Antioxidant and antiplatelet activities of flavonoid-rich fractions of three citrus fruits from Korea

3 Biotech, Apr 2016

Three different fractional (methanol, ethyl acetate and hexane) extracts from yuzu (Citrus junos Sieb ex Tanaka), hallabong [(C. unshiu Marcov × C. sinensis Osbeck) × C. reticulata Blanco] and orange (C. sinensis) were evaluated for their antioxidant activity and antiplatelet effects. 2,2-Diphenyl-1-Picrylhydrazyl (DPPH), cupric reducing antioxidant capacity (CUPRAC) and ferric-reducing antioxidant power (FRAP) methods were used for the antioxidant activity tests. Total flavonoids and total phenolics were also evaluated spectrophotometrically. The ethyl acetate fraction contained the highest total flavonoid and total phenolic content and exhibited the highest antioxidant activities (185.2 ± 14.5 and 208.7 ± 17.5 mg/g dry extract for FRAP and CUPRAC values, respectively). The total phenolic and total flavonoid content ranged from 58.2 ± 1.4 to 102.4 ± 8.6 and 19.6 ± 0.5 to 64.3 ± 0.8 mg/g dry extract, respectively. The results of DPPH assay showed that ethyl acetate fractions had the least IC50 values (0.12 ± 0.002, 0.04 ± 0.0006, in mg/mL for orange and hallabong samples, respectively) followed by the hexane fraction (0.19 ± 0.007 mg/mL) of the orange sample. For all fractions, the antioxidant activity and contents of total phenolics and total flavonoids correlated well with each other. In vitro investigation of the antiplatelet effect showed that ethyl acetate fraction has the highest inhibition (84.3 ± 5.8 to 96.1 ± 1.8 %). Hexane and MeOH fractions of hallabong and orange samples inhibited platelet aggregations by less than or equal to 41 %.

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Antioxidant and antiplatelet activities of flavonoid-rich fractions of three citrus fruits from Korea

Biotech Antioxidant and antiplatelet activities of flavonoid-rich fractions of three citrus fruits from Korea Awraris Derbie Assefa 0 1 Eun Young Ko 0 1 So Hyun Moon 0 1 Young-Soo Keum 0 1 0 Department of Bioresources and Food Science, Konkuk University , Seoul 143-701 , South Korea 1 Awraris Derbie Assefa Three different fractional (methanol, ethyl acetate and hexane) extracts from yuzu (Citrus junos Sieb ex Tanaka), hallabong [(C. unshiu Marcov 9 C. sinensis Osbeck) 9 C. reticulata Blanco] and orange (C. sinensis) were evaluated for their antioxidant activity and antiplatelet effects. 2,2-Diphenyl-1-Picrylhydrazyl (DPPH), cupric reducing antioxidant capacity (CUPRAC) and ferric-reducing antioxidant power (FRAP) methods were used for the antioxidant activity tests. Total flavonoids and total phenolics were also evaluated spectrophotometrically. The ethyl acetate fraction contained the highest total flavonoid and total phenolic content and exhibited the highest antioxidant activities (185.2 ± 14.5 and 208.7 ± 17.5 mg/ g dry extract for FRAP and CUPRAC values, respectively). The total phenolic and total flavonoid content ranged from 58.2 ± 1.4 to 102.4 ± 8.6 and 19.6 ± 0.5 to 64.3 ± 0.8 mg/g dry extract, respectively. The results of DPPH assay showed that ethyl acetate fractions had the least IC50 values (0.12 ± 0.002, 0.04 ± 0.0006, in mg/mL for orange and hallabong samples, respectively) followed by the hexane fraction (0.19 ± 0.007 mg/mL) of the orange sample. For all fractions, the antioxidant activity and contents of total phenolics and total flavonoids correlated well with each other. In vitro investigation of the antiplatelet effect showed that ethyl acetate fraction has the highest inhibition (84.3 ± 5.8 to 96.1 ± 1.8 %). Hexane and MeOH fractions of hallabong and orange samples FRAP; DPPH phenolics; Citrus fruits - inhibited platelet aggregations by less than or equal to 41 %. Introduction Platelet aggregation Total Because of the unhealthy lifestyle and aged population, mortality associated with cancer and cardiovascular diseases has increased (Fuentes et al. 2013a) . Reactive oxygen species (ROS) are known to play a dual role, being either harmful or beneficial to living systems. ROS have beneficial effect in defense against infectious agents, in the function of a number of cellular signaling systems and in the induction of a mitogenic response at low concentrations. At high concentrations, ROS can be important mediators of damage to cell structures, including lipids and membranes, proteins and nucleic acids (Valko et al. 2006) . ROS play an important role in degenerative or pathological processes such as cancer (Valko et al. 2006) , coronary heart disease (Watt et al. 2012) , neurodegenerative disorders (Roy et al. 2008) , atherosclerosis (Patel et al. 2000) and inflammation (Kielland et al. 2009) . The development of cardiovascular diseases such as acute myocardial infarction, cerebrovascular disease and peripheral arterial thrombosis is related to the interaction process of atherosclerotic lesions and thrombus formation. This interaction is basically established with the participation of platelets (Fuentes et al. 2013b) . Platelets adhere to endothelial cells and contribute to the recruitment of leukocytes involved in the local vascular inflammation and thrombosis formation (Jackson et al. 2009). Several synthetic drugs such as aspirin and triflusal are used to inhibit the aggregation of platelets (Cruz et al. 1998; Yu et al. 2011) . However, it has been reported that synthetic drugs are accompanied by several adverse side effects. Low-dose aspirin increases the risk of major bleeding and intestinal ulceration (Schror 1997). The term ‘aspirin resistance’ has been used to describe the failure of aspirin to inhibit platelet activity. Between 8 and 45 % of patients who suffered an ischemic stroke or cardiovascular disease are aspirin resistant (Ohmori et al. 2006) . Therefore, the development of safe, alternative therapeutic agents with antiplatelet activity is crucial. Reports show that plant-derived foods like spinach, which is rich in nitrates, apple rich in flavonoids and tomato in lycopene exhibit antiplatelet activity (Bondonno et al. 2012; Fuentes et al. 2013a) . Plants are believed to be valuable sources of antioxidants. Epidemiological studies suggest that consumption of fruits and vegetables is associated with significantly lower risk of coronary artery disease and stroke (Hu 2003). The health benefits of citrus fruits are largely attributed to the presence of the relatively high concentrations of antioxidant compounds, such as ascorbic acid and phenolic compounds, mainly flavonoids (Gonza´lez-Molina et al. 2010; Kawaii et al. 1999) . Various studies showed that extracts from citrus peels exhibited antiplatelet activities besides their antioxidant activity (Sagdic et al. 2011; Yu et al. 2005) . Although it is believed that citrus fruits have several be (...truncated)


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Awraris Derbie Assefa, Eun Young Ko, So Hyun Moon, Young-Soo Keum. Antioxidant and antiplatelet activities of flavonoid-rich fractions of three citrus fruits from Korea, 3 Biotech, 2016, pp. 109, Volume 6, Issue 1, DOI: 10.1007/s13205-016-0424-8