目的： 建立热炎宁颗粒 HPLC 特征图谱和多成分同时定量的分析方法,为评价其质量提供依据。方法： 采用 Welch Ultimate Phenyl-Ether 色谱柱（250 mm×4.6 mm,5 μm）,以乙腈（A）-0.15% 磷酸水溶液（B）为流动相,梯度洗脱,流速为 1.0 mL·min-1,柱温为 30 ℃,检测波长为 290 nm,进样量 10 μL。采用“中药色谱指纹图谱相似度评价系统（2012 年版）”进行相似度评价,结合聚类分析（CA）、主成分分析（PCA）以及正交偏最小二乘-判别分析（OPLS-DA）对热炎宁颗粒进行质量评价。结果： 建立了热炎宁 颗粒的 HPLC 特征图谱,共标定了 15 个共有峰,并指认了其中 6 个共有峰,分别为单咖啡酰酒石酸、虎杖苷、野黄芩苷、菊苣酸、白藜芦醇、大黄素;16 批样品相似度为 0.957~0.992。CA、PCA 将样品分为 3 类,结 合 OPLS-DA 发现导致不同批次样品之间差异性的 6 个标志性化合物。定量分析的 4 个成分（虎杖苷、野 黄芩苷、白藜芦醇、大黄素）分离度 >1.5,精密度、稳定性（30 h）和重复性试验的 RSD 均< 2.2%,平均加样回收率在 96.4%~98.2% 之间;16 批热炎宁颗粒中虎杖苷、野黄芩苷、白藜芦醇、大黄素的含量范围分别为 2.516~6.190、0.454 6~1.950、0.225 8~0.629 7、0.183 0~0.713 5 mg·g-1。结论： 所建立的 HPLC 特征图谱及多成分同时定量的方法准确、重复性好,可为热炎宁颗粒的质量控制和评价提供依据。
Objective： To establish the HPLC characteristic chromatogram and simultaneous determination method of multi-components of Reyanning granules,so as to provide the basis for effective quality evaluation. Methods： The analysis was performed on Welch Ultimate Phenyl-Ether（250 mm×4.6 mm,5 μm）column with mobile phase consisting of acetonitrile and 0.15% phosphoric acid at a flow rate of 1.0 mL·min-1 with gradient elution. The column temperature was 30 ℃. The detection wavelength was set at 290 nm,and the injection volume was 10 μL. The similarity of Reyanning granules was evaluated by the Traditionai Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System（2012 edition）. Then,the further quality evaluation of the drug was carried out by cluster analysis（CA）,principal component analysis（PCA）and orthogonal partial least squares discriminant analysis（OPLS-DA）. Results： The HPLC characteristic chromatogram of Reyanning granules was established and 15 common peaks were found,6 of them were indentified as monocaffeyltartaric acid,polydatin, scutellarin,chicoric acid,resveratrol and emodin,respectively. The similarities of 16 batches of samples were 0.957-0.992. The samples were classified into three categories by CA and PCA,and six components were main markers that caused differences in the different batches of samples by OPLS-DA. The resolution of 4 components of quantitative qnalysis（polydatin,scutellarin,resveratrol and emodin）was more than 1.5,the RSDs of precision, stability（30 h）and reproducibility tests were lower than 2.2%,and the average recovery rates of four compounds were between 96.4% and 98.2%. The contents of polydatin,scutellarin,resveratrol and emodin in 16 batches of Reyanning granules were 2.516-6.190,0.454 6-1.950,0.225 8-0.629 7 and 0.183 0-0.713 5 mg·g-1, respectively. Conclusion： The method of HPLC characteristic chromatogram analysis and multi-index components determination was accurate and reproducible,which could be used for the quality control and evaluation of Reyanning granules.
[1] 贾红慧, 勾倞, 包涵, 等.热炎宁颗粒的抑菌、抗炎和解热作用研究[J].中药药理与临床, 2007, 23(2):66
JIA HH, GOU J, BAO H, et al.Study on bacteriostatic, anti-inflammatory and antipyretic effects of Reyanning granule[J].Pharm Clin Chin Mater Med, 2007, 23(2):66
[2] 中华人民共和国药典 2020 年版.一部[S].2020:1453 ChP 2020.Vol Ⅰ[S].2020:1453
[3] 李桂荣, 姜志辉, 刘晓丽 .HPLC 法测定热炎宁颗粒中大黄素的含量[J].山东医药工业, 2003, 22(2):22
LI GR, JIANG ZH, LIU XL.Determination of emodin in Reyanning granules by HPLC[J].J Pharm Res, 2003, 22(2):22
[4] 陈冬利, 龙洪 . 热炎宁颗粒质量标准研究[J].中医药导报, 2009, 15(2):72
CHEN DL, LONG H.Study on the quality standard of Reyanning granule[J].Guid J Tradit Chin Med Pharm, 2009, 15(2):72
[5] 王冬, 雷勇胜, 王晨, 等 . 超高效液相色谱法测定热炎宁颗粒中大黄素、咖啡酸、绿原酸、野黄芩苷、木犀草素和虎杖苷[J].现代药物与临床, 2014, 29(12):1365
WANG D, LEI YS, WANG C, et al.Determination of emodin, caffeic acid, chlorogenic acid, scutellarin, luteolin, and polydatin in Reyanning granules by UPLC[J].Drugs Clin, 2014, 29(12):1365
[6] 刘恺, 刘海涛, 梁俊, 等 . 高效液相色谱法测定热炎宁颗粒中野黄芩苷的含量[J].临床合理用药, 2018, 11(2):18
LIU K, LIU HT, LIANG J, et al.Determination of scutellaria baicalensis georgi in Reyanning granules by high performance liquid chromatography[J].Chin J Clin Ratio Drug Use, 2018, 11(2):18
[7] 李静, 李耀磊, 于健东, 等 . 一测多评法结合特征图谱的新疆紫草质量控制研究[J].药物分析杂志, 2020, 40(7):1209
LI J, LI YL, YU JD, et al.Quality control of Arnebiae Radix based on characteristic chromatogram and QAMS[J].Chin J Pharm Anal, 2020, 40(7):1209
[8] 谭志滨, 徐俊鸿, 邓卓燕, 等 . 补肾解毒颗粒中4种成分测定及HPLC 特征图谱建立[J].中成药, 2020, 42(6):1426
TAN ZB, XU JH, DENG ZY, et al.Determination of four constituents and establishment of characteristic HPLC chromatograms for Bushen Jiedu Granules[J].Chin Tradit Pat Med, 2020, 42(6):1426
[9] 田芳, 何小芳, 谭梓君, 等 . 心枳壳 HPLC 特征图谱及6 个黄酮苷类成分含量测定[J].药物分析杂志, 2020, 40(6):1090
TIAN F, HE XF, TAN ZJ, et al .Study on characteristic chromatogram and quantitation method of six components in Aurantii Fructus[J].Chin J Pharm Anal, 2020, 40(6):1090
[10] 王康华, 张英涛, 杨秀伟, 等 . 妇科千金胶囊 HPLC 特征图谱和多指标成分定量分析[J].中国中药杂志, 2019, 44(8):1564
WANG KH, ZHANG YT, YANG XW, et al.Characteristic fingerprint and multi-components quantitative determination for Fuke Qianjin capsules by HPLC[J].China J Chin Med, 2019, 44(8):1564
[11] 何艳, 胡小祥. 银黄胶囊HPLC 特征图谱和6 个成分含量测定[J].中国现代应用药学, 2020, 37(11):1314
HE Y, HU XX.HPLC characteristic chromatogram and content determination of six components for Yinhuang capsules[J].Chin J Mod Appl Pharm, 2020, 37(11):1314
[12] 李昕, 王瑞, 李肖莉, 等 . 不同加工方法乌梅 UPLC 特征图谱及模式识别研究[J].中国中医药信息杂志, 2020, 27(11):76
LI X, WANG R, LI XL, et al.Study on UPLC fingerprints and pattern recognition of Mume Fructus with different processing methods[J].Chin J Inf Tradit Chin Med, 2020, 27(11):76
[13] YUAN ZY, L I J, Z HOU X J, e t al .H S-GC-IMS-based metabonomics study of Baihe Jizihuang Tang in a rat model of chronic unpredictable mild stress[J].J Chromatogr B, 2020, 1148(1):122
[14] 范兴, 安昌, 陈鸣, 等.肝宝胶囊HPLC指纹图谱建立及5种成分同时测定[J].中成药, 2019, 41(4):752
FAN X, AN C, CHEN M, et al.Establishement of HPLC fingerprints of Ganbao capsules and simultaneous determination of five constituents[J].Chin Tradit Pat Med, 2019, 41(4):752
[15] 李静, 张青, 肖春霞, 等 .HPLC 波长切换法同时测定排毒养颜胶囊中10种成分[J].中草药, 2018, 49(20):4824
LI J, ZHANG Q, XIAO CX, et al.Simultaneous determination of 10 components in Paidu Yangyan capsules by HPLC wavelength change method[J].Chin Tradit Herb Drugs, 2018, 49(20):4824
[16] 兰瑞容, 刘燕, 苏英 .HPLC 同时测定蒲公英颗粒中的单咖啡酰酒石酸和菊苣酸[J].华西药学杂志, 2018, 33(2):197
LAN RR, L IU Y, SU Y .Sim ultaneous determination of monocaffeyltartaric acid and chicoric acid in Pugongying granules by HPLC[J].West China J Pharm Sci, 2018, 33(2):197
[17] 刘志浩, 宋平顺, 杨平荣, 等 . 高效液相色谱法同时测定北败酱草中 6 个成分的含量[J].药物分析杂志, 2020, 40(6):1104
LIU ZH, SONG PS, YANG PR, et al.Simultaneous determination of six components in Sonhi Arvensis Herba by HPLC[J].Chin J Pharm Anal, 2020, 40(4):607
[18] 陈黎, 黄良永, 叶方, 等 .UPLC-MS/MS 法同时测定虎杖中9种蒽醌类和2种二苯乙烯类化学成分的含量[J].中南药学, 2020, 18(7):1143
CHEN L, HUANG LY, YE F, et al.Simultaneous determination of 9 anthraquinones and 2 stilbenes in Polygonum cuspidatum Sieb.et Zucc. by UPLC-MS/MS[J].Cent South Pharm, 2020, 18(7):1143
