高效液相色谱法测定重组人干扰素α-2b注射液中乙二胺四乙酸二钠含量  

HPLC Method for the Determination of Disodium EDTA in Recombinant Human Interferon α-2b Injection

作　　者：李增礼[1] 周乐春[1] 赵辉[1,2] 储成风[1] 倪晓燕[1] 王荣海[1,2] 宋礼华[1,2] 

机构地区：[1]安徽安科生物工程(集团)股份有限公司,安徽合肥230088 [2]安徽省生物研究所,安徽合肥230088

出　　处：《生物技术通讯》2012年第3期419-422,共4页Letters in Biotechnology

基　　金：国家"重大新药创制"科技重大专项(2008ZX09203-003)

摘　　要：目的：利用高效液相色谱（HPLC）法测定重组人干扰素α-2b注射液中EDTA二钠（乙二胺四乙酸二钠）的含量。方法：将EDTA二钠与氯化铁溶液于70℃水浴中反应20 min左右;色谱柱为SunFire C18（250 mm×4.6 mm,5μm,Waters）,流动相为5%甲醇＋95%0.64 g/L四丁基溴化铵和4.1 g/L三水合乙酸钠混合液,用冰乙酸调pH值至4.0,流速1 mL/min,检测波长254 nm。结果：该测定方法线性范围为0.025-0.5 g/L,线性关系良好（r=0.9997）,加样回收率为98.27%（n=9,RSD=2.55%）。结论：本方法准确、快速、可靠,可用于重组人干扰素α-2b注射液中EDTA二钠含量的测定。Objective： To develop a HPLC method for the determination of disodium EDTA in recombinant human interferon α-2b injection. Methods： Disodium EDTA was derivatized with ferric chloride solution by heating 70℃ in water bath for about 20 rain. The chromatographic column was SunFire C18（250 mm×4.6 mm, 5 μm,Waters）. The mobile phase consisting of 5% methanol and 95% of 0.64 g/L solution of tetra butyl ammonium bromide and 4.1 g/L solution of sodium acetate trihydrate in water. The pH was adjusted to 4.0 with acetic acid glacial. The flow rate was 1.0 mL/min. The UV determination was measured at 254 nm. Results： Disodium EDTA has good linearity in the range of 0.025-0.5 g/L（r=0.9997）. The average recovery was 98.27%（n=9, RSD=2.55%）. Conclusion： This method is accurate, rapid and credible, and can be used to determinate disodium EDTA in recombinant human interferon α-2b injection.

关 键 词：重组人干扰素α-2b注射液 高效液相色谱 乙二胺四乙酸二钠 NAFEEDTA 

分 类 号：Q502[生物学—生物化学]