超声波辅助酶解法制备宝藿苷Ⅰ

Optimization of Ultrasonic-Assisted Enzymatic Hydrolysis Technology for Baohuoside Ⅰ from Icariin

摘要:

目的: 优选宝藿苷Ⅰ的超声波辅助酶解工艺。方法: 采用HPLC测定宝藿苷Ⅰ含量,色谱条件为Diamond C₁₈色谱柱(4.60 mm×250 mm,5 μm),流动相乙腈-水(25:75),流速1.0 mL·min^-1,柱温30℃,检测波长270 nm。以宝藿苷Ⅰ转化率为指标,在单因素试验基础上,通过正交试验考察超声功率、pH、温度、酶用量及反应时间对超声酶解工艺的影响。结果: 最佳酶解条件为超声功率200 W,pH 5.0,温度40℃,酶用量5%,反应时间4 h;宝藿苷Ⅰ转化率97.4%。结论: 超声波辅助酶解法制备宝藿苷Ⅰ具有转化率高、反应温度低、反应时间短、酶用量少等优点,优选的工艺简单稳定,适合工业化生产。

Abstract:

Objective: To optimize ultrasonic-assisted enzymatic hydrolysis technology of baohuosideⅠ. Method: The content of baohuosideⅠ was determined by HPLC,chromatographic conditions were as follows:Diamond C₁₈ column(4.60 mm×250 mm,5 μm),mobile phase of acetonitrile-water(25:75),flow rate 1.0 mL·min^-1,column temperature 30℃,detection wavelength 270 nm.Taking bioconversion rate of baohuosideⅠ as index,based on single factor tests,effects of ultrasonic power,pH,temperature,enzyme dosage and reaction time on preparation technology were investigated L₉(3⁴) orthogonal design. Result: Optimal enzymatic conditions were:ultrasonic power of 200 W,pH of 5.0,reaction temperature at 40℃,reaction time of 4 h,enzyme dosage 5%.Under these conditions,conversion rate of baohuosideⅠ was 97.4%. Conclusion: Ultrasonic-assisted enzymatic method could improve conversion rate of baohuosideⅠ with low reaction temperature,short reaction time,little enzyme and other advantages,optimized process was stable,simple and suitable for industrial production.