不同配比补肾活血方HPLC-UV-ELSD指纹图谱研究
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摘要
目的:建立不同配比补肾活血方HPLC-UV-ELSD指纹图谱,并对各峰来源进行归属。方法:采用HPLC法,色谱柱为Inertsil OSD-SP C_(18)(250 mm×4.6 mm,5μm);流动相为乙腈-0.05%甲酸水溶液(梯度洗脱),紫外检测器:检测波长为250 nm,蒸发光散射检测器:漂移管温度为105℃,空气体积流量为3.2 L/min;进样量20μL;流速为1 mL/min。运用《中药色谱指纹图谱相似度评价系统(2008版)》对12批不同配比补肾活血方指纹图谱进行相似度计算,并对各共有峰进行药材来源归属及成分指认。结果:建立了不同配比补肾活血方指纹图谱;在其HPLC-UV指纹图谱中标定了23个共有峰,其中1～15、17、21号峰来源于葛根,18～20、22、23号峰来源于丹参,16号峰为丹参和葛根共有。在其HPLC-ELSD指纹图谱中标定了7个共有峰,其中1’～4’号峰来源于葛根,5’号峰来源于丹参,6’、7’号峰来源于人参。通过与对照品对照指认出复方中4个成分,分别是6/3’号峰为葛根素,22/5’号峰为丹酚酸B,10号峰为大豆苷,6’号峰为人参皂苷Rb1。结论:建立了不同配比补肾活血方HPLC-UV-ELSD指纹图谱,为筛选补肾活血方最佳配比、确定其防治DR药效物质奠定基础。
Objective:Different Ratios HPLC-UV-ELSD fingerprint was developed to assign the main characteristic peaks in Bushen Huoxue Prescription.Methods:High performance liquid chromatography,Inertsil OSD-SP C_(18) column(250 mm×4.6 mm,5 μm)was used with a gradient mobile phase system of acetonitrile-water containing 0.05% formic acid.Ultraviolet detector:The wavelength was 250 nm.Evaporative light scattering detector:The temperature of drift tube was maintained at 105 ℃ and the flow rate of air was 3.2 L/min.The injection volume was 20 μL.The flow rate was 1 mL/min.The similarities between the HPLC-UV-ELSD fingerprints of 12 batches of different ratios of Bushen Huoxue Prescription were calculated by similarity evaluation software and the common peaks were assigned and identified simultaneously.Results:The fingerprint of different ratios of Bushen Huoxue Prescription was established.The HPLC-UV fingerprint of it included 23 mutual peaks,of which seventeen mutual peaks were from P.Ohwi,five peaks from S.miltiorrhiza,one peak from S.miltiorrhiza and P.Ohwi.The HPLC-ELSD fingerprint of it included 7 mutual peaks,of which four mutual peaks were from P.Ohwi,one peak from S.miltiorrhiza,two peaks from P.ginseng.Four components were identified by comparison with the reference substance,which were Salvianolic acid B,puerarin and ginsenoside Rb1.Conclusion:The HPLC-UV-ELSD fingerprint of different ratios of Bushen Huoxue Prescription was established in order to select the best ratio of Bushen Huoxue Prescription and establish the foundation for its prevention and treatment of drug substance of DR.
Objective:Different Ratios HPLC-UV-ELSD fingerprint was developed to assign the main characteristic peaks in Bushen Huoxue Prescription.Methods:High performance liquid chromatography,Inertsil OSD-SP C_(18) column(250 mm×4.6 mm,5 μm)was used with a gradient mobile phase system of acetonitrile-water containing 0.05% formic acid.Ultraviolet detector:The wavelength was 250 nm.Evaporative light scattering detector:The temperature of drift tube was maintained at 105 ℃ and the flow rate of air was 3.2 L/min.The injection volume was 20 μL.The flow rate was 1 mL/min.The similarities between the HPLC-UV-ELSD fingerprints of 12 batches of different ratios of Bushen Huoxue Prescription were calculated by similarity evaluation software and the common peaks were assigned and identified simultaneously.Results:The fingerprint of different ratios of Bushen Huoxue Prescription was established.The HPLC-UV fingerprint of it included 23 mutual peaks,of which seventeen mutual peaks were from P.Ohwi,five peaks from S.miltiorrhiza,one peak from S.miltiorrhiza and P.Ohwi.The HPLC-ELSD fingerprint of it included 7 mutual peaks,of which four mutual peaks were from P.Ohwi,one peak from S.miltiorrhiza,two peaks from P.ginseng.Four components were identified by comparison with the reference substance,which were Salvianolic acid B,puerarin and ginsenoside Rb1.Conclusion:The HPLC-UV-ELSD fingerprint of different ratios of Bushen Huoxue Prescription was established in order to select the best ratio of Bushen Huoxue Prescription and establish the foundation for its prevention and treatment of drug substance of DR.
引文
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[3] 王紫燕,李杨,姚红娥,等.基于视网膜Müller细胞的人参皂苷类成分调节糖基化终末产物干预下HIF-1α表达的谱效关系研究[J].中药药理与临床,2017,33(2):91-94.
[4] 李杨,徐秒,陈换飞,等.丹参提取物对糖基化终末产物/缺氧条件下视网膜Müller细胞低氧诱导因子-1α 作用的谱效关系研究[J].中草药,2017,48(11):2231-2236.
[5] 王紫燕,张梅,李春沁,等.补肾活血方与其组方药材指纹图谱的相关性[J].中成药,2016,38(2):341-345.
[6] 陈萌,李爱平,李科,等.防己黄芪汤HPLC-UV/ELSD指纹图谱研究[J].中草药,2017,48(13):2653-2659.
[7] 方开泰.均匀设计与均匀设计表[M].北京:科学出版社,1994:47-49.
[8] 李红伟,孟祥乐.地黄化学成分及其药理作用研究进展[J].药物评价研究,2015,38(2):218-228.
[9] 李易强.丹参化学成分与药理作用的研究进展[J].科技视界,2017,9(32):265,240.
[10] 姚梦杰,吕金朋,张乔,等.人参化学成分及药理作用研究[J].吉林中医药,2017,37(12):1261-1263.
[11] 李昕,潘俊娴,陈士国,等.葛根化学成分及药理作用研究进展[J].中国食品学报,2017,17(9):189-195.
[12] 范欣生,段金廒,王中越,等.中药量效关系特征问题的探讨[J].中华中医药杂志,2009,24(3):270-274.