双波长-UPLC法同时测定胡黄连中4个环烯醚萜类成分的含量

目的：建立同时测定中药材胡黄连中环烯醚萜类成分胡黄连苷Ⅰ、Ⅱ、Ⅲ和Ⅳ含量的双波长-UPLC法。方法：采用Waters ACQUITY UPLC BEH C₁₈色谱柱（2.1 mm×100 mm，1.7 μm），柱温：28℃；流动相：乙腈-0.5%乙酸水溶液，梯度洗脱；流速：0.4 mL·min^-1；检测波长分别为275 nm（胡黄连苷Ⅰ和Ⅱ）、295 nm（胡黄连苷Ⅲ和Ⅳ）。结果：胡黄连苷Ⅰ、Ⅱ、Ⅲ和Ⅳ在质量浓度为4.44~88.72 μg·mL^-1（r=1.000）、17.23~344.61 μg·mL^-1（r=1.000）、1.97~39.35 μg·mL^-1（r=1.000）、1.98~39.51 μg·mL^-1（r=1.000）范围内与峰面积线性相关，平均回收率分别为102.4%、98.8%、96.5%、101.9%，RSD分别为1.0%、2.3%、1.1%、2.0%。结论：该方法操作简便、快速、重复性好、结果准确可靠，适用于胡黄连的质量控制。

Objective: To establish a dual-wavelength UPLC method for simultaneous determination of iridoids including picrosideⅰ,picrosideⅱ,picrosideⅲ and picrosideⅣ in Chinese herbal medicine Picrorhizae Rhizoma. Methods: The separation was performed on a Waters ACQUITY UPLC BEH C₁₈ (2.1 mm×100 mm,1.7 μm) and the column temperature was set at 28℃. The mobile phase was composed of acetonitrile and 0.5% acetic acid with gradient elution at a flow rate of 0.4 mL·min^-1. The detection wavelength was set at 275 nm for picrosideⅠ and picrosideⅡ and at 295 nm for picrosideⅢ and picrosideⅣ. Results: The calibration curves were linear within the range of 4.44-88.72 μg·mL^-1 (r=1.000) for picrosideⅠ,17.23-344.61 μg·mL^-1 (r=1.000) for PicrosideⅡ,1.97-39.35 μg·mL^-1 (r=1.000) for picrosideⅢ,1.98-39.51 μg·mL^-1 (r=1.000) for picrosideⅣ. The average recovery rates of picrosideⅠ,picrosideⅡ,picrosideⅢ and picrosideⅣ were 102.4%,98.8%,96.5% and 101.9% with RSD values of 1.0%,2.3%,1.1% and 2.0%,respectively. Conclusion: The method is simple,rapid,reproducible,accurate and reliable,and can be used for quality control of Picrorhizae Rhizoma.

[1] 中华人民共和国药典2015年版.一部[S].2015:242 ChP 2015.Vol Ⅰ[S].2015:242
[2] 陈炜伟,李鹏,姚仲青,等.胡黄连苷-Ⅰ和胡黄连苷-Ⅱ研究进展[J].中药材,2015,38(8):1756 CHEN WW,LI P,YAO ZQ,et al.Progress of study on picrosideⅠ and picrosideⅡ[J].J Chin Med Mater,2015,38(8):1756
[3] 朱仝飞,陈日荣,孙庆文,等.西藏胡黄连化学成分研究[J].中草药,2017,48(2):263 ZHU TF,CHEN RR,SUN QW,et al.Chemical constituents from Picrorhiza scrophulariiflora[J].Chin Tradit Herb Drugs,2017,48(2):263
[4] LI C,LIU Y,ABDULLA R,et al.Characterization and identification of chemical components in Neopicrorhiza scrphulariiflora roots by liquid chromatography-electrospray ionization quadrupole time-of-flight tandem mass spectrometry[J].Anal Methods,2014,6(11):3634
[5] GIRISH C,KONER BC,JAYANTHY S,et al.Hepatoprotective activity of picroliv,curcumin and ellagic acid compared to silymarin on paracetamol induced liver toxicity in mice[J].Fundam Clin Pharmacol,2009,23(6):735
[6] KAWOOSA T,SINGH H,KUMAR A,et al.Light and temperature regulated terpene biosynthesis:hepatoprotective monoterpene picroside accumulation in Picrorhiza kurrooa[J].Funct Integr Genomics,2010,10(3):393
[7] GUO Y,XU X,LI Q,et al.Anti-inflammation effects of picroside 2 in cerebral ischemic injury rats.[J].Behav & Brain Func,2010,6(1):43
[8] CAO Y,LIU JW,YU YJ,et al.Synergistic protective effect of picroside II and NGF on PC12 cells against oxidative stress induced by H₂O₂[J].Chin J Clin Pharmaco Ther,2007,59(5):573
[9] 周广正.胡黄连苷对大鼠脑缺血再灌注损伤后JNK信号通路的调节作用机制[D].青岛:青岛大学,2016. ZHOU GZ.PicrosideⅡ regulates JNK signal transduction to protect the cerebral ischemic injury in rats[D].Qingdao:Qingdao University,2016
[10] JOY KL,RAJESHKUMAR NV,KUTTAN G,et al.Effect of Picrorrhiza kurroa extract on transplanted tumours and chemical carcinogenesis in mice[J].J Ethnopharmacol,2000,71(1):261
[11] NAJARI A,SACHIN BS,SHARMN SC,et al.Modulation of P-glycoprotein ATPase activity by some phytoconstituents[J].Phytotherapy Res,2010,24(3):454
[12] 黄劲梅,郑清瑗,梁惠珍.胡黄连中香草酸、阿魏酸和肉桂酸的含量测定[J].中药材,2002,25(12):881 HUANG JM,ZHENG QY,LIANG HZ.Determination of vanillic acid,ferulic acid and cinnamic acid in Picrorhiza scrophulariiflora[J].J Chin Med Mater,2002,25(12):881
[13] 曹玉华,汪云.毛细管电泳电化学检测法测定胡黄连中香草酸和阿魏酸的含量[J].分析测试学报,2003,22(6):95 CAO YH,WANG Y.Determination of vanillic acid and ferulic acid content by CE-ED[J].J Instrum Anal,2003,22(6):95
[14] 张永文.试谈中药新药质量标准制定的整体思路[J].中国新药杂志,2017,26(18):2185 ZHANG YW.Strategic discussion on the overall thinking of developing quality standards of new Chinese medicinal prescriptions[J].Chin J New Drugs,2017,26(18):2185
[15] 张可佳,孙宁宁,杨雪,等.UPLC结合化学计量学方法的胡黄连指纹图谱研究[J].中草药,2018,49(3):685 ZHANG KJ,SUN NN,YANG X,et al.Fingerprint study of Picrorhiza scrophulariiflora by UPLC coupled with chemometrics method[J].Chin Tradit Herb Drugs,2018,49(3):685