RP-HPLC测定胰岛素龙血竭纳米颗粒中胰岛素的含量

RP-HPLC determination of insulin encapsulated in insulin-loaded Sanguis Draxonis nanoparticles

目的:建立反相高效液相色潜(RP-HPLC)法测定胰岛素龙血竭纳米颗粒(ISDN)中胰岛素的含量。方法:ISDN 先用80%的乙醇水溶液(pH 3.0)溶解制样,样品的检测采用 Phenomenex C_(18)色谱柱(4.6 mm×250 mm,5μm);流动相:0.1 mol·L^(-1)磷酸二氢钠溶液(以磷酸调 pH 为3.0)-乙睛,梯度洗脱(0～15 min,比例为72:28;16～40 min,比例由72:28→30:70),流速为1.0 mL·min^(-1);检测波长为210 nm,柱温为30℃。结果:胰岛素在5～200μg·mL^(-1)范围内,峰面积与浓度呈良好线性关系(r=0.9999);高、中、低3种浓度水平的回收率(n=3)分别为99.6%,98.9%,98.7%,RSD 分别为0.3%,0.8%,0.4%;日内、日间精密度的 RSD 分别为0.2%,0.5%,0.5%和0.4%,1.0%,1.2%。结论:该方法样品处理过程简单,分离效果和重复性均良好,结果准确可靠,可用于该制剂中胰岛素含量的检测,同时也为龙血竭包裹其他蛋白质类药物的分析提供了可靠依据。

Objective ：To develop a simple RP - HPLC method for the determination of insulin encapsulated in insulin- loaded Sanguis Draxonis nanoparticles （ISDN）. Methods：The ISDN separation was performed on a Phenomenex C18 （4. 6 mm × 250 mm,5 μm）column using a mobile phase consisting of a solution of 0. 1 mol · L^-1sodium dihydrogen phosphate （ adjusted to pH 3.0 with phosphoric acid） - acetonitrile （ gradient elution ：0 - 15 min,72 ：28 ;16 -40 min,72： 28→30： 70）at 30 ℃ ,with the flow rate at 1.0 mL · min^-1 and the detection wavelength at 210 nm. Results：It showed that mass concentration and peak area had linearity from 5 μg · mL^-1 to 200 μg · mL^-1（ r = 0. 9999）. The average recoveries（ n = 3 ） of the three groups（ concentration ： 140,80,40 μg · mL^-1, respectively）were 99.6% （RSD =0. 3% ） ,98.9% （RSD =0. 8% ）and 98.7% （ RSD =0. 4% ） ,respectively. The intra - day and inter - day RSDs of the three concentrations were 0.2% ,0.5% ,0.5% and 0.4% ,1.0% ,1.2%. Conclusion： Being convenient, sensitive, accurate, and reproducible, the method is available for the determination of insulin encapsulated in ISDN.