成分分析

基于质量标志物有效性、特有性及可测性理念的天葵子Q-marker成分预测*

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  • 1.安徽中医药大学第一附属医院,合肥 230031;
    2.安徽中医药大学,合肥 230031;
    3.中国中医科学院 中药资源中心 道地药材国家重点实验室培育基地,北京 100700
第一作者 Tel:13856069856;E-mail:46784412@ qq.com
** 杨 俊 Tel:13866661936;E-mail:1094704506@ qq.com
孟 楣 Tel:15375512726;E-mail:ahzyymm@ 163.com

收稿日期: 2023-09-04

  网络出版日期: 2024-05-31

基金资助

* 中央本级重大增减支项目,名贵中药资源可持续利用能力建设项目,天葵子、儿茶和败酱草药材商品规格等级标准研制(2060302-1906-23);全国中医药特色技术传承人才培训项目(T20184828005)

Q-marker component prediction of Semiaquilegiae Radix based on the concept of effectiveness, specificity and testability of quality markers*

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  • 1. The First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei 230031, China;
    2. Anhui University of Chinese Medicine, Hefei 230031, China;
    3. State Key Laboratory of Dao-di Herbs Breeding Base, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China

Received date: 2023-09-04

  Online published: 2024-05-31

摘要

目的:桥连网络药理学和特征图谱技术,预测天葵子质量标志物,为天葵子全面的质量控制体系建立提供参考。方法:文献及成分-靶点-通路网络可视化研究,定位并预测天葵子干预疾病的潜在质量标志物;采用Agilent Eclipse Plus C18 (250 mm×4.6 mm, 5 μm)色谱柱,柱温30 ℃,以乙腈(A)-含15%乙腈的0.1%磷酸水溶液(每100 mL含SDS 0.2 g)(B)为流动相,梯度洗脱(0~5 min,0%A;5~10 min,0%A→27%A;10~33 min,27%A→29%A;33~38 min,29%A→37%A;38~55 min,37%A→38%A;55~72 min,38%A),流速0.8 mL·min-1,检测波长为260 nm (0~6 min)、220 nm(6~10 min)、260 nm(10~29 min)、220 nm(29~34 min)、260 nm(34~37 min)、220 nm(37~72 min);对收集的38批天葵子药材进行HPLC特征图谱定性定量分析,运用OPLS-DA筛选出造成组间差异的主要标志性成分;根据质量标志物的有效性、特有性及可测性预测天葵子潜在的Q-marker。结果:锁定了天葵子质量标志物的主要来源范围,网络药理学预测了其干预疾病的3个潜在质量标志物;成功建立了7个共有峰的特征图谱,标定了4个特征峰,其中3个差异性代谢产物,且格列风内酯、木兰花碱和小檗红碱质量浓度分别在2.22~44.4、14.2~284、3.8~76 μg·mL-1范围内均与峰面积积分值呈良好的线性关系,平均加样回收率分别为98.8%、98.8%和99.6%,RSD分别为0.64%、1.7%和1.9%;38批样品中格列风内酯、木兰花碱和小檗红碱的含量范围分别为0.046~0.757、0.391~2.796、0.077~0.572 mg·g-1。根据质量标志物的有效性、特有性及可测性,预测天葵子中格列风内酯和小檗红碱2个成分为天葵子的Q-marker成分。结论:本研究预测了天葵子的Q-marker成分,同时建立了简便、准确、可靠的天葵子特征图谱定性定量方法,为天葵子全面质量控制体系的建立提供了参考,研究结果将有利于市场上天葵子药材质量评价。

本文引用格式

陈莉, 王盛, 刘柳珊, 杨俊, 孟楣, 詹志来 . 基于质量标志物有效性、特有性及可测性理念的天葵子Q-marker成分预测*[J]. 药物分析杂志, 2023 , 43(10) : 1672 -1683 . DOI: 10.16155/j.0254-1793.2023.10.05

Abstract

Objective: To establish a comprehensive quality control system for Semiaquilegiae Radix. To predict the quality markers of Semiaquilegiae Radix by a bridged network pharmacology and feature map technology. Methods: Based on literature research and network pharmacology research, potential quality biomarkers of Semiaquilegiae Radix were predicted. Qualitative and quantitative analysis of HPLC characteristic chromatogram was carried out on 38 batches of Semiaquilegiae Radix. Agilent Eclipse Plus C18 chromatographic column (250 mm×4.6 mm, 5 μm) was used. The mobile phase was acetonitrile(A) and a 0.1% phosphoric acid aqueous solution containing 15% acetonitrile(containing 0.2 g of SDS per 100 mL)(B), with gradient elution(0-5 min, 0%A; 5-10 min, 0%A→27%A; 10-33 min, 27%A→29%A; 33-38 min, 29%A→37%A; 38-55 min, 37%A→38%A; 55-72 min, 38%A) at flow rate of 0.8 mL·min-1. The column temperature was 30 ℃, and the detection wavelengths were 260 nm for 0-6 min, 220 nm for 6-10 min, 260 nm for 10-29 min, 220 nm for 29-34 min, 260 nm for 34-37 min, and 220 nm for 37-72 min. OPLS-DA was used to screen the main marker components causing the difference between groups. According to the effectiveness, specificity and testability of quality markers, the potential Q-marker of Semiaquilegiae Radix was predicted. Results: The main source range of quality markers of Semiaquilegiae Radix was locked, and three potential quality markers for disease intervention were predicted by network pharmacology. The characteristic chromatogram with 7 common peaks was successfully established, and 3 differential metabolites were screened. Contents of glifepristone, magnoflorine, and berberine were between 2.22 and 44.4, respectively μg·mL-1, 14.2-284 μg·mL-1 and 3.8-76 μg·mL-1. There was a good linear relationship between the concentration range and the peak area integral value in each of them, with average sample recovery rates of 98.8%, 98.8%, and 99.6%, and RSD of 0.64%, 1.7%, and 1.9%, respectively. The content ranges of glifepristone, magnoflorine, and berberine in 38 batches of samples were 0.046-0.757 mg·g-1, 0.391-2.796 mg·g-1, and 0.077-0.572 mg·g-1. According to the effectiveness, specificity and testability of quality markers, it was predicted that griffonilide and berberrubine were Q-marker components of Semiaquilegiae Radix. Conclusion: This study predicts the Q-marker composition of Semiaquilegiae Radix, and establishes a simple, accurate and reliable qualitative and quantitative method for characteristic chromatogram of Semiaquilegiae Radix, which provides a reference for the establishment of Semiaquilegiae Radix total quality control system. The research results will be conducive to the quality evaluation of Semiaquilegiae Radix in the market.

参考文献

[1] 中华人民共和国药典2020年版. 一部[S]. 2020: 60
ChP 2020. Vol Ⅰ[S]. 2020: 60
[2] 牛锋. 天葵子的化学成分及质量标准研究[D]. 沈阳: 沈阳药科大学, 2006
NIU F. Study on Chemical Constituents and Quality Standard of Sunflower Seed[D]. Shenyang: Shenyang Pharmaceutical University, 2006
[3] 孙建. 天葵子总生物碱化学成分及对肿瘤炎性微环境的干预作用研究[D]. 北京: 中国中医科学院, 2020
SUN J. Study on the Chemical Constituents of Total Alkaloids of Tiankuizi and the Intervention Effect of Tumor Inflammation Microenvironment[D]. Beijing: China Academy of Chinese Medical Sciences, 2020
[4] 关频, 王建农. 天葵子化学成分和抗肿瘤活性的初步研究[J]. 时珍国医国药, 2011, 22(1):225
GUAN P, WANG JN. A preliminary study on the chemical constituents and anti-tumor activity of Tiankuizi[J]. Lishizhen Med Mater Med Res, 2011, 22(1):225
[5] 徐冉, 肖海涛, 王建塔, 等. 天葵化学成分及其药理作用研究进展[J]. 天然产物研究与开发, 2014, 26(7):1154
XU R, XIAO HT, WANG JT, et al. Research progress on chemical constituents and pharmacological effects of Tiankui[J]. Nat Prod Res Dev, 2014, 26(7):1154
[6] 彭任, 陆兔林, 胡立宏, 等. 中药饮片质量标志物(Q-marker)研究进展[J]. 中草药, 2020, 51(10):2603
PENG R, LU TL, HU LH, et al. Progress on Q-marker research of traditional Chinese medicine decoction pieces[J]. Chin Tradit Herb Drugs, 2020, 51(10):2603
[7] 张铁军, 白钢, 刘昌孝. 中药质量标志物的概念、核心理论与研究方法[J]. 药学学报, 2019, 54(2):187
ZHANG TJ, BAI G, LIU CX. The concept, core theory and research methods of Chinese medicine quality markers[J]. Acta Pharm Sin, 2019, 54(2):187
[8] 刘学勇, 姬志勤. 天葵子化学成分及抑菌活性研究[J]. 天然产物研究与开发, 2019, 31(7):1177
LIU XY, JI ZQ. Studies on chemical constituents and antibacterial activity of Semiaquilegia adoxoides[J]. Nat Prod Res Dev, 2019, 31(7):1177
[9] 冯沼润. 天葵子对D-半乳糖致衰老小鼠抗氧化活性的初步研究[D]. 重庆: 重庆医科大学, 2018
FENG ZR. Preliminary Study on Antioxidant Activity of Tiankuizi on D-Galactose-Induced Aging Mice[D]. Chongqing: Chongqing Medical University, 2018
[10] 张海彬, 程海波, 沈卫星, 等. 中药天葵子的化学成分研究进展[J]. 南京中医药大学学报, 2018, 34(2), 205
ZHANG HB, CHENG HB, SHEN WX, et al. Research progress on chemical constituents from Radix Semiaquilegiae[J]. J Nanjing Univ Tradit Chin Med, 2018, 34(2), 205
[11] HAN QB, JIANG B, MEI SX, et al. Constituents from the roots of Semiaquilegia adoxoides[J]. Fitoterapia, 2001, 72(1), 86
[12] 邹建华, 杨峻山. 天葵的化学成分研究[J]. 中国药学杂志, 2004, 39(4):256
ZOU JH, YANG JS. Study on chemical constituents isolated from Semiaquilegia adoxoides[J]. Chin Pharm J, 2004, 39(4):256
[13] 范汇森, 闫秋莹, 张海彬, 等. UPLC-MS/MS法测定大鼠血浆中紫草氰苷、格列风内酯浓度[J]. 南京中医药大学学报, 2018, 34(1):91
FAN HS, YAN QY, ZHANG HB, et al. Determination of lithospermoside and griffonilide in rat plasma by UPLC-MS/MS[J]. J Nanjing Univ Tradit Chin Med, 2018, 34(1):91
[14] NIU F, XIE GB, CUI Z, et al. Chemical constituents from roots of Semiaquilegia adoxoides[J]. J Chin Pharm Sci, 2006, (4):251
[15] LEE CL, HWANG TL, PENG CY, et al. Anti-neutrophilic inflammatory secondary metabolites from the traditional Chinese medicine, Tiankuizi[J]. Nat Prod Commun, 2012, 7(12):1623
[16] 闫秋莹, 程海波, 张海彬, 等. 中药天葵子醇提部位的化学成分分离与分析[J]. 中华中医药杂志, 2016, 31(10):4172
YAN QY, CHENG HB, ZHANG HB, et al. Separation and analysis on chemical components from the ethanol extract of Semiaquilegiae Radix[J]. China J Tradit Chin Med Pharm, 2016, 31(10):4172
[17] 陈健龙, 张玉玲, 董宇, 等. 小檗碱的酶反应动力学及其代谢酶表型和代谢产物研究[J].中草药, 2013, 44(23):3334
CHEN JL, ZHANG YL, DONG Y, et al. Enzyme reaction kinetics, metabolic enzyme phenotype, and metabolites of berberine[J]. Chin Tradit Herb Drugs, 2013, 44(23):3334
[18] MA ST, FENG CT, DAI GL, et al. In silico target fishing for the potential bioactive components contained in Huanglian Jiedu Tang(HLJDD) and elucidating molecular mechanisms for the treatment of sepsis[J]. Chin J Nat Med, 2015, 13(1):30
[19] CAO SJ, XU PX, YAN JK, et al. Berberrubine and its analog,hydroxypropyl-berberrubine, regulate LDLR and PCSK9 expression via the ERK signal pathway to exert cholesterol-lowering effects in humanhepatoma HepG2 cells[J]. J Cell Biochem, 2018: 1
[20] 匡海学. 中药化学[M]. 北京: 中国中医药出版社, 2002: 348
KUANG HX. Chinese Medicine Chemistry[M]. Beijing: China Press of Traditional Chinese Medicine, 2002: 348
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