高效液相色谱法测定复方制剂中的大黄酸、大黄素、大黄酚、大黄素甲醚和二苯乙烯苷的含量

Simultaneous determination of rhein,emodin,chryophanol,physcion and stibene glucoside in compound preparations by HPLC

目的建立高效液相色谱法(high efficiency liquid chromatography,HPLC)同时测定明目上清片中4种化学成分(大黄酸、大黄素、大黄酚和大黄素甲醚)的含量以及降脂宁片中二苯乙烯苷的含量。方法明目上清片采用Kromasil C_(18)柱(250 mm×4.6 mm,5μm),以甲醇-四氢呋喃-1%磷酸(84∶7∶9)为流动相,流速1.0 m L/min,检测波长254 nm,进样量20μL;降脂宁片采用Kromasil C_(18)柱(250 mm×4.6 mm,5μm),以甲醇-0.1%磷酸为流动相等度洗脱,流速1.0 m L/min,检测波长320 nm,进样量20μL。结果大黄酸、大黄素、大黄酚和大黄素甲醚分别在0.500~4.00(r=0.999 3)、0.370~2.96(r=0.999 0)、1.10~8.80(r=0.999 1)、1.00~8.00(r=0.999 3)ng范围内呈良好线性关系,回归方程分别为Y=8.60×10~3+9.62×10~4X、Y=-1.16×10~4+1.20×10~5X、Y=7.41×10~3+1.20×10~5X、Y=4.13×10~3+6.91×10~4X;二苯乙烯苷在0.218~1.31μg范围内线性关系良好(r=0.998 8),回归方程为Y=-6.33+3.18×10~3X。明目上清片含量测定的平均回收率分别为97.8%、97.8%、98.7%和99.5%,相对标准差(relative standard deviation,RSD)(n=6)分别为1.2%、0.89%、1.7%和1.7%;降脂宁片的平均回收率为96.6%,RSD(n=6)=0.65%。结论该方法简便快速,准确度高,专属性好,可以用于明目上清片以及降脂宁片的质量控制。

Objective To develop an analysis method for Rhein,Emodin,Chrysophanol and Physcion in Mingmu Shangqing tablets and stibene glucoside in Jiangzhining tablets by high efficiency liquid chromatography（ HPLC）. Methods The four constituents of Mingmu Shangqing tablets were separated on Kromasil C_（18）column（ 250 mm × 4. 6 mm,5 μm） using a mobile phase consisted of methanol-tetrahydrofuran-1% phosphoric acid in water at the detection wavelength of 254 nm. The constituent stibene glucoside in Jiangzhining tablets was separated on Kromasil C_（18）column（ 250 mm ×4. 6 mm,5 μm） using a mobile phase consisted of methanol 0. 1% phosphoric acid in water at the detection wavelength of 320 nm. Results Rhein,Emodin,Chrysophanol and Physcionhad had a good liner relationship within the scope of 0. 500 to 4. 00（ r = 0. 999 3）,0. 370 to 2. 96（ r = 0. 999 0）,1. 10 to 8. 80（ r = 0. 999 0）,1. 00 to 8. 00（ r = 0. 999 0） ng,respectively. And their regression equation wereY = 8. 60 ×10~3＋ 9. 62 ×10~4 X,Y =- 1. 16 ×10~4＋ 1. 20 ×10~5 X,Y = 7. 41 ×10~3＋ 1. 20 ×10~5 X,Y =4. 13 ×10~3＋ 6. 91 ×10~4 X,respectively. The average recoveries（ n = 6） were 97. 8%,97. 8%,98. 7%,99. 5% and with relative standard deviation（ RSD）（ n = 6） of 1. 2%,0. 89%,1. 7%,1. 7%. Stibene glucoside had a good liner relationship within the scope of 0. 218 to 1. 31 ng（ r = 0. 999 0）. The regression equation was Y =- 6. 33 ＋ 3. 18 ×10~3 X. The average recoveries were 96. 6% and with RSD of 0. 65%. Conclusion The method is rapid,simple,accurate and with good reproducibility and can be used in quality control of Mingmu Shangqing tablets and Jiangzhining tablets.