HPLC法测定心无忧片中洋川芎内酯H、洋川芎内酯I、洋川芎内酯A、藁本内酯、丹参素、丹酚酸B和丹参酮Ⅱ_A
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摘要
目的建立高效液相色谱梯度洗脱法同时测定心无忧片中洋川芎内酯H、洋川芎内酯I、洋川芎内酯A、藁本内酯、丹参素、丹酚酸B和丹参酮Ⅱ_A的方法。方法采用Hydrosphere C18色谱柱(250 mm×4.6 mm,5μm);流动相A:甲醇–乙腈(2∶1),流动相B:0.2%冰醋酸溶液,梯度洗脱(0~11 min,45.0%A;11~26 min,45.0%→68.0%A;26~39 min,68.0%→82.0%A;39~45 min,82.0%→45.0%A);检测波长:280 nm;体积流量:0.9 m L/min;柱温:30℃;进样量为10μL。结果洋川芎内酯H、洋川芎内酯I、洋川芎内酯A、藁本内酯、丹参素、丹酚酸B和丹参酮Ⅱ_A的线性范围分别为2.08~41.60μg/m L(r=0.999 3),3.36~67.20μg/m L(r=0.999 8),4.29~85.80μg/m L(r=0.999 9),6.26~125.20μg/m L(r=0.999 6),3.49~69.80μg/m L(r=0.999 2),49.08~981.60μg/m L(r=0.999 7),7.17~143.40μg/m L(r=0.999 5);平均加样回收率分别为96.81%、98.19%、99.24%、98.93%、97.39%、100.19%、98.63%,RSD值分别为1.51%、1.24%、0.98%、1.12%、0.84%、0.74%、1.44%。结论建立的HPLC梯度洗脱法同时测定心无忧片中心无忧片中洋川芎内酯H、洋川芎内酯I、洋川芎内酯A、藁本内酯、丹参素、丹酚酸B和丹参酮Ⅱ_A 7个成分的测定方法,供试品溶液处理简便,溶液稳定性好,检测方法快捷、准确、灵敏度高,为心无忧片质量标准的提高提供了参考。
Objective To develop HPLC gradient elution method for simultaneous determination of senkyunolide H, senkyunolide I, senkyunolide A, ligustilide, danshensu, salvianolic acid B, and tanshinoneⅡ_A in Xinwuyou Tablets. Methods The determination was carried out on Hydrosphere C18 column(250 mm × 4.6 mm, 5 μm). The mobile phase was methanol-acetonitrile(2∶1) and 0.2% acetic acid glacial solution with gradient elution. The elution procedures were as following: 0 — 11 min with 45.0% A, 11 — 26 min with 45.0%→68.0% A, 26 — 39 min with 68.0%→82.0% A, and 39 — 45 min with 82.0%→45.0% A. The detection wavelengths were set at 280 nm. The flow rate was 0.9 m L/min, temperature of column was set at 30 ℃, and volume of injection was 10 μL. Results Senkyunolide H, senkyunolide I, senkyunolide A, ligustilide, danshensu, salvianolic acid B, and tanshinoneⅡ_A had good linearities in the range of 2.08 — 41.60 μg/m L(r=0.999 3), 3.36 — 67.20 μg/m L(r=0.999 8), 4.29 — 85.80 μg/m L(r=0.999 9), 6.26 — 125.20 μg/m L(r=0.999 6), 3.49 — 69.80 μg/m L(r=0.999 2), 49.08 — 981.60 μg/m L(r=0.999 7), and 7.17 — 143.40 μg/m L(r=0.999 5), respectively. The average recoveries were 96.81%, 98.19%, 99.24%, 98.93%, 97.39%, 100.19%, and 98.63%, respectively. And the corresponding RSD values were 1.51%, 1.24%, 0.98%, 1.12%, 0.84%, 0.74%, and 1.44%, respectively. Conclusion The established method of HPLC gradient elution method for simultaneous determination of senkyunolide H, senkyunolideI, senkyunolide A, ligustilide, danshensu, salvianolic acid B, and tanshinone Ⅱ_A in Xinwuyou Tablets is simple for the solution treatment, stable for the solution, and fast for the detection method. The method is accurate and sensitive which can be applied to reference for the quality control of Xinwuyou Tablets.
Objective To develop HPLC gradient elution method for simultaneous determination of senkyunolide H, senkyunolide I, senkyunolide A, ligustilide, danshensu, salvianolic acid B, and tanshinoneⅡ_A in Xinwuyou Tablets. Methods The determination was carried out on Hydrosphere C18 column(250 mm × 4.6 mm, 5 μm). The mobile phase was methanol-acetonitrile(2∶1) and 0.2% acetic acid glacial solution with gradient elution. The elution procedures were as following: 0 — 11 min with 45.0% A, 11 — 26 min with 45.0%→68.0% A, 26 — 39 min with 68.0%→82.0% A, and 39 — 45 min with 82.0%→45.0% A. The detection wavelengths were set at 280 nm. The flow rate was 0.9 m L/min, temperature of column was set at 30 ℃, and volume of injection was 10 μL. Results Senkyunolide H, senkyunolide I, senkyunolide A, ligustilide, danshensu, salvianolic acid B, and tanshinoneⅡ_A had good linearities in the range of 2.08 — 41.60 μg/m L(r=0.999 3), 3.36 — 67.20 μg/m L(r=0.999 8), 4.29 — 85.80 μg/m L(r=0.999 9), 6.26 — 125.20 μg/m L(r=0.999 6), 3.49 — 69.80 μg/m L(r=0.999 2), 49.08 — 981.60 μg/m L(r=0.999 7), and 7.17 — 143.40 μg/m L(r=0.999 5), respectively. The average recoveries were 96.81%, 98.19%, 99.24%, 98.93%, 97.39%, 100.19%, and 98.63%, respectively. And the corresponding RSD values were 1.51%, 1.24%, 0.98%, 1.12%, 0.84%, 0.74%, and 1.44%, respectively. Conclusion The established method of HPLC gradient elution method for simultaneous determination of senkyunolide H, senkyunolideI, senkyunolide A, ligustilide, danshensu, salvianolic acid B, and tanshinone Ⅱ_A in Xinwuyou Tablets is simple for the solution treatment, stable for the solution, and fast for the detection method. The method is accurate and sensitive which can be applied to reference for the quality control of Xinwuyou Tablets.
引文
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[11]刘金亮,范巧佳,郑顺林,等.HPLC测定不同采收期川芎药材中5种药效成分的含量[J].中国中药杂志,2014,39(9):1650-1655.
[12]刘洁,余李敏,翟帆.高效液相色谱-质谱法同时测定川芎提取液中7种有效成分[J].中成药,2010,32(8):1380-1382.
[2]李剑平,范小芬.心无忧片治疗冠心病心绞痛40例观察—附复方丹参片治疗40例对照[J].浙江中医杂志,2001,36(5):230.
[3]中华人民共和国卫生部药典委员会.卫生部颁药品标准(中药成方制剂第十三册)[S].1997:46.
[4]中国药典[S].一部.2015:40-41,76-77.
[5]满喜霞,黄占海.HPLC测定心无忧片中丹参酮ⅡA的含量[J].海峡药学,2010,22(7):65-67.
[6]龚青.高效液相色谱法测定心无忧片中丹参酮ⅡA的含量[J].中国现代应用药杂志,2003,20(7):51-52.
[7]曹建敏,王宗花,丁明玉,等.反相高效液相色谱法同时测定川芎中的四种内酯类化合物[J].色谱,2005,23(5):531-533.
[8]翟学佳,徐锦凤.高效液相色谱法同时测定丹参药材水溶性和脂溶性成分的含量[J].医药导报,2009,28(10):1345-1348.
[9]程沛,韩东岐,胡伟慧,等.高效液相色谱法同时测定丹参中10种水溶性和4种脂溶性成分的含量[J].药物分析杂志,2015,35(6):991-996.
[10]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):1357-1361.
[11]刘金亮,范巧佳,郑顺林,等.HPLC测定不同采收期川芎药材中5种药效成分的含量[J].中国中药杂志,2014,39(9):1650-1655.
[12]刘洁,余李敏,翟帆.高效液相色谱-质谱法同时测定川芎提取液中7种有效成分[J].中成药,2010,32(8):1380-1382.