氯化钠含量对人凝血因子Ⅷ冻干品结晶形态的影响  

Effect of sodium chloride content on crystalline form of freeze-dried human coagulation factor Ⅷ

作　　者：蒋桂香 张伟 朱光祖 吕应年[2] 

机构地区：[1]广东双林生物制药有限公司广东省血液制品工程中心,广东湛江524005 [2]广东医学院广东天然药物研究与开发重点实验室,广东湛江524023

出　　处：《中国生物制品学杂志》2015年第2期134-137,141,共5页Chinese Journal of Biologicals

基　　金：国家863计划子课题项目(2012AA021903-6);广东省企业重点实验室建设项目(2012A0616000)

摘　　要：目的研究人凝血因子Ⅷ(FⅧ)中氯化钠含量与冻干品结晶形态的关系,为FⅧ冻干品的质量控制提供依据。方法采用阴离子交换凝胶柱层析法制备高纯度的FⅧ溶液,分别加入氯化钠至低中高浓度(40、80、160 mmol/L)后,真空冻干,并与添加160 mmol/L氯化钠和100 mmol/L甘氨酸的冻干品进行对比。目测法观察冻干品外观,扫描电镜(scanning electron microscope,SEM)下观察结晶形态;差示扫描量热法(differential scanning calorimeter,DSC)检测热力学性质;参考《中国药典》三部(2010版)检测FⅧ活性、含水量、复溶时间。结果 FⅧ溶液随着盐浓度增加,冻干品外观依次变差,由疏松网孔状结构变致密片层状结构,共晶点降低,凝血活性降低,复溶时间延长,含水量增加;添加甘氨酸作为保护剂的冻干品外观明显变好,呈疏松网孔状结构,共晶点高,凝血活性强,复溶时间短,含水量低。结论氯化钠含量对冻干品结晶形态有显著影响,可在溶液配制阶段调节氯化钠含量及添加保护剂,以控制冻干品的结晶形态。Objective To investigate the relationship between sodium chloride content and crystalline form of freeze-dried human coagulation factor Ⅷ（FⅧ）and provide a basis for the quality control of FⅧ. Methods Highly purified FⅧ was prepared by anion-exchange chromatography, then added with sodium chloride to low（40 mmol / L）, moderate（80 mmol / L）and high（160 mmol / L） concentrations respectively and lyophilized in vacuum, using the freeze-dried F Ⅷ containing160 mmol / L sodium chloride and 100 mmol / L glycine as control. The appearance of freeze-dried FⅧ was examined visually, while the crystalline form was observed by scanning electron microscopy（SEM）, the thermodynamic property was evaluated by differential scanning calorimeter（DSC）, and the activity, moisture content and reconstitution time according to the requirements in Chinese Pharmacopoeia（Volume Ⅲ, 2010 edition）. Results Along with the increasing sodium chloride concentration, the appearance of FⅧ changed from loose hole-net structure to dense lamellar structure, the eutectic point was lowered, and clotting activity decreased, and time for reconstitution increased, and the moisture content in-creased. However, the appearance of FⅧ using glycine as a stabilizer showed a loose hole-net structure, of which the eutectic point and clotting activity increased, and the time for reconstitution and moisture content decreased. Conclusion The crystalline form of lyophilized FⅧ was distinctly influenced by sodium chloride content. The sodium chloride content should be adjusted while the stabilizer should be added during formulation of F Ⅷ solution to control the crystalline form of freeze-dried FⅧ.

关 键 词：凝血因子 冻干 氯化钠 结晶 

分 类 号：R554.1[医药卫生—血液循环系统疾病] R392-33[医药卫生—内科学]