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

Quantitation of polysorbate 20 in protein solutions using mixed-mode chromatography and charged aerosol detector

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(2.1mm×20 mm,30μm),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30μL;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 iso- propanol containing 2 % （v/v） formic acid ] and water [ containing 2 % （v/v） formic acid ] with gradient elution ; the injection volume was 30 IxL;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 ac- curate, which can be used in the quantitative determination of polysorbate 20 in protein pharmaceuticals.