大孔树脂纯化黑骨藤总黄酮工艺研究

Technique Study on Macroporous Resin Purifying Total Flavonoids in Gardneria Angustifolia

目的:探讨大孔树脂分离纯化黑骨藤总黄酮工艺,比较6种类型大孔树脂吸附率和解析率,筛选最佳树脂,单因素分析最佳纯化工艺路线。方法:采用NaNO2-Al (NO)3-NaOH法定量,比较6种不同型号大孔吸附树脂对黑骨藤中总黄酮的吸附和解析效果,筛选出适合分离纯化黑骨藤中总黄酮的树脂,并对其吸附和解析条件进行考察和优化,应用单因素实验,确定最终优化工艺。结果:AB-8树脂的吸附量明显高于NKA-9和S-8树脂,吸附率明显高于X-5、D101、NKA-9、S-8树脂,而解析率明显高于X-5、D101、NKA-9、S-8、HPD100树脂,差异均有统计学意义(P<0.05),故选择AB-8为最佳树脂。上样液浓度1.308 mg/mL,上样流速为3 BV/h,pH为3.24,上样体积2 BV,以4 BV 75%乙醇在流速2 BV/h下洗脱,得黄酮的纯度为70.31%。结论:AB-8树脂用于分离纯化黑骨藤总黄酮,工艺稳定可靠。

Objective:To discuss the technique of macroporous resin separating and purifying total flavonoids in Gardneria angustifolia,compare the adsorption rate and the desorption rate of six types of macroporous resin,filter and select the optimal resin and conduct a single-factor analysis on the optimal process of purification technique.Methods:By means of Na NO2-Al(NO)3-NaOH method for quantification,compared the results of six types of macroporous resin with different models in adsorbing and desorbing total flavonoids in Gardneria angustifolia;filtered and selected the resin which was suitable for separating and purifying total flavonoids in Gardneria angustifolia,examined and optimized the corresponding requirements for adsorption and desorption;determined the ultimate optimal technical process via single-factor experiment.Results:The adsorption capacity of AB-8 resin was evidently higher than that of X-5,D101,NKA-9,S-8 and HPD100 resin,and its desorption rate was evidently higher than that of X-5,D101,NKA-9,S-8 and HPD100 resin,differences being significant(P<0.05).Therefore,AB-8 was considered as the optimal resin.The concentration of loading buffer was 1.308 mg/m L,the sampling velocity was 3 BV/h,the value of p H was 3.24 and the sampling volume was 2 BV;the purity of flavonoid was70.31% when eluted with 75% ethanol in 4 BV at the velocity of 2 BV/h.Conclusion:AB-8 resin may well be applied for separating and purifying total flavonoids in Gardneria angustifolia owing to its stable and reliable technical process.