萃取法去除低分子肝素中细菌内毒素研究  

Study on Removal of Bacterial Endotoxins from Low-molecular-mass Heparins by Extraction

作　　者：郑华淦 廖冬 曲和之 ZHENG Hua-gan;LIAO Dong;QU He-zhi(Shenzhen Sciprogen Bio-Pharmaceutical Co.,Ltd.,Shenzhen 518129,China)

机构地区：[1]深圳赛保尔生物药业有限公司研发部,深圳518129 [2]深圳赛保尔生物药业有限公司,深圳518129

出　　处：《药物生物技术》2022年第5期466-469,共4页Pharmaceutical Biotechnology

基　　金：深圳市工信局扶持项目(No.20200619145347736)。

摘　　要：采用正丁醇-乙酸丁酯萃取法去除低分子肝素中细菌内毒素。以达肝素钠为例,通过对萃取前后供试品中细菌内毒素含量、抗Xa因子效价、分子量及分布进行测定,表明正丁醇-乙酸丁酯萃取法可有效去除达肝素钠粗品溶液中细菌内毒素,对产品效价及分子量无影响。经3次萃取,达肝素钠粗品溶液中细菌内毒素含量可由6000~10000 EU/mL下降至100 EU/mL以下,内毒素去除率大于98%,抗Xa因子效价收率大于96%,重均分子量与分子量分布基本不变。最终产品符合EP标准要求,其中细菌内毒素小于0.01 EU/IU。可见正丁醇-乙酸丁酯萃取法去除达肝素钠中细菌内毒素具有效率高、重复性好、操作方便等优点,可能开发为其他低分子肝素中去除细菌内毒素的方法。The main raw material of low-molecular-mass heparins is originated from animal tissue extraction,so it is easily contaminated by microorganisms in the early preparation process,resulting in bacterial endotoxins.In this paper,a quick and easy method to remove bacterial endotoxins from the low-molecular-mass heparin crude solutions was attempted,the n-butanol-butyl acetate extraction method was used to remove bacterial endotoxins that may exist in the preparation process.Taking dalteparin sodium as an example,through the determination of bacterial endotoxins content,anti-Xa factor potency,mass-average relative mole-cular mass and distribution in the test sample before and after extraction,it shows that the n-butanol-butyl acetate extraction method can effectively remove bacterial endotoxins in the crude solutions of Dalteparin sodium,and has no effect on the anti-factor Xa potency and the mass-average relative molecular mass of the product.After three times of extraction,the bacterial endotoxins content in the crude dalteparin sodium solutions can be reduced from 6000~10000 EU/mL to below 100 EU/mL,the bacterial endotoxins removal rate is greater than 98%,and the yield of anti-Xa factor potency is greater than 96%,the mass-average relative molecular mass and the molecular mass distribution are basically unchanged.The final product meets the requirements of EP standard,in which bacterial endotoxins is less than 0.01 EU/IU.It can be seen that the n-butanol-butyl acetate extraction has the advantages of high efficiency,good repea-tability and convenient operation to remove bacterial endotoxins from dalteparin sodium,and it may be developed as a method for removing bacterial endotoxins from other heparins.In addition,both n-butanol and butyl acetate are classified into the third class solvents of common organic solvents in ICH,which are low toxicity and less harm to the human body,the n-butanol and butyl acetate introduced during extraction,can be easily removed during the ultrafiltration and alcohol precipitation,and

关 键 词：正丁醇 乙酸丁酯 萃取 细菌内毒素 低分子肝素 达肝素钠 

分 类 号：R973[医药卫生—药品]