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
樊梦妮 1谭澄莹 1段颖 1陈秋 1秦国宏1
作者信息
- 1. 南京正大天晴制药有限公司 芳华药物研究院,江苏南京 210046
- 折叠
摘要
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,3 μm),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40 μL,激发波长320 nm,发射波长400 nm.结果:在1～100 μg/mL,葡聚糖硫酸钠与峰面积线性关系良好.定量限为0.5 μg/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.
Abstract
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,3 μm)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 40 μL,an excitation wavelength of 320 nm,and an emission wavelength of 400 nm.Results:At 1～100 μg/mL,dextran sodium sulfate has a good linear relationship with peak area.The limit of quantification is 0.5 μg/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.
关键词
高效液相色谱法/葡聚糖硫酸钠/细胞抗结团剂/细胞培养Key words
high performance liquid chromatography/dextran sulfate sodium/cell anti-agglomeration agent/cell culture引用本文复制引用
出版年
2024
NETL
NSTL
万方数据