基于UPLC指纹图谱结合化学模式识别的冠心丹参胶囊质量控制研究
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摘要
目的采用UPLC法建立冠心丹参胶囊的指纹图谱,并结合化学模式识别技术对其进行系统、全面和科学的质量评价。方法采用Waters UPLC超高效液相色谱仪,Acquity UPLC?HSS T3色谱柱,以乙腈-0.1%甲酸水为流动相进行梯度洗脱,检测波长为256 nm,建立10批次冠心丹参胶囊的指纹图谱。通过相似度分析并结合聚类分析(CA)、主成分分析(PCA)及正交偏最小二乘法-判别分析(OPLS-DA)等模式识别技术对冠心丹参胶囊的总体质量进行分析评价。结果建立的指纹图谱共标定75个共有峰,经对照品进行化学指认共鉴定了其中的13个色谱峰,分别是甜菜碱、琥珀酸、丹参素、原儿茶醛、咖啡酸、芦丁、木犀草苷、迷迭香酸、丹酚酸B、丹酚酸A、二氢丹参酮I、隐丹参酮、丹参酮IIA。10批供试品的相似度均大于0.97,表明该药物总体质量较为稳定;但通过CA及PCA均发现不同批次药物质量之间存在微小差异,且主要分为2类,最后进一步采用OPLS-DA筛选出了导致批次药物质量差异的3种主要成分,分别为甜菜碱、芦丁和丹酚酸B。结论本研究建立的分析方法科学、准确、可靠且简便,指纹图谱结合化学模式识别技术可更加系统、全面地评价冠心丹参胶囊的药物质量,同时为今后中药制剂更进一步的质量控制研究奠定了理论基础。
Objective To establish the UPLC fingerprint of Guanxin Danshen Capsule(GDC) and conduct a systemic, comprehensive, and scientific quality evaluation of GDC using a chemical pattern recognition method. Methods Ultra-high performance liquid chromatography instrument and Acquity UPLC? HSS T3 chromatographic column was employed, the separation was performed with the mobile phase consisting of acetonitrile and 0.1% formic acid aqueous solution, and the detection wavelength was set at 256 nm to establish the UPLC fingerprint of ten batches of GDC. Then, the further quality assessment of the drug was carried out by similarity evaluation, cluster analysis(CA), principal component analysis(PCA), and orthogonal partial least squares discriminant analysis(OPLS-DA). Results In this research, 75 peaks were recognised as common peaks in the fingerprint, 13 peaks were identified using standard references, they were betaine, succinic acid, tanshinol, protocatechualdehyde, caffeic acid, rutin, galuteolin, rosmarinic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone I, cryptotanshinone, and tanshinone IIA. The similarity values of the drugs were all above 0.97, indicating a relatively stable quality of the drugs. Little difference was then discovered between the batches of the drug by CA and PCA. Finally, glycinebetaine, rutin, and salvia acid B were recognised as the quality makers using a OPLS-DA method. Conclusion The analysis method established in this study was scientific, accurate, reliable, and simple; The drug quality of GDC could be evaluated systematically and comprehensively using a fingerprint combined with chemical pattern recognition technique. Moreover, it will also lay a solid theoretical basis for the further quality control of traditional Chinese medicine and its preparations at the same time.
Objective To establish the UPLC fingerprint of Guanxin Danshen Capsule(GDC) and conduct a systemic, comprehensive, and scientific quality evaluation of GDC using a chemical pattern recognition method. Methods Ultra-high performance liquid chromatography instrument and Acquity UPLC? HSS T3 chromatographic column was employed, the separation was performed with the mobile phase consisting of acetonitrile and 0.1% formic acid aqueous solution, and the detection wavelength was set at 256 nm to establish the UPLC fingerprint of ten batches of GDC. Then, the further quality assessment of the drug was carried out by similarity evaluation, cluster analysis(CA), principal component analysis(PCA), and orthogonal partial least squares discriminant analysis(OPLS-DA). Results In this research, 75 peaks were recognised as common peaks in the fingerprint, 13 peaks were identified using standard references, they were betaine, succinic acid, tanshinol, protocatechualdehyde, caffeic acid, rutin, galuteolin, rosmarinic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone I, cryptotanshinone, and tanshinone IIA. The similarity values of the drugs were all above 0.97, indicating a relatively stable quality of the drugs. Little difference was then discovered between the batches of the drug by CA and PCA. Finally, glycinebetaine, rutin, and salvia acid B were recognised as the quality makers using a OPLS-DA method. Conclusion The analysis method established in this study was scientific, accurate, reliable, and simple; The drug quality of GDC could be evaluated systematically and comprehensively using a fingerprint combined with chemical pattern recognition technique. Moreover, it will also lay a solid theoretical basis for the further quality control of traditional Chinese medicine and its preparations at the same time.
引文
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[2]杜建平,施之琪.HPLC法同时测定冠心丹参胶囊中丹参酮IIA和橙花叔醇的含量[J].江西中医药,2011,42(2):53-54.
[3]梁文仪,袁永兵,陈文静,等.一测多评法测定不同丹参制剂中4个酚酸成分[J].药物评价研究,2018,41(5):828-835.
[4]周霖,姜晓芳,左莉华,等.基于质谱-主成分分析法的冠心丹参胶囊中10个活性成分含量测定及质量评价研究[J].药物分析杂志,2017,37(10):1824-1831.
[5]张福君,张冬璇.UPLC法同时测定冠心丹参胶囊中紫草酸和迷迭香酸的含量[J].中国药房,2016,27(18):2560-2562.
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[7]刁璇,肖丽和,林亚珠,等.多波长UPLC法同时测定冠心丹参胶囊中三七皂苷R1等5种有效成分的含量[J].沈阳药科大学学报,2015,32(6):445-450.
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[9]钱霞,张洁,范玫玫,等.冠心丹参制剂药理临床及质量标准研究进展[J].中草药,2009,40(6):1007-1009.
[10]刘亚东,刘灿群,曾荣添,等.HPLC法测定冠心丹参胶囊中隐丹参酮、丹参酮I和丹参酮IIA含量[J].今日药学,2014,24(8):584-586.
[11]孙志,侯朋艺,周霖,等.基于超高效液相色谱-四级杆/静电场轨道阱高分辨质谱的益心舒片中多种成分定量测定研究[J].中草药,2017,48(10):1977-1982.
[12]周霖,姜晓芳,孙志,等.UPLC-MS/MS法结合模式识别同时测定芪参益气滴丸中11种成分[J].中成药,2017,39(10):2064-2068.
[13]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):4085-4094.
[14]陈林伟,秦昆明,徐雪松,等.中药指纹图谱数据库的研究现状及展望[J].中草药,2014,45(21):3041-3047.
[15]褚延斌,苏小琴,李德坤,等.中药注射剂质量控制研究进展[J].药物评价研究,2018,41(3):345-353.
[16]戴云,叶艳青,冯伟博,等.化学模式识别在中药质量控制中的应用[J].云南民族大学学报:自然科学版,2007,(4):334-337.
[17]孙如煜,李鹏跃,杜守颖,等.基于中药指纹图谱结合模式识别的清开灵硬胶囊批次间稳定性控制研究[J].中华中医药杂志,2017,32(10):4629-4633.
[18]王元清,韩彬,向荣,等.总量统计矩结合聚类分析与主成分分析评价虎杖饮片一致性与差异性[J].中草药,2015,46(19):2863-2869.
[19]吴燕.中药质量控制中的中药全息指纹图谱模式识别方法的研究及应用[D].武汉:中南民族大学,2013.
[2]杜建平,施之琪.HPLC法同时测定冠心丹参胶囊中丹参酮IIA和橙花叔醇的含量[J].江西中医药,2011,42(2):53-54.
[3]梁文仪,袁永兵,陈文静,等.一测多评法测定不同丹参制剂中4个酚酸成分[J].药物评价研究,2018,41(5):828-835.
[4]周霖,姜晓芳,左莉华,等.基于质谱-主成分分析法的冠心丹参胶囊中10个活性成分含量测定及质量评价研究[J].药物分析杂志,2017,37(10):1824-1831.
[5]张福君,张冬璇.UPLC法同时测定冠心丹参胶囊中紫草酸和迷迭香酸的含量[J].中国药房,2016,27(18):2560-2562.
[6]梁文仪,袁永兵,陈文静,等.一测多评技术在3种丹参制剂质量控制中的可行性分析[J].中草药,2016,47(23):4179-4185.
[7]刁璇,肖丽和,林亚珠,等.多波长UPLC法同时测定冠心丹参胶囊中三七皂苷R1等5种有效成分的含量[J].沈阳药科大学学报,2015,32(6):445-450.
[8]汤杰,宋冰娜,郑盼,等.HPLC测定冠心丹参胶囊中主成分的含量[J].中国现代中药,2015,17(10):1078-1082.
[9]钱霞,张洁,范玫玫,等.冠心丹参制剂药理临床及质量标准研究进展[J].中草药,2009,40(6):1007-1009.
[10]刘亚东,刘灿群,曾荣添,等.HPLC法测定冠心丹参胶囊中隐丹参酮、丹参酮I和丹参酮IIA含量[J].今日药学,2014,24(8):584-586.
[11]孙志,侯朋艺,周霖,等.基于超高效液相色谱-四级杆/静电场轨道阱高分辨质谱的益心舒片中多种成分定量测定研究[J].中草药,2017,48(10):1977-1982.
[12]周霖,姜晓芳,孙志,等.UPLC-MS/MS法结合模式识别同时测定芪参益气滴丸中11种成分[J].中成药,2017,39(10):2064-2068.
[13]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):4085-4094.
[14]陈林伟,秦昆明,徐雪松,等.中药指纹图谱数据库的研究现状及展望[J].中草药,2014,45(21):3041-3047.
[15]褚延斌,苏小琴,李德坤,等.中药注射剂质量控制研究进展[J].药物评价研究,2018,41(3):345-353.
[16]戴云,叶艳青,冯伟博,等.化学模式识别在中药质量控制中的应用[J].云南民族大学学报:自然科学版,2007,(4):334-337.
[17]孙如煜,李鹏跃,杜守颖,等.基于中药指纹图谱结合模式识别的清开灵硬胶囊批次间稳定性控制研究[J].中华中医药杂志,2017,32(10):4629-4633.
[18]王元清,韩彬,向荣,等.总量统计矩结合聚类分析与主成分分析评价虎杖饮片一致性与差异性[J].中草药,2015,46(19):2863-2869.
[19]吴燕.中药质量控制中的中药全息指纹图谱模式识别方法的研究及应用[D].武汉:中南民族大学,2013.