反相高效液相色谱法测定多糖包覆脂质体中胰岛素  

RP - HPLC Determination of Insulin in Polysaccharide - coated Insulin Liposomes

作　　者：吴正红[1] 平其能[1] 张磊[1] 郭建新[1] 魏毅[1] 赖家明[1] 

机构地区：[1]中国药科大学药剂教研室,南京210009

出　　处：《药物分析杂志》2002年第6期425-427,共3页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金(No.39930200)

摘　　要：目的:建立反相高效液相色谱法测定多糖包覆的胰岛素脂质体含量和包封率。方法:采用逆相蒸发-超声法制备胰岛素脂质体;用高效液相色谱法和超速离心法测定胰岛素脂质体含量与包封率;采用反相色谱柱Lichrospher ODS-C_(18)(250 mm×4.6 mm,5μm),以0.025 mol·L^(-1)硫酸盐缓冲液(pH 2.0)-己腈(72:28)为流动相,流速1.0 mL·min^(-1),柱温40℃,检测波长214nm。结果:线性范围为2～10 μg·mL^(-1)。在空白壳聚糖包覆脂质体中胰岛素加样回收率为94.65％～100.5％,日内、日间精密度的RSD分别为0.92％～1.6％和4.0％～5.6％。在空白海藻酸钠包覆脂质体中胰岛素加样回收率为96.12％～101.1％,日内、日间精密度的RSD分别为1.7％～2.4％和4.0％～4.7％。壳聚糖包覆的胰岛素脂质体和海藻酸钠包覆的胰岛素脂质体包封率分别为73.6％和68.7％,胰岛素总含量分别为0.682 mg·mL^(-1)和0.713mg·mL^(-1)。结论:本方法便捷、灵敏、准确、重现性好,可用于测定多糖包覆的胰岛素脂质体含量与包封率。Objective: A RP - HPLC method was developed for determining the contents of insulin in polysaccha-ride -coated insulin liposomes and the entrapment efficiencies of the liposomes. Methods: Insulin - liposomes were prepared by reversed - phase evaporation and sonication. The entrapment efficiencies of the liposomes and the contents of insulin in the liposomes were analyzed by HPLC and ultracentrifuge. Chromatographic conditions included Lichrospher ODS - C18 (250 mm ×4. 6 mm, 5μm) and mobile phase consisting of a mixture of acetoni-trile - sulfate buffer solution (0. 025 mol · L-1 , pH 2. 0). The flow rate was 1. 0 mL· min-1 . The temperature of column was 40℃ . Detection wavelength was 214 nm. Results: The calibration curve was linear in the range of 2 - 10μg· mL-1. The average recovery, the RSDs of within -day and day -to -day of insulin in blank chitosan - coated liposomes were 94. 65% - 100. 5% , 0. 92% - 1. 6% and 4. 0% - 5. 6% , those in blank alginate -coated liposomes were 96. 12% - 101. 1% , 1.7% -2.4% and 4.0% -4.7% respectively. The entrapment efficiency of chitosan - coated insulin liposomes and the content of insulin in the liposomes were 73. 6% and 0. 682 mg· mL-1 respectively. The entrapment efficiency of alginate - coated insulin liposomes and the content of insulin in the liposomes were 68.7% and 0.713 mg· mL-1 respectively. Conclusions: The method was convenient, sensitive, accurate, reproducible, and suitable for determining the contents of insulin in polysaccharide - coated liposomes and the entrapment efficiencies of the liposomes.

关 键 词：反相高效液相色谱法 多糖 胰岛素脂质体 包封率 含量测定 

分 类 号：R927.2[医药卫生—药学]