Box-Behnken Design响应面法优化银杏叶总黄酮柱层析纯化工艺

Optimization of the column chromatographic purification of total flavonoids extracted from Ginkgo biloba using Box-Behnken design and response surface method

为得到质量分数较高的银杏叶总黄酮提取物,在单因素试验基础上,以上样质量浓度、乙醇体积分数、洗脱液接收体积和洗脱流速为考察因素,以银杏叶总黄酮质量分数、收率为考察指标,采用BBD(Box-Behnken Design)响应面法对银杏叶总黄酮柱层析纯化工艺进行优化。得到最佳工艺为上样质量浓度3.1 mg/mL,乙醇体积分数71%,洗脱液接收体积2 BV(BV为装柱体积)及洗脱流速2 BV/h。在最佳工艺条件下,完成三批验证实验,银杏叶总黄酮平均质量分数可达到35.77%,收率为93.30%,银杏总内酯质量分数达到《中国药典》标准。利用BBD响应面法优化银杏叶总黄酮柱层析纯化工艺是科学、合理、可行的,可为工业化大生产提供参考。

The study’s aim was to obtain Ginkgo biloba extract with a high mass fraction of total flavonoids. On the basis of single-factor test, the sample concentration, ethanol volume fraction, eluate volume, and elution flow rate were used as the investigation factors and the mass fraction and yield of total flavonoids extracted from G. biloba were used as the investigation indicators. Box-Behnken design(BBD) and response surface method were used to optimize the column chromatography process for purifying the total flavonoids extracted from G. biloba. Optimal purification was achieved by using a sample concentration of 3.1 mg/mL, ethanol volume fraction of 71%, eluate volume of 2 BV(BV is the column void volume), and elution flow rate of 2 BV/h. Under the optimal conditions, three batches of verification experiments were conducted. The average mass fraction of total flavonoids in the G. biloba extract reached 35.77%, and the yield was 93.30%. The mass fraction of total lactones in the G. biloba extract reached the standards of The Pharmacopoeia of the People′s Republic of China. Using BBD and response surface method for optimizing the column chromatography process for the purification of total flavonoids is scientific, reasonable, and feasible.This study can be used as a reference for the industrial production of total flavonoids.