大孔树脂联合Sephadex LH-20葡聚糖凝胶纯化蜂胶总黄酮工艺研究  

Purification of Total Flavonoids from Propolis by Macroporous Resin Combined with Sephadex LH-20 Dextran Gel

作　　者：李治 汤毛毛 李景 常相伟 刘劲松 彭代银 王国凯 桂双英 LI Zhi;TANG Mao-mao;LI Jing;CHANG Xiang-wei;LIU Jin-song;PENG Dai-yin;WANG Guo-kai;GUI Shuang-ying(School of Pharmacy, Anhui University of Chinese Medicine, Anhui Hefei 230012, China;Anhui Province Key Laboratory of Pharmaceutical Preparation Technology and Application, Anhui Hefei 230012, China;Institute of Pharmaceutics, Anhui Academy of Chinese Medicine, Anhui Hefei 230012, China;Engineering Technology Research Center of Modernized Pharmaceutics, Education Office of Anhui Province, Anhui Hefei 230012, China;Key Laboratory of Chinese Medicinal Formula of Anhui Province, Anhui Hefei 230012, China)

机构地区：[1]安徽中医药大学药学院,安徽合肥230012 [2]药物制剂技术与应用安徽省重点实验室,安徽合肥230012 [3]安徽省中医药科学院药物制剂研究所,安徽合肥230012 [4]安徽省教育厅现代药物制剂工程技术研究中心,安徽合肥230012 [5]中药复方安徽省重点实验室,安徽合肥230012

出　　处：《安徽中医药大学学报》2021年第4期93-98,共6页Journal of Anhui University of Chinese Medicine

基　　金：国家自然科学基金项目(81873019);安徽高校协同创新项目(GXXT-2020-025)。

摘　　要：目的优选大孔树脂联合Sephadex LH-20葡聚糖凝胶分离纯化蜂胶总黄酮的最佳工艺参数。方法采用硝酸铝-醋酸钾比色法定量,通过比较9种大孔吸附树脂对蜂胶总黄酮的吸附率和解吸率,筛选出最佳大孔吸附树脂型号,通过动态吸附与解吸试验优选大孔树脂最佳纯化工艺参数,采用Sephadex LH-20葡聚糖凝胶进一步纯化。结果优选大孔树脂型号为HPD-722型。最佳纯化工艺:上样液浓度2.5 mg/mL,比上样量0.43 g/g(药材/湿树脂),上样容积流量为每小时2倍床体积(bed volume,BV),径高比1∶9;先以30%乙醇3 BV除杂,再用80%乙醇4 BV洗脱,洗脱容积流量1 BV/h,洗脱液浓缩、干燥,得蜂胶总黄酮Ⅰ,纯度为41.42%;取蜂胶总黄酮Ⅰ1.2 g,用少量甲醇超声溶解,上样于Sephadex LH-20层析柱,以甲醇洗脱,洗脱流速0.75 mL/min,7.5 mL作为一个流份收集,以三氯化铝为显色剂,聚酰胺薄层色谱检识各流份,合并、干燥得蜂胶总黄酮Ⅱ,纯度为68.48%。结论HPD-722型大孔吸附树脂联合Sephadex LH-20葡聚糖凝胶可有效纯化蜂胶总黄酮。Objective To investigate the optimal process parameters for the separation and purification of total flavonoids from propolis using macroporous resin combined with Sephadex LH-20 dextran gel.Methods The aluminum nitrate-potassium acetate colorimetric method was used for quantification,and the adsorption and desorption rates of nine types of macroporous adsorption resins for total flavonoids from propolis were compared to screen out the best type of macroporous adsorption resin.Dynamic adsorption and desorption tests were used to determine the optimal purification process parameters of macroporous resin,and Sephadex LH-20 dextran gel was used for further purification.Results The optimal type of macroporous resin was HPD-722 and the optimal purification process was a concentration of the loading solution of 2.5 mg/mL,a specific loading volume of 0.43 g/g(medicinal material/wet resin),a loading volume flow rate of 2 BV/h,and a diameter-to-height ratio was 1∶9.First,3 BV of 30%ethanol was used to remove impurities,and then 4 BV of 80%ethanol was used for elution,with a volume flow rate of 1 BV/h.The eluate was concentrated and dried to obtain total flavonoids from propolisⅠwith a purity of 41.42%,and then ultrasonic dissolution with a small amount of methanol was performed for 1.2 g of the total flavonoids from propolisⅠ,which was loaded to a Sephadex LH-20 column and eluted with methanol at a flow rate of 0.75 mL/min,and 7.5 mL was collected as a fraction.With aluminum trichloride as the chromogenic reagent,all fractions were identified by polyamide thin-layer chromatography and were then mixed and dried to obtain total flavonoids from propolisⅡwith a purity of 68.48%.Conclusion HPD-722 macroporous adsorbent resin combined with Sephadex LH-20 dextran gel can effectively purify the total flavonoids from propolis.

关 键 词：蜂胶 大孔吸附树脂 葡聚糖凝胶 总黄酮 纯化工艺 

分 类 号：R94[医药卫生—药剂学]