牛源肝素钠特性与病毒灭活研究

Study on Characteristics and Virus Inactivation of Bovine Heparin Sodium

对牛源肝素钠特性及制备工艺的病毒去除/灭活能力进行研究。采用分子排阻色谱法、羊血浆法、SAX-HPLC法等对牛源肝素钠与猪源肝素钠的分子量、效价、双糖等特性进行分析研究;通过核磁共振(NMR)波谱法对牛源肝素钠、猪源肝素钠进行分析,并对获得的^(1)H-NMR谱和^(13)C-NMR谱进行对比;采用指示病毒,验证高温、高pH、离子交换、氧化处理等步骤的病毒去除/灭活能力。相对于猪源肝素钠,牛源肝素钠重均分子量(M_(r))低约5%,效价低约23%,其主要双糖含量、NMR波谱图也有一定差别;病毒去除/灭活验证表明,处理前后病毒感染性滴度下降值达4个Log_(10)值以上。特性研究表明牛源肝素钠与猪源肝素钠存在一定差别,病毒去除/灭活验证研究证明制备工艺能够有效去除/灭活病毒。

The objective of this article is to study the characteristics of bovine heparin sodium and the virus removal/inactivation ability of the preparation process.The molecular weight,potency and disaccharide of bovine heparin sodium and porcine heparin sodium was analyzed by the methods of molecular exclusion chromatography,sheep plasma and SAX-HPLC.Bovine heparin sodium and porcine heparin sodium was analyzed by nuclear magnetic resonance(NMR)spectroscopy and then the obtained^(1)H-NMR and^(13)C-NMR spectra were compared.The process steps including high temperature,high pH,ion exchange,and oxidation treatment were validated for the virus removal/inactivation capability with indicator virus.Compared with porcine heparin sodium,the weight-average molecular weight(M_(r))of bovine heparin sodium was 5%lower,and the potency was 23%lower.The contents of main disaccharides and NMR spectra were also different.Virus removal/inactivation validation showed that the titer of infection decreased by more than 4 Log_(10)values before and after treatment.There are some differences between bovine heparin sodium and porcine heparin sodium.Virus removal/inactivation validation study proves that the preparation process can effectively remove/inactivate virus.