目的:建立正心降脂片的HPLC指纹图谱,为该制剂的质量控制提供参考依据。方法: 采用Kromasil C18色谱柱(4.6 mm × 150 mm, 3.5 μm)进行色谱分离,使用乙腈-0.1%甲酸流动相系统进行梯度洗脱,流速0.8 mL·min-1,柱温25 ℃,检测波长254 nm。采用国家药典委员会颁发的中药色谱指纹图谱相似度评价软件(2012版)分析评价所构建的10批正心降脂片指纹图谱相似度。采用Q-Exactive轨道阱高分辨质谱技术和对照品比对鉴定共有峰成分。结果:构建了正心降脂片的指纹图谱共有模式并确认了其中的32个共有指纹峰,去除芦丁积分前后计算得到的10批正心降脂片指纹图谱与对照指纹图谱的相似度分别大于0.99和0.95。采用高分辨质谱技术共鉴定了26个共有峰,其中13个共有峰采用对照品比对进一步确认为3'-羟基葛根素、葛根素、3'-甲氧基葛根素、大豆苷、芦丁、木犀草素-7-O-葡萄糖醛酸苷、橙皮苷、大豆苷元、丹酚酸A、槲皮素、异鼠李素、橙黄决明素、大黄素。通过与组方中各单味药以及阴性样品的比对,对共有峰进行峰归属,32个共有峰涵盖了组方中全部8味药材。结论:本文所建立的正心降脂片HPLC指纹图谱方法简便、可靠,专属性强,结合共有峰的高分辨质谱定性分析,为正心降脂片的质量评价及药效物质基础研究奠定了基础。
Objective: To establish the HPLC fingerprint of Zhengxin Jiangzhi tablets for its quality evaluation. Methods: The separation was performed on a Kromasil C18 column (4.6 mm×150 mm, 3.5 μm) with acetonitrile-0.1% formic acid solution as mobile phase. The flow rate was 0.8 mL·min-1. The detection wavelength was 254 nm and the column temperature was 25 ℃. The similarity evaluation of 10 batches of Zhengxin Jiangzhi tablets was conducted by “Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 edition)”. The common peaks were identified by Q-Exactive Orbitrap HRMS and the reference substance. Results: The fingerprint of Zhengxin Jiangzhi tablets was established and a total of 32 common peaks were marked. The similarities between the 10 batches of Zhengxin Jiangzhi tablets and the control fingerprint were greater than 0.99 after integrating all the 32 common peaks, while the similarities between the 10 batches of Zhengxin Jiangzhi tablets and the control fingerprints were greater than 0.95 after removing the rutin integral. A total of 26 common peaks were identified by Q-Exactive Orbitrap HRMS. By comparing with reference substance, 13 of the identified peaks by HRMS were further confirmed as 3'-hydroxy puerarin, puerarin, 3'-methoxy puerarin and daidzein, rutin, luteolin-7-O-glucuronide, hesperidin, daidzein, salvianolic acid A, quercetin, isorhamnetin, aurantio-obtusin, emodin. The adscription of the 32 common peaks was analyzed by comparing the Zhengxin Jiangzhi tablets sample with each single herb and negative sample, and the 32 common peaks covered all the 8 herbs in the prescription. Conclusion: The established method is simple, stable and specific, which will lay a foundation for the quality evaluation and substance basis study of Zhengxin Jiangzhi tablets.
[1] 中华人民共和国药典2020年版. 一部[S].2020: 789
ChP 2020.Vol Ⅰ[S].2020:789
[2] 蓝日春,韦作干,樊立勇. HPLC测定正心降脂片中丹参酮ⅡA的含量[J].内科, 2012, 7(4): 406
LAN RC, WEI ZG, FAN LY. Determination of tanshinone ⅡA in Zhengxin Jiangzhi tablets by HPLC[J].Intern Med China, 2012, 7(4): 406
[3] 李志平,王加良,刘世娟,等. 基于一测多评法对正心降脂片中7种成分的质量控制研究[J].西北药学杂志, 2020, 35(4): 510
LI ZP, WANG JL, LIU SJ, et al. Quality control for 7 components in Zhengxin Jiangzhi tablets based on quantitative analysis of multi-components wiith a single marker[J].Nothwest Pharm J, 2020, 35(4): 510
[4] 张慧,陈燕,汪佳楠,等. 指纹图谱技术在中药配方颗粒质量评价及过程控制中的应用[J].中国中药杂志, 2018, 43(19): 3822
ZHANG H,CHEN Y,WANG JN, et al. Application of fingerprint technology in quality evaluation and process control of traditional Chinese medicine formula granules[J].China J Chin Mater Med, 2018, 43(19): 3822
[5] 刘东方,赵丽娜,李银峰,等. 中药指纹图谱技术的研究进展及应用[J].中草药, 2016, 47(22): 4085
LIU DF, ZHAO LN, LI YF, et al. Research progress and application in fingerprint technology on Chinese materia medica[J].Chin Tradit Herbal Drugs, 2016, 47(22): 4085
[6] 蒋爱,陈健,韩加伟,等. 小儿退热洗剂HPLC指纹图谱及指标成分测定[J].中草药, 2019, 50(13): 3077
JIANG A, CHEN J, HAN JW, et al. HPLC fingerprint and multi-index content determination of children antifebrile lotion[J].Chin Tradit Herbal Drugs, 2019, 50(13): 3077
[7] 黄萌萌,王琪,李晓琦,等. 基于全-精细指纹图谱与多元统计分析的栀子炒制前后差异评价[J].中草药, 2020, 51(9): 2460
HUANG MM, WANG Q, LI XQ, et al. Comparative study on differences of Gardeniae Fructus and its stir-baked prepared slices based on whole and partial fingerprint and multivariate statistical analysis[J].Chin Tradit Herbal Drugs,2020,51(9): 2460
[8] 李晓明,杨滨,黄璐琦. 高效液相色谱-质谱联用分析鉴别葛根的异黄酮成分[J].中国中药杂志, 2008, 33(11): 1337
LI XM, YANG B, HUANG LQ. Analysis and identification of isoflavones in Pueraria lobata by HPLC-MS[J].China J Chin Mater Med, 2008, 33(11): 1337
[9] 孙冬梅,董玉娟,胥爱丽,等. 葛根枳椇软胶囊中黄酮类化合物的UPLC/Q-TOF-MS快速分析[J].中草药, 2015, 46(7): 970
SUN DM, DONG YJ, XU AL, et al. Rapid analysis on flavonoids in Gegen Zhiju Soft Capsule by UPLC/Q-TOF-MS[J].Chin Tradit Herbal Drugs, 2015, 46(7): 970
[10] 孙梦佳,王焕军,丁富娟,等. 基于UPLC-Q-Exactive Orbitrap MS和质量亏损过滤技术的三叶青黄酮类化合物定性分析[J].质谱学报, 2020, 41(4): 359
SUN MJ, WANG HJ, DING FJ, et al. Qualitative analysis of flavonoids in Tetrastigma hemsleyanum based on UPLC-Quadrupole/exactive orbitrap mass spectrometryand mass defect filter method[J].J Chin Mass Spectr Soc, 2020, 41(4): 359
[11] 祝婧,袁恩,牟俊雍,等. 基于UPLC-Q/TOF-MS分析江西特色炮制工艺对枳壳化学成分的影响[J].中国实验方剂学杂志, 2020, 26(16): 142
ZHU J, YUAN E, MOU JY, et al. Analysis of influence of Jiangxi characteristic processing technology on chemical compositions of Aurantii Fructus based on UPLC-Q/TOF-MS[J].Chin Exp Tradit Med Form, 2020, 26(16): 142
[12] 张驰,张彩娟,邱敏懿,等. 丹参多化学成分的UPLC-LTQ Orbitrap MS快速表征解析[J].中医药学报, 2018, 46(3): 14
ZHANG C, ZHANG CJ, QIU MY, et al. Quick feature analyzation of chemical constituents of Radix et Rhizoma Salviae Miltiorrhizae by UPLC-LTQ Orbitrap MS[J].Acta Chin Med Pharm, 2018, 46(3):14
[13] 刘瑞,王瑞,裴科,等. UHPLC-Q-Orbitrap HRMS快速鉴定羊红膻药材的化学成分[J].中国实验方剂学杂志, 2020, 26(5): 145
LIU R, WANG R, PEI K, et al. Rapid identification of chemical constituents of Pimpinella thellungiana by UHPLC-Q-orbitrap HRMS[J].Chin Exp Tradit Med Form, 2020, 26(5): 145
[14] 孙宁宁,张可佳,耿婉丽,等. 基于UPLC-Q-TOF-MS的加参片提取物化学成分分析[J].中草药,2018,49(2): 293
SUN NN, ZHANG KJ, GENG WL, et al. Analysis of chemical constituents of Jiashen tablets extract by UPLC-Q-TOF-MS[J].Chin Tradit Herbal Drugs, 2018, 49(2): 293
[15] 骆宜,张乐,王卫华,等. 高效液相色谱-离子阱-飞行时间质谱鉴定决明子化学成分[J].药物分析杂志,2015,35(8): 1408
LUO Y, ZHANG L, WANG WH, et al. Components identification in Cassiae Semen by HPLC-IT-TOF MS[J].Chin J Pharm Anal, 2015, 35(8): 1408
