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目的:优选大孔树脂纯化4种新疆山楂果实总黄酮的工艺条件,评价其抗氧化活性。方法:以总黄酮纯度为评价指标,比较13种大孔树脂对4种山楂果实总黄酮的吸附分离效果,考察上样流速、洗脱剂浓度对4种山楂总黄酮大孔树脂纯化工艺的影响。以清除2,2’-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)和二苯代苦味酰基(DPPH)自由基的方法测定样品的抗氧化活性,利用HPLC测定样品芦丁和金丝桃苷的含量,流动相乙睛-0.4%甲酸溶液(16∶84),检测波长360 nm。结果:选择HPD722型大孔树脂纯化总黄酮,最佳纯化工艺条件为上样流速0.84 m L·min-1,总黄酮经树脂充分吸附后,加40%乙醇动态洗脱。经HPD722型大孔吸附树脂纯化后,4种山楂的总黄酮、芦丁、金丝桃苷质量分数和抗氧化活性分别提高了1.9~3.3,2.4~5.1,2.6~3.7,2.0~4.6倍。结论:HPD722型大孔树脂可有效地对4种新疆山楂果实总黄酮进行富集纯化,为这4种药材资源的充分利用提供参考。
Objective: To optimize the technological conditions for the purification of total flavonoids from four species of hawthorn fruit by macroporous resin and evaluate its antioxidant activity. Methods: The total flavonoids purity as the evaluation index, the comparison of 13 kinds of macroporous resin hawthorn fruit flavonoids adsorption and separation efficiency, study the sample flow rate, eluent concentration on the four kinds of Hawthorn macroporous resin macroporous resin purification process . The antioxidant activity of the samples was determined by a method of eliminating 2,2’-azino-di (3-ethylbenzothiazole-6-sulfonic acid) diammonium salt (ABTS) and dibenzyl picrate (DPPH) The contents of rutin and hyperoside were determined by HPLC. The mobile phase was acetonitrile - 0.4% formic acid solution (16:84) and the detection wavelength was 360 nm. Results: The total flavonoids were purified by HPD722 macroporous resin. The optimum purification conditions were as follows: the flow rate of sample was 0.84 m L · min-1, the total flavonoids were fully adsorbed by resin and then eluted with 40% ethanol. After purified by macroporous resin HPD722, the content of total flavonoids, rutin, hyperoside and antioxidant activity of four hawthorn were increased by 1.9-3.3, 2.4-5.1, 2.6-3.7, 2.0-4.6 times, respectively. CONCLUSION: HPD722 macroporous resin can effectively enrich and purify the total flavonoids of four species of hawthorn fruits in Xinjiang and provide a reference for the full utilization of these four medicinal materials.