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Natural Antioxidant Properties of Selected Wild Mangifera Species in Malaysia

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Natural Antioxidant Properties of Selected Wild Mangifera Species in Malaysia J. Trop. Agric. and Fd. Sc. 44(1)(2016): 63 – 72 Mirfat, A.H.S., Salma, I. and Razali, M. Natural antioxidant properties of selected wild Mangifera species in Malaysia Mirfat, A.H.S.,1 Salma, I.2 and Razali, M.1 1Agrobiodiversity and Environmental Research Centre, Persiaran MARDI-UPM, 43400 Serdang, Selangor, Malaysia 2Gene Bank and Seed Centre, MARDI Headquarters, Persiaran MARDI-UPM, 43400 Serdang, Selangor, Malaysia Abstract Many wild fruit species found in Malaysia are not well known and are underutilised. Information on their health benefits is critical in efforts to promote these fruits. This study was conducted to evaluate the antioxidant potential of seven species of wild Mangifera (mango) in Malaysia: M. caesia (binjai), M. foetida (bacang), M. pajang (bambangan), M. laurina (mempelam air), M. pentandra (mempelam bemban), M. odorata (kuini) and M. longipetiolata (sepam). The results were compared to those obtained from a popular mango, M. indica. Among the mangoes, M. caesia was found to be the most potential source of antioxidant as evidenced by its potent radical scavenging activity (92.09 ± 0.62%), ferric reducing ability (0.66 ± 0.11 mm) and total flavonoid content (550.67 ± 19.78 mg/100 g). Meanwhile, M. pajang showed the highest total phenolic (7055.65 ± 101.89 mg/100 g) and ascorbic acid content (403.21 ± 46.83 mg/100 g). In general, from the results obtained, some of the wild mango relatives were found to have strong antioxidant potential that is beneficial to health. This study provides a better understanding of the nutraceutical and functional potential of underutilised Mangifera species. The information is very useful for genetic enhancement of the Mangifera species in the future and justified the need of its conservation. Keywords: wild Mangifera, antioxidant activity, total phenolic, total flavonoid, ascorbic acid Introduction villages. On the other hand, wild fruits are Malaysia has a rich diversity of tropical naturally distributed in the forests, and they fruits grown naturally in the region of are the indigenous fruits of Malaysia. Peninsular Malaysia, Sabah and Sarawak. Mangifera belongs to the Sapindales Currently, it has been estimated that 370 order in the Anacardiaceae family. Its tropical fruit species occur in Malaysia centre of origin and diversity is firmly (Rukayah 2002). Raziah and Salma (2006) established in Southeast Asia (Bompard and categorised these tropical fruits as major, Schnell 1997). The genus Mangifera was minor, rare and wild fruits. Major fruits mostly restricted to tropical Asia, with the are those that are commonly consumed, highest diversity occurs in Malaysia, Java commercially grown and of economic and Sumatra (Kostermans and Bompard importance. Minor and rare fruits are usually 1993). Salma et al. (2006) have listed 30 grown in the orchards, home gardens and (cultivated and wild) Mangifera species Article history Authors’ full names: Mirfat Ahmad Hasan Salahuddin, Salma Idris and Razali Mirad Received: 29.11.2013 Accepted: 8.5.2015 E-mail: [email protected] ©Malaysian Agricultural Research and Development Institute 2016 63 Antioxidant properties of wild Mangifera occurring in Malaysia, with three species are M. laurina (mempelam air), M. pentandra endemic. Of these 30 species, only 16 bear (mempelam bemban), M. odorata (kuini) and edible fruits. Twelve species were reported M. longipetiolata (sepam) with the intention to be cultivated, of which eight species to promote the conservation and utilisation are found in the wild (Kostermans and on these fruits species in the future. Bompard 1993). Mangifera indica L., the common Materials and methods mango originated from India and Myanmar, Mangifera foetida, M. caesia, M. pajang, is widely cultivated throughout the world. M. laurina, M. pentandra, M. odorata Mango is a popular and economically (kuini) and M. longipetiolata (sepam) were important tropical fruit due to its excellent collected from MARDI field genebanks in eating quality and nutritional composition Serdang and also from various locations in (vitamins, minerals, fibre and other Peninsular Malaysia, Sabah and Sarawak. phytochemical compounds). Mango was Only ripe Mangifera fruits were used in also reported to contain various classes of this study. polyphenols, carotenoids and ascorbic acid Fruit samples were washed with demonstrating different health-promoting running tap water before being weighed for properties, mainly from their antioxidant their whole and edible portion parts. Then, activities (Talcott et al. 2005). Polyphenols the edible portions were cut into small are one of the major antioxidants that pieces and freeze-dried using a bench-top scavenge free radicals and very useful in freeze dryer (Virtis, USA). Then, the freeze- reducing the harmful effects of oxidative dried samples were finely ground and kept damage which contribute to health in airtight containers prior to extraction. enhancement and disease prevention (Liu et The samples were extracted using methanol al. 2000). (1:10) and shaken for approximately 1 h Most studies on Mangifera were before centrifuged for 10 min at 10,000 rpm. extensively focused on the common mango. The residue was separated from the However, other species, which include wild supernatant and the procedure was repeated mango relatives, have not been explored and twice. The two resulting supernatants were received much attentions. Among the wild mixed together to obtain the crude extracts and underutilised Mangifera that produce which were stored at –80 °C prior to edible fruits are M. caesia Jack, M. foetida analysis. Lout., M. kemanga Bl., M. laurina Bl., For ascorbic acid analysis, the M. odorata Griff., M. pajang Kostermans, extraction was performed according to M. sylvatica Roxb, M. horsefieldia, Patric et al. (2006) and Yurena et al. (2006) M. lagenifera and M. torquenda (Tanaka with some modifications. The freeze- 1976; Bompard 1992). Wild mango relatives dried fruit samples were weighed and are currently being threatened to genetic loss mixed with extract buffer (100 µg/ml Tris due to their lack of popularity among local (2-carboxyethyl)-phosphine hydrochloride communities and also lack of information (TCEP-HCl), 3% MPA, 8% acetic acid on their health benefits. From ethnobotanical and 1 mm ethylenediaminetetraacetic acid reports, wild fruits could also play an disodium salt (EDTA). The mixture was important role in various health promoting homogenised in a Virtishear homogeniser benefits. The most commonly studied health (Virtis, USA) in ice for 1 min and then benefit of fruits is the antioxidant effect. centrifuged at 10,000 rpm (refrigerated This study was conducted to evaluate at 4 °C) for 10 min. The supernatant the antioxidant activity of selected wild was filtered through a 0.45 µm cellulose mango relatives: M. foetida (bacang), membrane and stored at –80 °C until M. caesia (binjai), M. pajang (bambangan), further use. 64 Mirfat, A.H.S., Salma, I. and Razali, M. The antioxidant activity 2,2-diphenyl- total of 7 µl of sample and 20 µl of distilled 1-picrylhydrazyl (DPPH) assay was water were added to 200 µl of FRAP reagent determined through scavenging activity of and incubated at 37 °C for 4 min. Standards the fruit extracts on DPPH radicals which of known Fe2+ concentrations were run were was assayed according to Molyneux using several concentrations ranging from (2004) with some modifications. Various 100 – 1000 mm. All analyses were run in concentrations of the crude extracts in triplicate and the absorbance was measured methanol were prepared to get a final at 593 nm. A standard of known Fe2+ (100 – volume of 7 µl and were mixed with 280 µl 1000 mm) was used to produce a calibration of methanolic solution containing DPPH curve. The final results were expressed as (Sigma, USA) radicals resulting in a final the concentration of antioxidant having a concentration of 0.06 mm. The mixture ferric reducing ability in mm. was vigorously shaken and left to stand Total phenolic content (TPC) of the for 30 min in the dark. The absorbance extracts was estimated by a colorimetric was measured at 517 nm and ascorbic acid assay as described by Singleton and Rossi (Sigma, USA) was used as the positive (1965) with some modifications. Briefly, control. The assays were carried out in 50 µl of the crude extracts were mixed with triplicate and the results were expressed 100 µl of Folin Ciocalteau’s phenol reagent as mean values ± standard deviations. The (Merck, Germany). After 3 min, 100 µl of scavenging effect on DPPH free radicals 10% sodium carbonate (Na2CO3) (Sigma- was calculated as follows: Aldrich, USA) was added to the reaction mixture and allowed to stand in the dark A − A t for 60 min. The absorbance was measured Inhibition (%) = ––––––––––––––––Control Extrac x 100% at 725 nm and TPC was obtained from a [ AControl ] calibration curve using gallic acid (0 – 10 µg/ml) as a standard reference. Estimation of AControl is the absorbance of the control TPC was carried out in triplicate. The results reaction (containing all reagents except were mean values ± standard deviations the test compound), and AExtract is the and expressed as mg gallic acid per 100 g absorbance of the test compound. The samples. All procedures were carefully results were expressed as IC50 value carried out with minimum exposure of light. (mg/ml), where IC50 is the inhibitory Determination of total flavonoid concentration at which DPPH radicals were content (TFC) was based on the method scavenged by 50% and was obtained by described by Kim et al. (2003) with some interpolation from linear regression analysis. modifications. An aliquot of 100 µl of All procedures were carefully carried out fruit extract was diluted with 400 µl of with minimum exposure of light. distilled water. Afterwards, 30 µl of 5% Ferric reducing antioxidant power sodium nitrite (NaNO2), (Sigma, USA) (FRAP) assay was determined based on the solution was added and allowed to react reduction of Fe3+-TPTZ to a blue coloured for 5 min.
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