胶束絮凝反溶剂重结晶法纯化落叶松木中二氢槲皮素工艺优化

Optimization of Purification Process of Dihydroquercetin from Larch by Micellar Flocculation Anti-Solvent Recrystalization

摘要: 为解决二氢槲皮素工业化纯化效率低、环境污染大、成本高的问题，本研究以落叶松木粉为原料，构建绿色胶束絮凝–反溶剂重结晶集成纯化工艺，重点优化工艺参数并明确纯化核心机制。通过筛选表面活性剂与絮凝剂组合，确定茶皂素与壳聚糖为最优组合，絮凝率达85.3%，絮凝后二氢槲皮素纯度为27.8%；以重结晶纯度和收率为响应值，采用单因素实验结合响应面法优化关键参数，确定最佳工艺为乙醇与水体积比1:10、结晶温度16℃、结晶时间48 h，此条件下二氢槲皮素一次重结晶纯度达94.88%，收率为38.92%。通过Zeta电位和荧光探针技术验证了胶束絮凝的电荷中和与疏水包载协同作用机制，为该工艺的工业化应用提供理论支撑。本方法简化了分离步骤，减少有害有机溶剂使用，兼具环保性与经济性，为二氢槲皮素的规模化纯化提供了可行方案，也为天然产物绿色的纯化技术提供了新思路。

Abstract: To address the challenges of low efficiency, high environmental pollution, and high cost in the industrial purification of dihydroquercetin, this study developed an integrated purification process combining green micellar flocculation and anti-solvent recrystallization using larch wood powder as the raw material. The process parameters were optimized, and the core purification mechanism was elucidated. By screening combinations of surfactants and flocculants, tea saponin and chitosan were identified as the optimal pair, achieving a flocculation rate of 85.3% and a dihydroquercetin purity of 27.8% after flocculation. Using recrystallization purity and yield as response values, single-factor experiments combined with response surface methodology were employed to optimize key parameters. The optimal conditions were determined as an ethanol-to-water volume ratio of 1:10, a crystallization temperature of 16˚C, and a crystallization time of 48 hours. Under these conditions, the purity of dihydroquercetin after a single recrystallization reached 94.88%, with a yield of 38.92%. The synergistic mechanism of charge neutralization and hydrophobic encapsulation in micellar flocculation was verified through Zeta potential and fluorescence probe techniques, providing theoretical support for the industrial application of this process. This method simplifies the separation steps, reduces the use of harmful organic solvents, and is both environmentally friendly and economically viable. It offers a feasible solution for the large-scale purification of dihydroquercetin and provides new insights into green purification technologies for natural products.