生物技术进展 ›› 2014, Vol. 4 ›› Issue (2): 113-117.DOI: 10.3969/j.issn.2095-2341.2014.0 .

• 研究论文 • 上一篇    下一篇

苯扎氯铵对单克隆抗体亲和层析填料的杀菌效果及其残留量的检测

阮乾坤,韩志刚*,余军阳,杜伊达,杨大军   

  1. 北京三元基因工程有限公司, 北京 102600
  • 收稿日期:2013-12-25 出版日期:2014-03-25 发布日期:2014-01-23
  • 通讯作者: 韩志刚,硕士,主要从事发酵和蛋白纯化的研究。E-mail:836254508@qq.com
  • 作者简介:阮乾坤,硕士,主要从事发酵和蛋白纯化的研究。E-mail:451522361@qq.com。

Benzalkonium Chloride's Bacteria-killing Effect on Affinity Chromatography Gel and Residues Detection

RUAN Qian-kun, HAN Zhi-gang, YU Jun-yang, DU Yi-da, YANG Da-jun   

  1. Beijing Tri-Prime Genetic Engineering Co., Ltd., Beijing 102600, China
  • Received:2013-12-25 Online:2014-03-25 Published:2014-01-23

摘要: 通过菌悬液定量杀菌实验和亲和层析凝胶蛋白吸附量测定,最终从众多杀菌剂中筛选出一种既能有效杀死亲和层析介质中感染的细菌,又不影响其蛋白吸附量的理想杀菌剂苯扎氯铵;建立了利用反相离子对高效液相色谱测定层析柱中苯扎氯铵残留量的方法。采用C18色谱柱,流动相为乙腈-0.2 mol/L庚烷磺酸钠,于210 nm紫外波长下检测,检出限为0.000 25 mg/mL,实际样品中苯扎氯铵含量测定结果的相对标准偏差为0.63%,本方法具有良好的灵敏度和精密度。

关键词: 苯扎氯铵, 蛋白吸附量, 检出限, 反相离子对高效液相色谱法

Abstract: An ideal bactericide benzalkonium chloride which can effectively kill the bacteria infected in affinity chromatography gel and not affect the protein adsorption capacity was eventually selected from several bactericides by quantitative bactericidal test on bacterial suspension and affinity chromatography gel protein adsorption quantity determination. An ion pair reversed phase high performance liquid chromatography (RP-IPHPLC) method was developed for the determination of benzalkonium chloride residue in affinity chromatography gel. Separation was carried out on C18 column, using acetonitrile and 0.2mol/L sodium heptanesulfonate solution as the mobile phase in an isocratic elution.The detection wavelength was 210 nm; the detection limit was 0.000 25 mg/mL; relative standard deviation of benzalkonium chloride determination test in practical samples was 0.63%. This method presents excellent sensitivity and precision.

Key words: benzalkonium chloride, protein adsorption capacity, detection limit, ion pair revered phase high-performance liquid chromatography