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高效液相紫外色谱法同时测定抗耐药脂质体中的表阿霉素、氨氯地平和地喹氯铵

门萤, 王小星, 居瑞军, 田玮, 应雪, 姚红娟, 张燕, 李若婧, 吕万良*   

  1. 北京大学医学部 天然药物及仿生药物国家重点实验室; 药学院 药剂系, 北京 100191
  • 收稿日期:2010-03-09 修回日期:2010-06-10 出版日期:2010-07-15 发布日期:2010-07-15
  • 通讯作者: 吕万良*

Development of an HPLC-UV method for the simultaneous determination of epirubicin, amlodipine and dequalinium in anti-resistant liposomes

Ying Men, Xiao-Xing Wang, Rui-Jun Ju, Wei Tian, Xue Ying, Hong-Juan Yao, Yan Zhang, Ruo-Jing Li, Wan-Liang Lu*   

  1. State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
  • Received:2010-03-09 Revised:2010-06-10 Online:2010-07-15 Published:2010-07-15
  • Contact: Wan-Liang Lu*

摘要:

为克服白血病的内源性耐药, 以表阿霉素为抗肿瘤药物, 氨氯地平为凋亡诱导剂, 地喹氯铵为靶向分子, 构建了一种新的表阿霉素抗耐药性脂质体。研究目的是建立一种高效液相方法可同时测定该脂质体中表阿霉素、氨氯地平和地喹氯铵等三种成分。室温条件采用ODS色谱柱进行分离, 流动相为乙腈-0.02 M磷酸二氢钠-三乙胺 (34:66:0.3, v/v/v, 用磷酸调pH值至4.0), 检测波长为240 nm, 流速为1.0 mL/min。在1~50 µg/mL 浓度范围内, 表阿霉素、氨氯地平与地喹氯铵的浓度与峰面积之间分别呈现良好的线性关系 (r2 = 0.9999)。表阿霉素, 氨氯地平与地喹氯铵的平均回收率分别为95.86%~ 97.52%, 97.17%~98.92% 和 98.04%~101.13%; 三批载药脂质体中表阿霉素、氨氯地平和地喹氯铵含量范围分别为(564.2~606.1) µg/mL、(641.0~704.0) µg/mL、(816.0~898.0) µg/mL。脂质体中表阿霉素和氨氯地平的包封率分别为90%左右, 地喹氯铵的修饰率大约70 µg/µmol磷脂。该方法简便、重现性好, 可用于同时检测该脂质体中的表阿霉素、氨氯地平和地喹氯铵。

关键词: 表阿霉素, 氨氯地平, 地喹氯铵, 脂质体, 高效液相紫外色谱法

Abstract: Novel anti-resistant liposomes have been developed to overcome intrinsic resistance in leukemia. Anticancer agent epirubicin and apoptotic inducer amlodipine were encapsulated into the liposome aqueous core, and the surface of the liposome was modified using dequalinium. The objective of the present study was to establish a high performance liquid chromatography (HPLC) method for the determination of epirubicin, amlodipine and dequalinium in the liposomes. Analysis was performed on an ODS column with an isocratic elution at ambient temperature. Mobile phase was consisted of acetonitrile, 0.02 M NaH2PO4 and triethylamine (34:66:0.3, v/v/v, pH 4.0). The detection wavelength was set at 240 nm and the flow rate was 1.0 mL/min. The results showed that the calibration curves of epirubicin, amlodipine and dequalinium were linear in the range of (1-50) µg/mL (r2 = 0.9999), respectively. The mean recoveries of epirubicin, amlodipine, and dequalinium were in the range of 95.86%-97.52%, 97.17%-98.92% and 98.04%-101.13%, respectively. The contents of epirubicin, amlodipine and dequalinium in the liposomes were in the range of (564.2-606.1) µg/mL, (641.0-704.0) µg/mL, and (816.0-898.0) µg/mL, respectively. The encapsulation efficiencies of epirubicin and amlodipine were around 90%, and the modification rate of dequalinium was approximate 70 µg/µmol lipids. The proposed HPLC method was simple and accurate for the simultaneous determination of epirubicin, amlodipine and dequalinium in newly developed anti-resistant liposomes.

Key words: Epirubicin, Amlodipine, Dequalinium, Liposomes, HPLC-UV

中图分类号: 

Supporting:

Foundation items: Beijing Natural Science Foundation (Grant No. 7091005) and National Natural Science Foundation of China (Grant No. 30772664).
*Corresponding author. Tel./fax: 86-10-82802683