微柱凝胶离心-HPLC测定酮康唑醇质体包封率

Determination of Entrapment Efficiency of Ketoconazole Ethosomes with Dextarn Gel Minicolumn Centrifugation-HPLC

关键词:  酮康唑醇质体  包封率  微柱凝胶离心法  高效液相色谱法

Key words:  ketoconazole ethosomes  entrapment efficiency  minicolumn gel centrifugation  HPLC

摘要:

目的 建立酮康唑醇质体包封率的微柱凝胶离心-HPLC测定方法。方法 药物含量采用HPLC测定，醇质体与游离药物的分离采用葡聚糖凝胶微柱离心法。色谱条件为：色谱柱Sinochrom ODS-BP(4.6 mm×200 mm，5 μm)，流动相乙腈-水-三乙胺(60∶40∶0.5，磷酸调pH 8.6)，流速1.0 mL·min^-1，检测波长243 nm，进样量20 μL。分离条件为：以2 000 r·min^-1离心3 min制备葡聚糖凝胶微柱，测样量0.2 mL，以2 400 r·min^-1离心5 min，再用0.2 mL蒸馏水洗脱，重复3次。结果 微柱对酮康唑的平均吸附率>99.30%，对空白醇质体洗脱率>99.49%。结论 葡聚糖凝胶微柱离心-HPLC可作为醇质体包封率测定的简单快捷方法。

Abstract:

OBJECTIVE To develop a determination method for the entrapment efficiency of ketoconazole ethosomes. METHODS The content of ketoconazole was determined by HPLC. The minicolumn of filling dextran gel was adopted to separate the free ketoconazole from ethosome dispersions by centrifugation. Chromatographic conditions were as follows: column was Sinochrom ODS-BP(4.6 mm×200 mm, 5 μm) and the mobile phase was acetonitrile-water-triethylamine (60∶40∶0.5, adjust to pH 8.6 using phosphate) with the flow rate of 1.0 mL·min^-1, and detection wavelength was 243 nm. The Dextran G-50 minicolumn was spinned at 2 000 r·min^-1 for 3 min for removing excess amount of water. The 0.2 mL sample added on prepared column was centrifuged at 2 400 r·min^-1 for 5 min, and then washed for 3 times by 0.2 mL of distilled water. RESULTS By flowing through the Dextran G50 minicolumn, the free ketoconazole was adsorbed more than 99.30%, while the average recovery of blank ethosomes was more than 99.49%. CONCLUSION This method is technically simple and rapid to determine entrapment efficiency of ketoconazole ethosomes.