目的 建立并验证SIBP-A01单克隆抗体（单抗）中聚山梨酯20（polysorbate 20，PS20）残留量的高效液相色谱（high performance liquid chromatography，HPLC）-蒸发光散射检测器（evaporative light-scattering detector，ELSD）检测方法。方法 应用HPLC-ELSD方法检测SIBP-A01单抗中PS20残留量，并对该方法的专属性、精密度、线性、准确度、定量限和耐用性进行验证。结果 稀释液以及不含PS20的制剂缓冲液无干扰峰；3批单抗供试品重复检测结果、不同人员检测结果、同一实验人员不同时间检测结果的相对标准偏差均< 5%；PS20含量在12.5~100.0 μg/mL范围内时，峰面积对数与含量对数线性关系良好，决定系数为0.996 8；加标回收率在103.5%~109.2%之间；定量限为5.0 μg/mL；当色谱柱温度为（30±3） ℃、供试品于10 ℃放置24 h或使用不同品牌PS20时，检测差<5%。结论 成功建立了SIBP-A01单抗中PS20残留量的HPLC-ELSD检测方法，该方法的专属性、精密度、线性、准确度和耐用性良好。

Objective To establish and validate a high performance liquid chromatography (HPLC) method coupled with an evaporative light-scattering detector (ELSD) for the determination of residual polysorbate 20 (PS20) in the monoclonal antibody SIBP-A01.Methods The HPLC-ELSD method was used to detect the residual PS20 in SIBP-A01. The method was validated in terms of specificity, precision, linearity, accuracy, limit of quantitation (LOQ), and robustness.Results No interfering peaks were observed in the diluent or the formulation buffer without PS20. For the three batches of SIBP-A01 test samples, the relative standard deviations (RSD) of repeated detection results, detection results obtained by different operators, and detection results obtained by the same operator at different time were all less than 5%. When the concentration of PS20 ranged from 12.5 to 100.0 μg/mL, a good linear relationship was observed between the logarithm of the peak area and the logarithm of the concentration, with a coefficient of determination of 0.996 8. The spiked recovery rates were in the range of 103.5% to 109.2%. The LOQ was 5.0 μg/mL. When the column temperature was set at (30±3) ℃, the samples were stored at 10 ℃ for 24 h, or different brands of PS20 were used, the difference of the detection results was less than 5%.Conclusions An HPLC-ELSD method for the determination of residual PS20 in SIBP-A01 has been successfully established. This method shows good specificity, precision, linearity, accuracy, and robustness.