HPLC方法同时测定清热利胆片中10个成分含量

目的：建立同时测定清热利胆片中绿原酸、迷迭香酸、黄芩苷、芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚、和厚朴酚、厚朴酚10个指标成分含量的HPLC方法。方法：采用WondaSil C₁₈色谱柱（4.6 mm×250 mm，5 μm），以0.3%磷酸水溶液（A）-甲醇（B）为流动相进行梯度洗脱，流速1 mL·min^-1，柱温37℃，检测波长分别为327 nm（绿原酸、迷迭香酸）、280 nm（黄芩苷）、254 nm（芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚）和294 nm（和厚朴酚、厚朴酚）。结果：清热利胆片中10个指标成分可实现完全分离，在一定的浓度范围内线性关系良好。方法的精密度、稳定性、重复性符合《中华人民共和国药典》2015年版中药品质量标准分析方法验证指导原则（四部9101）的要求（RSD ≤ 2.0%）；平均回收率（n=6）在94.8%~99.5%范围内，RSD在0.93%~2.1%范围内。3批清热利胆片中绿原酸、迷迭香酸、黄芩苷、芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚、和厚朴酚、厚朴酚的含量范围分别为0.281~0.320、0.252~0.281、1.876~2.090、0.127~0.144、0.178~0.198、0.146~0.163、0.492~0.565、0.122~0.142、0.630~0.673、0.954~1.014 mg·片^-1。结论：该方法能够用于同时检测清热利胆片中10个化学成分。

Objective:To develop an HPLC method for the determination of chlorogenic acid, rosmarinic acid, baicalin, aloe-emodin, rhein, emodin, chrysophanol, physcion, honokiol and magnolol in Qingre Lidan tablets. Methods:The analysis was performed on a WondaSil C₁₈ column (4.6 mm×250 mm, 5 μm) eluted with mobile phase of 0.3%phosphoric acid aqueous solution (A) and methanol (B) in a linear gradient mode. The flow rate was kept at 1 mL·min^-1 and the column temperature was set at 37℃. The detector wavelengths were 327 nm (chlorogenic acid, rosmarinic acid), 280 nm (baicalin), 254 nm (aloe-emodin, rhein, emodin, chrysophanol, physcion) and 294 nm (honokiol, magnolol). Results:10 selected components in Qingre Lidan tablets were separated satisfactorily. All the components exhibited good linear correlation in the given concentration range (r=0.999 9-0.999 3). The precision, stability and repeatability were all fit to the requirement of the guiding principle about the methodology validation in Chinese Pharmacopoeia (Edition 2015) (RSD ≤ 2.0%). The average recoveries (n=6) of the 10 components were 94.8%-99.5%with the RSD of 0.93%-2.1%. Three batches of samples were tested and the contents of ten components were between 0.281-0.320, 0.252-0.281, 1.876-2.090, 0.127-0.144, 0.178-0.198, 0.146-0.163, 0.492-0.565, 0.122-0.142, 0.630-0.673 and 0.954-1.014 mg per tablet, respectively.Conclusion:This method is suitable for simultaneous determination of 10 contents in Qingre Lidan tablets.

[1] 天津市医疗机构制剂规范.2016年版[S].2016:274 Preparation Standards for Medical Institutions in Tinjin[S].2016:274
[2] 杨爱华.茵陈蒿汤药效部位筛选及配伍机理初步研究[D].南京:南京中医药大学,2013 Yang ah.Effective Fraction Screen and Preliminary Study on Compatibility Mechanism of Yinchenhao Decoction[D].nanjing:Nanjing University of Chinese Medicine,2013
[3] 候金燕,窦志华.茵陈蒿汤保肝作用研究进展[J].中医药导报,2015,21(19):88 Hou jy,dou zh.Research progress on hepatoprotective effect of Yinchenhao decoction[J].guid j tradit chin med pharm,2015,21(19):88
[4] Liu J,Hou LL,Zhao CY.Effect of YHHJ on the expression of the hepatocellular bile acid transporters multidrug resistance-associated protein 2 and bile salt export pump in ethinylestradiol-induced cholestasis[J].Exp Ther Med,2018,15(4):3699
[5] Yang Q,Yang F,Tang X,et al.Chlorpromazine-induced perturbations of bile acids and free fatty acids in cholestatic liver injury prevented by the Chinese herbal compound Yin-Chen-Hao-Tang[J].BMC Complement Altern Med,2015,15:122
[6] Huang W,Zhang J,Moore DD.A traditional herbal medicine enhances bilirubin clearance by activating the nuclear receptor CAR[J].J Clin Invest,2004,113(1):137
[7] 华圆,冯健,李范珠.茵陈蒿汤利胆退黄物质基础的研究进展[J].中华中医药学刊,2011,29(7):1520 Hua y,feng j,li fz.Progress in study on cholagogue active components of Yinchenhao decoction[J].Clin Arch Tradit chin med,2011,29(7):1520
[8] Cheng F,Wang Y,Li J,et al.Berberine improves endothelial function by reducing endothelial microparticles-mediated oxidative stress in humans[J].Int J Cardiol,2013,167(3):936
[9] 唐彬,宋振梅,肖琦凡.黄芩苷对高脂-脂肪肝模型大鼠肝脏炎症改善及炎性因子表达的影响[J].青岛大学医学院学报,2017,53(6):659 Tang B,song zm,xiao qf.Effect of balcalin on the improvement of liver inflammation and the expression of inflammatory factors in a rat model of hyperlipidemia-fatty liver disease[J].J Qingdao Med Univ,2017,53(6):659
[10] 王韵,赵亮,柴逸峰,等.RP-HPLC法同时测定茵陈蒿汤中14个成分[J].药物分析杂志,2017,37(3):393 Wang y,zhao l,chai yf,et al.Simultaneous determination of fourteen constituents in Yin-Chen-Hao-Tang by RP-HPLC[J].chin J Pharm Anal,2017,37(3):393
[11] 杨佩磊,蔡咏梅.HPLC法同时测定厚朴排气合剂中和厚朴酚、厚朴酚、大黄素和大黄酚的含量[J].药物分析杂志,2017,37(5):922 Yang pl,cai ym.Simultaneous determination of honokiol,magnolol,emodin and chrysophanol in Houpopaiqi mixture by HPLC[J].chin J Pharm Anal,2017,37(5):922
[12] 郭琪,王海波,李向阳,等.HPLC法同时测定利胆排石片中14个成分的含量[J].药物分析杂志,2017,37(4):631 Guo q,wang hb,li xy,et al.Simultaneous determination of 14 components in Lidanpaishi tablets by HPLC[J].chin J Pharm Anal,2017,37(4):631
[13] 杨跃华,胡春丽,张洪霞,等.RP-HPLC法同时测定利胆排石片中5种有效成分的含量[J].沈阳药科大学学报,2012,29(5):373 YANG YH,HU CL,ZHANG HX,et al.Simultaneous determination of five constituents in Lidanpaishi tablets by RP-HPLC[J].J Shenyang Pharm Univ,2012,29(5):373
[14] 郝乘仪,郭淑英,朱鹤云,等.高效液相色谱法同时测定栀子金花丸中9个成分的含量[J].药物分析杂志,2014,34(9):1571 HAO CY,GUO SY,ZHU HY,et al.Simultaneous determination of nine compounds in Zhizi Jinhua pills by HPLC wavelength switching method[J].chin J Pharm Anal,2014,34(9):1571