目的 建立并验证检测阿达木单抗电荷变异体的阳离子交换高效液相色谱法。方法 采用以羧基官能团的弱阳离子为固定相的交换柱（4 mm×250 mm），以0.01 mol/L 磷酸氢二钠溶液、pH 7.5为流动相 A，以0.01 mol/L 磷酸氢二钠+0.5 mol/L 氯化钠溶液、pH 5.5为流动相 B进行梯度洗脱。按面积归一法计算赖氨酸变异体和酸性组分含量，并对该方法进行专属性、重复性、中间精密度、线性、准确度和耐用性验证。结果 该方法检测阿达木单抗注射液可见溶剂峰和目标峰；重复检测6次各组分相对标准偏差均＜2%；不同实验员、不同实验设备检测各组分的相对标准偏差均＜2%；赖氨酸变异体之间的峰的定量限（信噪比＞10）为100 μg/mL、检测限（信噪比＞3）为30 μg/mL；在企业质量标准限度范围内检测不同浓度各组分线性良好，决定系数均＞0.99；第1酸区峰、赖氨酸变异体之间的峰和赖氨酸变异体2峰准确度回收率均在80%～115%，第2酸区峰、主峰和赖氨酸变异体1峰准确度回收率均在85%～110%；不同流动相pH、不同流动相氯化钠浓度、不同流速与柱温、不同批次色谱柱条件下检测结果符合要求，一致性较好。结论 建立的方法具有良好的专属性、重复性、中间精密度、线性、准确度和耐用性，适用于阿达木单抗电荷变异体的测定。

Objective To establish and validate a cation exchange chromatography detection method for adalimumab charge variants.Methods An exchange column (4 mm×250 mm) with weak cations of carboxylic functional groups was used as the stationary phase. 0.01 mol/L disodium hydrogen phosphate solution, pH 7.5 was used as mobile phase A, and 0.01 mol/L disodium hydrogen phosphate + 0.5 mol/L sodium chloride solution, pH 5.5 was used as mobile phase B for gradient elution. The content of lysine variants and acidic components were calculated by area normalization method. Specificity, repeatability, intermediate precision, linearity, accuracy, and robustness of the method were validated.Results Adalimumab target peak and solvent peak were observed. For 6 repeated tests, relative standard deviations (RSDs) of all components were all ＜2%. With different experimenters and test equipment, RSDs of each component were all ＜ 2%. The quantification limit (signal-to-noise ratio＞10) and detection limit (signal-to-noise ratio＞3) of peaks between lysine variants were 100 μg/mL and 30 μg/mL, respectively. Within the quality standard limits of Wuhan Institute of Biological Products Co., Ltd., the linearities of all components at different concentrations were good, with coefficients of determination all＞0.99. Recovery rates were within 80%-115% for the first acidic region peak, the intermediate peak between lysine variants, and lysine variant 2 peak, and 85%-110% for the second acidic region peak, main peak, and lysine variant 1 peak. Analytical results met the specified requirements and exhibited acceptable consistency under different mobile phase pH and NaCl concentration, flow rate, column temperature, and chromatographic column lot numbers.Conclusion The method established has good specificity, repeatability, intermediate precision, linearity, accuracy, and robustness, and is suitable for the determination of the purity of adalimumab charge variants.