聚合物基质高效液相色谱填料在白细胞介素-2分离中的应用  

Separation of Interleukin-2 by High Performance Liquid Chromatographic Column Packed with Polymer Resin

作　　者：魏荣卿[1] 张婷婷[1] 邵勇军[1] 刘晓宁[1] 

机构地区：[1]南京工业大学制药与生命科学学院,南京210009

出　　处：《分析化学》2007年第4期505-510,共6页Chinese Journal of Analytical Chemistry

基　　金：科技部创新基金(No.05C26213200578);江苏省十五攻关项目基金(No.BE2001040)资助

摘　　要：聚合物型苯乙烯-二乙烯基苯（PS-DVB）微球装填的麦科菲反相高效液相色谱柱（MKF-RP-HPLC）,用于白细胞介素-2（IL-2）分离的最佳色谱条件：流动相A：0.1%三氟乙酸,流动相B：80%乙腈＋0.1%三氟乙酸。B液在30min内从0%线性增大至100%,流速1.0mL/min;检测波长：280nm。在该色谱条件下进行系统性实验,结果表明：MKF-RP色谱柱分离IL-2的柱效、分离度、重复性和拖尾因子均能达到药典要求,且柱效和分离度与SOURCE色谱柱相当;MKF-RP色谱柱的pH适用范围为1-14;柱压与SOURCE色谱柱相当,且低于Hypersil C8色谱柱和Polaris C18色谱柱的柱压,可在更高流速下操作;MKF-RP色谱柱的非极性与SOURCE色谱柱相当;MKF-RP色谱柱的超载性能优于SOURCE色谱柱,其对IL-2进样液的最大载样量为300μg。The optical separate condition of Interleukin-2 （IL-2） by MKF reversed phase high performance liquid chromatography （RP-HPLC）column packed with monodisperse polystyrene-divinylbenzene （PS-DVB） microspheres was that the mobile phase B（0.1% trifluoroacetic acid ＋ 80% acetonitrile） was linear increased from 0 % to 100% in 30 min while the mobile phase A was 0.1% trifluoroacetic acid. The MKF-RP（ MKF reverse-phase） column was evaluated by the Systemic Test according to the Chinese pharmacopoeia on the optical condition. The results showed that the column efficiency, resolution, reproducibility and asymmetry of MKF-RP column could meet the requirement of Pharmacopoeia, and the column efficiency, resolution, backpressure and polarity of MKF-RP column were equivalent to those of SOURCE column. The application range of pH for MKF-RP column was 1 - 14. Backpressure and non-polarity of MKF-RP column were lower than those of Hypersil C8 and Polaris C18. The result of overloading test showed that the MKF-RP column was more suitable for scaling up than the SOURCE column, and the maximum injection of IL-2 on MKF-RP column was 300 μg.

关 键 词：白细胞介素-2 苯乙烯-二乙烯基苯 反相高效液相色谱 麦科菲反相色谱柱 

分 类 号：R392[医药卫生—免疫学]