J. Korean Soc. Appl. Biol. Chem. 52(1), 92-95 (2009) Short Communication

Free Radical Scavenging Effect of yet. In the present study, the antioxidative activities of the Ethanol Extract from catechu ethanol extracts obtained from selected medicinal , that are frequently used as food and herb medicine, have been evaluated by measuring their DPPH radical- Byung-Yong Ahn* scavenging activities. Here we report a herb species, which has an exceptionally high antioxidative activity Department of Oriental Medicine Resources, Chenbuk effective for the elimination of the free radicals. Natioal University, Iksan, 570-749, Republic of Korea The medicinal plants used in the present study were Received September 8, 2008; Accepted December 5, 2008 purchased from the Korea Medicine Herbs Association (Jeonju, Korea). The plants were ground into 80 mesh and stored at 4°C for use as specimen for extraction. Key words: Areca catechu L., DPPH radical, ethanol DPPH, L-ascorbic acid, BHA, and BHT were purchased extract, scavenging activity, vitamin C from Sigma Chemical Co. (St. Louis, MO). Approximately 10 g of each medicinal was separately put into 100 Free radicals play detrimental roles in peroxidation of mL of 100% ethanol, extracted repeatedly at 37°C two lipid, denaturation of protein, tumor, transformation, times for 12 h each time. The solution was vacuum- mutation, aging, and cancer [Simic, 1988]. To maintain filtrated twice through a filter paper (Whaman, No. 2), healthy life and prevent quality deterioration of food by concentrated, and stored at −20oC until use. peroxidation of lipid, effective prevention of various DPPH radical-scavening assay. The scavenging diseases caused by free radicals is necessary. Researches effects of the ethanol extracts against the DPPH radicals are on-going for the development of antioxidants that were determined according to the method of Yosida et al. inhibits the generation and activity of free radicals. BHA [1989]. In brief, after dissolving the herbal ethanol extract and BHT, which are synthetic phenol derivatives, have with MeOH to concentrations of 120, 80.0, 40.0, 20.0, been widely used for commercial foods owing to their 10.0, 5.0 and 2.50 μg/mL, the prepared herb methanol outstanding antioxidative activities; however, they are solutions were each mixed with 1 mL of 0.15 mM DPPH known through animal experiments to have various solution. After standing for 30 min, the optical densities effects on, among others, the liver, lung, kidney, circulatory of the solutions were measured at 517 nm. The scavenging system, and reproductive system [Choe and Yang, 1982]. effects against the DPPH radicals were expressed as IC50 Thus, demands for plant-originated natural antioxidants values. with more stability and better antioxidation effects are Scavenging effects of medicinal plants on DPPH increasing due to the rejection of synthetically produced radicals. Table 1 shows DPPH radical-scavenging activities ones by the consumers. Cha et al. [2000] reported the of the ethanol extracts of 70 medicinal plants expected to antioxidative activities of 30 species of the medicinal have physiological activity. The medicinal plants with plants marketed in Korea. The antioxidative activities of IC50 below 100 μg/mL were 11 kinds including Acorus 86 species of domestic native plants, 118 medicinal gramineus Solander, those with IC50 below 50 μg/mL plants, 28 medicinal plants, and 32 medicinal plants were were 21 kinds including Arctium lappa L. Areca catechu studied by Lim et al. [1996], Jeong et al. [2004], Kim et L., Cassia tora L., Santalum album L., Terminalia al. [1995], and Kim et al. [1995], respectively. The chebula Retz, and Ulmus macrocarpa Hance, which have antioxidative activities of four mushroom species were high radical scavenging activities, showed IC50 4.5, 8.0, studied by Kwon et al. [2008]. In spite of these studies, 8.0, 7.0, and 6.0 μg/mL. Among these plant extracts, the there still exist numerous frequently used medicinal ethanol extract of A. catechu L. showed the highest plants, whose antioxidant activities have not been studied DPPH radical-scavenging activity. Kim et al. [1993] previously reported that methanol *Corresponding author extract of Cullen corylifolium Medik. showed the Phone: +82-63-850-0743; Fax:+82-63-850-0741 E-mail: [email protected] strongest antioxidant activity among the methanol extracts of 120 medicinal plants by measuring lipid Abbreviations: BHA, butylated hydroxyanisole; BHT, butylated peroxides in a modeling system. The strong antioxidant hydroxytoluene; DPPH, 1,1-diphenyl-2-picrylhydrazyl;. IC50, activities of the methylene chloride extract of Scutellaria concentration required for the 50% reduction of DPPH radicals baicalensis Georgi, 75% ethanol extract of Caesalpinia doi:10.3839/jksabc.2009.016 sappan L. [Lim et al., 1996], 80% methanol extracts of Antioxidant activity of Areca catechu 93

Table 1. Scavenging effects of ethanol extracts of medicinal plants on DPPH radicals

1) Scientific name Part used IC50(μg/mL) Achyranthes japonica Nakai Radix >100 Aconitum carmichaeli Debx Radix >100 Acorus gramineus Solander Rhizoma 95.0 Akebia quinata Decne Lignum 67.5 Angelica gigus Nakai Radix >100 Anselica tenuissima Nakai Radix >100 Arctium lappa L. Fructus 22.0 Areca catechu L. Semen 4.5 Artemisia lavandulaefolia DC. Folium 40.5 Artemisia messerschmidtiana Besser Herba 21.0 Asarum heterotropoides Maekawa Radix >100 Astraglus membranaceus Bunge Radix >100 Atractyodes japonica Kitag Rhizoma >100 Bupleurum falcatum L. Radix >100 Cassia tora L. Semen 8.0 Chaenomelesl agenaria Koidz Fructus >100 Cibotium borometz J. Sm Rhizoma >100 Cinnamomum cassia Presl. Cortex 82.5 Cinnamoum cassia Blume Ramulus 42.5 Citrus nobilis Loun Cortex >100 Clematis mandshurica Maximowicz Radix >100 Codonopsis lanceolata Bentham et Hooker Radix >100 Crataegus pinnatifida Bunge Fructus 22.0 Dendrobium moniliforme Swartz Herba 41.0 Epimedium koreanum Nakai Herba >100 Eucommia ulmoides Oliver Cortex >100 Forsythia korean Nakai Fructus 31.0 Gardenia jasminoides Ellis Fructus 45.0 Gasfrodia elata Blume Rhizoma >100 Gleditschia horrida Makino Spina 25.0 Kalopanax pictus Nakai Cortex 57.0 Leonurus sibiricus L. Herba >100 Ligusticum jeholense Kitag Rhizoma  100 Lnula helenium L. Radix >100 Lycium chinense Miller Fructus >100 Menth arvensis var. piperascens Malinv. Herba 30.0 Morinda officinalis How Radix >100 Morus bombycis Koitzumi Cortex 57.0 Ophiopogon japonicus Ker-Gawler Radix 93.0 Orobanche coerulescens Willdenow Caulis 63.0 Ostericum koreanum(Max) Kitagwa Radix >100 Paeonia japonica Miyabe et Takeda Radix 16.0 Perilla frutescens var. acuta Kudo Folium 68.0 Phellodendron amurense Rupr. Cortex 75.0 Pinellia ternata Breit. Tuber >100 Platycodom grandiflorum A. DC. Radix >100 Polygonum multiflorium Thunb Radix >100 Polygonum multiflorium Thunb Radix >100 Poncirus trifoliata Rafiiensque Fructus >100 94 Byung-Yong Ahn

Table 1. Continued

1) Scientific name Part used IC50(μg/mL) Poria cocos Wolf Poria >100 Prunus armeniaca L. var. ansu Max Semen >100 Pueraria hirsuta Matsum Flos >100 Pueraria thunbergiana Benth. Radix 87.0 Rehmannia glutinosa (Gaertner) Liboshitz Rhizoma >100 Rheum undulatum L. Rhizoma 11.0 Ribe diacantha Pallas Herba 30.0 Rosa laevaigata Michx Fructus 35.0 Rubia cordifolia L. Radix 20.0 Rubus coreanus Miq. Fructus 25.0 Salvia miltriorrhiza Bunge Radix 20.0 Santalum album L. Flos 8.0 Scutellaria baicalensis Georgi Radix 38.0 Sophora flanescens Aiton Radix 66.0 Retz Fructus 7.0 Thyja orientalis L. Folium 20.0 Ulmus macrocarpa Hance Radix 6.0 Valeriana fauriei Briquet Herba 66.0 Xanthium strmarium L. Flos 21.0 Zanthoxylum piperitum DC. Semen 15.0 Zingiber officinale Ros. Rhizoma 29.5 L-ascorbic acid 7.5 BHA 4.3 BHT 13.0 1)The values indicate 50% decrease of DPPH radical and are the means of triplicate data. hull and Ulmus davidiana Planchon var. ability than hot water extract [Lim et al., 2000]. japonica Nakai [Lim et al., 2000], 80% methanol extract Antioxidants are classified into fat-soluble antioxidants, of Sophora japonica L. [Jeong et al., 2004], and 100% such as tocopherol, ubiqunone, and carotenoid, and methanol extract of Salvia miltriorrhiza Bunge [Kwon et water-soluble antioxidants, including ascobic acid and al., 2008] have been previously reported. In my research, glutathion [Kim et al., 1995], implying that A. catechu L. the ethanol extracts of Salvia miltriorrhiza Bunge and may be used as a fat-soluble antioxidant, leading to the Ulmus davidiana Planchon var. japonica Nakai also possibility of developing natural antioxidant material to showed strong DPPH radical-scavenging activity, as be used for deterring oxidation and acidification of fat. previously reported by Lim et al. [2000] and Kwon et al. In view of the result that medicinal plants have high [2008]. content of antioxidizing phenol ingredients, and particularly A. catechu L showed a stronger antioxidative activity the higher the contents of phenol and in A. catechu than S. miltriorrhiza Bunge and U. davidiana Planchon L. methanol extract, the higher the anti-oxidation ability, var. japonica Nakai, with IC50 of 20 and 6.0 μg/mL, the antioxidation-active ingredients are presumed to be respectively, whereas IC50 was 4.5 μg/mL for A. catechu phenolic and tannic compounds with hydroxyl groups. L. In the case of Scutellaria baicalensis Georgi reported Comparison of the DPPH radical elimination ability of A. by Kim et al. [1995] as having strong DPPH radical catechu L. ethanol extract with those of ascorbic acid, elimination ability, the IC50 was 38 μg/mL, which is not BHA, and BHT, the ethanol extract of A. catechu L. was an indication of a strong radical-scavenging activity. shown to have stronger radical elimination ability than A. catechu L. possesses insecticidal, anti-biosis, anti- ascorbic acid and BHT. Further research should be mildew, anti-infection, and antioxidation effects. Upon followed to isolate and identify the active ingredients comparison methanol extract showed higher antioxidation with strong antioxidationability in A. catechu L. Antioxidant activity of Areca catechu 95

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