INDONESIAN MEDICINAL PLANTS EXTRACT AS ANTI-OBESITY:

INHIBITORY EFFECT ON THE ACTIVITY OF PANCREATIC LIPASE

BY IN VITRO AND IN VIVO METHOD
1
DYAH ISWANTINI, 2MIN RAHMINIWATI, 3ASEP WAHYUDIN, 4TRIVADILA, 5WARAS
NURCHOLIS
1,2,5
Biopharmaca Research Center, Bogor Agricultural University,Bogor, Jl.Taman Kencana no 3 Bogor, Indonesia.
1,3,4
Department of Chemistry, Faculty of Mathematics and Natural Sciences, Bogor Agricultural, University,
Gedung Kimia, Jl Agathis, IPB Darmaga,Bogor, Indonesia.
E-mail: 1dyahprado@gmail.com

Abstract - Indonesian medicinal plants such as: Asam gelugur (Garcinia atroviridis) and kunci pepet (Kaempferia
angustifolia) have been used as herbal medicine traditionally to reduce body weight. By in vitro method, the single extract of
these plants had been inhibited the activity of pancreatic lipase. The objective of the research was to evaluate the activity of
the combination of ethanol extracts towards human pancreatic lipase activity by in vitro and in vivo method. In vitro assay
was performed at pH : 8.0, incubation time : 45 minutes, and temperature : 40 C. In vivo assay was conducted using
female rats by investigating the change of their body weight and feed consumption. The result of in vitro assay indicated
that the combination of ethanol extract of asam gelugurkunci pepet had the highest inhibition activity towards human
pancreatic lipase (75.69%). Result of in vivo assay showed that the formula could decrease the body weight of rats group up
to 11.22% decreasing their appetite. It was concluded that the formula of ethanol extract of asam gelugurkunci pepet has
efficacy as anti-obesity by in vitro and in vivo inhibition toward pancreatic lipase.

Keywords - Anti-obesity, asam gelugur (Kaempferia angustifolia), kunci pepet (Garcinia atroviridis), pancreatic lipase, in
vitro, in vivo

I. INTRODUCTION the activity of pancreatic lipase in vitro. The results of

Overweight or better known as obesity occurs
because of the imbalance in the amount of food
intake and expenditure energy by the body. Obesity
can cause several serious diseases such as diabetes,
coronary heart disease, high blood pressure, and
constriction of blood vessels. Diseases caused need
patterns requiring some treatments and preventions.
There are two ways someone can do to reduce body
weight and makes an ideal return, which is naturally
and using aid. Natural treatment can be done by
rearranging the diet and exercise. Given this
treatment takes a long time, then only little of the
obese people who do this way. The second way is
using a slimming drug.
in vivo study, described that a combination of OB-
200G extract from Garcinia cambogia fruit,
Commiphora mukul resin gum, Zingiber officinale
rhizome, Piper longum fruit, Gymnema sylvestre
leaves [4], and orange peel extract, black tea, caffeine
[5] could reduce rat body weight significantly
compared to negative control by inhibiting pancreatic
lipase activity.
Herbal plants which are currently investigated their
potential as a slimming drug are kunci pepet and
asam gelugur. The result of our previous study
indicated that kunci pepet rhizome pepet could
inhibit pancreatic lipase activity in vitro by 65.25%
and asam gelugur fruit was 86.30% .

One of slimming drug mechanism is inhibits the
absorption of fatty acids through the inhibition of
pancreatic lipase activity. Pancreatic lipase is an
enzyme that functions to hydrolyze fat in the intestine
into glycerol and fatty acids. The increasement of
pancreatic lipase activity will increase the absorption
of fatty acids and leading to obesity.
Tetrahydrolipstatin (orlistat) is a slimming drug with
a mechanism to inhibit the activity of synthetic
pancreatic lipase [1].
The use of slimming drugs that are traditionally made
from selected herbs are more selected by obese
people because it is safe to health. Based on the study
on saponin active compounds from Kochia scoparia
plant [2], the polyphenol herbal extracts of berry fruit
that contain many kinds of tannins [3] could inhibit
Proceedings of ISER 9th International Conference, Berlin, Germany, 30th October 2015, ISBN: 978-93-85832-26-0
27
In this study, combination extracts of asam gelugur
fruit and kunci pepet rhizome in inhibiting pancreatic
lipase activity in vitro were tested. Efficacy of the
kunci pepet rhizome and asam gelugur fruit extract
combination as slimming were conducted further tests
in vivo on rats, observed from changes in body weight
and feed intake.
II. DETAILS EXPERIMENTAL
2.1. Materials and Procedures Materials
The asam gelugur fruit were collected from Aceh
(North Sumatra, Indonesia). The kunci pepet
rhizomes were obtained from Pati (Central Java,
Indonesia). The albumin, sesame oil, and pancreatic
lipase were purchased from Sigma chemical Co.,
Indonesian Medicinal Plants Extract as Anti-Obesity: Inhibitory Effect on the Activity of Pancreatic Lipase by in Vitro and in Vivo Method
USA. The experiment was used the rats of the
Sprague-Dawley strain.
2.2. Procedures Preparation of Plant
The rhizomes of kunci pepet and the fruit of asam
gelugur were separated and dried at 50 C,
powdered, sieved and stored prior to futher use.
Preparation of plant extract
Extraction was used maceration method of National
Agency of Drug and Food Control of the Republic of
Indonesia [6]. One hundred grams of the dried
samples were macerated using 10 x 100 mL ethanol
70% and water for 1 x 24 h. The filtrate obtained was
concentrated under vacuum on a rotary evaporator at
40 50C and then stored at 4C for further use.
2.3. Inhibition assay for pancreatic lipase activity by
in vitro method
The inhibition assay for pancreatic lipase activity was
determined by in vitro method as described by Xu et
al. (2005) [7] with slight modification. The
experiment used ethanol and water combination
extract between asam gelugur and kunci pepet.
Substrate (15 L) was premixed with the extract at
various concentrations (10-30 ppm). About 10 L
10% albumin was added to the mixture. The mixture
was added pH 7 phosphate buffer to 1 mL. The
pancreatic lipase (100 L) was added in the tube
mixture sample. Subsequently the mixture sample
was homogenized in vortex. The reaction was
incubated at 40C for 45 minutes. After incubation
complete, the mixture sample was added 3 mL
chloroform and homogenized and separated using
centrifuge for 5 minutes. One mL chloroform phase
was obtained and added another tube. Three mL
chloroform phase was obtained and added another
tube. Subsequently the tube was added 0.25 mL Na-
diethyldithiocarbamate. The absorbance was then
measured at 435 nm.
2.4. Inhibition assay for pancreatic lipase activity by
in vivo method
The inhibition assay for pancreatic lipase activity was
determined by in vivo method as described by Kaur
& Kulkarni (2000) [4] with slight modification. The
experiment was used combination extract between
asam gelugur and kunci pepet which the best activity
for in vitro inhibition of pancreatic lipase. Adult rats
of the Sprague-dawley strain (160-220 g) were
maintained in individual cages and divided into five
groups of twelve animals each. Group one, two, and
three were treated with combination extract between
asam gelugur and kunci pepet at certain dosages.
Groups four and five were positive and negative
control, respectively. The animal had free access to
Proceedings of ISER 9th International Conference, Berlin, Germany, 30th October 2015, ISBN: 978-93-85832-26-0
28
standard pellet diet and water for 30 days. The
administration of herbal extract combination was
started from the 15th day and was continued up to the
30th day of experiment.
III. RESULTS AND DISCUSSION
3.1. Extraction of plant materials
The result of the extraction of asam gelugur and
kunci pepet were used maceration with ethanol and
water solution. The weight of extract ethanol and
water of asam gelugur were 21.30% and 19.23%,
respectively (Table 1). The weight of extract ethanol
and water of kunci pepet were 15.75% and 18.13%,
respectively (Table 1).
Table 1. The weight of extract ethanol and water
of asam gelugur and kunci pepet
The weight of extract from plant materials dependent
was used polarity of solution. The water is polar
solution, so its can be extraction polar compound of
plant material. Whereas, the ethanol is polar at
hydroxyl group and non-polar at alkyl group, so its
can be extraction compounds at different polarity.
3.2. Inhibition assay for pancreatic lipase activity by
in vitro method
Pancreatic lipase plays an important role in lipid
metabolism and is one of the most widely studied
models for the evaluation of the potential efficacy of
plant medicinal as anti obesity agent. The all
combination extracts ethanol and water of asam
gelugur and kunci pepet were guided by their
pancreatic lipase inhibitory activity, using Xenical
as a positive control. The extract combination I and
III at concentration 15 to 30 ppm were the best
inhibitory of pancreatic lipase activity compared to
Xenical as a positive control (Figure 1). The
combination of ethanol extract of asam gelugur
kunci pepet had the highest inhibition activity
towards human pancreatic lipase (75.69%).
The extract combination was done for investigated
effect combination of secondary metabolites e.g.
flavonoid, saponin and tannin from kunci pepet and
hydroxide citrate acid (HCA) from asam gelugur. In
one study, it was concluted that the extract tannin and
flavonoid, combination extract flavonoid and tannin,
and combination extract steroid and tannin from
bangle has potent inhibitory of pancreatic lipase
Indonesian Medicinal Plants Extract as Anti-Obesity: Inhibitory Effect on the Activity of Pancreatic Lipase by in Vitro and in Vivo Method
activity [8]. The compound of antiobesity in any
medicinal plant is saponin [9] [10] [11] [3].
Fig.1. Inhibitory effects of extract ombination on pancreatic
lipase. KP: Xenical (control positive); K1: Combination I; K2:
Combination II; K3: Combination III; K4: Combination IV;
and S1: single water extract water.
3.3. Inhibition assay for pancreatic lipase activity by
in vivo method
There was a significant increase in body weight of
animals experiment in adaptation stage (Figure 2).
Treatment with combination extract ethanol asam
gelugur and extract water kunci pepet caused
significant decrease in body weight when compared
to another group animal. The combination extract
ethanol asam gelugur and extract water kunci pepet
could decrease the body weight of rats group up to
11.22% decreasing their appetite. The weight
reducing effect of the combination extract ethanol
asam gelugur and extract water kunci pepet may be
attributed to asam gelugur (contains Hydroxycitric
acid reported to inhibit lipogenesis [12] and the
metabolite secondary from kunci pepet. The
mechanism effect of the weight reducing with extract
combination of asam gelugur-kunci pepet was
predicted through inhibition of pancreatic lipase
activity and pushed down of appetite.
Fig. 2. Effect of the extract combination of asam gelugur-kunci
pepet on body weight changes
P1: treatment 1; P2: treatment 2; P3: treatment 3 ; KP:
positive control; KN: negative control
Proceedings of ISER 9th International Conference, Berlin, Germany, 30th October 2015, ISBN: 978-93-85832-26-0
29
CONCLUSIONS
The present study demonstrated that the combination
of ethanol extract of asam gelugurkunci pepet had
efficacy as anti-obesity significantly by in vitro and in
vivo inhibition toward pancreatic lipase.
ACKNOWLEDGMENTS
Author would like to thanks to Indonesian
Government who provided funding for this reasearch.
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