高效液相-柱后衍生化法同时测定双氢青蒿素和哌喹  

HPLC with post-column derivatization for simultaneous determination of dihydroartemisinine and piperaquine

作　　者：吴婧[1] 王锐利[1] 申薇薇[1] 张淑秋[1] 

机构地区：[1]山西医科大学药学院临床药学教研室,太原030001

出　　处：《中国药物与临床》2009年第3期178-181,共4页Chinese Remedies & Clinics

基　　金：国家自然科学基金(30572367);山西医科大学2007年度青年基金(02200704)

摘　　要：目的建立同时测定双氢青蒿素和哌喹的方法。方法采用反相高效液相-柱后衍生化-紫外检测法,流动相为乙腈:甲醇:醋酸钠缓冲液(65:20:15,pH4.0),流速0.8ml/min;衍生化试剂为1mol/LKOH的90%乙醇溶液,流速0.3ml/min;衍生化反应体系包括三通阀和6m长的聚醚醚酮管以及超级恒温水浴,流动相所载药物在柱后与衍生试剂经过三通实现混合后在70℃水浴的聚醚醚酮管中反应1min,然后进入检测器;柱温35℃,检测波长289nm。结果双氢青蒿素在3.0～300μmol/L范围内线性关系良好,以峰面积对双氢青蒿素浓度进行线性回归,方程为A=3544.3C+3846.4,r=0.9999;同时,哌喹在6.8～680μmol/L范围内线性关系良好,以峰面积对哌喹浓度进行线性回归,方程为A=7217.1C+28115,r=0.9996;对两种药物测定的日内、日间精密度相对平均偏差(RSD)均小于2%;平均回收率均在98%～102%之间。结论该方法的灵敏度、精密度、准确性及重现性均符合药典要求,可为各种复方制剂中双氢青蒿素及哌喹的质量控制提供依据。Objective To establish a method for simultaneous determination of dihydroartemisinine and piperaquine using HPLC with post-column derivatization and UV detection. Methods HPLC method was developed with a mobile phase consisted of acetonitrile, methanol and 0.1 mol/L acetate buffer （pH 4.0） （65：20：15）, delivered at a flow-rate of 0.8 ml/min. 1.0 mol/L potassium hydroxide in 90% ethanol as post-column derivatization reagents, was delivered at a flow- rate of 0.3 ml/min and mixed with mobile phase through a T-valve post column. The derivative reaction was rendered at 70℃ for 1 min. Column temperature was kept at 35℃, and UV detection wave length was set at 289 nm. Results Caliberation curve was linear over the concentration range of 3 -300 μmol/L for dihydroartemisinine and 6.8-680 μLmol/L for piperaquine. Linear regression between peak area and concentration yielded equation A =3544.3C＋3846.4 （r=0.9999） for dihydroartemisinine and A=7217.1 C＋28115 for piperaquine （r=0.9996）. With regard to both of dihydroartemisinin and piperaquine, the inter-day and intra-day RSDs were less than 2 %. Mean recovery were 98%-102%. Conclusion The accuracy, precision and reproducibility of this method appeared consistent with the requirements of pharmacopedia, and may provide a basis for quality control of dihydroartemisinine and piperaquine preparations.

关 键 词：药用制剂 色谱法高效液相 双氢青蒿素 哌喹 

分 类 号：R286[医药卫生—中药学] R927[医药卫生—中医学]