HPLC方法同时测定清热利胆片中10个成分含量
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摘要
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 C_(18)色谱柱(4.6mm×250 mm,5μm),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 C_(18) 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.
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 C_(18) 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.
引文
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[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):88HOU JY,DOU ZH.Research progress on hepatoprotective effect of Yinchenhao decoction[J].Guid J Tradit Chin Med Pharm,2015,21(19):88
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[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):659TANG 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].JQingdao 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):393WANG 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):922YANG 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):631GUO Q,WANG HB,LI XY,et al.Simultaneous determination of14 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):373YANG YH,HU CL,ZHANG HX,et al.Simultaneous determination of five constituents in Lidanpaishi tablets by RP-HPLC[J].JShenyang 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):1571HAO 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
[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,2013YANG 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):88HOU 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-HaoTang[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):1520HUA 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):659TANG 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].JQingdao 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):393WANG 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):922YANG 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):631GUO Q,WANG HB,LI XY,et al.Simultaneous determination of14 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):373YANG YH,HU CL,ZHANG HX,et al.Simultaneous determination of five constituents in Lidanpaishi tablets by RP-HPLC[J].JShenyang 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):1571HAO 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