Determination of antioxidant activity and tannin content from Aristolochia longa leaves and roots using liquid-liquid fractionation method

Soumia Attou, Yamina Maizi, A. Belmimoun, Khadidja SIDE LARBI, B. Meddah, Aicha Tir touil
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Abstract

Aristolochia longa has been known by its potentiel to cure various diseases.This study was aimd to estimate the antioxidant activity and tannin content of A.longa leaves and roots. Liquid-liquid fractionation by using three solvants was experimented for extraction of tannin from leaves and roots.Then the fractions were oven-dried and powdered to determine the condensed tannin content and antioxidant activity by acidic vanillin, DPPH radical scavenging activity and ferric reducing antioxidant power (FRAP) methods.The qualitative analysis was releazed by high-performance liquid chromatography coupled to mass spectrometry (HPLC DAD-MS).The highest extraction yield was obtained by roots N-butanol fraction (5.15±0.47%).The maximum tannin content value was noted in roots N-butanol fraction (3.90±0.24 mg QUE/g DM) followed by leaves ethyl acetate fraction (3.03±0.01 mg QUE/g DM).The IC50 values of DPPH radical scavenging activity varied significantly from IC50=0.123±0.009mg/ml to 3.737±0.56 mg/ml. Since, the FRAP values ranged from EC50=0.066±0.002 mg/ml to 4,00±0,2 mg/ml. The highest antioxydant ability was observed in leaves and roots N-butanol fractions .Also, the identification by HPLC analysis showed that the roots contains some tannin compounds than leaves such as vescalagin, Roburin D and 1,3,4-tri- O -galloylquinic acid.Our study confirmed the use of A.longa to prevent and control the oxidative stress.
液液分馏法测定长马兜铃叶和根的抗氧化活性和单宁含量
马兜铃因其治疗各种疾病的潜力而闻名。本研究旨在测定长叶和根的抗氧化活性和单宁含量。采用三种溶剂对单宁进行液液分馏提取。焙烧干燥后粉碎,采用酸性香兰素法、DPPH自由基清除法和铁还原抗氧化能力法测定浓缩单宁含量和抗氧化活性。采用高效液相色谱-质谱联用(HPLC - DAD-MS)进行定性分析。根茎正丁醇萃取率最高(5.15±0.47%)。单宁含量最高的部位为根正丁醇部位(3.90±0.24 mg QUE/g DM),其次为叶乙酸乙酯部位(3.03±0.01 mg QUE/g DM)。DPPH自由基清除活性的IC50值从0.123±0.009mg/ml到3.737±0.56 mg/ml变化显著。因此,FRAP值范围为EC50=0.066±0.002 mg/ml至4000±0.2 mg/ml。抗氧化能力最强的部位是叶和根的正丁醇部位。HPLC分析表明,根中含有的单宁类化合物有鳞状胶质蛋白、红毛蕊素D和1,3,4-三- O -没食子酰奎宁酸。我们的研究证实了龙舌兰对氧化应激的预防和控制作用。
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