高效液相色谱-荧光检测器法检测单克隆抗体注射液中吐温80的含量

Determination of Tween80 Content in Monoclonal Antibody Injection by HPLC-FLD

目的:采用高效液相色谱-荧光检测器法(HPLC-FLD)测定单克隆抗体注射液中吐温80的含量,并进行方法学验证。方法:按设定的色谱条件对检测单克隆抗体注射液中吐温80含量的HPLC-FLD法进行方法学验证,包括专属性、精密度、准确度、线性和范围、耐用性,并采用该方法对不同单克隆抗体注射液进行检测。结果:经验证,该方法具有专属性;重复性验证试验中,保留时间和含量的RSD值分别为0.5%和0.4%;中间精密度验证试验中,保留时间和含量的RSD值分别为0.4%和2.2%;准确度验证试验中,回收率分别为105%、98%和108%;线性相关系数为0.9991,检测范围为0.05~0.80 mg/mL;不同批次反应线圈对检测结果无影响,不同品种单克隆抗体注射液吐温80含量检测结果偏差较小。结论:该方法具有专属性,线性关系良好,精密度和准确度较好,测定结果稳定可靠,适用于单克隆抗体注射液中吐温80含量的检测。

Objective:To establish a method for the determination of Tween 80 in monoclonal antibody injection by High-performance liquid chromatography fluorescence detector.Methods:Under designed condition,the method was verified for specificity,precision,accuracy,linearity,range and durability.The method was also used to determination of Tween 80 content of different monoclonal antibody injections.Results:The HPLC-FLD method showed good specificity.In reproducibility test,the RSD of retention time and content were 0.5%and 0.4%respectively.In intermediateprecision test,the RSD of retention time and content were 0.4%and 2.2%respectively.The recovery rates were 105%,98%and 108%respectively.The correlation Coefficient was 0.9991 and the range was from 0.05 to 0.8 mg/mL.The different batches of reaction coils had no effect on the detection results.The Tween 80 contents in different mAb injections showed little deviation.Conclusion:The HPLC-FLD method showed good specificity,precision,accuracy and linearity,and the test result was stable and reliable,which was suitable for determination of Tween 80 content of different monoclonal antibody injections.