罗汉松叶中总黄酮的提取与纯化富集工艺研究

Extraction and purification of total flavonoids from the leaves of Podocarpus macrophyllus by D101 macroporous resin

目的优选提取罗汉松叶总黄酮的乙醇体积分数及D101型大孔吸附树脂分离纯化罗汉松叶总黄酮的工艺条件。方法以酪氨酸酶抑制率为指标确定提取罗汉松叶总黄酮的乙醇体积分数;以罗汉松叶总黄酮质量分数为指标,研究D101型大孔吸附树脂分离纯化罗汉松叶总黄酮的工艺条件,包括上样流速、洗脱速率、上样浓度和乙醇洗脱体积。结果体积分数70%乙醇的罗汉松叶提取物对酪氨酸酶抑制率最高;大孔树脂分离纯化罗汉松叶总黄酮的最佳工艺条件为:上样浓度4.00 mg/m L,上样流速2.0m L/min,4 BV体积分数80%乙醇以2.0 m L/min速率洗脱,回收率为60.80%。结论 D101型大孔吸附树脂可富集罗汉松叶中的总黄酮,是纯化前的2.85倍,该工艺简单且成本低,可为进一步开发罗汉松叶总黄酮提供依据。

Objective To optimize the ethanol concentration in extraction and purification process of total flavonoids from the leaves of P. macrophyllus by D101 macroporous resin. Methods The ethanol concentration was screened with inhibitory activity against tyrosinase as index. Purification conditions including loading flow rate, eluting velocity, loading concentration and elution volume were optimized with the contents of total flavonoids as index. Results The inhibitory activity of the extract by 70% ethanol against tyrosinase were up to maximum. The optimum purification conditions were as follows： the sample concentration of 4.00 mg/mL, loading flow rate of 2. 0 mL/min , eluted by 80% alcohol of 4BV at the velocity of 2. 0 mL/min. The yield of total flavonoids was 60. 80%. Conclusion The content of total flavonoids was 2.85 fold after purification by D101 macroporous resin. The method is simple and low cost, which could provide the basis for further development and utilization of the leaves of P. macrophyllus.