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[摘要]
目的 研究宝藿苷I的制备工艺。方法 采用纤维素酶水解淫羊藿苷制备宝藿苷I,以转化率为指标,通过单因素考察pH值、温度、底物的浓度、酶用量、反应时间及金属离子对转化率的影响L9(34),正交试验优化制备工艺;采用MS,1H-NMR、13C-NMR鉴定水解产物。结果 酶解反应的最适条件为温度50℃、反应介质pH 5.2醋酸-醋酸钠缓冲液,底物浓度为10 mg/mL,酶与底物质量比1∶1,反应时间48 h,钠离子、钙离子、镁离子、锌离子、对酶解反应无显著影响(P>0.05),铁离子对酶解反应有抑制作用(P<0.01);反应产物相对分子质量为514,核磁图谱证实产物为宝藿苷I。结论 纤维素酶水解淫羊藿苷制备宝藿苷Ⅰ,工艺简单可靠,反应条件温和,适合工业化生产。
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[Abstract]
Objective The hydrolysis method of icariin by cellulase was studied to prepare baohuoside.Methods Taking the bioconversion rate as the index,cellulase was used for enzymolysis of icariin and the effects of pH value,temperature, reaction time,dosage of enzyme,concentration of icariin,and metal ion on conversion rate of baohuoside I were studied.L9(34) Orthogonal design was used to optimize the preparation condition.Hydrolysis product was identified by MS,1H-NMR,and 13C-NMR.Results The optimum enzymolysis with cellulase was achieved at 50 ℃,a pH 5.2 acetic acidsodium acetate(1∶1) buffer solution for 48 h.The substrate concentration is 10 mg/mL.The effects of sodium ion,calcium ion,magnesium ion,and zinc ion on cellulase were slightly(P0.05) but the inhibition of iron ion on enzymolysis was significant(P0.01);It's structure was elucidated on the basis of its spectral evidences.Conclusion It is feasible to prepare baohuoside I by enzymolysis with cellulase,the condition is moderate and it is suitable for industrialization.
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[基金项目]
江苏省医药高技术计划项目(BG2007614);江苏省中医药领军人才项目(2006)