经典名方芍药甘草汤UPLC指纹图谱的建立
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摘要
目的:建立古代经典名方芍药甘草汤UPLC指纹图谱检测方法。方法:采用UPLC方法,利用Waters Xselect HSS T3色谱柱(2. 1 mm×100 mm,1. 8μm),以乙腈(A)-醋酸-醋酸铵缓冲溶液(pH=4. 5)(B)作为流动相进行梯度洗脱,柱温40℃,流速0. 2 m L·min~(-1),全波长扫描,选取甘草酸为参照峰,分析15批次芍药甘草汤水煎液的UPLC指纹图谱,并使用中药色谱指纹图谱相似度评价系统对指纹图谱相似度进行评价。结果:在芍药甘草汤水煎液指纹图谱中标定了72个共有峰,指认了7个共有峰,15批样品指纹图谱相似度均> 0. 97,不同批次间差异较小。结论:该方法灵敏度高、稳定性强,数据准确可靠,基本体现了芍药甘草汤的整体化学成分特征,可用于古代经典名方芍药甘草汤开发的质量控制。
Objective: To establish the chromatographic fingerprint for Shaoyao gancao decoction by UPLC.Methods: The fingerprint was determined on Waters Xselect HSS T3 column (2. 1 mm × 100 mm,1. 8 μm),with acetonitrile (A)-acetate-ammonium acetate buffer solution (pH = 4. 5) (B) as mobile phase for gradient elution at a flow rate of 0. 2 m L·min~(-1). The temperature of column was 40 ℃. Full wavelength scanning was conducted and glycyrrhizic acid was selected as the as reference peak. The UPLC fingerprints of 15 batches of Shaoyao gancao decoction were analyzed and the similarity was evaluated with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine. Results: There were 72 common peaks in the UPLC fingerprints of Shaoyao gancao decoction and 7 common peaks were determined by the relative time of the standard substance. The similarities of 15 batches were all more than 0. 97 and there was little difference among these batches. Conclusion:This method is accurate, stable and reliable, which will basically reflect the overall chemical composition characteristics and can be taken as an effective method for the quality control in the development of Shaoyao gancao decoction.
Objective: To establish the chromatographic fingerprint for Shaoyao gancao decoction by UPLC.Methods: The fingerprint was determined on Waters Xselect HSS T3 column (2. 1 mm × 100 mm,1. 8 μm),with acetonitrile (A)-acetate-ammonium acetate buffer solution (pH = 4. 5) (B) as mobile phase for gradient elution at a flow rate of 0. 2 m L·min~(-1). The temperature of column was 40 ℃. Full wavelength scanning was conducted and glycyrrhizic acid was selected as the as reference peak. The UPLC fingerprints of 15 batches of Shaoyao gancao decoction were analyzed and the similarity was evaluated with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine. Results: There were 72 common peaks in the UPLC fingerprints of Shaoyao gancao decoction and 7 common peaks were determined by the relative time of the standard substance. The similarities of 15 batches were all more than 0. 97 and there was little difference among these batches. Conclusion:This method is accurate, stable and reliable, which will basically reflect the overall chemical composition characteristics and can be taken as an effective method for the quality control in the development of Shaoyao gancao decoction.
引文
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[2]朱广伟,张贵君,汪萌,等.中药芍药甘草汤基原及药效组分和药理作用研究概况[J].中华中医药杂志,2015,30(8):2865-2869.
[3]范桂滨,黄志刚.大剂量芍药甘草汤治疗普通型肌肉痛性痉挛68例[J].中医药临床杂志,2007,19(3):248-248.
[4]张颖,柯尊记,杨坚,等.芍药甘草汤及其活性成分的脑保护作用机制研究概述[J].现代生物医学进展,2014,14(34):6773-6777.
[5]谢兰兰,陈思思,王燕鸿,等.芍药甘草汤加味联合运动训练治疗脑卒中后偏瘫肘关节屈曲痉挛疗效观察[J].现代中西医结合杂志,2016,25(7):750-752.
[6]洪嘉琪,史家欣,敖欢,等.芍药苷通过抑制脊髓Akt-NF-k B信号通路及小胶质细胞激活缓解炎症性疼痛[J].中国生物化学与分子生物学报,2018,34(3):325-333.
[7]刘华岩,王军.芍药苷对Aβ_(1-42)诱导的小胶质细胞炎性反应和趋化性的影响[J].中国神经免疫学和神经病学杂志,2017,24(4):276-280.
[8]韩瑶聃,王彬,王政雨,等.甘草酸药理作用的研究进展[J].中国新药杂志,2012,21(21):2499-2505.
[9]傅延龄,宋佳,张林.经方本原剂量问题研究[M].北京:科学出版社,2015.
[10]杨玲娟,赵晓莉,狄留庆,等.HPLC法测定芍药甘草汤总有效部位中4个活性成分的含量[J].中华中医药学刊,2007,25(3):522-524.
[11]刘瑞新,施钧瀚,张璐,等.基于HPLC,PCA与相似度评价的芍药甘草汤传统汤剂与配方颗粒汤剂的差异规律分析[J].中国实验方剂学杂志,2014,20(24):1-5.
[12]方玉婷,董洁.反相高效液相色谱法测定大鼠灌胃芍药甘草汤血浆中甘草酸的含量[J].现代中药研究与实践,2010,24(6):39-41.
[13]高海洋.基于β_2-AR色谱的芍药甘草汤抗急性哮喘物质基础研究[D].西安:西北大学,2017.
[14]朱广伟,张贵君,汪萌,等.中药芍药甘草汤基原及药效组分和药理作用研究概况[J].中华中医药杂志,2015(8):2865-2869.
[15]陈雪梅,蔡秋杰,张华敏.日本汉方药概况及其对我国中医古代经典名方制剂研发的启示[J].中国中医药图书情报杂志,2018,42(2):1-4.
[16]陈畅,程锦堂,刘安.经典名方研发策略[J].中国中药杂志,2017,42(9):1814-1818.
[17]王智民,刘菊妍,刘晓谦,等.谈经典名方的化学、生产和质量控制研发和监管[J].中国中药杂志,2017,42(10):1819-1824.