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Many diseases correlate with antioxidant deficiencies. Garcinia mangostana L rind (GMR) belong to waste product, contains xanthones which are antioxidant compounds. The aim of this study was to determine antioxidant properties of its ethanolic extract, hexane, ethylacetate, butanol, and water fractions in DPPH scavengingactivity, level of SOD and total antioxidant (TAS) compared against α-mangostin. Extract and all of these fractions had high DPPH trapping activity while α-mangostin had low activity. Level of SOD was highest in GMR water fraction while TAS level was highest in GMR ethylacetate fraction. It was concluded that GMR products had potential antioxidant properties
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Procedia Chemistry 13 ( 2014 ) 198 203
1876-6196 © 2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/3.0/).
Peer-review under responsibility of the School of Pharmacy, Bandung Institute of Technology
doi: 10.1016/j.proche.2014.12.027
ScienceDirect
International Seminar on Natural Product Medicines, ISNPM 2012
Antioxidant Properties of Garcinia mangostana L
(Mangosteen) Rind
Susy Tjahjania*, Wahyu Widowatia, Khie Khionga, Adrian Suhendraa, Rita
Tjokropranotoa
aFaculty of Medicine, Maranatha Christian University, Bandung-40164, Indonesia
Abstract
Many diseases correlate with antioxidant deficiencies. Garcinia mangostana L rind (GMR) belong to waste product, contains
xanthones which are antioxidant compounds. The aim of this study was to determine antioxidant properties of its ethanolic
extract, hexane, ethylacetate, butanol, and water fractions in DPPH scavengingactivity, level of SOD and total antioxidant (TAS)
compared against α-mangostin. Extract and all of these fractions had high DPPH trapping activity while α-mangostin had low
activity. Level of SOD was highest in GMR water fraction while TAS level was highest in GMR ethylacetate fraction. It was
concluded that GMR products had potential antioxidant properties
© 2014 The Authors. Published by Elsevier B.V.
Peer-review under responsibility of the School of Pharmacy, Bandung Institute of Technology.
Keywords: Garcinia mangostana L (mangosteen) rind, DPPH, SOD, TAS
* Corresponding author. Tel.: + 62-022-2012186, Fax.: +62-22-2005914
E-mail address: susy_tjahjani@yahoo.com
1. Introduction
Antioxidant deficiency correlates with many diseases. Lipid peroxidation is induced by oxidative stress and is a
key process in many pathological events. The destruction of membrane lipid caused by unsaturated lipid oxida tion
producing malondialdehyde as breakdown products is mutagenic and carcinogenic1. G.mangostana rind, as waste
product, contains a lot of water soluble antioxidant2. Various kinds of xanthones in G.mangostana rind had been
proven to have strong antioxidant activity included alpha mangostin3. According to Zarena and Udaya Sankar
(2009), antioxidant activity among these rind extracts was different each other deepending on the solvent:
G.mangostana rind extract especially acetone and ethyl acetate extract was effective to inhibit lipid peroxidation
induced by ferrous sulphate-ascorbate system in the linolenic acid medium as lipid phase model system4. Besides
© 2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/3.0/).
Peer-review under responsibility of the School of Pharmacy, Bandung Institute of Technology
Available online at www.sciencedirect.com
Susy Tjahjani et al. / Procedia Chemistry 13 ( 2014 ) 198 – 203 199
containing antioxidant activities, it has antitumoral, anti-inflamatory, antiallergy, antibacterial, antiviral , and
antifungal activities5,6. According to Palakawong et al. in their study, antioxidant activity of G.mangostana rind
extracted with 50 % ethanol, which IC50 was 5.94 μg/ml, was the strongest compared to its bark and leaves as well7.
The aim of this study was to evaluate the antioxidant properties of G. mangostana L (GMR) rind 96 % and 70 %
ethanolic extract and hexane, ethyl acetate, butanol, and water fraction by measuring DPPH scavenging activity,
SOD activity, and total antioxidant status of these samples
2. Experiments
2.1. Material
Ripe G.mangostana L which had purple colour collected from Subang (West Java), destilated water, 96 % and 70
% ethanol, hexane, ethyl acetate, butanol, methanol solution, DPPH solution (Sigma Aldrich), Cayman’s Superoxide
Dismutase Assay Kit, and Superoxide Dismutase Assay Kit
2.2. Extraction and`fractionation
Mangosteen fruit rind was washed as clean as possible, dried at 37 C, pounded, and macerated using 96% and
70% ethanol, until the filtrate becoming no colour anymore. The filtrate was evaporated until becoming pasta like
extract. Parts of the 96% ethanolic extracts was macerated again using hexane-water mixture in 1:1 ratio until this
filtrate becoming colourless again and then this filtrate was evaporated also becoming hexane fraction. The last
residu was treatedsuch as before consecutively using ethyl acetate-water mixture 1:1, butanol-water mixture 1:1, and
water only to get ethyl acetate fraction, butanol fraction, and water fraction.
2.3. DPPH scavenging activity assay
The DPPH assay was carried out as described by Unlu et al.8. To obtain the IC50 value, 50 μL of each extract and
fractions of GMR at a range of various final concentrations 100; 50; 25; 12.5; 6.25; 3.125; 1.563; 0.781; 0.391 and
0.195 μg/mL in methanol solution were pippeted and put into the microplate and then added with 200 μL of 0.077
mmol/L DPPH (Sigma Aldrich) in methanol.The reaction mixtures were shaken vigorously and kept in the dark for
30 minutes at room temperature, furthermore DPPH scavenging activity was determined by microplate reader at 517
nm.
The radical scavenging activity of each sample was determined by the ratio of DPPH absorption decrease against
the absorption of DPPH solution in the absence of test sample (negative control) using the following equation.
AS: absorbance of samples, AC: negative control absorbance (without sample)
2.4. Superoxide Dismutase (SOD) activity assay
The SOD assay was carried out using Cayman’s Superoxide Dismutase Assay Kit. 10 μL of three series of
concentrations including 500; 125; 31.25 μg/mL of GMRextract and fractions were pippeted into sample wells, then
200 μL diluted radical detector was added into each wells. For SOD standard wells, 200 μL diluted radical detector
and 10 μL standard was pippeted into the wells (tubes A-G). To intitiate the reactions, 20 μL diluted xanthine
oxidase was added into each wells. This well plate was shaked for few seconds for mixing and incubated on shaker
for 20 minutes at room temperature. Absorbance of each well was read at 450 nm curve and linearized rate (LR).
The SOD level in U/ mL was determinate by the equation below.
100(%) x
AC
AsAc
Scaveninge
200 Susy Tjahjani et al. / Procedia Chemistry 13 ( 2014 ) 198 – 203
dilutionsamplex
Slope
mLxerceptysampleLR
SOD..
23.0.).int.(
2.5. Total Antioxidant Status (TAS) assay
The TAS assay was carried out using Cayman’s Antioxidant Assay Kit. Into the each of the sample wells, 10 μL
of three series of concentrations including 500; 125; 31.25 μg/mL of GMRextract and fractions, 10 μL
metmyoglobin, and 150 μL chromogen were added. For Trolox standard wells, 10 μL trolox standard (tubes A-G),
metmyoglobin, and 150 μL chromogen were put. To intitiate the reactions, 40 μL hydrogen peroxide was added into
all wells. The plate was incubated on a shaker for 5 minutes at room temperature. Absorbance was read at 750 nm
using Elisa Reader. The TAS was calculated using the linear regression equation of the standard curve written in the
following equation3.
dilutionx
slope
erceptyabsorbance
mMTAS.
int
)(
(3)
3. Results and Discussion
To assure the quality of this crude drugs, proximate analysis was done as shown in Table 1 below.
Table 1. Proximate Analysis of G.mangostanaL Rind Crude Drug
Because of water content of this simplicia is only 10,31 %, this material is dry enough to protect from fungus
contamination. DPPH scavenging activity of GMR extract and its various fractions and alpha mangostin is shown in
the Figure1.
Figure 1. DPPH Scavenging Activity of GMR Extract, Fractions, and Alpha Mangostin at 500 μg/ mL * means significant differenc e between
these groups
No
Proximate Analysis
%
1.
Water content
10.31
2.
Ash
20.54
3.
Protein
3.43
4.
Rough fiber
25.53
5.
lipid
0.54
6.
Carbohydrat e
49.96
Susy Tjahjani et al. / Procedia Chemistry 13 ( 2014 ) 198 – 203 201
According to DPPH free radical scavenging activity, the IC50 of these materials is shown in the Table 2 below.
Table 2. IC50 of DPPH Scavenging Activity of GMR and Alpha Mangostin
Extract/ Fraction
IC50 (μg/mL)
96 % alcohol extract
7,48 ± 0,19
70 % alcohol extract
6.563 ± 0.311
Hexane fraction
3,62 ± 0,04
Ethyl acetate fraction
13,29 ± 0,12
Butanol fraction
12,23 ± 0,13
Water fraction
10,31 ± 0,04
Alpha mangostin
66,63 ± 34,65
IC50 of DPPH scavenging activity of GMR96% and 70 % ethanolic extracts were (7,48 ± 0,19)% and (6.563 ±
0.311)%. These IC50 is higher, that’s meant less DPPH scavenging activity, than reported by Palakawong et al.7
which was 5.94 μg/mL although they used 50% ethanol for the extraction. The different region of G.mangostana L
collection and degree of the fruit ripeness might play a role to cause it.
Similar results were concluded about different antioxidant activity in using various extraction solution for sample
extraction by Zarena and Udaya Sankar4. This difference could be caused by different extraction capacity of each
solution to extract GMR polar active compund4. Phenolic compound content of GMR extract such as tannin, alpha
mangostin, epicatechin was different if solvent extraction was different. Therefore DPPH and hydroxyl radical
scavenging and also lipid peroxidation inhibition activity was different in several extraction solution.Water extract
had stronger capacity than methanol extract whilemethanol extract had stronger capacitythan hexane extract9. Our
study had the similar results with that ones.
Total antioxidant status/ total antioxidant capacity (mM Trolox) of these GMR extracts, fractions, and alpha
mangostin is shown in Figure 2.
Fig. 2. Total Antioxidant Status (Total Antioxidant Capacity) of GMR (G.mangostana L rind 96 % ethanolic extract), Fractions, Alpha
Mangostin (p < 0,05). GMRE =, *= means significant difference between these groups.
At the same concentration, 500 μg/ mL, TAS of ethyl acetate fraction was the highest among other materials,
followed by alpha mangostin. GMR extract showed same TAS as hexane fraction and water fraction as well, while
butanol fraction showed the lowest (p < 0,05).
SOD activity of GMR 96% ethanolic extract, hexane, ethyl acetate, butanol, and water fraction is shown in the
following figure.
202 Susy Tjahjani et al. / Procedia Chemistry 13 ( 2014 ) 198 – 203
Fig. 3. SOD Activity of GMR 96% Ethanolic Extract, Hexane, Ethyl Acetate, Butanol, and Water Fraction * means significant difference
between each group. (p< 0,05)
At concentration 500 μg/ mL,comparing to other materials, GMR extract and also water fraction had the strongest
DPPH scavenging activity, in contrast against alpha mangostin which had the lowest DPPH scavenging activity. But
alpha mangostin had higher TAS than GMR extract (p< 0,05), it may be caused by other antioxidant mechanism of
alpha mangostin besides free radical scavenging such as other enzymatic mechanism besides SOD. GMR extract and
water fraction had same TAS and DPPH scavenging activity although water fraction, also butanol fraction, had the
most SOD activity (p< 0,05). Other antioxidants beside SOD activity could play more role in GMRextract. Although
butanol fraction had the strongest SOD activity, lack of other antioxidant properties might cause its lowest TAS.
Ethyl acetate fraction had the highest TAS even though in lower concentration i.e. 125 μg/mL comparing to other
samples (p< 0,05).
4. Conclusion
In clonclusion,Garcinia mangostana L rind products had potential antioxidant properties.
Acknowledgement
Authors would like to thanks Directorat General of Higher Education of Indonesian Education Ministry for the
research grant of this research program
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The antioxidant and antimicrobial activities of the extracts from peel, leaves, and bark of mangosteen (Garcinia mangostana L.), and some essential oils such as cinnamon and citrus were investigated. The antioxidant activities (IC50) of peel, leaves, and bark extracted, which were evaluated by DPPH method, were 5.94, 9.44, and 6.46 μg/ml, respectively. Either cinnamon or citrus essential oil showed no antioxidant activities with DPPH. A broth dilution method was employed to evaluate the antimicrobial activity against some Gram-positive bacteria (L. monocytogenes and S. aureus) and Gram-negative bacteria (E. coli and Salmonella sp.). The minimum inhibitory concentration (MIC) values of peel, leaves, and bark extracted against Gram-positive bacteria were ranged from 0.025-0.78 mg/ml. While the minimum bactericidal concentration (MBC) values were between 0.05-0.39 mg/ml. MIC and MBC values of cinnamon against S. aureus, E. coli and Salmonella sp. were 3.13 and 6.25 mg/ml, respectively. Citrus oil showed effect on only S. aureus with MIC and MBC values of 6.25 and 12.50 mg/ml, respectively.
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The antioxidative activity of natural plant productslacinilene A, naringin, galangin, and rutinwas examined using lipid peroxidation systems consisting of either ethyl linoleate, ethyl linolenate, or ethyl arachidonate plus Fenton's reagent. Inhibitory activity of plant products toward malonaldehyde (MA) formation from lipids was measured using gas chromatography. Lacinilene A, which showed the strongest antioxidative acitivity among the chemicals tested, inhibited MA formation from ethyl linolenate and ethyl arachidonate by 100% at the levels of 3.0 and 0.5 μmol, respectively. Natural flavonoid compounds naringin, galangin, and rutin exhibited appreciable antioxidative activities at doses lower than 0.125 μmol. Rutin, which exhibited the strongest activity among the three flavonoids, inhibited MA formation from ethyl arachidonate by 70% at the level of 0.125 μmol. These flavonoids exhibited only slight inhibition of MA formation at levels higher than 0.5 μmol from the ethyl esters of the three fatty acids. Keywords: Antioxidants; lipid peroxidation; malonaldehyde; plant components
Article
Antioxidative, skin protective activities, and cytotoxicity of three extracts (water, methanol, and hexane) from the fruit hull of mangosteen (Garcinia mangostana Linn. (Guttiferae)) and their phenolic constituents such as alpha-mangostin, epicatechin, and tannin, were evaluated. The amounts of alpha-mangostin, total flavonoid, and total tannin were different among the three extracts, except those of total tannin in methanol and hexane extracts. For the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free-radical scavenging, hydroxyl radical-scavenging, and inhibition of lipid peroxidation experiment, the water extract showed higher activity than the methanol extract and hexane extract. alpha-Mangostin, epicatechin, and tannin also revealed these antioxidant and free radical-scavenging activities. When added simultaneously with H(2)O(2) (200 microM) to keratinocyte cells, the water extract (50 microg/mL), epicatechin (200 microM), and tannin (200 microM) effectively protected cells from oxidative damage, but the methanol extract, hexane extract, and alpha-mangostin did not. The methanol extract and hexane extract exhibited moderate cytotoxicity, whereas alpha-mangostin showed strong cytotoxicity. The present study provides the evidence that Garcinia mangostana extracts, especially the G. mangostana water extract, act as antioxidants and cytoprotective agents against oxidative damage, which is at least partly due to its phenolic compounds in mangosteen.
Article
The essential oil, obtained by using a Clevenger distillation apparatus, and water-soluble (polar) and water-insoluble (nonpolar) subfractions of the methanol extract of Thymus pectinatus Fisch. et Mey. var. pectinatus were assayed for their antimicrobial and antioxidant properties. No (or slight) antimicrobial activity was observed when the subfractions were tested, whereas the essential oil showed strong antimicrobial activity against all microorganisms tested. Antioxidant activities of the polar subfraction and the essential oil were evaluated using 2,2-diphenyl-1-picrylhydrazyl, hydroxyl radical, superoxide radical scavenging, and lipid peroxidation assays. The essential oil, in particular, and the polar subfraction of the methanol extract showed antioxidant activity. The essential oil was analyzed by GC/MS, and 24 compounds, representing 99.6% of the essential oil, were identified: thymol, gamma-terpinene, p-cymene, carvacrol, and borneol were the main components. An antimicrobial activity test carried out with fractions of the essential oil showed that the activity was mainly observed in those fractions containing thymol, in particular, and carvacrol. The activity was, therefore, attributed to the presence of these compounds. Other constituents of the essential oil, such as borneol, gamma-terpinene, and p-cymene, could be also taken into account for their possible synergistic or antagonistic effects. On the other hand, thymol and carvacrol were individually found to possess weaker antioxidant activity than the crude oil itself, indicating that other constituents of the essential oil may contribute to the antioxidant activity observed. In conclusion, the results presented here show that T. pectinatus essential oil could be considered as a natural antimicrobial and antioxidant source.
Article
As part of ongoing research on cancer chemopreventive agents from botanical dietary supplements, Garcinia mangostana L. (commonly known as mangosteen) was selected for detailed study. Repeated chromatography of a CH2Cl2-soluble extract of the pericarp led to the isolation of two new highly oxygenated prenylated xanthones, 8-hydroxycudraxanthone G (1) and mangostingone [7-methoxy-2-(3-methyl-2-butenyl)-8-(3-methyl-2-oxo-3-butenyl)-1,3,6-trihydroxyxanthone, 2], together with 12 known xanthones, cudraxanthone G (3), 8-deoxygartanin (4), garcimangosone B (5), garcinone D (6), garcinone E (7), gartanin (8), 1-isomangostin (9), alpha-mangostin (10), gamma-mangostin (11), mangostinone (12), smeathxanthone A (13), and tovophyllin A (14). The structures of compounds 1 and 2 were elucidated by spectroscopic data analysis. Except for compound 2, which was isolated as a minor component, the antioxidant activities of all isolates were determined using authentic and morpholinosydnonimine-derived peroxynitrite methods, and compounds 1, 8, 10, 11, and 13 were the most active. Alpha-mangostin (10) inhibited 7,12-dimethylbenz[alpha]anthracene-induced preneoplastic lesions in a mouse mammary organ culture assay with an IC50 of 1.0 microg/mL (2.44 microM).
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The present study was aimed to investigate the activity of Thai medicinal plants on inflammation caused by Propionibacterium acnes in terms of free radical scavenging and cytokine reducing properties. P. acnes have been recognized as pus-forming bacteria triggering an inflammation in acne. Antioxidant activity was determined by DPPH scavenging and NBT reduction assay. The result showed that Garcinia mangostana possessed the most significant antioxidant activity and reduced reactive oxygen species production. Houttuynia cordata, Eupatorium odoratum, and Senna alata had a moderate antioxidant effect. In addition, Garcinia mangostana extracts could reduce the TNF-alpha production as determined by ELISA. Garcinia mangostana was highly effective in scavenging free radicals and was able to suppress the production of pro-inflammatory cytokines. This study has identified the promising source of anti-inflammatory agent which could be useful in treatment of acne vulgaris.
Effect of Garcinia mangostana on inflammation caused by Propionibacterium acnes
  • M T Chomnawang
  • S Surassmo
  • V S Nukoolkarn
  • W Gritsapanan
Chomnawang MT, Surassmo S, Nukoolkarn VS, Gritsapanan W. Effect of Garcinia mangostana on inflammation caused by Propionibacterium acnes. Fitoter 2007; 78: 401-8.