International Food Research Journal 17: 583-589 (2010)

Antioxidant and antimicrobial activities of crude extracts from

mangosteen (Garcinia mangostana L.) parts and some essential oils

1,
*Palakawong, C., 1Sophanodora, P., 2Pisuchpen, S. and 3Phongpaichit, S.
1
Department of Food Technology, Faculty of Agro-Industry, Prince of Songkla University,
Hat Yai, Songkhla 90112, Thailand.
2
Department of Material Product Technology, Faculty of Agro-Industry, Prince of
Songkla University, Hat Yai, Songkhla 90112, Thailand.
3
Department of Microbiology, Faculty of Science, Prince of Songkla University, Hat Yai,
Songkhla 90112, Thailand

Abstract: 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.

Keywords: Mangosteen, Garcinia mangostana, antioxidant, antimicrobial, MIC, DPPH

Introduction reduce these health hazards and to extend the shelf-

life of processed food.
Plants extracted have been added to many Mangosteen (Garcinia mangostana L.) is
kinds of food to improve their flavor and organoleptic one of the most famous fruits in Thailand. Previous
properties for many years. Especially, the extracts studies have shown that the extracts from various
from herbs and spices are known as antimicrobial parts contain varieties of secondary metabolites such
and antioxidation potential. The extracted plants as prenylated and oxygenated xanthones. Xanthones
were classified as natural compounds, which were or xanthen-9H-ones is a secondary metabolite found
the secondary metabolites. These metabolites have in some higher plant that involves mangosteen (Peres
demonstrated biological activities and received et al., 2000). Xanthones could be isolated from peel,
particular attention as potential natural agents whole fruit, bark, and leaves of mangosteen. Several
for food preservation. According to the current studies have shown that obtained xanthones from
consumers, more natural and fresh-like foods with mangosteen have remarkable biological activities
fewer synthetic additives but increase safety and such as antioxidant, antitumoral, anti-inflammatory,
shelf-life are needed (Negi et al., 2008). Resulting antiallergy, antibacterial, antifungal, and antiviral
from those demands, plants have emerged as popular activities (Suksamrarn et al., 2006; Pedraza-Chaverri
ingredients and have a tendency of replacing synthetic et al., 2008).
antimicrobial and antioxidant agents (Mayachiew Other secondary metabolites extracted from plants
and Devahastin, 2008). The use of natural products that play an important role in biological activities are
as antimicrobial and antioxidant compounds seem to essential oils. Essential oils from aromatic plants,
be an interesting way to support this tend in order to spices, and herbs have been used historically in

*Corresponding author.
Email: [email protected] © All Rights Reserved
Tel: +66-74-286330 ; Fax: +66-74-212889
584 Palakawong, C., Sophanodora, P., Pisuchpen, S. and Phongpaichit, S.

the pharmaceutical, food, and perfume industries Plant material and extraction method
because of their antibacterial, culinary and fragrant The whole peel (outer and inner peels), leaves,
properties (Salehi et al., 2005). In addition, essential and bark of G. mangostana were harvested from
oils have been used for preventing food spoilage and Songkhla province in April, 2009. The samples
deterioration, and also for extending shelf-life of food were first cleaned to remove any residual compost
since ancient time. Cinnamon and citrus essential and washed thoroughly to remove impurities. After
oils have been shown the possession of antimicrobial washing, the samples were chopped into small pieces
activities and could serve as a source of antimicrobial (0.5 × 1.0 cm2) and dried overnight in a tray dryer at
agents against food pathogen. Several references 45 °C. Then chopped samples were ground with a
on the antimicrobial efficiency are available in the grinder to make powder (around 18 meshes).
literature reviews such as cinnamon (Lopez et al., All ground samples were placed in 70 °C distilled
2005; Tzortzakis, 2009) and citrus (O’Bryan et al., water for 15 min at the ratio of sample powder:water
2008; Fisher and Phillips, 2006) of 1:4. The mixtures were boiled 4 times or until no
Because of consumer awareness, food content of tannin was found by dropping with 2%
processors have desired to reduce the use of gelatin solution in the mixtures (Weecharangsan et al.,
synthetic chemicals in food products. Then common 2006). The mixtures were filtrated, the residues were
culinary extracted plant such as ethanolic extract then dried at 40-45 °C in the hot air oven. The dried
from mangosteen and essential oils extracted from powder was macerated at room temperature for 7 days
cinnamon or citrus could be the sources of natural with 50% ethanol. In order to know the exact weight,
alternatives. the crude extracts were filtered and evaporated to
Although the extracts from mangosteen and obtain the dried crude extracts. The obtained extracts
essential oils from cinnamon and citrus contain potent were stored in a desiccator containing dry silica gel
antimicrobials and antioxidation activities, such prior using in each experiment.
extracts have not been sufficiently tested for their
activities. Even if extracted plants are considered to Microbial cultures
be safe (GRAS), however their uses are often limited Escherichia coli DMST 15537, Salmonella
by organoleptic criteria. For this reason, it will be sp. DMST 4464, Listeria monocytogenes DMST
necessary to determine the minimum concentration 17303, and Staphylococcus aureus DMST 6512
to inhibit the growth of pathogenic bacteria without were obtained from the Department of Medical
affecting the sensory quality of the food. The Sciences, Ministry of Public Health, Thailand. The
objectives of the present study were to investigate microorganisms were maintained in TSA at 5 °C.
the minimum inhibitory concentration (MIC), the Stock culture of microbial was grown in MHB (E.
minimum bactericidal concentration (MBC) against coli, S. aureus and Salmonella spp.) and TSB (L.
four food-borne pathogens (Gram-positive and Gram- monocytogenes) at 37+2 °C in a shaker incubator for
negative) and the antioxidant activity of mangosteen 3.5 h at 110 rpm (cells in early stationary phase). The
extracted from peel, leaves, and bark, and some bacterial suspension was subsequently adjusted to
essential oils such as cinnamon and citrus. 106 CFU/ml using MHB and TSB.

Materials and Methods Antioxidant activity

The scavenging of DPPH free radicals was
Chemicals used for measuring the antioxidant activity of the
Ascorbic acid was purchased from Riedel- extracts according to the method of Weecharangsan
de-Hae (Seelze, Germany). DPPH, resazurin salt et al. (2006) with slightly modification. Briefly,
and ethanol were purchased from Sigma-Aldrich stock solutions of the crude extracts were prepared
(St. Louis, MO, USA), while cinnamon and citrus by dissolving 0.1 g of dry extracts in 50 ml 50%
essential oils (0.05%) were purchased from Flavor- ethanolic solution. The stock solution was diluted with
Focus (Bangkok, Thailand). Gelatine powder was 50% ethanolic solution to obtain sample solutions at
purchased from Ajax Finechem (Auckland, New the concentrations of 1, 10, 50, and 100 µg/ml. The
Zealand). Cyprofloxacin and vancomycin were sample solutions were thoroughly mixed with freshly
obtained from BJC Healthcare (Bangkok, Thailand). prepared 0.05% DPPH ethanolic solutions at the
For antimicrobial tests, tryptic soy broth (TSB) and ratio of 1:1, and kept for 30 min in the dark at room
Mueller-Hinton broth (MHB) were purchased from temperature. The amount of the reaction mixture was
HiMedia Laboratories (Mumbai, India). determined by UV-VIS spectrophotometer at 517
nm. Neutralisation of DPPH radical was calculated

International Food Research Journal 17: 582-589

Antioxidant and antimicrobial activities of crude extracts from mangosteen (Garcinia mangostana L.) parts and some essential oils 585

using the equation: DPPH inhibition (%) = 100 × (A0 experiment of Weecharangsan et al. (2006). In those
–AS)/A0, where A0 is the absorbance of the control experiments, researchers used the same extraction
(containing all reagents except the test compound), method that was used in this study. However, the
and AS is the absorbance of the tested sample. same methods were used, the different results were
The antioxidant activity of the crude extract was reported. It is possible that because the different in
expressed as IC50, defined as the concentration of the the maturity stage of raw material was used. This
crude extract required to inhibit DPPH radicals by study used raw material in stage 3 of maturity but
50%, using the linear regression analysis. Ascorbic stage 5 or 6 by the experiment of Weecharangsan
acid was used as a standard antioxidant. et al. (2006). From Table 1, this can be concluded
that the extract from peel of mangosteen showed the
Antimicrobial activity best antioxidant activity among these extracts when
Minimum inhibitory concentration (MIC) and compared to L-ascorbic acid.
minimum bactericidal concentration (MBC) were Okonogi et al. (2007) reported that the antioxidant
performed by a broth dilution technique, using 96- activity (IC50) of the extract from peel of mangosteen
well microtitre plates according to Kuete et al. (2008) was 0.023 µg/ml, which was less than the IC50 found
and Saker et al. (2007). The bacterial inoculate in this experiment (5.94 µg/ml), for the reason of
applied contained approximately 1.0×105 cells in a different extraction methods of this study. This
final volume of 100 µl/well. implies that the different extraction methods could
The crude extracts were dissolved in 10% effect on the activity, agreed to the study of Liu et
dimethylsulfoxide (DMSO) in sterile MHB to obtain al. (2008).
a stock concentration of 12.50 mg/ml. Serial two-fold Among these extracts from mangosteen parts,
dilutions of each sample needed to be evaluated were each part (peel, leaves, and bark) showed antioxidant
made with MHB to yield volumes of 100 μl/well activity with different IC50 values. Extracted peel
with final concentrations ranging from 0.05-12.50 exhibited lowest IC50, followed by bark and leaves
mg/ml. MHB was used as a negative control while with IC50 5.94, 6.46 and 9.44 µg/ml, respectively.
ciprofloxacin and vancomycin were used as positive Zadernowski et al. (2009) reported that rind (inner
controls (0.5-2.0 µg/ml). The cultured microplates peel) of mangosteen had composed more phenolic
were incubated for 24 h at 37 °C. content than peel (outer peel) and aril parts.
The MIC of samples was detected following the Tachakittirungrod et al. (2007) reported that the active
addition of 10 μl resazurin (0.75%). Viable bacteria compounds in higher plant were located in different
reduced the blue dye to a pink color. Microbial parts with different contents. This is the answer why
growth was determined by observing the change of extracts of peel, leaves and bark showed the activity
color in the wells (blue when there is no growth and with different IC50. In addition, Maisuthisakul et al.
pink when there is growth). MIC was defined as the (2008), Mayachiew and Devahastin (2008), and Liu
lowest sample concentration that had prevented this et al. (2008) reported that the antioxidant activity
change. The minimum bactericidal concentration was correlated with phenolic content. Moreover,
(MBC) was determined by subcultivation of 50 µl of Mayachiew and Devahastin (2008) reported that
each blue well in plates containing Mueller-Hinton crude extract with different chemical compositions
agar and then further incubation for 24 h at 37 °C. can effect on the activity. These literatures may be
The lowest concentration with no visible growth was concluded that extracted peel composes more phenolic
defined as the MBC. content than those of bark and leaves, respectively.
Table 1 showed not only cinnamon but also citrus
Results and Discussion essential oil that exhibited no activity with DPPH. This
result agrees with Poloteo et al. (2006) who reported
Antioxidant activity that cinnamon essential oil had had low activity with
Extracted peel of G. mangostana expressed the DPPH. Eyob et al. (2008) reported that the activity
strongest activity (IC50 = 5.94 µg/ml) while bark and of essential oil from korarima was very low, for the
leaves extracted showed moderate activities (IC50 = reason that the OH group in aromatic ring of phenolic
6.46 and 9.44 µg/ml, respectively). Cinnamon oil compound was replaced by some functional group as
showed no activity and the citrus oil did not reach 50% a result of their hydrogen donating ability. This may
of DPPH-neutralisation at the highest concentration be used to support for the result of this study that why
applied (Table 1). cinnamon and citrus essential oil used in this study
In this experiment, the study found that the had low activity.
extracted peel showed IC50 value less than the

International Food Research Journal 17: 583-589

586 Palakawong, C., Sophanodora, P., Pisuchpen, S. and Phongpaichit, S.

Table 1. Antioxidant activity (IC50, µg/ml) of G. mangostana extracted from

peel, leaves, and bark and some essential oils (cinnamon and citrus).

IC50

Peel 5.94+0.14
Leaves 9.44+0.39
Bark 6.46+0.36
Cinnamon no activity
Citrus no activity
L-Ascorbic acid 4.30+0.14

Data are expressed as means ± standard deviation of three trials.

Antimicrobial activity sensitivity of the Gram-positive bacteria was higher

The MIC and MBC of the tested extracts than Gram-negative bacteria could be attributed to
ranged from 0.05 to 6.25 mg/ml against the 4 their differences in cell membrane constituents and
tested microorganisms (Table 2). This experiment arrangement (Negi et al., 2008). The Gram-positive
confirmed the strong antibacterial activity of extracted bacteria contains an outer peptidoglycan layer, which
mangosteen on Gram-positive bacteria but no activity is an ineffective permeability barrier (Scherrer and
on Gram-negative bacteria. The lowest MIC value Gerhardt, 1971). The resistance of Gram-negative
(0.025-0.05 mg/ml) was observed with extracts of bacteria towards antibacterial substances may be
peel and bark on L. monocytogenes and S. aureus. due to outer phospholipidic membrane carrying the
Cinnamon oil was moderately active against E. coli, structural lipopolysaccharide components, which
Salmonella sp. and S. aureus (MIC 3.13 mg/ml, MBC makes it impermeable to lipophilic solutes and porins
6.25 mg/ml) while citrus oil was active only against constitute of a selective barrier to the hydrophilic
S. aureus (MIC 6.25 mg/ml, MBC 12.5 mg/ml). solutes (Nikaido and Vaara, 1985).
Xanthone, an active compound found in all parts Cinnamon and citrus essential oil showed that the
of mangosteen. More than 20 xanthones were found effect against 4 microorganisms were different from
in mangosteen. Both of α- and β-mangostin were those extracts of mangosteen. Generally, the study
mainly found in this fruit (Furukawa et al., 1996; found that the essential oils had higher MIC or MBC
Chen et al., 2008), especially in peel of mangosteen. than extracted mangosteen. Citrus can inhibit against
Xanthone can inhibit several microorganisms. IInuma only Gram-positive, especially S. aureus. Not only S.
et al. (1996) and Sakagami et al. (2005) reported aureus but also all of Gram-negative were inhibited
that the MIC value of α-mangostin against S. aureus by cinnamon.
was lower than β-mangostin (6.25 and >100 µg/ml, From literature, main components in citrus oil
respectively). From these literatures and results of were limonene, linalool, and citral. Among these
this study, these can conclude that the efficiency of components, limonene showed the most abundant
microbial inhibition might be resulted directly from content, followed by linalool and citral, respectively.
α-mangostin. Because peel and bark were composed Fisher and Phillips (2006) reported that limonene
more of α-mangostin than leaves, so this study found had showed lowest effect against microorganisms.
that the extracts from peel and bark can inhibit both Inhibition effect against microorganisms was
of L. monocytogenes and S. aureus with lower MIC resulted from linalool rather than citral or limonene.
than extracted leaves. In addition, the study also Gram-negative was tolerant with citrus because of
found that only Gram-positive was susceptibility to the lipopolysaccharide present in outer membrane
extracted mangosteen. which provided protection against different agents
Canillac and Mourey (2001) reported that if the (Mahboubi and Haghi, 2008; Oussalah et al. 2007).
MBC/MIC ratio was found to be less than or equal to Cinnamaldehyde and eugenol were found to be a
4, the bacteria was considered to be susceptible; on main chemical composition in cinnamon essential oil.
the other hand, if this ratio was greater than 4, it was Oussalah et al. (2007) reported that cinnamaldehyde
considered to be tolerant. From these results, MBC/ was found mainly in bark or branch of cinnamon
MIC ratios of all extracts on Gram-positive bacteria tree while eugenol was found in leaves. Besides this,
were less than 4, so all of them were considered to eugenol was lower activity against microorganism
be sensitive to these extracts. The reason that the than cinnamaldehyde. Hussain et al. (2008) reported

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Antioxidant and antimicrobial activities of crude extracts from mangosteen (Garcinia mangostana L.) parts and some essential oils 587

Table 2. Minimum inhibitory and minimum bactericidal concentrations (MIC and MBC) of
G. mangostana extracted from peel, leaves, and bark and some essential oils (cinnamon and
citrus).

MIC/ Mangosteen parts Cinna- Cipro- Vanco-

Bacteria Citrus
MBC (mg/ml) mon floxacin mycin
(mg/
Peel Leaves Bark (mg/ml) (µg/ml) (µg/ml)
ml)

L. monocytogenes MIC 0.05 0.78 0.025 >12.5 >12.5 0.78 0.78

MBC 0.10 1.56 0.05 - - 0.78 0.78

S. aureus MIC 0.025 0.20 0.05 3.13 6.25 <.05 0.78

MBC 0.05 0.39 0.10 6.25 12.50 - 0.78

E. coli MIC >3.13 >3.13 >3.13 3.13 >12.5 <.05 ND

MBC - - - 6.25 - - ND

Salmonella sp. MIC >3.13 >3.13 >3.13 3.13 >12.5 <.05 ND

MBC - - - 6.25 - - ND

ND = Not Detected

that the different chemical composition in an essential 2. Therefore, S. aureus was considered to be sensitive
oil could result in different biological activity. Lopez to both of essential oils while E. coli and Salmonella
et al. (2005) and Inouye et al. (2001) found that sp. were considered to be sensitive to cinnamon
cinnamon with eugenol had showed higher activity essential oil.
against Gram-negative than Gram-positive. Cinnamon
with cinnamaldehyde showed higher activity against Conclusions
Gram-positive than Gram-negative. From the result
of this study, cinnamon can inhibit both of Gram- These sample data received show the inhibition
positive and Gram-negative. This result agreed with potency on extracts of peel, cinnamon, and the citrus
Oussalah et al. (2007) who reported that both of them oil that can be considered as preservative agents for
were inhibited by this essential oil. It is possible that both antibacterial and antioxidant activities. This
cinnamon in the present study composes of both of study provides the important baseline information
eugenol and cinnamaldehyde. for the use of extracted peel from G. mangostana as
These differences in the susceptibility of the test well as the essential oils as the preservative agents for
organisms to essential oil could bring to the conclusion some types of food. Then in vivo experiment should
of the variation in the rate of essential oil constituent’s be conducted for further study. The antibacterial and
penetration through the cell wall and cell membrane antioxidant studies of these peel and essential oils
structures. The ability of essential oil to disrupt the are being considered as the future objective for the
permeability barrier of cell membrane structures and planning of using these potential preservation agents
the accompanying loss of chemiosmotic control are in fresh-cut mangosteen.
the most likely reasons for its lethal action (Cox et
al., 2000). Nedorostova et al. (2009) reported that Acknowledgements
the leakage of intracellular metabolites due to their
activity on cell membranes seemed to be the main This research was supported by the grants
mechanism or mode of action. On the other hand, under the program Strategic Scholarship for Frontier
essential oils can interact with intracellular sites and Research Network for the Join Ph.D. Program Thai
cause death of the cell e.g., by alteration of protein Doctoral Degree from the Commission on Higher
structures after penetration into the cells. Education and the Graduate School, Prince of Songkla
The MBC/MIC ratio of citrus on S. aureus and the University, Thailand.
MBC/MIC ratio of cinnamon on E. coli, Salmonella
sp. and S. aureus were less than 4 according to Table

International Food Research Journal 17: 583-589

588 Palakawong, C., Sophanodora, P., Pisuchpen, S. and Phongpaichit, S.

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