分子排阻色谱法测定德谷胰岛素高分子蛋白含量  

Quantitation of High Molecular Weight Proteins in Insulin Degludec by Size Exclusion Chromatography

作　　者：杨庆 马鲁南 刘忠 张贵民 YANG Qing;MA Lunan;LIU Zhong;ZHANG Guimin(Shandong New Time Pharmaceutical Co.,Ltd.,Shandong Provincial Engineering Laboratory of Protein Pharmaceutical,Linyi 273400,China;Lunan Pharmaceutical Group Co.,Ltd.,State Engineering Laboratory of High Expression of Mammalian Cells,Linyi 276006,China)

机构地区：[1]山东新时代药业有限公司,山东省蛋白类药物工程实验室,山东临沂273400 [2]鲁南制药集团股份有限公司,哺乳动物细胞高效表达国家工程实验室,山东临沂276006

出　　处：《中国现代应用药学》2020年第23期2879-2882,共4页Chinese Journal of Modern Applied Pharmacy

摘　　要：目的建立德谷胰岛素高分子蛋白质的定量分析测定方法。方法采用Insulin HMWP色谱柱(7.8 mm×300 mm,10mm),以冰醋酸-异丙醇-水(4∶3∶10)为流动相,流速为0.5 mL·min^-1,检测波长为280 nm,利用分子排阻色谱法分离德谷胰岛素单体以及高分子蛋白质,并对高分子蛋白质进行定量分析。结果该方法的检测限和定量限分别为0.18和0.47μg;仪器精密度RSD为0.51%,德谷胰岛素在0.125~4.0mg·mL^-1内与峰面积呈良好的线性关系,回归方程为Y=20.864 1X-0.125 9,R^2=1.000 0;德谷胰岛素主峰与高分子蛋白质峰的分离度>2.0。结论建立的方法操作简便,灵敏度高,可有效地对德谷胰岛素高分子蛋白质进行定量分析。OBJECTIVE To establish a quantitative analysis method of the high molecular weight proteins in insulin degludec. METHODS The column was Insulin HMWP column(7.8 mm×300 mm, 10 mm). The mobile phase was acetic acid-isopropanol-water(4∶3∶10). The flow rate was 0.5 mL·min^-1. The detection wavelength was set at 280 nm. The high molecular weight proteins were separated and accurately determined by size exclusion chromatography. RESULTS The detection limit and quantitative detection limit were 0.18, 0.47 μg, respectively. The RSD of instrument precision test was 0.51%. The concentration of insulin degludec at the range of 0.125-4.0 mg·mL^-1 showed good linear relationship to the peak area, of which the regression equation was Y=20.864 1 X-0.125 9, with R^2=1.000 0. The resolution of main peak and molecular protein peak was >2.0. CONCLUSION The established method is simple, sensitive and can effectively quantitatively analyze the high molecular protein of insulin degludec.

关 键 词：德谷胰岛素 高分子蛋白质 分子排阻色谱法 

分 类 号：R927.2[医药卫生—药学]