Global journal of medicinal plant research, 2(1) January 2014, Pages: 6-10 AENSI Journals Global journal of medicinal plant research ISSN:2074-0883 Journal home page: http://www.aensiweb.com/gjmpr.html Characterization of Antioxidant, Antimicrobial, Anticancer Property and Chemical Composition of Garcinia Mangostana Rind Extract 1 Lee Seong Wei, 2Wendy Wee, 3Julius Yong Fu Siong and 3Desy Fitrya Syamsumir 1 Faculty of Agro Based Industry, Universiti Malaysia Kelantan Jeli Campus, 17600, Jeli, Kelantan, Malaysia. Department of Fisheries Science, Faculty of Fisheries and Aqua-Industry, Universiti Malaysia Terengganu Kuala Terengganu, 21030, Terengganu, Malaysia.. 3 Institute of Marine Biotechnology, Universiti Malaysia TerengganuKuala Terengganu, 21030, Terengganu, Malaysia. 2 ARTICLE INFO Article history: Received 28 January 2014 Received in revised form 19 March 2014 Accepted 26 March 2013 Available online 10 April 2014 Keywords: antioxidant, anticancer, antimicrobial, chemical compound, Garcinia mangostana ABSTRACT This study was carried out to characterize antimicrobial, antioxidant and anticancer activities of Garcinia mangostana rind extract as well as its chemical composition. The main objective of the present study is to reveal the potential of G. mangostana rind in medicinal uses. Antimicrobial property of G. mangostana rind extract was revealed by using two fold microdilution method whereas antioxidant activity of the extract was determined with α, α-diphenyl-β-picrylhydrazyl (DPPH) radical scavenging method. The anticancer property of the plant extract was revealed through Colorimetric MTT (tetrazolium) assay against human breast adenocarcinoma (MCF-7). Chemical compounds of the plant extract were screening and identified by using gas chromatography-mass spectrometry (GC-MS). The minimum inhibitory concentration (MIC) values of the plant extract against the tested bacterial isolates ranged from 31.26 to 125 mg/l in which the plant extract was found can inhibit all the tested bacterial isolates namely A. hydrophila, E. tarda, E. coli, Flavobacterium sp., Klebsiella sp., P. aeruginosa, Salmonella sp., V. alginolyticus, V. cholerae and V. parahaemolyticus. The value of IC50 of G. mangostana rind extract against DPPH and MCF-7 cell was 2.67 ± 0.33 ppt and 1.98 ± 0.12 µg/ml, respectively. A total of 33 chemical compounds was successfully identified with Cyclopenta[d]antrhacene-6, 8, 11-trione, 1, 2, 3a, 4, 5, 6a, 7, 8, 11, 12-dodecahydro-3- (1-methylethyl) 12.63 % was the major compound in the plant extract. In conclusion, the potential of G. mangostana rind as antimicrobial, antioxidant and anticancer agents are promising. © 2014 AENSI Publisher All rights reserved. To Cite This Article: Lee Seong Wei, 2Wendy Wee, 3Julius Yong Fu Siong and 3Desy Fitrya Syamsumir., Characterization of Antioxidant, Antimicrobial, Anticancer Property and Chemical Composition of Garcinia Mangostana Rind Extract. Glob. j. Med. Plant Res., 2(1): 6-10, 2014 INTRODUCTION Garcinia mangostana is a member of family Guttiferae. It is popular for its succulent fruit in most of the Asia countries like Malaysia. The mangosteen tree can reach up to 25 m [18,21] and produce dark purple fruit [11,21]. The rinds of the fruit were widely used as a traditional medicine in Thailand for the treatment of trauma, diarrhea and skin infections [19,6]. Furthermore, this plant was reported used in treatment for various types diseases such as dysentery, eczema, trush [18], haemorrhoids, food allergies, arthritis, wounds, skin infections [23], tuberculosis [27], inflammation, ulcers, micosis, [9], mouth apthae, fever [4], abdominal pain, suppuration and leucorrhoea [16]. Although many researches have been done to reveal the biological properties of G. mangostana, however, no study were conducted to study the potential medicinal properties of Malaysian G. mangostana. Therefore, in the present study, antimicrobial, antioxidant, anticancer activities of G. mangostana rind were characterized as well as its chemical composition to reveal the medicinal potential of this plant. MATERIALS AND METHODS Plant material: The plant sample was purchased from herbal nursery located at Pasir Puteh, Kelantan, Malaysia. The fresh plant sample was oven dried at 37 °C for 4 days. Next, the plant sample was freeze dried prior to extraction using 70% methanol and concentrated at 1 g/ml. Finally, the plant extraction was kept in -20 °C until further use. Corresponding Author: Lee Seong Wei, Faculty of Agro Based Industry, Universiti Malaysia Kelantan Jeli Campus, 17600, Jeli, Kelantan, Malaysia. E-mail: leeseongwei@yahoo.com 7 Lee Seong Wei et al, 2014 Global journal of medicinal plant research, 2(1) January 2014, Pages: 6-10 Bacteria isolates: All bacterial isolates were provided by Universiti Malaysia Kelantan namely Aeromonas hydrophila, Escherichia coli, Edwardsiella tarda, Flavobacterium spp., Klebsiella pneumonia, Salmonella typhi, Vibrio alginolyticus, V. parahaemolyticus, V. cholerae and Pseudomonas aeruginosa. These bacteria were isolated from various aquatic animals and kept in tryptic soy agar (TSA) for further uses. Minimum inhibitory concentration (MIC) determination The MIC values of G. mangostana rind extract against bacterial isolates were determined through a twofold broth micro dilution method [12,13]. The bacterial isolates were cultured in tryptic soy broth for 24 h at room temperature and the concentration of these cultures were adjusted to 10 9 CFU mL-1 by using physiological saline. The concentration was cross check with a Biophotometer (Eppendorf, Germany). The bacterial suspensions were then inoculated into a microtiter plate that contained a serial dilution of G. mangostana rind extract and positive control. The microplate was then incubated at room temperature for 24 h. The MIC values were defined as the lowest concentration of the G. mangostana rind extract and positive control in the wells of the microtiter plate that showed no visible turbidity after 24 h incubation. Determination of antioxidant activity with α, α-diphenyl-β-picrylhydrazyl (DPPH) radical scavenging method DPPH radical scavenging method was conducted as described by Blois [2], Yen and Duh [31], BrandWilliam et al. [3] and Gadow et al. [8] with some modifications. The assay was carried in a 96 wells elisa plate with three replicates. 5 µl of the sample (0.5 mg/ml) solution was added into the well followed by 200 µl DPPH. The absorbance of the sample was recorded by using ELISA reader for ever interval 6 s. The percentage inhibition of DPPH radical was calculated based on the absorbance. Cancer cell lines: The human breast adenocarcinoma (MCF-7) cell line was derived from Institute of Marine Biotechnology, Universiti Malaysia Terengganu. All the cells were grown in standard cell medium (RPMI 1640) supplemented with 5 % fetal bovine serum in a 5 % CO2 atmosphere. The cells was then transferred into microplate at the concentration of 1 X 102 cells per well for cytotoxicity test of the plant extract. At 48 h, proliferation was measured by the MTT colorimetric assay. The IC 50 value was calculated from the following formula as described Adebayo et al. (2010): IC50 = 10log10(IC50) Where: I H I L C H C L : Low drug concentration : : : I% I% High above below drug 50% 50% concentration Colorimetric MTT (tetrazolium) assay Colorimetric MTT (3-(4, 5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (Sigma, USA) assay was carried out as described by Mosmann (1983). 10 µl of MTT solution (5 mg/ml) was added to all wells of 96 wells micro plate followed by 4 h incubation at 37 oC. Acid isopropanol was added to all wells for dissolving the dark blue crystals. The microplate plate was then read on an ELISA reader at wavelength 570 nm within 1 h after adding isopropanol. Bioactive compound characterization: The chromatographic procedure was carried out using a Shimadzu QP2010-GC-MS with autosampler. The sample was diluted 25 times with acetone and 1 μl of sample was injected into a column. A fused silica capillary column HP5-MS (30 m x 0.32 mm, film thickness 0.25 μm) was used. Helium was the carrier gas, and a split ratio of 1/100 was used. The oven temperature used was maintained at 60 oC for 8 min. The temperature was then gradually raised at a rate of 3 oC per min to 180 oC and maintained at 180 oC for 5 min. The temperature at the injection port was 250 oC. The components of the test solution were identified by comparing the spectra with those of known compounds stored in internal library. 8 Lee Seong Wei et al, 2014 Global journal of medicinal plant research, 2(1) January 2014, Pages: 6-10 RESULTS AND DISCUSSION The MIC values of the plant extract against the tested bacterial isolates ranged from 31.26 to 125 mg/l (Table 1). At the concentration of 31.26 mg/l of the plant extract, E. tarda, E. coli, Flavobacterium sp., P. aeruginosa and V. cholerae were failed to grow whereas the plant extract was able to control the growth of A. hydrophila, Klebsiella sp., Salmonella sp. and V. alginolyticus at the concentration of 62.5 mg/l. At the concentration of 125 mg/l of the plant extract was also can inhibit the growth of V. parahaemolyticus. The value of IC50 of G. mangostana rind extract against DPPH and MCF-7 cell was 2.67 ± 0.33 ppt and 1.98 ± 0.12 µg/ml, respectively. A total of 33 chemical compounds was successfully identified with Cyclopenta[d]antrhacene-6, 8, 11-trione, 1, 2, 3a, 4, 5, 6a, 7, 8, 11, 12-dodecahydro-3- (1-methylethyl) 12.63 % was the major compound in the plant extract (Table 2). This was followed by 1, 2-Benzenediol 7.42 %, Acetic acid 6.28 %, Propanoic acid 5.67 %, 2-Propanone, 1-hydroxy- 5.48 %, Sitosterol 5.32 %, Benzeneacetic acid 4.86 %, Vitamin E 4.63 %, Phenol 3.51 %, Octaethylene glycol 2.82 %, 2-Furanmethanol 2.31 %, 1-Hexanamine, 3, 5, 5-trimethyl- 2.29 %, 4, 5-Octanediol, 2, 7-dimethyl- 2.21 %, 2, 4-Dihydroxy-2, 5-dimethyl-3(2H)-furan-3-one 1.91 %, Eicosanoic acid 1.89 %, A-Norcholestan-3-one, 5-ethenyl-, (5 β)- 1.69 %, 6-Thioguanoside 1.54 %, Glycerin 1.30 %, 13Docosenamide 1.18 %, Pentaethylene glycol 1.14 %, Benzenecarbothioic acid, hydrazine 1.03 %, 1, 2Cyclopentanedione 1.00 %, Dimethyl sulfoxide 0.92 %, Octadecadienoic acid 0.90 %, Hexagol 0.86 %, Oxalic acid 0.83 %, 2, 4, 5-Trihydroxypyrimidine 0.54 %, Cyclooctanone, 2-bromo- 0.49 %, Nonanoic acid 0.46 %, Tocopherol 0.47 %, Copaene 0.23 %, Maltol 0.17 %, Phytol 0.15 % and 12 unidentified compounds 15.87 %. Many studies were reported the antimicrobial property of G. mangostana in the literature. For instance, Sundaram et al. [28] claimed that α-mangostin isolated from G. mangostana was found can inhibit Staphylococcus aureus, P. aeruginosa, S. thypimurium and Bacillus subtilis. Furthermore, the study of Phongpaichit et al. [22] also found that α-mangostin derived from G. mangostana can inhibit the growth of 49 species of methicillin – resistant Staphylococcus aureus (MRSA) and 13 species of Enterococcus spp. Other studies that revealed the antimicrobial property of G. mangostana were Chanarat et al. [5], Suksamrarn et al. [26], Chomnawang et al. [7], Sakagami et al. [24] and Voravuthikunchai and Kitpipit [29]. Although many studies worked on the antimicrobial activity of G. mangostana, however, all of the studies emphasized on clinical bacteria. Therefore, the present study verified the antimicrobial property of G. mangostana in which the plant extract was successfully inhibited all the tested bacterial isolates from various types of aquatic animals. Furthermore, the finding of the present study revealed that the plant extract contain chemical compounds such as Acetic acid, Propanoic acid, Benzeneacetic acid, Eicosanoic acid and Oxalic acid that responsible to the antimicrobial activity of the plant extract. Thus, we can make a conclusion that the plant extract possesses the huge potential as antimicrobial agent not only for human uses but also animal. In terms of antioxidant activity, the findings of the present study indicated that G. mangostana can be a good antioxidant agent. This was supported by several studies such as Yoshikawa et al. [32], Leong and Shui [14], Weecharangsan et al. [30] and Chomnawang et al. [7] in which they also found that G. mangostana possesses scavenging activity against DPPH. Furthermore, the chemical compounds such as Acetic acid, Propanoic acid, Sitosterol, phenol and phytol that found in the plant extract of the present study were responsible to the activity of antioxidant property of the plant extract. From the literature survey, we found that G. mangostana showed the positive response to gastric and lung cancer cells [10], human leukemia HL60 cells [15], human breast cancer SKBR3 cells [16] and human colon cancer DLD-1 cells [15,20]. However, no report on the activity of the plant extract against MCF-7 cells. Therefore, this study was the first report on the positive activity of G. mangostana extract against this breast cancer cells. Thus, the anticancer activity of the G. mangostana extracts is undoubtedly. Furthermore, the finding of chemical compound such as phytol that exists in the plant extract of the present study was responsible to the anticancer activity of the plant extract. In conclusion, G.mangostana rind possesses the high medicinal values. However, clinical test should be carried out to evaluate the effectiveness of this plant as medicinal drug before it is introduce into market. Table 1: Minimum inhibition concentration (MIC) of Garcinia mangostana rind extract against bacterial isolates Bacterial isolates MIC (mg/l) Aeromonas hydrophila 62.5 Edwardsiella tarda 31.26 Escherichia coli 31.26 Flavobacterium sp. 31.26 Klebsiella sp. 62.5 Pseudomonas aeruginosa 31.26 Salmonella sp. 62.5 Vibrio alginolyticus 62.5 Vibrio cholera 31.26 Vibrio parahaemolyticus 125 9 Lee Seong Wei et al, 2014 Global journal of medicinal plant research, 2(1) January 2014, Pages: 6-10 Table 2: Compound composition of Garcinia mangostana rind extract Compound Cyclopenta[d]antrhacene-6, 8, 11-trione, 1, 2, 3a, 4, 5, 6a, 7, 8, 11, 12-dodecahydro-3-(1methylethyl) 1, 2-Benzenediol Acetic acid Propanoic acid 2-Propanone, 1-hydroxySitosterol Benzeneacetic acid Vitamin E Phenol Octaethylene glycol 2-Furanmethanol 1-Hexanamine, 3, 5, 5-trimethyl4, 5-Octanediol, 2, 7-dimethyl2, 4-Dihydroxy-2, 5-dimethyl-3(2H)-furan-3-one Eicosanoic acid A-Norcholestan-3-one, 5-ethenyl-, (5 β)6-Thioguanoside Glycerin 13-Docosenamide Pentaehtylene glycol Benzenecarbothioic acid, hydrazine 1, 2-Cyclopentanedione Dimethyl sulfoxide Octadecadienoic acid Hexagol Oxalic acid 2, 4, 5-Trihydroxypyrimidine Cyclooctanone, 2-bromoNonanoic acid Tocopherol Copaene Maltol Phytol 12 unidentified compounds Total Compound Composition (%) 12.63 7.42 6.28 5.67 5.48 5.32 4.86 4.63 3.51 2.82 2.31 2.29 2.21 1.91 1.89 1.69 1.54 1.30 1.18 1.14 1.03 1.00 0.92 0.90 0.86 0.83 0.54 0.49 0.46 0.47 0.23 0.17 0.15 15.87 100.00 ACKNOWLEDGEMENT This project was funded by (R/FRGS/A07.00/00387A/005/2013/00107) Malaysia Fundamental Research Grant Scheme REFERENCES 1. 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