基于超高效液相色谱法(UPLC)测定芪丹利心丸中丹酚酸B的含量
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摘要
目的:验证超高效液相色谱法(UPLC),测定芪丹利心丸中丹酚酸B的含量的可靠性与可行性,并与高效液相色谱法(HPLC)的测定结果进行比较。方法:UPLC:色谱柱:Acquity UPLC HSS T3 1.8μm,2.1×100mm Column;流动相:乙腈-0.05%磷酸水溶液;流速:0.4 mL/min;梯度洗脱;检测波长:286nm;柱温:35℃;样品室:10℃。HPLC:色谱柱:SunFire C18 (4.6×150mm,5μm);流动相:乙腈:0.05%磷酸水溶液(40:60);流速:1.0 mL/min;检测波长:286 nm;柱温:35℃。结果:在所建立的UPLC色谱条件下,芪丹利心丸中丹酚酸B的含量在考察的浓度范围内呈良好的线性关系(R=1.0000),精密度、重复性、稳定性及加样回收率结果均符合含量测定的要求;采用UPLC和HPLC测定芪丹利心丸中丹酚酸B的含量,其结果基本一致,但UPLC法分析时间更短,分析效果更佳,溶剂消耗更少,灵敏度更高。结论:测定芪丹利心丸中丹酚酸B的含量,采用UPLC定量分析方法较HPLC法更加快速、简便,可以用于芪丹利心丸中丹酚酸B的含量测定,为其质量控制提供了一种新的技术手段
Objective:To verify the reliability and feasibility of determining the content of salvianolic acid B in Qidanlixin pill by ultra-performance liquid chromatography(UPLC),and to compared with the result of determining by high performance liquid chromatography(HPLC). Methods:By UPLC:chromatography column was Acquity UPLC HSS T3 1.8μm 2.1×100 mm column;mobile phase was acetonitrile-0.05% phosphoric acid water solution;flow rate was 0.4 mL/min;gradient elution;detection wavelength was 286 nm;column temperature was 35;sample room was 10. By HPLC:chromatography column was SunFire C18 column(4.6×150 mm,5μm);mobile phase was acetonitrile-0.05% phosphoric acid water solution(40∶60);flow rate was 1.0 mL/min;detection wavelength was 286 nm;column temperature was 35. Results:Under the established chromatographic condition of UPLC,the content of salvianolic acid B in Qidanlixin pill showed a good linear relationship(R=1.0000)in the investigated range of concentration;and the precision,repeatability,stability and recovery rate all were met the requirements of determination. The results of determining content of salvianolic acid B in Qidanlixin pill by UPLC and HPLC were basically consistent,but UPLC had shorter analysis time,better analysis effect,less solvent consumption and higher sensitivity. Conclusion:In determining content of salvianolic acid B in Qidanlixin pill,the quantitative analysis by UPLC is faster and simpler than by HPLC,can be used for determinating content of salvianolic acid B in Qidanlixinpill,and provides a new technical means for quality control of Qidanlixin pill.
Objective:To verify the reliability and feasibility of determining the content of salvianolic acid B in Qidanlixin pill by ultra-performance liquid chromatography(UPLC),and to compared with the result of determining by high performance liquid chromatography(HPLC). Methods:By UPLC:chromatography column was Acquity UPLC HSS T3 1.8μm 2.1×100 mm column;mobile phase was acetonitrile-0.05% phosphoric acid water solution;flow rate was 0.4 mL/min;gradient elution;detection wavelength was 286 nm;column temperature was 35;sample room was 10. By HPLC:chromatography column was SunFire C18 column(4.6×150 mm,5μm);mobile phase was acetonitrile-0.05% phosphoric acid water solution(40∶60);flow rate was 1.0 mL/min;detection wavelength was 286 nm;column temperature was 35. Results:Under the established chromatographic condition of UPLC,the content of salvianolic acid B in Qidanlixin pill showed a good linear relationship(R=1.0000)in the investigated range of concentration;and the precision,repeatability,stability and recovery rate all were met the requirements of determination. The results of determining content of salvianolic acid B in Qidanlixin pill by UPLC and HPLC were basically consistent,but UPLC had shorter analysis time,better analysis effect,less solvent consumption and higher sensitivity. Conclusion:In determining content of salvianolic acid B in Qidanlixin pill,the quantitative analysis by UPLC is faster and simpler than by HPLC,can be used for determinating content of salvianolic acid B in Qidanlixinpill,and provides a new technical means for quality control of Qidanlixin pill.
引文
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[18]宋延平,孔令姗,吴静,等.丹酚酸A与丹酚酸B改善大鼠心肌缺血作用比较[J].中国中医药信息杂志,2007,14(9):36-38.
[19]沈盼盼,谢晓梅,张圣龙,等.UPLC法测定宣木瓜中齐墩果酸、熊果酸[J].南京中医药大学学报,2013,29(3):271-273.
[20]刘建海,崔维利,陈涛,等.HPLC-ELSD法测定新生脉散液中黄芪甲苷的含量[J].世界科学技术-中医药现代化★技术应用研究,2013,15(1):137-140.
[21]胡晓琴,李进,陈涛,等.HPLC法测定新生脉散片中丹参素钠的含量[J].天津中医药,2012,29(2):181-183.
[2]袁杨,毛静远,王贤良,等.益气活血利水治疗慢性心力衰竭伴低血压验案2则[J].中西医结合心脑血管病杂志,2015,13(4):557-558.
[3]黄芪,张运,郑颖.新生脉散片对射血分数正常心力衰竭患者心血管危险因素的影响[J].中西医结合心脑血管病杂志,2014,12(1):20-21.
[4]张宇.新生脉散片治疗射血分数保留的心力衰竭临床观察[J].中国中医药信息杂志,2011,18(5):75-76.
[5]庞磊,毛静远,万蝉俊,等.新生脉散优化方对阿霉素致小鼠心肌损伤保护作用的研究[J].吉林中医药,2012,32(7):712-715.
[6]赵志强,毛静远,王贤良,等.病证结合治疗慢性心力衰竭理法方药刍议[J].中医杂志,2013,54(7):561-563.
[7]G.T.Liu,T.M.Zhang,B.E.Wang,et al.Protective action of seven natural phenolic compounds against peroxidative damage to biomembranes[J].Biochem.Pharmacol,1992(43):147-152.
[8]C.P.Chen,T.Yokozawa,H.Y.Chung.Inhibitory effect of caffeic acid analogues isolated from Salviae miltiorrhizae radix against 1,1-diphenyl-2-picrylhydrazyl radical[J].Exp.Toxicol.Pathol,1999(51):59-63.
[9]X.J.Wu,Y.P.Wang,W.Wang,et al.Free radical scavenging and inhibition of lipid peroxidation by magnesium lithospermate B[J].Acta Pharmacol.Sin,2000(21):855-858.
[10]S.I.Jang,S.I.Jeong,K.J.Kim,et al.Tanshinone IIA from Salvia miltiorrhiza inhibits inducible nitric oxide synthase expression and production of TNF-α,IL-1βand IL-6 in activated RAW264.7 cells[J].Planta Med,2003(69):1 057-1 059.
[11]G.R.Zhao,H.M.Zhang,T.X.Ye,et al.Characterization of the radical scavenging and antioxidant activities of danshensu and salvianolic acid B[J].Food Chem.Toxicol,2008(46):73-81.
[12]T.O.Cheng.Cardiovascular effects of Danshen[J].Int.J.Cardiol,2007(121):9-22.
[13]于金玲.丹酚酸B药理作用研究进展[J].天津药学,2012,24(3):64-68.
[14]邵静萍,张善飞,陈渊成,等.丹酚酸B在心肌细胞和血管内皮细胞内的摄取特性[J].中国药科大学学报,2009,40(3):263-268.
[15]林超,刘兆国,钱星,等.丹酚酸B在心血管疾病中药理作用研究进展[J].中国药理学通报,2015,31(4):449-452.
[16]胡旭光,相湘,杨超燕,等.丹酚酸B对实验性大鼠急性心肌缺血的保护作用[J].广东医学,2010,31(3):334-336.
[17]高枫,孙桂波,任小宇,等.丹酚酸B对大鼠离体心脏缺血再灌注损伤的保护作用[J].中国中药杂志,2012,37(3):358-361.
[18]宋延平,孔令姗,吴静,等.丹酚酸A与丹酚酸B改善大鼠心肌缺血作用比较[J].中国中医药信息杂志,2007,14(9):36-38.
[19]沈盼盼,谢晓梅,张圣龙,等.UPLC法测定宣木瓜中齐墩果酸、熊果酸[J].南京中医药大学学报,2013,29(3):271-273.
[20]刘建海,崔维利,陈涛,等.HPLC-ELSD法测定新生脉散液中黄芪甲苷的含量[J].世界科学技术-中医药现代化★技术应用研究,2013,15(1):137-140.
[21]胡晓琴,李进,陈涛,等.HPLC法测定新生脉散片中丹参素钠的含量[J].天津中医药,2012,29(2):181-183.