在线、离线结合快速筛选杨梅叶中黄酮类抗氧化活性成分

Rapid screening of flavonoids antioxidant components from Bayberry leaves by on-line and off-line assays

目的:建立一种2,2-联苯基-1-苦基肼基高效液相色谱(2,2-diphenyl-1-picrylhydrazyl-high performance liquid chromatography,DPPH-HPLC)法在线筛选杨梅叶中黄酮类抗氧化活性成分,并对抗氧化活性进行检测和评价。方法:采用Shimadzu C18柱(150 mm×4.6 mm,5μm)反相HPLC梯度洗脱法分离杨梅叶中黄酮类成分,通过DPPH-HPLC法快速筛选出具有抗氧化活性的化合物,并以2,2-联苯基-1-苦基肼基(DPPH)自由基清除实验、ABTS自由基清除实验及铁离子还原(FRAP)实验,抗氧化能力检测方法评价其抗氧化活性。结果:在HPLC条件下可以检测出杨梅叶中所含的杨梅苷、杨梅素、槲皮素和山柰酚四种黄酮类化合物并达到完全分离。四种化合物都具有抗氧化活性,其中槲皮素抗氧化能力最强,其次为杨梅素,且两者抗氧化能力都强于L-抗坏血酸,而杨梅苷和山柰酚抗氧化能力较弱。结论:所建立的DPPH-HPLC法可筛选复杂体系中的抗氧化活性成分。

Objective:To establish a 2,2-diphenyl-1-picrylhydrazyl-high performance liquid chromatography(DPPH-HPLC)method for screening the antioxidant active components of flavonoids in the Bayberry leaves,and to detect and analyze antioxidant active components.Methods:Flavonoids in Bayberry leaves were determined by high performance liquid chromatography(HPLC)with the Shimadzu C18(150 mm×4.6 mm,5μm).Compounds with high antioxidant activity were screened by DPPH-HPLC,and their antioxidant activities were evaluated by 2,2-al phenyl-1-bitter base hydrazine(DPPH),2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS)and ferric reducing antioxidant power(FRAP)experiments.Results:Four flavonoids including myricitrin,myricetin,quercetin and kaempferol were detected by HPLC and separated completely.All the four compounds had antioxidant activities,among which quercetin had the strongest antioxidant capacity,followed by myricetin,and both of them have stronger antioxidant capacity than L-ascorbic acid,while myricitrin and kaempferol have weaker antioxidant capacity.Conclusion:The established DPPH-HPLC method could screen the antioxidant active components in the complex system.