HPLC-波长切换法同时测定强肾片中8种活性成分的含量
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摘要
目的建立强肾片中没食子酸、5-羟甲基糠醛、莫诺苷、马钱苷、芍药苷、丹酚酸B、丹皮酚和丹参酮ⅡA8种成分含量的测定方法。方法采用HPLC-波长切换法,Diamonsil C_(18)色谱柱(250 mm×4.6 mm,5μm),以乙腈(A)-0.05%甲酸水溶液(B)为流动相进行梯度洗脱,检测波长(0～10 min,270 nm; 10～30 min,240 nm; 30～65 min,270 nm),柱温35℃,流速1.0 m L·min~(-1),进样体积20μL。结果强肾片中没食子酸、5-羟甲基糠醛、莫诺苷、马钱苷、芍药苷、丹酚酸B、丹皮酚和丹参酮ⅡA质量浓度分别在6.06～30.30μg·m L-1、4.96～24.80μg·m L-1、10.31～51.55μg·mL-1、4.915～24.58μg·mL~(-1)、10.01～50.05μg·mL~(-1)、24.02～120.1μg·mL~(-1)、20.02～100.1μg·mL~(-1)及1.622～8.112μg·mL~(-1)内与峰面积呈良好的线性关系(r≥0.999 8),平均回收率(n=9)分别为99.7%、97.6%、100.8%、99.9%、98.6%、101.1%、101.1%及101.5%。结论所建立的方法稳定可靠,灵敏度高,重复性好,可同时测定强肾片中8种成分的含量,可为强肾片质量评价提供依据。
OBJECTIVE To determinate the contents of gallic acid, 5-hydroxymethyl furfural, morroniside, loganin,paeoniflorin,salvianolic acid B,paeonol and tanshinone ⅡAin Qiangshen tablet by HPLC,and provide references for its quality control. METHODS The HPLC analysis was carried out by Diamonsil C_(18) column( 250 mm× 4.6 mm,5 μm) with acetonitrile( A)-0.05% formic acid solution( B) as the mobile phase for gradient elution. The detection wavelength was set at 270 nm for 0-10 min,240 nm for 10-30 min and 270 nm for 30-65 min. In addition,the flow rate was 1. 0 m L·min-1,column temperature was 35 ℃ and injection volumn was 20 μL. RESULTS Gallic acid,5-hydroxymethyl furfural,morroniside,loganin,paeoniflorin,salvianolic acid B,paeonol and tanshinone ⅡAshowed good linearity in the range of 6.06-30.30 μg·mL~(-1),4.96-24.80 μg·mL~(-1),10.31-51.55 μg·mL~(-1),4.915-24.58 μg·mL~(-1),10.01-50. 05 μg·mL~(-1),24. 02-120. 1 μg·mL~(-1),20. 02-100. 1 μg·mL~(-1) and 1. 622-8. 112 μg·mL~(-1)( r≥0.999 8),respectively.The recovery of those 8 components were 99.7%,97.6%,100.8%,99.9%,98.6%,101.1%,101.1% and101.5%,respectively. CONCLUSION The established method can be used to determine the contents of those 8 compounds. Such stable,reliable and repeatable analytical mothed can provide a reference for the quality evaluation and its control on Qiangshen tablet.
OBJECTIVE To determinate the contents of gallic acid, 5-hydroxymethyl furfural, morroniside, loganin,paeoniflorin,salvianolic acid B,paeonol and tanshinone ⅡAin Qiangshen tablet by HPLC,and provide references for its quality control. METHODS The HPLC analysis was carried out by Diamonsil C_(18) column( 250 mm× 4.6 mm,5 μm) with acetonitrile( A)-0.05% formic acid solution( B) as the mobile phase for gradient elution. The detection wavelength was set at 270 nm for 0-10 min,240 nm for 10-30 min and 270 nm for 30-65 min. In addition,the flow rate was 1. 0 m L·min-1,column temperature was 35 ℃ and injection volumn was 20 μL. RESULTS Gallic acid,5-hydroxymethyl furfural,morroniside,loganin,paeoniflorin,salvianolic acid B,paeonol and tanshinone ⅡAshowed good linearity in the range of 6.06-30.30 μg·mL~(-1),4.96-24.80 μg·mL~(-1),10.31-51.55 μg·mL~(-1),4.915-24.58 μg·mL~(-1),10.01-50. 05 μg·mL~(-1),24. 02-120. 1 μg·mL~(-1),20. 02-100. 1 μg·mL~(-1) and 1. 622-8. 112 μg·mL~(-1)( r≥0.999 8),respectively.The recovery of those 8 components were 99.7%,97.6%,100.8%,99.9%,98.6%,101.1%,101.1% and101.5%,respectively. CONCLUSION The established method can be used to determine the contents of those 8 compounds. Such stable,reliable and repeatable analytical mothed can provide a reference for the quality evaluation and its control on Qiangshen tablet.
引文
[1]国家药典委员会.中华人民共和国药典[M].1部.北京:中国医药科技出版社,2015:1637-1638.
[2]关剑锋,张萍,赵岩.高效液相色谱法测定强肾片的含量[J].辽宁医药,2001,16(3):12-15.
[3]徐木根,朱山寅.高效液相色谱法测定强肾片中芍药苷的含量[J].医药导报,2005,24(1):69-70.
[4]郭强,宋汉敏,李振国.HPLC法同时测定强肾片中补骨脂素与异补骨脂素的含量[J].中医研究,2011,24(7):24-27.
[5]丁霞,王明艳,余宗亮,等.山茱萸中5-羟甲基糠醛的分离鉴定及生物活性研究[J].中国中药杂志,2008,33(4):392-396.
[6]王喜军,张宁,孙晖,等.六味地黄丸血中移行成分对氢化可的松致大鼠肾虚动物模型的保护作用[J].中国实验方剂学杂志,2008,14(2):33-37.
[7]唐锦辉,占成业.丹参酮ⅡA对大鼠肾间质纤维化的抑制作用[J].内科急危重症杂志,2008,14(1):16-18.
[8]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.
[9]李向阳,屠万倩,张留记.RP-HPLC法测定不同产地的牡丹皮中芍药苷和丹皮酚的含量[J].中药新药与临床药理,2011,22(5):563-565.
[10]杨正生,彭振辉,姚青海,等.丹皮酚的药理作用研究进展[J].中国药物与临床,2011,11(5):545-547.
[11]李肖玲,崔岚,祝德秋.没食子酸生物学作用的研究进展[J].中国药师,2004,7(10):767-769.
[12]董建华,王德秀,赵延武,等.六味地黄制剂化学成分的定量研究进展[J].今日药学,2008,18(1):74-78.
[13]蔡银燕,石婷婷,黄巧玲.HPLC测定六味地黄浓缩丸中5-HMF的含量[J].中国现代应用药学,2012,29(4):356-358.
[14]陈宇,赵芝兰,朱河水.不同提取方法对复方丹参片中丹参酮ⅡA和丹酚酸B含量测定的影响[J].中国农学通报,2007,23(5):134-138.
[15]李娴,王本杰,袁桂艳,等.六味地黄丸中四种活性成分的HPLC法测定[J].中国医药工业杂志,2010,41(2):126-128.
[16]刘潇潇,杨立伟,于江泳,等.HPLC法测定参芪扶正注射液中5-羟甲基糠醛的含量及其来源的初步探讨[J].药物分析杂志,2012,32(4):639-642.
[2]关剑锋,张萍,赵岩.高效液相色谱法测定强肾片的含量[J].辽宁医药,2001,16(3):12-15.
[3]徐木根,朱山寅.高效液相色谱法测定强肾片中芍药苷的含量[J].医药导报,2005,24(1):69-70.
[4]郭强,宋汉敏,李振国.HPLC法同时测定强肾片中补骨脂素与异补骨脂素的含量[J].中医研究,2011,24(7):24-27.
[5]丁霞,王明艳,余宗亮,等.山茱萸中5-羟甲基糠醛的分离鉴定及生物活性研究[J].中国中药杂志,2008,33(4):392-396.
[6]王喜军,张宁,孙晖,等.六味地黄丸血中移行成分对氢化可的松致大鼠肾虚动物模型的保护作用[J].中国实验方剂学杂志,2008,14(2):33-37.
[7]唐锦辉,占成业.丹参酮ⅡA对大鼠肾间质纤维化的抑制作用[J].内科急危重症杂志,2008,14(1):16-18.
[8]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.
[9]李向阳,屠万倩,张留记.RP-HPLC法测定不同产地的牡丹皮中芍药苷和丹皮酚的含量[J].中药新药与临床药理,2011,22(5):563-565.
[10]杨正生,彭振辉,姚青海,等.丹皮酚的药理作用研究进展[J].中国药物与临床,2011,11(5):545-547.
[11]李肖玲,崔岚,祝德秋.没食子酸生物学作用的研究进展[J].中国药师,2004,7(10):767-769.
[12]董建华,王德秀,赵延武,等.六味地黄制剂化学成分的定量研究进展[J].今日药学,2008,18(1):74-78.
[13]蔡银燕,石婷婷,黄巧玲.HPLC测定六味地黄浓缩丸中5-HMF的含量[J].中国现代应用药学,2012,29(4):356-358.
[14]陈宇,赵芝兰,朱河水.不同提取方法对复方丹参片中丹参酮ⅡA和丹酚酸B含量测定的影响[J].中国农学通报,2007,23(5):134-138.
[15]李娴,王本杰,袁桂艳,等.六味地黄丸中四种活性成分的HPLC法测定[J].中国医药工业杂志,2010,41(2):126-128.
[16]刘潇潇,杨立伟,于江泳,等.HPLC法测定参芪扶正注射液中5-羟甲基糠醛的含量及其来源的初步探讨[J].药物分析杂志,2012,32(4):639-642.