毛细管区带电泳法测定重组人干扰素α-1b滴眼液中间甲酚含量

Determination of M-cresol in rhIFNα1-b Eye Drops by Capillary Zone Electrophoresis

目的:运用毛细管区带电泳法(CZE)检测重组人干扰素α^(-1)b(rh IFNα^(-1)b)滴眼液中间甲酚含量。方法:优化的CZE方法参数为采用无涂层的熔融石英毛细管(60 cm×50μm),电泳缓冲液为50 mmol·L^(-1)磷酸盐缓冲液(p H 8.0),压力进样(1.0 psi),分离电压25 k V,检测波长195 nm,毛细管温度25℃。结果:间甲酚在0.05~0.8 mg·ml^(-1)浓度范围内与峰面积呈良好线性关系(r=0.997 3),最低检出限0.002 mg·ml^(-1)。3批rh IFNα1b滴眼液中间甲酚的相对含量分别为1.94,1.89和1.97 mg·ml^(-1),平均回收率分别为94.3%(RSD=1.57%,n=9)。结果与HPLC方法结果基本一致。结论:新建毛细管区带电泳法简便,分析速度快,试剂消耗量小,结果准确,可以用于重组人干扰素α1b滴眼液中间甲酚含量的检测。

Objective： To determine the content of M-cresol in rhlFNα-1 b eye drops by capillary zone electrophoresis （CZE）. Methods： The optimized CZE parameters were as follows： an uncoated fused-silica capillary （60 em x 50 Ixm） was used, 50 mmol· L-1 phosphate buffer （pH 8.0） was used as the electrophoresis buffer, pressure injection （1.0 psi） was adopted, the running voltage was 25 kV, the detection wavelength was 195 nm, and the capillary temperature was set at 25 ℃. Results： The content of M-cresol had good linear relationship with peak area within the Concentration range of 0.05-0.8 mg · ml^-1 （ r = 0.9986 ）. The detection limit was 0.002 mg · ml^-1. The content of M-cresol in three batches of rhlFNotlb eye drops was 1.94 mg · ml^-1 , 1.89 mg · ml^-1 and 1.97 mg · ml^-1, and the average recovery was 94.3% （RSD = 1.57% , n =9） , respectively, which were basically consistent with the test re- sults of HPLC. Conclusion： The developed CZE method is simple and quick in operation, reliable in determination results and low in reagent comsuption, which can be used for the content determination of M-cresol in rhlFNα-1b eye drops.