HPLC-荧光法测定贝伐珠单抗注射液中聚山梨醇酯20的含量  

Determination of polysorbate 20 content in Bevacizumab injection by high performance liquid chromatography-fluorescence assay

作　　者：赵威 郭圣荣[1] 黄馨 ZHAO Wei, GUO Sheng-rong, HUANG Xin(School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, Chin)

机构地区：[1]上海交通大学药学院,上海200240

出　　处：《中国生物制品学杂志》2018年第3期294-298,共5页Chinese Journal of Biologicals

摘　　要：目的采用HPLC-荧光法测定贝伐珠单抗注射液中聚山梨醇酯20的含量。方法对HPLC色谱条件进行优化后,检测贝伐珠单抗注射液中聚山梨醇酯20含量,并进行方法的专属性、线性、准确度、精密度、检测限、耐用性验证。用建立的方法检测3批贝伐珠单抗注射液中聚山梨醇酯20的含量。结果该方法专属性良好;聚山梨醇酯20浓度在0.20~0.60 mg/mL范围内,线性关系良好,R^2=0.999 9;不同浓度的聚山梨醇酯20溶液检测结果的准确度在98%~100%范围内,RSD为0.8%;精密度各考察项RSD均不高于2%;检测限为0.002 0 mg/mL;耐用性各考察项与原始条件的相对百分比均在100%~102%范围内。3批贝伐珠单抗注射液中聚山梨醇酯20的含量分别为0.40、0.39和0.39 mg/mL,分别为标示量(0.40 mg/mL)的100%、98%和98%。结论建立的HPLC-荧光法快速、准确、可靠,可用于测定贝伐珠单抗注射液中聚山梨醇酯20的含量。Objective To determine the polysorbate 20 content in Bevacizumab injection by high performance liquid chromatography（HPLC）-fluorescence assay. Methods The condition for HPLC was optimized, under which the polysorbate 20 content in Bevacizumab injection was determined. The method was verified for specificity,linearity,accuracy, precision, minimum detection limit and robustness. The polysorbate 20 contents in three batches of Bevacizumab injection were determined by the method. Results The method showed high specificity,of which the linear range was 0. 20 ~ 0. 60 mg/mL,with a correlation coefficient（R^2） of about 0. 999 9. The average recovery rates of polysorbate 20 at various concentrations were 98% ~ 100%,with a RSD of 0. 8%. The RSDs of various indexes in precision test were not more than 2%. The minimum detection limit was 0. 002 0 mg/mL. The relative percentages of determination results of various indexes in robustness test under the optimized condition to those under original condition were 100% ~ 102%. The contents in polysorbate 20 in three batches of Bevacizumab injection were 0. 40,0. 39 and0. 39 mg/mL,which were 100%,98% and 98% of the stated amounts,respectively. Conclusion The developed HPLC-fluorescence assay was rapid,accurate and reliable,which might be used for determination of the polysorbate 20 content in Bevacizumab injection.

关 键 词：聚山梨醇酯20 贝伐珠单克隆抗体 高效液相色谱 荧光法 N-苯基-1-萘胺 

分 类 号：Q657.12[生物学—生物物理学] R392-33[医药卫生—免疫学]