高效液相色谱法测定抗体中葡聚糖硫酸钠含量

Determination of Dextran Sulfate Sodium Content in Antibodies by High Performance Liquid Chromatography

目的:建立高效液相色谱法测定葡聚糖硫酸钠含量的方法。方法:采用Sepax Zenix SEC-150色谱柱(300 mm×7.8 mm,3μm),以0.01 mol/L的十二水合磷酸氢二钠、0.09 mol/L的二水合磷酸二氢钠为流动相,等度洗脱,柱温50℃,流速1.2 mL/min,进样体积40μL,激发波长320 nm,发射波长400 nm。结果:在1~100μg/mL,葡聚糖硫酸钠与峰面积线性关系良好。定量限为0.5μg/mL,加标回收率在100.5%~110.0%,重复性实验的相对标准偏差为2.67%,稳定性实验的相对标准偏差为1.07%。结论:建立的高效液相色谱方法准确、可靠,可用于抗体中葡聚糖硫酸钠含量的测定。

Objective:To establish a method for the determination of dextran sulfate sodium content by high performance liquid chromatography.Methods:Sepax Zenix SEC-150 Column(300 mm×7.8 mm,3μm)are employed,0.01 mol/L sodium hydrogen phosphate dodecahydrate and 0.09 mol/L sodium dihydrogen phosphate dihydrogen dihydrate are used as mobile phases,with an isocratic elution at 50℃,a flow rate of 1.2 mL/min,an injection volume of 40μL,an excitation wavelength of 320 nm,and an emission wavelength of 400 nm.Results:At 1~100μg/mL,dextran sodium sulfate has a good linear relationship with peak area.The limit of quantification is 0.5μg/mL,the recovery rate is 100.5%~110.0%,the relative standard deviation of the repeatability experiment is 2.67%,and the relative standard deviation of the stability experiment is 1.07%.Conclusion:The established high performance liquid chromatography method is accurate and reliable,and can be used for the determination of dextran sodium sulfate content in antibodies.