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该实验采用纤维素酶水解朝藿定B以制备箭藿苷B。以转化率为指标,通过单因素考察pH、温度、反应时间、酶用量、底物的浓度对转化率的影响,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。结果表明,酶解反应的最适条件为温度50℃,反应介质(pH 5.6)醋酸-醋酸钠缓冲液,底物质量浓度20 g·L-1,酶与底物质量比3∶5,反应时间24 h,反应产物相对分子质量646.23,核磁图谱证实产物为箭藿苷B。该工艺简单可靠,反应条件温和,适合工业化生产。
This experiment uses cellulase to hydrolyze Attapagin B to prepare Icariin B. The effects of pH, temperature, reaction time, enzyme dosage and substrate concentration on the conversion rate were investigated by single factor analysis. The orthogonal design was used to optimize the preparation process. MS, 1H-NMR and 13C- NMR identified hydrolyzate. The results showed that the optimum conditions for enzymolysis reaction were temperature 50 ℃, reaction medium (pH 5.6) acetate-sodium acetate buffer, substrate concentration 20 g · L-1, mass ratio of enzyme to substrate 3: 5 Time 24 h, the relative molecular mass of the reaction product was 646.23, and the product was confirmed to be icariin B by NMR. The process is simple and reliable, mild reaction conditions, suitable for industrial production.