丹参波长叠加指纹图谱及多指标成分测定研究
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摘要
对丹参波长叠加指纹图谱和7种丹参指标成分同时定量进行研究。采用Agilent Eclipse Plus C_(18)(4. 6 mm×100 mm,3. 5μm)色谱柱,以0. 1%甲酸的水溶液(A)-乙腈(B)为流动相梯度洗脱(0～5 min,10%～20%B; 5～20 min,20%～30%B; 20～25 min,30%～50%B; 25～40 min,50%～65%B; 40～45 min,65%～80%B; 45～46 min,80%～10%B; 46～50 min,10%B),流速1mL·min~(-1),柱温30℃,检测波长250,280,310,340 nm。运用波长叠加技术建立指纹图谱,并选取主要成分建立多指标测定方法。波长叠加指纹图谱体现了丹参250,280,310,340 nm特征吸收波长的指纹信息,17个不同来源的丹参样品与指纹图谱之间具有良好的相似性,相似度在0. 828～0. 936。确定了15个共有峰,并指认了其中7个色谱峰,分别是迷迭香酸、丹酚酸B、丹酚酸A、二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ和丹参酮ⅡA,其方法学考察结果良好。建立了丹参波长叠加谱指纹图谱,该方法简便可靠、耐用性好,可用于丹参的质量控制。
This project was launched to and establish the wavelength overlapping HPLC fingerprint for Danshen and determine the contents of 7 component in Danshen. The chromatographic fingerprints were built by using Agilent Eclipse Plus C_(18)( 4. 6 mm×100 mm,3. 5 μm) as stationary phase and 0. 1% formic acid solution( A)-acetonitrile( B) as mobile phase with gradient elution( 0-5 min,10%-20% B; 5-20 min,20%-30% B; 20-25 min,30%-50% B; 25-40 min,50%-65% B; 40-45 min,65%-80% B; 45-46 min,80%-10% B; 46-50 min,10% B) at a flow rate of 1 mL·min~(-1). The column temperature was maintained at 30 ℃ and the detection wavelength was set at 250,280,310 and 340 nm. The technique of wavelength overlapping fingerprint was established and the contents of 7 indicative compounds have been determined in this method. The results of wavelength overlapping HPLC fingerprint showed all-around information of the fingerprints at 250,280,310 and 340 nm,and the similarity among 17 batches of Danshen was over 0. 828-0. 936. In wavelength overlapping HPLC fingerprint,15 common peaks were selected as the common peaks,and 7 contents of them were indentified as rosmarinic acid,salvianolic acid B,salvianolic acid A,phenolic acid,dihydrotanshinone Ⅰ,tanshinone Ⅰ and tanshinone ⅡA. The results of methodological study demonstrated that the method met the requirements of the determination. The method established in this study is simple,convenient and durable,which can provide a scientific basis for the quality control of Danshen.
This project was launched to and establish the wavelength overlapping HPLC fingerprint for Danshen and determine the contents of 7 component in Danshen. The chromatographic fingerprints were built by using Agilent Eclipse Plus C_(18)( 4. 6 mm×100 mm,3. 5 μm) as stationary phase and 0. 1% formic acid solution( A)-acetonitrile( B) as mobile phase with gradient elution( 0-5 min,10%-20% B; 5-20 min,20%-30% B; 20-25 min,30%-50% B; 25-40 min,50%-65% B; 40-45 min,65%-80% B; 45-46 min,80%-10% B; 46-50 min,10% B) at a flow rate of 1 mL·min~(-1). The column temperature was maintained at 30 ℃ and the detection wavelength was set at 250,280,310 and 340 nm. The technique of wavelength overlapping fingerprint was established and the contents of 7 indicative compounds have been determined in this method. The results of wavelength overlapping HPLC fingerprint showed all-around information of the fingerprints at 250,280,310 and 340 nm,and the similarity among 17 batches of Danshen was over 0. 828-0. 936. In wavelength overlapping HPLC fingerprint,15 common peaks were selected as the common peaks,and 7 contents of them were indentified as rosmarinic acid,salvianolic acid B,salvianolic acid A,phenolic acid,dihydrotanshinone Ⅰ,tanshinone Ⅰ and tanshinone ⅡA. The results of methodological study demonstrated that the method met the requirements of the determination. The method established in this study is simple,convenient and durable,which can provide a scientific basis for the quality control of Danshen.
引文
[1]梁勇,羊裔明,袁淑兰.丹参酮药理作用及临床应用研究进展[J].中草药,2000,31(4):304.
[2] Chen Y H,Du G H,Zhang J T. Salvianolic acid B protects brain against injuries caused by ischemia-reperfusion in rats[J]. Acta Pharmacol Sin,2000,21(5):463.
[3] Liu J,Shen H M,Ong C N. Salvia miltiorrhiza inhibits cell growth and induces apoptosis in human hepatoma HepG(2)cells[J]. Cancer Lett,2000,153(1/2):85.
[4]林敬阳,王海清.丹参多酚酸盐减少PCI相关性心肌梗死的初步研究[J].中国现代应用药学,2018,35(6):884.
[5]丛梦雨,龚彦溶,梁莎碧,等.HPLC-ELSD指纹图谱分析提取与炮制对柴胡中化学成分的影响[J].中国实验方剂学杂志,2018,24(7):13.
[6]吴婉莹,果德安.中药整体质量控制标准体系构建的思路与方法[J].中国中药杂志,2014,39(3):351.
[7]李华,刘国雄.北枳椇子与枳椇子UPLC指纹图谱的建立及特征成分分析[J].今日药学,2015,25(12):824.
[8]王志强,陈勇,刘双双,等.基于形态、显微、多成分测定和指纹图谱的牡丹皮质量评价系统研究[J].中国中药杂志,2018,43(14):2899.
[9]陈亮元,程轩轩,唐晓敏,等.指纹图谱、模式识别结合一测多评法在广金钱草质量评价中的应用研究[J].中国中药杂志,2018,43(16):3322.
[10]罗红群,刘绍璞.多波长叠加近红外吸收光谱法直接测定酒精饮料中的乙醇[J].分析化学,1998,26(1):97.
[11]刘根起,韩玲,张小玲,等.三波长叠加分光光度法测定微量钯的研究[J].光谱学与光谱分析,2004,24(11):1422.
[12]尚永辉,李华,孙家娟.双波长叠加法测定样品中槲皮素含量[J].分析科学学报,2011,27(4):495.
[13]刘蕖,吴江艳,李浩,等.双波长叠加吸收光谱法测定药物中的酒石酸美托洛尔[J].分析测试学报,2017,36(10):1250.
[14]吴征真,程乙真,曾庆瑞,等.波长叠加可见吸收光谱法测定食品中的Cu[J].分析试验室,2018,37(1):58.
[15]庞向东,丁世敏,李浩,等.双波长叠加吸收光谱法测定奥扎格雷钠[J].现代化工,2018,38(2):219.
[16]王涛,张慧,古咸杰,等.川芎多波长叠加指纹图谱初步研究[J].药物分析杂志,2017,37(9):1648.
[17]张慧,王涛,陈杰,等.白芍多波长叠加指纹图谱研究[J].中药材,2017,40(9):2121.
[18]王涛.诱导子处理下的丹参悬浮细胞生物活性成分、金属组和代谢组研究[D].雅安:四川农业大学,2016.
[19]中国药典.一部[S]. 2015.
[20]刘伟,周冰谦,王晓,等.高效生态间作模式对丹参生长及有效成分含量的影响[J].中药材,2018,41(5):1027.
[21]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.
[22] Liu A H,Li L,Xu M,et al. Simultaneous quantification of six major phenolic acids in the roots of Salvia miltiorrhiza and four related traditional Chinese medicinal preparations by HPLC-DAD method[J]. J Pharmaceut Biomed Anal,2006,41(1):48.
[23] Mulinacci N,Bardazzi C,Romani A,et al. HPLC-DAD and TLC-Densitometry for quantification of hypericin in Hypericum perforatum L. extracts[J]. Chromatographia,1999,49(3/4):197.
[24] Escarpa A,Gonzalez M C. Fast separation of(poly)phenolic compounds from apples and pears by high-performance liquid chromatography with diode-array detection[J]. J Chromatogr A,1999,830(2):301.
[25] Cui H,He C X,Zhao G W,Determination of polyphenols by high-performance liquid chromatography with inhibited chemiluminescence detection[J]. J Chromatogr A,1999,855(1):171.
[2] Chen Y H,Du G H,Zhang J T. Salvianolic acid B protects brain against injuries caused by ischemia-reperfusion in rats[J]. Acta Pharmacol Sin,2000,21(5):463.
[3] Liu J,Shen H M,Ong C N. Salvia miltiorrhiza inhibits cell growth and induces apoptosis in human hepatoma HepG(2)cells[J]. Cancer Lett,2000,153(1/2):85.
[4]林敬阳,王海清.丹参多酚酸盐减少PCI相关性心肌梗死的初步研究[J].中国现代应用药学,2018,35(6):884.
[5]丛梦雨,龚彦溶,梁莎碧,等.HPLC-ELSD指纹图谱分析提取与炮制对柴胡中化学成分的影响[J].中国实验方剂学杂志,2018,24(7):13.
[6]吴婉莹,果德安.中药整体质量控制标准体系构建的思路与方法[J].中国中药杂志,2014,39(3):351.
[7]李华,刘国雄.北枳椇子与枳椇子UPLC指纹图谱的建立及特征成分分析[J].今日药学,2015,25(12):824.
[8]王志强,陈勇,刘双双,等.基于形态、显微、多成分测定和指纹图谱的牡丹皮质量评价系统研究[J].中国中药杂志,2018,43(14):2899.
[9]陈亮元,程轩轩,唐晓敏,等.指纹图谱、模式识别结合一测多评法在广金钱草质量评价中的应用研究[J].中国中药杂志,2018,43(16):3322.
[10]罗红群,刘绍璞.多波长叠加近红外吸收光谱法直接测定酒精饮料中的乙醇[J].分析化学,1998,26(1):97.
[11]刘根起,韩玲,张小玲,等.三波长叠加分光光度法测定微量钯的研究[J].光谱学与光谱分析,2004,24(11):1422.
[12]尚永辉,李华,孙家娟.双波长叠加法测定样品中槲皮素含量[J].分析科学学报,2011,27(4):495.
[13]刘蕖,吴江艳,李浩,等.双波长叠加吸收光谱法测定药物中的酒石酸美托洛尔[J].分析测试学报,2017,36(10):1250.
[14]吴征真,程乙真,曾庆瑞,等.波长叠加可见吸收光谱法测定食品中的Cu[J].分析试验室,2018,37(1):58.
[15]庞向东,丁世敏,李浩,等.双波长叠加吸收光谱法测定奥扎格雷钠[J].现代化工,2018,38(2):219.
[16]王涛,张慧,古咸杰,等.川芎多波长叠加指纹图谱初步研究[J].药物分析杂志,2017,37(9):1648.
[17]张慧,王涛,陈杰,等.白芍多波长叠加指纹图谱研究[J].中药材,2017,40(9):2121.
[18]王涛.诱导子处理下的丹参悬浮细胞生物活性成分、金属组和代谢组研究[D].雅安:四川农业大学,2016.
[19]中国药典.一部[S]. 2015.
[20]刘伟,周冰谦,王晓,等.高效生态间作模式对丹参生长及有效成分含量的影响[J].中药材,2018,41(5):1027.
[21]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.
[22] Liu A H,Li L,Xu M,et al. Simultaneous quantification of six major phenolic acids in the roots of Salvia miltiorrhiza and four related traditional Chinese medicinal preparations by HPLC-DAD method[J]. J Pharmaceut Biomed Anal,2006,41(1):48.
[23] Mulinacci N,Bardazzi C,Romani A,et al. HPLC-DAD and TLC-Densitometry for quantification of hypericin in Hypericum perforatum L. extracts[J]. Chromatographia,1999,49(3/4):197.
[24] Escarpa A,Gonzalez M C. Fast separation of(poly)phenolic compounds from apples and pears by high-performance liquid chromatography with diode-array detection[J]. J Chromatogr A,1999,830(2):301.
[25] Cui H,He C X,Zhao G W,Determination of polyphenols by high-performance liquid chromatography with inhibited chemiluminescence detection[J]. J Chromatogr A,1999,855(1):171.
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