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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: To establish an RP-HPLC method for determination of polysaccharide-encapsulated insulin liposomes content and entrapment efficiency. METHODS: Insulin liposomes were prepared by reverse phase-evaporating-ultrasonication. The content of liposomes and entrapment efficiency of liposomes were determined by high performance liquid chromatography and ultracentrifugation. mm × 4.6 mm, 5 μm) with a flow rate of 1.0 mL · min -1 using 0.025 mol·L -1 sulfate (pH 2.0) and hexanenitrile (72:28) Temperature 40 ℃, detection wavelength 214nm. Results: The linear range was 2 ~ 10 μg · mL -1. The recoveries of insulin in the blank chitosan-coated liposomes ranged from 94.65% to 100.5%. The intra-day and inter-day RSDs were 0.92% -1.6% and 4.0% -5.6%, respectively. The recoveries of insulin in blank alginate-coated liposomes ranged from 96.12% to 101.1%. The intra-day and inter-day RSDs were 1.7% -2.4% and 4.0% -4.7%, respectively. The encapsulation efficiency of chitosan-coated insulin liposomes and sodium alginate-coated insulin liposomes were 73.6% and 68.7%, respectively, and the total contents of insulin were 0.682 mg · mL -1 and 0.713 mg · mL ~ (-1). Conclusion: The method is convenient, sensitive, accurate and reproducible. It can be used to determine the content and encapsulation efficiency of polysaccharide-coated insulin liposomes.