[关键词]
[摘要]
目的 采用超滤膜技术对脉络宁注射液废弃物进行资源化研究。方法 脉络宁注射液大生产废弃醇沉物中富含石斛多糖、牛膝多糖和金银花多糖,通过免疫活性筛选自废弃物中分离出富含多糖的有效部位。采用不同孔径超滤膜将废弃醇沉物分离所得粗多糖分为相对分子质量<3×103、3×103~1×104、1×104~3×104、3×104~1×105、1×105~3×105、>3×105共6个部位,分别命名为MFP1、MFP2、MFP3、MFP4、MFP5、MFP6;利用苯酚-硫酸比色法和考马斯亮蓝G-250染色法测定MFP1~MFP6中多糖和蛋白质的量;ELSD-HPLC法检测MFP1~MFP6中多糖组分;通过小鼠脾淋巴细胞增殖、转化和小鼠单核巨噬细胞RAW264.7释放NO实验对MFP1~MFP6进行免疫活性筛选。结果 MFP1~MFP6多糖定量测定结果发现MFP4多糖的量最高且免疫活性最优,多糖的量和得率分别为76.80%和10.81%;MFP5免疫活性次之,其量和得率分别为73.23%和14.73%;结论 MFP4和MFP5是免疫活性较优的部位,具开发利用价值。超滤膜技术可用于脉络宁注射液废弃物中多糖资源化利用,该工艺过程简便易行,成本低廉,在循环经济利用中具有独特优势。
[Key word]
[Abstract]
Objective In this study, ultrafiltration membrane process is employed first to separate the polysaccharides from industrialization waste in Mailuoning Injection and to promote the resources utilization of the waste. Methods The waste ethanol sediments in the production of Mailuoning Injection mainly include dendrobium polysaccharides, Achyranthes bidentata polysaccharides (ABPS), and honeysuckle polysaccharides, and the active components of polysaccharide with high content were selected through the immune activity. Six different relative molecular weight cut-off membranes (< 3 × 103, 3 × 103-1 × 104, 1 × 104-3 × 104, 3 × 104-1 × 105, 1 × 105-3 × 105, > 3 × 105) made of polyether sulfone (PES) were used. Six fractions were obtained and named as MFP1, MFP2, MFP3, MFP4, MFP5, and MFP6. Meanwhile, polysaccharides and protein content were determined by phenol-sulfuric acid colorimetry and Coomassie brilliant blue G-250 staining method; Polysaccharides species was analyzed by ELSD-HPLC; Immunocompetence of different fractions were investigated in vitro, including mouse spleen lymphocytes proliferation, transformation, and production of nitric oxide after acting on the macrophage RAW264.7. Results The results showed MFP4 (3 × 104-1 × 105) have the highest content and immunocompetent, MFP5 (1 × 105-3 × 105) immunocompetent followed, compared to others, purity, and recovery of MFP4 are 76.80% and 10.81%, purity and recovery of MFP5 are 73.23% and 14.73%, respectively. Conclusion The findings reveal MFP4 and MFP5 serves as our object of exploitation in the future; Moreover, separation of polysaccharides from "Mailuoning Injection" by the membrane is feasible with high recovery and low cost in the realization of circular economy.
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[基金项目]
江苏省科技厅产学研联合创新资金-前瞻性研究项目(BY2012036);江苏省六大人才高峰项目(2014-YY-014);江苏省中药资源产业化过程协同创新中心分离平台建设项目(ZDXM-1-6,ZDXM-1-8);江苏省高校优势学科建设工程资助项目;南京中医药大学校级创新团队资助;江苏省重点研发计划-社会发展面上项目(BE2016754)