市售丹参配方颗粒的UPLC指纹图谱及化学模式识别研究<sup>*</sup>

刘现磊, 李琨, 张辉, 李伟霞, 王晓艳, 冯科冉, 陈毓龙, 泥文娟, 唐进法

doi:10.16155/j.0254-1793.2022.11.17

药物分析杂志 >

2022 , Vol. 42 >Issue 11: 2000 - 2009

DOI: https://doi.org/10.16155/j.0254-1793.2022.11.17

质量分析

市售丹参配方颗粒的UPLC指纹图谱及化学模式识别研究^*

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1.河南中医药大学第一附属医院河南省中药临床应用、评价与转化工程研究中心河南省中药临床药学中医药重点实验室,郑州 450000;
2.河南中医药大学药学院,郑州 450046;
3.河南中医药大学呼吸疾病中医药防治省部共建协同创新中心,郑州 450046

第一作者 Tel:(0371)66613929;E-mail:liuxianlei0330@163.com

^** 李伟霞 Tel:(0371)66233612;E-mail:liweixia01@126.com
唐进法 Tel:(0371)66245104;E-mail:a0519@163.com

收稿日期: 2021-08-27

网络出版日期: 2024-06-25

基金资助

^* 国家自然科学基金(82004082);河南省中医药科学研究专项(20-21ZY2102,20-21ZY1001);河南省中医药拔尖人才培养项目(豫中医科教[2018]35号)

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UPLC fingerprint and chemical pattern recognition study on commercial available Danshen formula granules^*

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1. Henan Provincial Key Laboratory for Clinical Pharmacy of Traditional Chinese Medicine, Henan Province Engineering Laboratory for Clinical Evaluation Technology of Chinese Medicine, The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450000, China;
2. School of Pharmacy, Henan University of Chinese Medicine, Zhengzhou 450046, China;
3. Provincial and Ministerial Co-Construction Collaborative Innovation Center for Prevention and Treatment of Respiratory Diseases with Traditional Chinese Medicine of Henan University of Chinese Medicine, Zhengzhou 450046, China

Received date: 2021-08-27

Online published: 2024-06-25

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摘要

目的: 建立市售丹参配方颗粒超高效液相色谱指纹图谱和化学模式识别方法,为其质量一致性研究提供参考。方法: 采用 Waters symmetry C₁₈ Analytical色谱柱(150 mm×4.6 mm, 5 μm)进行检测,流动相为乙腈-0.05%磷酸水溶液,梯度洗脱,流速为0.3 mL·min^-1,柱温为30 ℃,检测波长为280 nm,进样量为2 μL。建立4个厂家市售的31批丹参配方颗粒指纹图谱,并进行相似度评价、层次聚类分析(HCA)和主成分分析(PCA),结合正交偏最小二乘-判别分析(OPLS-DA)筛查不同批次丹参配方颗粒的差异成分。结果: 建立的市售丹参配方颗粒UPLC指纹图谱共标定了13个共有峰。通过与对照品比对,共指认了10个成分,分别为丹参素(1号峰)、原儿茶醛(2号峰)、咖啡酸(3号峰)、丹酚酸D(4号峰)、丹酚酸E(5号峰)、迷迭香酸(6号峰)、紫草酸(8号峰)、丹酚酸B(9号峰)、丹酚酸A(11号峰)、丹参酮ⅡA(13号峰)。31批市售丹参配方颗粒指纹图谱相似度在0.877~0.999,HCA、PCA和OPLS-DA均将样品分为了4类,在OPLS-DA模型下,以VIP值大于1筛选出了导致各批次间产生差异的3个差异性标志物,分别为咖啡酸(3号峰)、12号峰和丹参酮Ⅱ_A(13号峰)色谱峰。结论: 该分析方法简单可行,具有良好的精密度、重复性和稳定性,适用于快速评定不同厂家市售不同批次丹参配方颗粒的稳定性和一致性,可为市售丹参配方颗粒整体的质量评价提供参考。

关键词： 丹参; 市售配方颗粒; 超高效液相色谱; 丹酚酸类; 丹参酮类; 指纹图谱; 化学模式识别; 质量评价

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刘现磊, 李琨, 张辉, 李伟霞, 王晓艳, 冯科冉, 陈毓龙, 泥文娟, 唐进法 . 市售丹参配方颗粒的UPLC指纹图谱及化学模式识别研究^*[J]. 药物分析杂志, 2022 , 42(11) : 2000 -2009 . DOI: 10.16155/j.0254-1793.2022.11.17

Abstract

Objective: To establish an UPLC fingerprint-chemical pattern recognition method for commercially available Danshen formula granules, and to provide a reference for the quality improvement and differential markers of the preparation. Methods: Waters symmetry C₁₈ Analytical (150 mm×4.6 mm, 5 μm) chromatographic column was used for detection, the mobile phase was acetonitrile-0.05% phosphoric acid aqueous solution for gradient elution at a flow rate of 0.3 mL·min^-1, andthe column temperature was 30 ℃.The wavelength was 280 nm, the injection volume was 2 μL to established the UPLC fingerprint of 31 batches of commercially available Danshen formula granules sold by 4 manufacturers. Then the further assessment of 31 batches of samples were carried out by similarity evaluation, hierarchical clustering analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) to find the different components of different batches of Danshen formula granule. Results: Thirteen common peaks were identified fromUPLC fingerprints of commercially available Danshen formula granules. By comparing with the reference substances, a total of 10 components were identified as Danshensu (Peak 1), protocatechualdehyde (Peak 2), caffeic acid (Peak 3), salvianolic acid D (Peak 4), and salvianolic acid E (Peak 5), rosmarinic acid (Peak 6), shikonicacid (Peak 8), salvian acid B (Peak 9), salvianolic acid A (Peak 11) and tanshinone ⅡA (Peak 13). The similarities of the fingerprints of 31 batches of Danshen formula granules were above 0.877. The HCA, PCA and OPLS-DA analysis classified samples into 4 clusters. Under the OPLS-DA model, the VIP value was greater than 1 to screen out that may lead to different batches. The three differential markers that produced differences between them were chromatographic peaks of caffeic acid (Peak 1), Peak 12, and tanshinoneⅡ_A (Peak 13). Conclusion: The analytical method is simple and feasible, with good precision, repeatability and stability. It is suitable for quick assessment ofthe stability and consistency of different batches of Danshen formula granules sold by different manufacturers, and can offer reference to evaluate the overall quality of commercial Danshen formula granules.

Key words： Salviae Miltiorrhizae Radix et Rhizoma; commercially available formula granules; UPLC; salvianolic acids; tanshinones; fingerprint; chemical pattern recognition; quality evaluation

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