UPLC法同时测定丹参片中11个水溶性和脂溶性成分的含量
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摘要
目的:采用UPLC法同时测定丹参片中11个水溶性和脂溶性成分丹参素钠、二氢丹参酮、隐丹参酮、丹参酮、丹参酮ⅡA、熊果酸、原儿茶醛、迷迭香酸、紫草酸、丹酚酸B和丹酚酸A的含量。方法:采用Waters ACQUITY UPLC C_8色谱柱(2.1 mm×100 mm,1.7μm),以0.1%磷酸水溶液(A)-乙腈(B)为流动相,梯度洗脱(0～13 min,5%B→22%B;13～16 min,22%B→40%B;16～20 min,40%B→53%B;20～39 min,53%B→80%B),流速0.3 mL·min~(-1);检测波长280 nm,柱温20℃,样品管理器温度4℃。结果:丹参素钠、二氢丹参酮、隐丹参酮、丹参酮、丹参酮ⅡA、熊果酸、原儿茶醛、迷迭香酸、紫草酸、丹酚酸B和丹酚酸A进样量分别在0.203 1～8.125、0.035 8～1.431、0.063 3～2.531、0.091 8～3.671、0.090 2～3.606、0.096 1～3.844、0.036 0～1.439、0.158 1～6.324、0.103 1～4.125、0.283 1～11.325和0.191 2～7.647μg的范围内与色谱峰峰面积呈良好线性关系;平均回收率(n=6)分别为99.2%、99.7%、99.8%、98.9%、99.5%、99.4%、99.6%、99.5%、98.7%、99.3%和98.9%,RSD分别为0.49%、0.44%、0.19%、0.61%、0.63%、0.29%、0.79%、0.48%、0.92%、0.40%和0.88%。8个厂家74批丹参片样品中上述11个成分含量范围分别为0.645～7.182、0.079～1.281、0～2.21、0.109～3.37、0.203～3.063、2.033～3.527、0～1.063、1.354～5.677、1.046～3.909、32.759～51.588、0.156～9.472 mg·g-1。结论:所建立的方法可作为丹参片的质量控制方法。
Objective:To establish an UPLC method for simultaneous determination of 11 water soluble and lipophilic components including danshensu sodium,dihydrotanshinone,cryptotanshinone,tanshinone,tanshinoneⅡA,ursolic acid,protocatechuic aldehyde,rosmarinic acid,lithospermic acid,salvianolic acid B and salvianolic acid A in Danshen tablets. Methods:The sepatation was performed on Waters ACQUITY UPLC C_8 column(2.1 mm×100 mm,1.7 μm)with the mobile phase consisting of 0.1% phosphoric acid aqueous(A)-acetonitrile(B)by gradient elution(0-13 min,5%B → 22%B;13-16 min,22%B → 40%B;16-20 min,40%B → 53%B;20-39 min,53%B was 4 ℃. The detection wavelength was 280 nm. Results:The linear ranges of danshensu sodium,dihydrotanshinone,cryptotanshinone,tanshinone,tanshinone ⅡA,ursolic acid,protocatechuic aldehyde,rosmarinic acid,lithospermic acid, salvianolic acid B and salvianolic acid A were 0.203 1-8.125,0.035 8-1.431,0.063 3-2.531,0.091 8-3.671,0.090 2-3.606,0.096 1-3.844,0.036 0-1.439,0.158 1-6.324,0.103 1-4.125,0.283 1-11.325 and 0.191 2-2.296 μg,respectively. The recoveries(n=6)of the above-mentioned components were 99.2%,99.7%,99.8%,98.9%,99.5%,99.4%,99.6%,99.5%,98.7%,99.3% and 98.9%,respectively,and the relative standard deviations were 0.49%,0.44%,0.19%,0.61%,0.63%,0.29%,0.79%,0.48%,0.92%,0.40% and 0.88%,respectively. The content ranges of abovementioned component in 74 batches of samples from 8 manufactures were 0.645-7.182,0.079-1.281,0-2.21,0.109-3.37,0.203-3.063,2.033-3.527,0-1.063,1.354-5.677,1.046-3.909,32.759-51.588 and 0.156-9.472 mg·g-1, respectively. Conclusion:The method can be used for quality control of Danshen tablets.
Objective:To establish an UPLC method for simultaneous determination of 11 water soluble and lipophilic components including danshensu sodium,dihydrotanshinone,cryptotanshinone,tanshinone,tanshinoneⅡA,ursolic acid,protocatechuic aldehyde,rosmarinic acid,lithospermic acid,salvianolic acid B and salvianolic acid A in Danshen tablets. Methods:The sepatation was performed on Waters ACQUITY UPLC C_8 column(2.1 mm×100 mm,1.7 μm)with the mobile phase consisting of 0.1% phosphoric acid aqueous(A)-acetonitrile(B)by gradient elution(0-13 min,5%B → 22%B;13-16 min,22%B → 40%B;16-20 min,40%B → 53%B;20-39 min,53%B was 4 ℃. The detection wavelength was 280 nm. Results:The linear ranges of danshensu sodium,dihydrotanshinone,cryptotanshinone,tanshinone,tanshinone ⅡA,ursolic acid,protocatechuic aldehyde,rosmarinic acid,lithospermic acid, salvianolic acid B and salvianolic acid A were 0.203 1-8.125,0.035 8-1.431,0.063 3-2.531,0.091 8-3.671,0.090 2-3.606,0.096 1-3.844,0.036 0-1.439,0.158 1-6.324,0.103 1-4.125,0.283 1-11.325 and 0.191 2-2.296 μg,respectively. The recoveries(n=6)of the above-mentioned components were 99.2%,99.7%,99.8%,98.9%,99.5%,99.4%,99.6%,99.5%,98.7%,99.3% and 98.9%,respectively,and the relative standard deviations were 0.49%,0.44%,0.19%,0.61%,0.63%,0.29%,0.79%,0.48%,0.92%,0.40% and 0.88%,respectively. The content ranges of abovementioned component in 74 batches of samples from 8 manufactures were 0.645-7.182,0.079-1.281,0-2.21,0.109-3.37,0.203-3.063,2.033-3.527,0-1.063,1.354-5.677,1.046-3.909,32.759-51.588 and 0.156-9.472 mg·g-1, respectively. Conclusion:The method can be used for quality control of Danshen tablets.
引文
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[8]何昱,成金乐,徐吉银,等.HPLC-DAD同时测定丹参中原儿茶醛、丹酚酸B、隐丹参酮、丹参酮ⅡA四种成分的含量[J].世界科学技术-中医药现代化,2011,13(4):688HE Y,CHENG JL,XU JY,et al.Simultaneous determination of four constituents of protocatechualdehyde,saivianolic acid B,cryptotaushinone,taushinoneⅡA in Salvia miltiorrhiza preparations by HPLC-DAD[J].World Sci Technol Mod Tradit Chin Med Mater Med,2011,13(4):688
[9]刘敏彦,赵韶华,盖潇桦,等.丹参不同提取方法中水溶性成分含量的比较研究[J].时珍国医国药,2007,18(5):1186LIU MY,ZHAO SH,GAI XH,et al.Study on determination of water-soluble active principles of Salvia miltiorrhiza by different extraction method[J].Lishizhen Med Mater Med Res,2007,18(5):1186
[10]周晓希,孔羽,魏宇昆,等.RP-HPLC-DAD同时测定丹参及丹参片中6种水溶性成分的含量[J].中药材,2014,37(2):337ZHOU XX,KONG Y,WEI YK,et al.Simultaneous determination of six water-soluble components in Salvia miltiorrhiza and Danshen tablets by RP-HPLC-DAD[J].J Chin Med Mater,2014,37(2):337
[11]许虎,刘训红,傅兴圣,等.高效毛细管电泳法同时测定丹参饮片中5种酚酸类成分[J].中国药学杂志,2012,47(20):1661XU H,LIU XH,FU XS,et al.Simultaneous determination of five phenolic acids in Salviae Miltiorrhizae Radix et Rhizoma by HPCE[J].Chin Pharm J,2012,47(20):1661
[12]崔晓红,奇海涛,生宁,等.HPLC法同时测定益肝康颗粒中没食子酸、丹参素、芍药苷、丹酚酸B和丹参酮ⅡA的含量[J].药物分析杂志,2014,34(3):432CUI XH,QI HT,SHENG N,et al.HPLC simultaneous determination of galic acid,tanshinol,paeoniflorin salvianolic acid Band tanshinoneⅡA in Yigankang granules[J].Chin J Pharm Anal,2014,34(3):432
[13]何昱,李晓如,李冰心,等.HPLC-DAD法同时测定丹参中丹酚酸B和隐丹参酮的含量[J].现代中药研究与实践,2010,24(5):78HE Y,LI XR,LI BX,et al.Simultaneous determination of salvianolic acid B and cryptotanshinone in Salvia miltiorrhiza preparations by HPLC-DAD[J].Chin Med J Res Pract,2010,24(5):78
[14]李桃,王春娥.高效液相色谱法测定丹参药材中丹参素的含量[J].中国医院药学杂志,2005,25(9):832LI T,WANG CE.Determination of danshensu in Salvia miltiorrhiza with HPLC[J].Chin Hosp Pharm J,2005,25(9):832
[15]周国军,李焱,秦民坚,等.高效液相色谱法快速测定丹参中5种活性成分的含量[J].药物分析杂志,2012,32(8):1357ZHOU GJ,LI Y,QIN MJ,et al.Fast determination of 5 constituents in Radix et Rhizoma Salviae Miltiorrhizae by HPLC[J].Chin JPharm Anal,2012,32(8):1357
[16]王伟,李焱,谢晓梅,等.高效液相色谱法同时测定丹参中4种酚酸类成分的含量[J].安徽医药,2010,14(11):1276WANG W,LI Y,XIE XM,et al.Simultaneous determination of four phenolic acids in roots of Salvia miltiorrhiza by HPLC[J].Anhui Med Pharm J,2010,14(11):1276
[17]程茜菲.高效液相色谱法同时测定丹参中6种化学成分的含量[J].西北药学杂志,2016,31(6):551CHENG QF.Simultaneous determination of six components in Radix Salviae Miltiorrhizae by HPLC[J].Northwest Pharm J,2016,31(6):551
[18]李耿,孟繁蕴,杨洪军,等.UPLC法同时测定丹参中11种成分的含量[J].中国药房,2014,25(19):1767LI G,MENG FY,YANG HJ,et al.Simultaneous determination of11 components in Salvia miltiorrhiza by UPLC[J].China Pharm,2014,25(19):1767
[2]周清芬,朱菲白,干倩,等.丹参片对慢性心力衰竭患者的疗效及预后影响[J].内科理论与实践,2011,6(6):428ZHOU QF,ZHU FB,GAN Q,et al.Eficacy of Danshen tablets and its impact on prognosis in treatment of patients with chronic heart failure[J].J Intern Med Concepts Pract,2011,6(6):428
[3]陈晓文,张凤如,吴春芳.丹参片对心力衰竭合并2型糖尿病病人炎症因子及预后的影响[J].中西医结合心脑血管病杂志,2008,6(1):12CHEN XW,ZHANG FR,WU CF.Danshen tablet on TNF-α,hsCRP and 6-MWT in patients with diabetes and heart failure[J].Chin J Integr Med Cardio-/Cerebrovasc Dis,2008,6(1):12
[4]姬晓鹏,蒋斌,潘琳,等.复方丹参片与丹参片对冠状动脉粥样硬化患者血小板活化抑制作用的比铰[J].中国药房,2010,21(19):1085JI XP,JIANG B,PAN L,et al.Comparison of inhibition effect of compound Danshen tablets on platelet activation and that of Danshen tablets in patients with coronary atherosclerosis[J].China Pharm,2010,21(19):1085
[5]孙棣,刘文英,梁彩,等.HPLC法同时测定丹参片中4种有效成分的含量[J].中国药科大学学报,2007,38(1):51SUN D,LIU WY,LIANG C,et al.Simultaneous quantitative determination of four active components in Salvia miltiorrhiza tablets by HPLC[J].J China Pharm Univ,2007,38(1):51
[6]侯媛媛,黄晓婧,晁若冰,等.HPLC同时测定丹参及其制剂中4种酚酸类成分的含量[J].中国药学杂志,2007,42(14):1108HOU YY,HUANG XJ,CHAO RB,et al.Simultaneous determination of four phenolic acids in Salvla miltiorrhiza Bge.and its preparations by HPLC[J].Chin Pharm J,2007,42(14):1108
[7]刘少华,金郁,周大勇,等.超高效液相色谱(UPLC)用于丹参药材水溶性成分指纹图谱研究[J].世界科学技术-中医药现代化,2007,9(6):46LIU SH,JIN Y,ZHOU DY,et al.Studying fingerprint chromatograms of water-soluble components in Radix Salviae Miltiorrhizae using UPLC[J].World Sci Technol Mod Tradit Chin Med Mater Med,2007,9(6):46
[8]何昱,成金乐,徐吉银,等.HPLC-DAD同时测定丹参中原儿茶醛、丹酚酸B、隐丹参酮、丹参酮ⅡA四种成分的含量[J].世界科学技术-中医药现代化,2011,13(4):688HE Y,CHENG JL,XU JY,et al.Simultaneous determination of four constituents of protocatechualdehyde,saivianolic acid B,cryptotaushinone,taushinoneⅡA in Salvia miltiorrhiza preparations by HPLC-DAD[J].World Sci Technol Mod Tradit Chin Med Mater Med,2011,13(4):688
[9]刘敏彦,赵韶华,盖潇桦,等.丹参不同提取方法中水溶性成分含量的比较研究[J].时珍国医国药,2007,18(5):1186LIU MY,ZHAO SH,GAI XH,et al.Study on determination of water-soluble active principles of Salvia miltiorrhiza by different extraction method[J].Lishizhen Med Mater Med Res,2007,18(5):1186
[10]周晓希,孔羽,魏宇昆,等.RP-HPLC-DAD同时测定丹参及丹参片中6种水溶性成分的含量[J].中药材,2014,37(2):337ZHOU XX,KONG Y,WEI YK,et al.Simultaneous determination of six water-soluble components in Salvia miltiorrhiza and Danshen tablets by RP-HPLC-DAD[J].J Chin Med Mater,2014,37(2):337
[11]许虎,刘训红,傅兴圣,等.高效毛细管电泳法同时测定丹参饮片中5种酚酸类成分[J].中国药学杂志,2012,47(20):1661XU H,LIU XH,FU XS,et al.Simultaneous determination of five phenolic acids in Salviae Miltiorrhizae Radix et Rhizoma by HPCE[J].Chin Pharm J,2012,47(20):1661
[12]崔晓红,奇海涛,生宁,等.HPLC法同时测定益肝康颗粒中没食子酸、丹参素、芍药苷、丹酚酸B和丹参酮ⅡA的含量[J].药物分析杂志,2014,34(3):432CUI XH,QI HT,SHENG N,et al.HPLC simultaneous determination of galic acid,tanshinol,paeoniflorin salvianolic acid Band tanshinoneⅡA in Yigankang granules[J].Chin J Pharm Anal,2014,34(3):432
[13]何昱,李晓如,李冰心,等.HPLC-DAD法同时测定丹参中丹酚酸B和隐丹参酮的含量[J].现代中药研究与实践,2010,24(5):78HE Y,LI XR,LI BX,et al.Simultaneous determination of salvianolic acid B and cryptotanshinone in Salvia miltiorrhiza preparations by HPLC-DAD[J].Chin Med J Res Pract,2010,24(5):78
[14]李桃,王春娥.高效液相色谱法测定丹参药材中丹参素的含量[J].中国医院药学杂志,2005,25(9):832LI T,WANG CE.Determination of danshensu in Salvia miltiorrhiza with HPLC[J].Chin Hosp Pharm J,2005,25(9):832
[15]周国军,李焱,秦民坚,等.高效液相色谱法快速测定丹参中5种活性成分的含量[J].药物分析杂志,2012,32(8):1357ZHOU GJ,LI Y,QIN MJ,et al.Fast determination of 5 constituents in Radix et Rhizoma Salviae Miltiorrhizae by HPLC[J].Chin JPharm Anal,2012,32(8):1357
[16]王伟,李焱,谢晓梅,等.高效液相色谱法同时测定丹参中4种酚酸类成分的含量[J].安徽医药,2010,14(11):1276WANG W,LI Y,XIE XM,et al.Simultaneous determination of four phenolic acids in roots of Salvia miltiorrhiza by HPLC[J].Anhui Med Pharm J,2010,14(11):1276
[17]程茜菲.高效液相色谱法同时测定丹参中6种化学成分的含量[J].西北药学杂志,2016,31(6):551CHENG QF.Simultaneous determination of six components in Radix Salviae Miltiorrhizae by HPLC[J].Northwest Pharm J,2016,31(6):551
[18]李耿,孟繁蕴,杨洪军,等.UPLC法同时测定丹参中11种成分的含量[J].中国药房,2014,25(19):1767LI G,MENG FY,YANG HJ,et al.Simultaneous determination of11 components in Salvia miltiorrhiza by UPLC[J].China Pharm,2014,25(19):1767