodeoxycholic acid were linear within the range of 26. 52-265. 20 μg·ml~(-1) (r = 0. 999 8) and 19. 84~(-1)98. 40 μg·ml~(-1) (r = 0. 999 1),respectively. The average recoveries were 97. 19% and 103. 33%,and the RSDs were 3. 23% and 4. 00% (n = 6),respectively. The two kinds of conjugated bile acids were detected in 30 batches of Hugan tablets from 16 manufacturers. Among them,the content of glycohyodeoxycholic acid in each tablet was between 3. 03 mg and 6. 10 mg,and the content of glycochenodeoxycholic acid was between 1. 54 mg and 3. 39 mg. Conclusion: The method is highly specific and accurate,and suitable for the quantitative analysis of bile acids in Hugan tablets,which provides reference for improving the quality standard of Hugan tablets.
引文
1中国药典[S].2015年版.一部.961-962
2龚梦鹃,巫圣乾,岳贺,等.基于血清和肝代谢组学研究护肝片的保肝作用[J].中国药房,2017,28(34):4776-4780
3王哲,王文波.超高效液相色谱质谱联用法快速测定护肝片中五味子醇甲和五味子醇乙的含量[J].化工时刊,2017,31(5):10-12
4伍国怡.HPLC同时测定护肝片中5种成分的含量[J].中国药师,2016,19(5):873-875
5孙黎明,梁军,王杰姝,等.护肝片中5种木脂素成分的高效液相-荧光检测研究[J].中医药学报,2015,43(3):13-16
6唐秋竹,王艺纯,孟庆妍,等.HPLC法同时测定护肝片中五味子醇甲、五味子甲素、五味子乙素的含量[J].中国药品标准,2012,13(4):258-261
7陈立群,杨燕飞,耿笑.HPLC-ELSD法同时测定护肝片中猪去氧胆酸和鹅去氧胆酸的含量[J].药学研究,2016,35(9):526-528+552
8石岩,耿颖,郑天骄,等.猪胆粉HPLC-ELSD全轮廓谱图主要胆汁酸测定及化学计量学分析[J].药物分析杂志,2017,37(2):283-289
9卫亚洁,张泽坤,关彦玲,等.护肝片高效液相色谱指纹图谱及多成分定量分析研究[J].中南药学,2018,16(1):31-34
2龚梦鹃,巫圣乾,岳贺,等.基于血清和肝代谢组学研究护肝片的保肝作用[J].中国药房,2017,28(34):4776-4780
3王哲,王文波.超高效液相色谱质谱联用法快速测定护肝片中五味子醇甲和五味子醇乙的含量[J].化工时刊,2017,31(5):10-12
4伍国怡.HPLC同时测定护肝片中5种成分的含量[J].中国药师,2016,19(5):873-875
5孙黎明,梁军,王杰姝,等.护肝片中5种木脂素成分的高效液相-荧光检测研究[J].中医药学报,2015,43(3):13-16
6唐秋竹,王艺纯,孟庆妍,等.HPLC法同时测定护肝片中五味子醇甲、五味子甲素、五味子乙素的含量[J].中国药品标准,2012,13(4):258-261
7陈立群,杨燕飞,耿笑.HPLC-ELSD法同时测定护肝片中猪去氧胆酸和鹅去氧胆酸的含量[J].药学研究,2016,35(9):526-528+552
8石岩,耿颖,郑天骄,等.猪胆粉HPLC-ELSD全轮廓谱图主要胆汁酸测定及化学计量学分析[J].药物分析杂志,2017,37(2):283-289
9卫亚洁,张泽坤,关彦玲,等.护肝片高效液相色谱指纹图谱及多成分定量分析研究[J].中南药学,2018,16(1):31-34