生物传感器技术在分离赤芍抗内毒素单体成分中的研究

目的 :利用生物传感技术从赤芍中分离抗内毒素单体成分。方法 :以LipidA包被生物传感器样品池表面 ,建立筛选靶点 ,并以传感器上回收物的紫外扫描结果为参照 ,通过跟踪测定硅胶柱层析、高效液相色谱 (HPLC)流出组分与LipidA的结合能力 ,分离抗内毒素的单体成分 ,并应用鲎试剂法测定单体成分与体外脂多糖 (LPS)的中和作用。结果 :从生物传感器回收到可与LipidA结合的组分 ,其紫外吸收峰分别为194、215、275nm波长。通过HPLC分离出与LipidA具有较高结合活性的单体成分为1 ,2 ,3 ,4 ,6—O—五没食子酰—β—D—葡萄糖 (PGG ) ,浓度为8、4、2μg/ml的PGG分别能够中和0 1EU/mlLPS活性的68 8 %、43 7 %、31 4 %。结论 :利用生物传感器技术 ,以LipidA为靶点分离赤芍抗内毒素单体成分是可行的 ,且具有高效、快速、准确等优点 ,适合于大规模地从中草药中分离抗内毒素的单体成分。

Study of Isolating Anti- Endotoxin Monomer Component from Radix Paeoniae Rubra Biosensor by Biosensor Technique

OBJECTIVE:To isolate anti-endotoxin monomer component from Radix Paeoniae Rubra by means of biosenˉsor technique.METHODS:The surface of biosensor cuvette was embedded by Lipid A;the screening target was established,tracking the silica gel column chromatogram and the binding ability of effluent component from HPLC with Lipid A with the ultraviolet scan result of the reclaimed material from biosensor as reference;anti-endotoxin monomer component was isolated;the component of monomer and the synthetic action of extrinsic lipopolysaccharides were also assayed by LAL test method.RESULTS:Components binding to Lipid A was reclaimed from cuvetee by biosensor technique,with the wavelength of UV absorption peak at194nm,215nm and275nm respectively.Anti-endotoxin monomers of higher binding activity with Lipid A isolated by HPLC method were1,2,3,4,6—O—pentagalloyl—β—D—glucose(PGG).PGG at concentration of8,4,2μg/ml respectively neutralized68.8%,43.7%and31.4%of LPS at an activity of0.1EU/ml respectively.CONCLUSION:It is feaˉsible to isolate anti-endotoxin monomer component from Radix Paeoniae Rubra by means of biosensor technique,which is a fast,accurate and efficient and can be used to isolate anti-endotoxin monomer component from Radix Paeoniae Rubra on a large scale.