Human Journals Research Article March 2020 Vol.:17, Issue:4 © All rights are reserved by Latifa Nasser A. Abdulqawi et al. Antioxidant Activity of Yemeni Plants, Myrtus communis L. and Flemingia grahamiana Wight & Arn. Keywords: Myrtus communis, Flemingia grahamiana, Yemeni plants, DPPH antioxidant activity ABSTRACT Latifa Nasser A. Abdulqawi*1, Syed Atheruddin Quadri2 The present study was aimed to investigate the antioxidant activity of methanol and aqueous extracts of leaves of Myrtus 1 Department of Zoology, Maulana Azad Collage, Dr. communis and pods (Warrus) of Flemingia grahamiana Wight Babasaheb Ambedkar Marathwada University, & Arn. Antioxidant activity of extracts was determined by1, 1- Aurangabad, India. Diphenyl-2-picryl hydrazyl (DPPH) method. Both extracts showed antioxidant activity in a concentration-dependent 2 Department of Zoology, Maulana Azad Collage Dr. manner. The methanolic and aqueous extract of Myrtus Rafiq Zakaria Campus, Aurangabad, India communis leaves showed a scavenging ability with an IC50 value of 2.79 and 2.65 µg/ml, respectively. And IC50 of Submission: 25 February 2020 Accepted: 2 March 2020 Published: 30 March 2020 www.ijppr.humanjournals.com Flemingia grahamiana Warrus extracts were 3.95 and 0.71 µg/ml, respectively. IC50 value for the standard ascorbic acid was 3.35 μg/ml. www.ijppr.humanjournals.com INTRODUCTION The distinguished geographical location of Republic of Yemen and the diversity of landscapes and climate coupled with the Socotra Archipelago gives the Yemeni plants a great degree of diversity. In spite of the introduction of the modern medicinal system during the twentieth century, folk herbal medicine keeping to play an important role in many areas of Yemen [1, 2]. Many of Yemeni medicinal plants have shown good biological activities, but there are Yemeni plants to need future studies which will supply new insights about pharmacological activities [2]. Medicinal and Aromatic plants are the source of natural antioxidants due to their main secondary metabolites [3]. Nowadays, the interest in naturally happening antioxidants has much increased for use in cosmetic, food and pharmaceutical products to exchange synthetic antioxidants that are being limited due to their carcinogenicity [4]. Furthermore, growing evidence related to the event of cancer to the oxidative damage to proteins, lipids, and DNA in the body caused by free radicals and other carcinogens [5]. And due to many plants are hugely free from adverse effects and have good pharmacological actions, Plants are a rich source of bioactive chemicals [6]. Myrtus communis L. is one of the important medicinal and aromatic species from the Myrtaceae family [3]. Which it has several antioxidants like tannins, flavonoids, and α-tocopherol [7]. Flemingia grahamiana Wight & Arn. Belongs to the Fabaceae family, the genus Flemingia has about 42 species. It is a perennial shrub [8]. The pods of F. grahamiana covered with red powder it's called Warrus. In Arabia, warrus is applied as a cosmetic and in the treatment of skin disease, anthelmintic, cough and chills [9]. There are a few studies have assured the pharmacological activity of Flemingia genus [9, 10, 11]. The main goal of this study is to evaluate the antioxidant activity of methanolic and aqueous extracts of medicinal Yemeni plants: Myrtus communis L. and Flemingia grahamiana Wight & Arn. Citation: Latifa Nasser A. Abdulqawi et al. Ijppr.Human, 2020; Vol. 17 (4): 805-811. 806 www.ijppr.humanjournals.com MATERIALS AND METHODS Plant Materials and Extracts The leaves of Myrtus communis and pods of Flemingia grahamiana Wight & Arn were collected from Yaffe Mountains of Yemen. The plant material was authenticated by Dr. Rafiuddin Nasser, Assistant professor, Department of Botany, Maulana Azad College, Aurangabad. The leaves and pods were dried in the shade. The leaves were crushed, and pods of F. grahamiana were rubbed and gently shake, to denude them of their outer hairy covering and get powder ‘Warrus’ (Figure 1). Leaves and powder ‘Warrus’ (50 g) were extracted with 500 ml methanol separately in a Soxhlet extractor, then the solvents were filtered through a filter paper Whatman No. 1 and evaporated by a rotary evaporator, then transferred to an incubator at 40°C for 24 h to get a dry mass. The extracts were put in sterile labeled bottles and kept in a refrigerator. Also, 50 g of leaves and powder ‘Warrus’ were taken separately, in a beaker, and 500 ml of distilled water was added. The mixture was stirred by a magnetic stirrer for 24 h. The mixture was clarified by filtration and then subjected to evaporation and kept at -20°C till used [12]. Figure No. 1: The pods of F. grahamiana covered by Warrus Citation: Latifa Nasser A. Abdulqawi et al. Ijppr.Human, 2020; Vol. 17 (4): 805-811. 807 www.ijppr.humanjournals.com DPPH free radical scavenging assay Antioxidant activity of methanolic and aqueous extracts of Myrtus communis (leaves) and Flemingia grahamiana (pods) was determined using 1, 1-Diphenyl-2-picryl hydrazyl radical (DPPH) method with minor modification [13]. 0.05 ml of the extracts dissolved in methanol were diluted to 1.0 ml, using ethanol to attain the concentrations of 1-200 μg/ ml, and were added to DPPH (Final concentration 200 μM, in 95% ethanol). The absorbance of the resulting solution was read at 515 nm after 20 min using a spectrophotometer. Ascorbic acid was used as a reference standard. The tests were performed in triplicate and the percentage inhibition was calculated by using the following formula: % Inhibition = [(Abscontrol - Abssample) / Abscontrol] × 100 Where Abscontrol is the absorbance of DPPH radical solution without extract, Abssample is the absorbance of DPPH radical solution mixed with the extract. IC50 values were calculated using Linear regression by plotting scavenging activity against sample concentrations using Microsoft Excel software. STATISTICAL ANALYSIS Experimental data were conducted in triplicates and results for each measured parameter were expressed as means ± Stander error. Statistical analyses were performed by one-way ANOVA. The difference was considered to be statistically significant when p < 0.05. Analysis was done using SPSS ver. 20.0 software. RESULTS AND DISCUSSION DPPH free radical scavenging assay Free radical scavenging (DPPH) properties of methanolic and aqueous extracts from M. communis and F. grahamiana are presented in (Table 1) and (Figure 2 & 3). All extracts showed highest antioxidant activity on DPPH radicals with IC50 value of 0.71, and 2.65 μg/mL for aqueous extracts of F. grahamiana and M. communis. And IC50 value of 2.79 and 3.95 μg/mL for methanolic extracts of M. communis and F. grahamiana. DPPH scavenging ability of the extracts of F. grahamiana and M. communis were higher than Citation: Latifa Nasser A. Abdulqawi et al. Ijppr.Human, 2020; Vol. 17 (4): 805-811. 808 www.ijppr.humanjournals.com synthetic antioxidant (Ascorbic acid with IC50 of 3.35 μg/mL) except methanolic extracts of F. grahamiana. There are several studies showed M. communis has a strong antioxidant activity on DPPH radical. Amensour, et al, [14] reported antioxidant activity of Moroccan Myrtus communis leaves and berries in which four solvents were used. In his results both leaves and barriers extracts showed antioxidant activity in the order of methanol > water> ethanol >ethyl acetate. Leaves extracts have more activity in compare to berries extracts. The aqueous and methanol extracts from Tunisian M. communis showed an important free radical DPPH with IC50 value 1.9 μg/ml and 6.5 μg/ml, respectively [15]. Babou et al, [16] had reported antioxidant of leaves, berries and seed extracts of myrtle at different times and the best IC50 was obtained from leaves harvested in September and December (8.29 μg/mL and 9.44 μg/mL), respectively, berries harvested in September (8.42 μg/mL) and seeds harvested in December (3.89μg/mL). Ascorbic acid used as control with IC50 of 6.93 μg/mL. Also, IC50value of three Myrtle extracts; hydroalcoholic, ethyl acetate and aqueous were 0.36, 2.27 and 2.88 µm, respectively [7]. Gumula et al., 2014 [17] reported that leaf extract showed an IC50 value of 5.9 μg/mL in a DPPH. Also, there are few antioxidant studies of genus Flemingia. According to Madanet al, [18], methanolic extract of root and leaf of F. strobilifera showed a very good radical scavenging activity (DPPH) with low IC50 values of 11.4 µg/ml and 38.0 µg/ml, respectively. Also, Hsieh et al, [6] have studied antioxidant of aqueous extracts of four flemingia species in Taiwan (Flemingia macrophylla, Flemingia prostrata, Flemingia lineata and Flemingia strobilifera) in which aqueous extract of Flemingia macrophylla had the strongest antioxidant activity with IC50 values of 36 µg/ml. Table No. 1: IC50 values of methanolic and aqueous extracts of M. communis and F. grahamiana in DPPH radical scavenging activity. Plants & positive controls M. communis F. grahamiana Ascorbic acid Extracts IC50 (μg/mL) - Methanolic 2.79 ±0.02 - Aqueous 2.65 ±0.03 - Methanolic 3.95 ±0.03 - Aqueous 0.71 ±0.02* 3.35 ±0.01 All values are expressed as Mean ± SE (n=3). *= Difference statistically significant (P<0.05) Citation: Latifa Nasser A. Abdulqawi et al. Ijppr.Human, 2020; Vol. 17 (4): 805-811. 809 www.ijppr.humanjournals.com Figure No. 2: DPPH radical scavenging activity of methanolic and aqueous extracts of M. communis at different concentrations Figure No. 3: DPPH radical scavenging activity of methanolic and aqueous extracts of F. grahamiana at different concentrations CONCLUSION The present study of Myrtus communis leaves and Flemingia grahamiana Warrus indicate that the methanolic and aqueous extracts possess a good scavenging activity. Further studies are recommended to isolate and identify antioxidant components in Myrtus communis and Flemingia grahamiana, which could be used as an easily accessible source of natural antioxidants; possible food supplement and in the pharmaceutical industry. Citation: Latifa Nasser A. Abdulqawi et al. Ijppr.Human, 2020; Vol. 17 (4): 805-811. 810 www.ijppr.humanjournals.com REFERENCES 1- Kilian N, Hein P and Hubaishan MA. New and noteworthy records for the flora of Yemen, chiefly of Hadhramout and Al-Mahra.Willdenowia 2002; 32(2):239-69. 2- Chhetri BK, Ali NA and Setzer WN. A survey of chemical compositions and biological activities of Yemeni aromatic medicinal plants. Medicines 2015; 2(2):67-92. 3- Aleksic V and Knezevic P. Antimicrobial and antioxidative activity of extracts and essential oils of Myrtuscommunis L. Microbiol. Res. 2014; 169(4):240-54. 4- Djeridane A, Yousfi M, Nadjemi B, Boutassouna D, Stocker P and Vidal N. 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