采用混合模式色谱与电雾式检测器测定蛋白药物中的聚山梨醇酯20

目的:建立混合模式色谱和电雾式检测器测定蛋白药物中的聚山梨醇酯20的含量。方法:色谱柱为Oasis MAX(2.1 mm×20 mm,30 μm),柱温30℃,流动相A为2%(体积分数)甲酸的异丙醇溶液,流动相B为2%(体积分数)甲酸的水溶液,梯度洗脱,进样体积30 μL;电雾式检测器,雾化室温度30℃,采样频率5 Hz。结果:聚山梨醇酯20的峰面积与浓度线性回归方程为Y=6.643X-0.0445(r=0.9994),线性范围为9.4~188.0 mg·mL^-1;峰面积和保留时间的RSD分别为0.66%和0.04%;平均回收率为98.3%;方法定量限(S/N=10)为1.1 mg·mL^-1。结论:该仪器分析方法重现性较好,样品检出限较低,可作为药物及制剂中聚山梨醇酯20的含量测定方法。

Objective: To develop a mixed-mode chromatography and charged aerosol detector(CAD) method for the quantitative determination of polysorbate 20 in protein pharmaceuticals.Methods: Oasis MAX column(2.1 mm×20 mm,30 μm) was used in the study and the column temperature was 30℃.The mobile phase was composed of isopropanol[containing 2%(v/v) formic acid] and water[containing 2%(v/v) formic acid] with gradient elution;the injection volume was 30 μL;CAD:nebulization temperature was 30℃ and collection frequency was 5 Hz.Results: The equation of linear regression was Y=6.643X-0.0445(r=0.9994),linear range was 9.4-188.0 mg·mL^-1;the RSD value of peak area and retention time was 0.66% and 0.04%,respectively;the average recovery was 98.29%;the LOQ(as S/N=10) was 1.1 mg·mL^-1.Conclusion: The established method is sensitive and accurate,which can be used in the quantitative determination of polysorbate 20 in protein pharmaceuticals.