HPLC法测定DNA纳米运输系统载药阿霉素的含量

HPLC analysis of doxorubicin loaded in DNA nanoparticle delivery system

目的:建立反相高效液相色谱法测定DNA四面体纳米运输系统中阿霉素的含量。方法:采用Agilent Extend C18色谱柱,以含0.05%三氟乙酸的水溶液-含0.05%三氟乙酸的乙腈溶液(72:28)为流动相,流速1.0 mL·min^-1,检测波长260 nm,柱温30℃。结果:阿霉素浓度在5~150μmol·L^-1范围内与峰面积呈现良好的线性关系(r=0.999 5);加样回收率为97.6%~104.2%(RSD<1.5%,n=6)。优化得到DNaseⅠ脱氧核糖核酸酶酶解DNA四面体释放阿霉素的最佳酶解浓度和最佳酶解时间分别为0.3 mg·mL^-1和30min。3批样品中DNA四面体结合阿霉素含量分别为131.6、131L4、132.1μmol·L^-1。结论:该方法为定量分析DNA载体系统中阿霉素的含量提供了有效方法。

Objective:To establish a RP-HPLC analysis method for the determination of doxorubicin in the DNA tetrahedrons.Methods:An Agilent Extend C18 column was used with a mobile phase of a mixture containing0.05% trifluoroacetic acid in an aqueous solution and 0.05% trifluoroacetic acid in acetonitrile(72:28).The flow rate was 1.0 mL·min^-1,the detection wavelength was 260 nm and the column temperature was 30 ℃.Results:The concentration of doxorubicin showed a good linear relationship within a range of 5^-150 μmol·L^-1(r=0.999 5).Recovery rates were ranged from 97.6% to 104.2%(RSD1.5%,n=6).The optimal enzymolysis concentration and the enzymolysis time for DNase I hydrolysis of DNA tetrahedron to release doxorubicin were 0.3 mg·mL^-1 and 30 min,respectively.The contents of 3 samples were 131,6,131.4,132.1 μmol·L^-1,respectively.Conclusion:This method provides an effective way to quantify doxorubicin in the DNA carrier system.