Assessment Of Anti-Inflammatory Activity Of Ethanolic Extract Of Asam Kandis (Garcinia Xanthochymus Hook. F. EX T. Anderson) Fruit
Online - 2455-3891
Print - 0974-2441
Vol 11, Issue 4, 2018
Research Article
ASSESSMENT OF ANTI-INFLAMMATORY ACTIVITY OF ETHANOLIC EXTRACT OF ASAM
KANDIS (GARCINIA XANTHOCHYMUS HOOK. F. EX T. ANDERSON) FRUIT
HANAFIS SASTRA WINATA1*, ROSIDAH ROSIDAH1, PANAL SITORUS2
1
Department of Pharmacology, Faculty of Pharmacy, University of Sumatera Utara, Medan, Indonesia. 2Department of Biological Pharmacy,
Faculty of Pharmacy, University of Sumatera Utara, Medan, Indonesia. Email: [email protected]
Received: 10 November 2017, Revised and Accepted: 15 December 2017
ABSTRACT
Objective: The objective of this study was to evaluate the anti-inflammatory activity in acute and subacute models of inflammation from ethanolic
fruit extract of Asam kandis (Garcinia xanthochymus Hook. f. ex T. Anderson) in animal (rats) models.
Methods: Pleliminary phytochemical screening was carried out by using standard procedures.. Assessment of acute and subacute models of
inflammation was using carrageenan-induced paw edema method and cotton pellet granuloma method using three dosage treatments; 200 mg/kg
BW, 400 mg/kg BW, and 800 mg/kg BW along with a negative control group (0.5% Na CMC) and positive control (Na diclofenac 2.25 mg/kg BW). The
inhibition period was observed at 30, 60, 90, 120, 150, and 180 min time intervals.
Result: The phytochemical screening showed that the ethanolic fruit extract from Asam kandis contain contains flavonoids, glycosides, steroids, and
triterpenoids. The anti-inflammatory result showed that the strongest inhibition produced by ethanolic fruit extract of Asam kandis occurred on the
dosage of 800 mg/kg BW compared to the other doses (200 and 400 mg/kg BW) throughout the observation period.
Conclusion: This finding indicated that ethanolic fruit extract of Asam kandis (G. xanthochymus Hook. f. ex T. Anderson) might become an interesting
candidate for treatment of inflammation.
Keywords: Asam kandis, Anti-inflammatory, Carrageenan, Paw edema, Cotton pellet.
© 2018 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.
org/licenses/by/4. 0/) DOI: http://dx.doi.org/10.22159/ajpcr.2018.v11i4.23578
INTRODUCTION
Inflammation is the defensive response of the body which is nonspecific toward the tissue injury and infection. Pain and inflammation
are two states associated with numerous diseased conditions [1].
There are many conditions that could trigger the inflammation such
as pathogens, chemical irritation, extreme temperature, distortion or
disturbances of cells, and abrasions [2-4]. There are five characteristics
of inflammation such as redness, swelling, the rise of temperature, pain,
and loss of function [5].
Non-steroidal anti-inflammatory drug (NSAID) is one of the most
common medications for inflammation, but unexpected bronchus,
gastric mucosa, cardiac, and kidney-related side effects have been
reported [6]. The utilization of natural sources to search for new
pharmacologically active agents has led to the discovery of many
clinically beneficial drugs that contributes to a key role in the treatment
of human disease [7]. Therefore, many researchers are interested
in drugs from the natural sources such as plants and herbs origin
due to their wide range of pharmacological activities, including antiinflammatory potential with minimum side effects [8].
Garcinia xanthochymus Hook. f. ex T. Anderson, also called as Asam
kandis, belongs to the Clusiaceae family. Garcinia species are common
in Southeast Asia. This species is rich of the phytochemicals compound
that includes xanthone, flavonoids, and phenolic acids. Due to its rich
pharmaceutical values, mostly the Garcinia species have been to treat
many diseases such as antifungal, antioxidant, antiprotozoal, and antiimmunosuppressive [9]. In vitro investigation reported that the ethanolic
fruit extract of G. xanthochymus has a good antioxidant effect (half
maximal inhibitory concentration [IC50]: 22 µg/mL) [10]. Therefore, from
the previous investigation, it can be concluded that G. xanthochymus has
a good antioxidant properties. In this regard, it makes great sense to
evaluate the anti-inflammatory activity of Asam kandis (G. xanthochymus
Hook. f. ex T. Anderson) fruit ethanolic extract using carrageenan-induced
paw edema method and cotton pellet method.
METHODS
Identification and authentication of plant materials
Asam kandis (G. xanthochymus Hook. f. ex T. Anderson) fruits were
obtained from region of Lubuk Minturun, Padang, West Sumatera,
Indonesia. Fresh fruits were dried and then grounded to obtain a finely
divided powder.
Plant extraction preparation and phytochemical screening
The powder of Asam kandis (G. xanthochymus Hook. f. ex T. Anderson)
fruits was extracted using a triple maceration process with ethanol
organic solvent. 900 g of powdered Asam kandis fruits is macerated
in advance with the ethanol solvent for 5 days, then filtered, do it
continuously until the filtrate obtained is clear and colorless [11].
Phytochemical screening carried out on Asam kandis fruits ethanolic
extract includes examining the chemical secondary metabolites of
alkaloids, flavonoids, glycosides, tannins, triterpenoids, and steroids.
The preliminary phytochemical screening was using widely used
methods [12-14].
Preparation of animals
Healthy adult male Wistar rats (150–200 g body weight) from animal
house of Faculty of Pharmacy, University of Sumatera Utara, were used
for the study. Rats were housed in a polycarbonate cages in a room
with 12 h day-night circle. They were fed on a standard pellet diet and
water ad libitum. The study was approved by Animal Research Ethics
Committees (AREC) of University of Sumatera Utara (AREC No: 120/
KEPH-FMIPA/2017) and the experiments were conducted according to
the ethical norms and AREC guidelines.
Winata et al.
Asian J Pharm Clin Res, Vol 11, Issue 4, 2018, 81-83
Anti-inflammatory activity
Table 1: Phytochemical screening result of ethanolic extract
from Asam kandis fruits
Carrageenan-induced paw edema
Healthy adult male Wistar rats were divided into five groups of six rats
each. Edema was induced by injecting 0.05 mL of 1 % carrageenan
suspension into the subplantar region of the right hind paw of the rats.
Control group rats received 0.5% (w/v) Na CMC and the reference
group of rats received 2.25 mg/kg BW sodium diclofenac, orally. The
test groups of rats were treated orally with 200, 400, and 800 mg/kg
BW ethanolic extract of Asam kandis fruits 40 min before carrageenan
injection. The paw volume was measured by plethysmometer before
carrageenan injection (V0) and 30, 60, 90, 120, 150, and 180 min after
(Vt). The inflammation was calculated as the increase in volume (mL)
of the paw after treatment subtracted of basal volume. Results were
expressed as percentage of inhibition of edema, calculated according
to the formula [15,16]:
Percentage
inhibition ( % )
No
Screening
Result
1
2
3
4
5
6
Alkaloids
Flavonoids
Glycosides
Tannins
Saponins
Triterpenoid/steroids
Negative
Positive
Positive
Negative
Negative
Positive
Mean paw inflammation of controlMean paw inflammation of test
=
×100%
Mean paw inflammation of control
Cotton pellet granuloma method
Wistar rats were divided into five groups of six rats each. Adsorbent
cotton was made into pellets weighing 20 ± 1 mg. The pellets were
sterilized in a hot air oven at 120°C for 2 h. The abdomen was shaved
cleanly, swabbed with 70% ethanol, and the sterilized cotton pellet
was implanted subcutaneously under light ether anesthesia. Test drugs
were administered once daily throughout the experimental period of
7 days. On the 8th day after implantation, rats were anesthetized and the
pellets were dissected and dried at 60oC for 6 h, weighed after cooling.
The mean weight of the cotton pellets of the control group as well as
of the test groups was calculated. The transudative weight and percent
granuloma inhibition of the test compound were calculated [17].
Fig. 1: Effect of various doses of Asam kandis fruit ethanolic
extract in carrageenan-induced paw edema in rats (change in
mean paw volume)
Statistical analysis
The significant difference of data between different groups was
compared by ANOVA followed by Duncan’s test.
RESULTS AND DISCUSSION
This study investigated the anti-inflammatory activity of Asam kandis
(G. xanthochymus Hook. f. ex T. Anderson) fruits ethanolic extract. The
phytochemical screening of ethanolic extract of Asam kandis fruits
revealed the presence of flavonoids, glycosides, steroids, and triterpenoids
(Table 1). These results are related to previous research [18] that shown
the presence of flavonoids, glycosides, steroids, and triterpenoids.
To evaluate the extract effect on anti-inflammatory activity, the ethanolic
extract of Asam kandis fruits was tested using carrageenan-induced paw
edema and cotton pellet granuloma method. In carrageenan-induced
paw edema method, the paw volumes and percentage of inhibition
of the control, standard, and test groups are shown in Figs. 1 and 2.
The ethanolic fruit extract of Asam kandis showed anti-inflammatory
activity compared to Na diclofenac. A dose of 800 mg/kg BW elicited a
greater percent of inhibition of inflammatory after 60–180 min. These
results showed that the ethanolic extract of Asam kandis fruit (dose
800 mg/kg BW) has the most potent anti-inflammatory activity.
The cotton pellet granuloma method has been widely used to access the
subacute inflammation. The results are shown in Fig. 3. As shown in Fig.
3, all the test groups have the inhibitory activity, but the best doses are
400 and 800 mg/kg BW. It can be shown by the percentage of inhibition
for 400 mg/kg BW is 58.10% and 35.89% for 800 mg/kg BW. These
results suggest that the ethanolic extracts exhibit antitransudative effect.
Drugs from plant sources have been used for the treatment of various
diseases since ancient times. Herbal drugs are inexpensive and effective
with fewer side effects [19]. Nowadays, the use of herbal drugs to cure
inflammation and pain is gaining popularity due to their effectiveness,
Fig. 2: Effect of various doses of Asam kandis fruit ethanolic
extract in carrageenan-induced paw edema in rats (% of
inhibition)
fewer side effects, low cost, and availability. NSAIDs act by inhibiting
cyclooxygenase and the production of prostaglandins. Diclofenac
offers relief from inflammation by suppressing the production of
prostaglandins and bradykinin. The presence of flavonoids has been
reported in Garcinia species, and flavonoids are known to inhibit
prostaglandin synthetase. Previous studies have shown similar
relationships between flavonoids and anti-inflammatory effects [20-29]
Therefore, it is possible that the anti-inflammatory action of Asam
kandis fruit extract may be related to the inhibition of prostaglandin
synthesis. The phytochemical profile of Asam kandis fruit extract may
be explored further to identify the active constituents responsible for
its anti-inflammatory activity.
CONCLUSIONS
The ethanolic extract of Asam kandis fruit (G. xanthochymus Hook. f. ex
T. Anderson) ethanoic extract at dose of 800 mg/kg BW had the most
82
Winata et al.
Asian J Pharm Clin Res, Vol 11, Issue 4, 2018, 81-83
9.
10.
11.
12.
13.
14.
15.
16.
Fig. 3: Effect of various doses of Asam kandis fruit ethanolic
extract in cotton pellet granuloma method
potent anti-inflammatory activity throughout the observation period.
These studies can be concluded that Asam kandis fruit (G. xanthochymus
Hook. f. ex T. Anderson) has the potential to be developed as an antiinflammatory agent.
ACKNOWLEDGMENTS
I express my thanks to Iksen, B. Pharm., M.Sc. (USU), for his never
ending guidance and suggestions throughout the preparation of this
research and article.
AUTHORS CONTRIBUTIONS
17.
18.
19.
20.
21.
All the author have contributed equally.
CONFLICT OF INTERESTS
Declared none.
22.
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Print - 0974-2441
Vol 11, Issue 4, 2018
Research Article
ASSESSMENT OF ANTI-INFLAMMATORY ACTIVITY OF ETHANOLIC EXTRACT OF ASAM
KANDIS (GARCINIA XANTHOCHYMUS HOOK. F. EX T. ANDERSON) FRUIT
HANAFIS SASTRA WINATA1*, ROSIDAH ROSIDAH1, PANAL SITORUS2
1
Department of Pharmacology, Faculty of Pharmacy, University of Sumatera Utara, Medan, Indonesia. 2Department of Biological Pharmacy,
Faculty of Pharmacy, University of Sumatera Utara, Medan, Indonesia. Email: [email protected]
Received: 10 November 2017, Revised and Accepted: 15 December 2017
ABSTRACT
Objective: The objective of this study was to evaluate the anti-inflammatory activity in acute and subacute models of inflammation from ethanolic
fruit extract of Asam kandis (Garcinia xanthochymus Hook. f. ex T. Anderson) in animal (rats) models.
Methods: Pleliminary phytochemical screening was carried out by using standard procedures.. Assessment of acute and subacute models of
inflammation was using carrageenan-induced paw edema method and cotton pellet granuloma method using three dosage treatments; 200 mg/kg
BW, 400 mg/kg BW, and 800 mg/kg BW along with a negative control group (0.5% Na CMC) and positive control (Na diclofenac 2.25 mg/kg BW). The
inhibition period was observed at 30, 60, 90, 120, 150, and 180 min time intervals.
Result: The phytochemical screening showed that the ethanolic fruit extract from Asam kandis contain contains flavonoids, glycosides, steroids, and
triterpenoids. The anti-inflammatory result showed that the strongest inhibition produced by ethanolic fruit extract of Asam kandis occurred on the
dosage of 800 mg/kg BW compared to the other doses (200 and 400 mg/kg BW) throughout the observation period.
Conclusion: This finding indicated that ethanolic fruit extract of Asam kandis (G. xanthochymus Hook. f. ex T. Anderson) might become an interesting
candidate for treatment of inflammation.
Keywords: Asam kandis, Anti-inflammatory, Carrageenan, Paw edema, Cotton pellet.
© 2018 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.
org/licenses/by/4. 0/) DOI: http://dx.doi.org/10.22159/ajpcr.2018.v11i4.23578
INTRODUCTION
Inflammation is the defensive response of the body which is nonspecific toward the tissue injury and infection. Pain and inflammation
are two states associated with numerous diseased conditions [1].
There are many conditions that could trigger the inflammation such
as pathogens, chemical irritation, extreme temperature, distortion or
disturbances of cells, and abrasions [2-4]. There are five characteristics
of inflammation such as redness, swelling, the rise of temperature, pain,
and loss of function [5].
Non-steroidal anti-inflammatory drug (NSAID) is one of the most
common medications for inflammation, but unexpected bronchus,
gastric mucosa, cardiac, and kidney-related side effects have been
reported [6]. The utilization of natural sources to search for new
pharmacologically active agents has led to the discovery of many
clinically beneficial drugs that contributes to a key role in the treatment
of human disease [7]. Therefore, many researchers are interested
in drugs from the natural sources such as plants and herbs origin
due to their wide range of pharmacological activities, including antiinflammatory potential with minimum side effects [8].
Garcinia xanthochymus Hook. f. ex T. Anderson, also called as Asam
kandis, belongs to the Clusiaceae family. Garcinia species are common
in Southeast Asia. This species is rich of the phytochemicals compound
that includes xanthone, flavonoids, and phenolic acids. Due to its rich
pharmaceutical values, mostly the Garcinia species have been to treat
many diseases such as antifungal, antioxidant, antiprotozoal, and antiimmunosuppressive [9]. In vitro investigation reported that the ethanolic
fruit extract of G. xanthochymus has a good antioxidant effect (half
maximal inhibitory concentration [IC50]: 22 µg/mL) [10]. Therefore, from
the previous investigation, it can be concluded that G. xanthochymus has
a good antioxidant properties. In this regard, it makes great sense to
evaluate the anti-inflammatory activity of Asam kandis (G. xanthochymus
Hook. f. ex T. Anderson) fruit ethanolic extract using carrageenan-induced
paw edema method and cotton pellet method.
METHODS
Identification and authentication of plant materials
Asam kandis (G. xanthochymus Hook. f. ex T. Anderson) fruits were
obtained from region of Lubuk Minturun, Padang, West Sumatera,
Indonesia. Fresh fruits were dried and then grounded to obtain a finely
divided powder.
Plant extraction preparation and phytochemical screening
The powder of Asam kandis (G. xanthochymus Hook. f. ex T. Anderson)
fruits was extracted using a triple maceration process with ethanol
organic solvent. 900 g of powdered Asam kandis fruits is macerated
in advance with the ethanol solvent for 5 days, then filtered, do it
continuously until the filtrate obtained is clear and colorless [11].
Phytochemical screening carried out on Asam kandis fruits ethanolic
extract includes examining the chemical secondary metabolites of
alkaloids, flavonoids, glycosides, tannins, triterpenoids, and steroids.
The preliminary phytochemical screening was using widely used
methods [12-14].
Preparation of animals
Healthy adult male Wistar rats (150–200 g body weight) from animal
house of Faculty of Pharmacy, University of Sumatera Utara, were used
for the study. Rats were housed in a polycarbonate cages in a room
with 12 h day-night circle. They were fed on a standard pellet diet and
water ad libitum. The study was approved by Animal Research Ethics
Committees (AREC) of University of Sumatera Utara (AREC No: 120/
KEPH-FMIPA/2017) and the experiments were conducted according to
the ethical norms and AREC guidelines.
Winata et al.
Asian J Pharm Clin Res, Vol 11, Issue 4, 2018, 81-83
Anti-inflammatory activity
Table 1: Phytochemical screening result of ethanolic extract
from Asam kandis fruits
Carrageenan-induced paw edema
Healthy adult male Wistar rats were divided into five groups of six rats
each. Edema was induced by injecting 0.05 mL of 1 % carrageenan
suspension into the subplantar region of the right hind paw of the rats.
Control group rats received 0.5% (w/v) Na CMC and the reference
group of rats received 2.25 mg/kg BW sodium diclofenac, orally. The
test groups of rats were treated orally with 200, 400, and 800 mg/kg
BW ethanolic extract of Asam kandis fruits 40 min before carrageenan
injection. The paw volume was measured by plethysmometer before
carrageenan injection (V0) and 30, 60, 90, 120, 150, and 180 min after
(Vt). The inflammation was calculated as the increase in volume (mL)
of the paw after treatment subtracted of basal volume. Results were
expressed as percentage of inhibition of edema, calculated according
to the formula [15,16]:
Percentage
inhibition ( % )
No
Screening
Result
1
2
3
4
5
6
Alkaloids
Flavonoids
Glycosides
Tannins
Saponins
Triterpenoid/steroids
Negative
Positive
Positive
Negative
Negative
Positive
Mean paw inflammation of controlMean paw inflammation of test
=
×100%
Mean paw inflammation of control
Cotton pellet granuloma method
Wistar rats were divided into five groups of six rats each. Adsorbent
cotton was made into pellets weighing 20 ± 1 mg. The pellets were
sterilized in a hot air oven at 120°C for 2 h. The abdomen was shaved
cleanly, swabbed with 70% ethanol, and the sterilized cotton pellet
was implanted subcutaneously under light ether anesthesia. Test drugs
were administered once daily throughout the experimental period of
7 days. On the 8th day after implantation, rats were anesthetized and the
pellets were dissected and dried at 60oC for 6 h, weighed after cooling.
The mean weight of the cotton pellets of the control group as well as
of the test groups was calculated. The transudative weight and percent
granuloma inhibition of the test compound were calculated [17].
Fig. 1: Effect of various doses of Asam kandis fruit ethanolic
extract in carrageenan-induced paw edema in rats (change in
mean paw volume)
Statistical analysis
The significant difference of data between different groups was
compared by ANOVA followed by Duncan’s test.
RESULTS AND DISCUSSION
This study investigated the anti-inflammatory activity of Asam kandis
(G. xanthochymus Hook. f. ex T. Anderson) fruits ethanolic extract. The
phytochemical screening of ethanolic extract of Asam kandis fruits
revealed the presence of flavonoids, glycosides, steroids, and triterpenoids
(Table 1). These results are related to previous research [18] that shown
the presence of flavonoids, glycosides, steroids, and triterpenoids.
To evaluate the extract effect on anti-inflammatory activity, the ethanolic
extract of Asam kandis fruits was tested using carrageenan-induced paw
edema and cotton pellet granuloma method. In carrageenan-induced
paw edema method, the paw volumes and percentage of inhibition
of the control, standard, and test groups are shown in Figs. 1 and 2.
The ethanolic fruit extract of Asam kandis showed anti-inflammatory
activity compared to Na diclofenac. A dose of 800 mg/kg BW elicited a
greater percent of inhibition of inflammatory after 60–180 min. These
results showed that the ethanolic extract of Asam kandis fruit (dose
800 mg/kg BW) has the most potent anti-inflammatory activity.
The cotton pellet granuloma method has been widely used to access the
subacute inflammation. The results are shown in Fig. 3. As shown in Fig.
3, all the test groups have the inhibitory activity, but the best doses are
400 and 800 mg/kg BW. It can be shown by the percentage of inhibition
for 400 mg/kg BW is 58.10% and 35.89% for 800 mg/kg BW. These
results suggest that the ethanolic extracts exhibit antitransudative effect.
Drugs from plant sources have been used for the treatment of various
diseases since ancient times. Herbal drugs are inexpensive and effective
with fewer side effects [19]. Nowadays, the use of herbal drugs to cure
inflammation and pain is gaining popularity due to their effectiveness,
Fig. 2: Effect of various doses of Asam kandis fruit ethanolic
extract in carrageenan-induced paw edema in rats (% of
inhibition)
fewer side effects, low cost, and availability. NSAIDs act by inhibiting
cyclooxygenase and the production of prostaglandins. Diclofenac
offers relief from inflammation by suppressing the production of
prostaglandins and bradykinin. The presence of flavonoids has been
reported in Garcinia species, and flavonoids are known to inhibit
prostaglandin synthetase. Previous studies have shown similar
relationships between flavonoids and anti-inflammatory effects [20-29]
Therefore, it is possible that the anti-inflammatory action of Asam
kandis fruit extract may be related to the inhibition of prostaglandin
synthesis. The phytochemical profile of Asam kandis fruit extract may
be explored further to identify the active constituents responsible for
its anti-inflammatory activity.
CONCLUSIONS
The ethanolic extract of Asam kandis fruit (G. xanthochymus Hook. f. ex
T. Anderson) ethanoic extract at dose of 800 mg/kg BW had the most
82
Winata et al.
Asian J Pharm Clin Res, Vol 11, Issue 4, 2018, 81-83
9.
10.
11.
12.
13.
14.
15.
16.
Fig. 3: Effect of various doses of Asam kandis fruit ethanolic
extract in cotton pellet granuloma method
potent anti-inflammatory activity throughout the observation period.
These studies can be concluded that Asam kandis fruit (G. xanthochymus
Hook. f. ex T. Anderson) has the potential to be developed as an antiinflammatory agent.
ACKNOWLEDGMENTS
I express my thanks to Iksen, B. Pharm., M.Sc. (USU), for his never
ending guidance and suggestions throughout the preparation of this
research and article.
AUTHORS CONTRIBUTIONS
17.
18.
19.
20.
21.
All the author have contributed equally.
CONFLICT OF INTERESTS
Declared none.
22.
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