214

AGRIVITA Journal of Agricultural Science. 2017. 39(2): 214-218

Phytochemical Screening and Antioxidant Activity of Selekop

(Lepisanthes amoena) Fruit
Heriad Daud Salusu1,2*), Farida Ariani2), Ernita Obeth3), Mark Rayment4), Edy Budiarso5),
Irawan Wijaya Kusuma6) and Enos Tangke Arung6)
1)
Faculty of Forestry Mulawarman University Samarinda East Kalimantan Indonesia
2)
Lab. of Wood Properties and Product Analysis Samarinda State Polytechnic of Agriculture Indonesia
3)
School of Processing Technology of Agricultural Product Samarinda State Polytechnic of Agriculture
Indonesia
4)
School of Environment Natural Resources and Geography Bangor University United Kingdom
5)
Lab. of Wood Preservative Faculty of Forestry Mulawarman University Indonesia
6)
Lab. of Forest Product Chemistry Faculty of Forestry Mulawarman University Indonesia
*)
Corresponding author E-mail:[email protected]

Received: February 19, 2016 /Accepted: March 9, 2017

ABSTRACT resin. Durian, mango, rambutan and mangosteen

are some of the most popular forest fruit species in
Selekop (Lepisanthes amoena (Hassk.) Leenh.)
Indonesia while more other species are generally
plant leaves are used by the Dayak tribe of East
known only by the local residents around the forest.
Kalimantan as traditional cosmetics. Selekop fruit is
Increasing the value and benefits of forest fruit or
also edible, but not well known. This study was con-
local fruit has now become a challenge following the
ducted to obtain the phytochemical content and an-
entry and domination of imported fruits in Indonesian
tioxidant assay in flesh, seed and pericarp extracts
markets. One effort that can be done to improve the
from the fruit of Selekop. Phytochemical analysis
image of forest fruits is by conducting more research
was conducted on ethanol extract for identification of
on the nutrients and bioactivity contained in the fruits.
flavonoid, alkaloid, tannin, saponin, triterpenoid and
Information on the benefits of certain forest fruits
steroid. The antioxidant activity was done by DPPH
has traditionally been known only by the locals. The
assay with ascorbic acid as positive control. The flesh
scientific research is not yet to be done. The Selekop
contained flavonoid, saponin, and tannin; the seed
fruit (Lepisanthes amoena (Hassk.) Leenh.) is a
contained flavonoid, alkaloid, saponin, triterpenoid,
species of forest fruit that is not well identified. This
and tannin; and the pericarp contained flavonoid, al-
fruit belongs to the family of Sapindaceae. In some
kaloid, saponin, triterpenoid, and tannin. Analysis of
regions in East Kalimantan it is known as Kokang
antioxidant activity revealed the following Inhibitory
or Kukang. In Western Java it is called Langir, while
Concentration (IC50 values): 122.51 ppm of flesh,
in South Kalimantan the locals call it Rembia. In
63.30 ppm of seed, 53.21 ppm of pericarp and 3.06
general, the Dayak tribe of East Kalimantan uses the
ppm of ascorbic acid. Based on these results, the
leaves as traditional cosmetic ingredients.
ethanol extract of the seed and the flesh had a phy-
To date, there has not been a single report
tochemical content and antioxidant activity which
on the benefits of this fruit except for its being an
was better than the flesh extract from Selekop fuit.
edible fruit. As a cosmetic ingredient, (by various
Keywords: antioxidant; flesh; pericarp; phytochemical; ethnic groups in Borneo) it is generally used in
seed; Selekop mixing powder, skin cleanser, shampoo and soap. It
is used by squeezing the leaves and mix them with
INTRODUCTION
a little water to produce lather like soap. But now,
Fruits obtained from forests are considered because of modern cosmetics, shampoo and soap
unimportant or non-profitable in comparison to are available to the public so that the leaves are
other forest products such as timber, rattan and rarely used.

Cite this as: Salusu, H. D., Ariani, F., Obeth, E., Rayment, M., Budiarso, E., Kusuma, I. W., & Arung, E. T. (2017).
Phytochemical screening and antioxidant activity of selekop (Lepisanthes amoena) fruit. AGRIVITA Journal of
Agricultural Science, 39(2), 214–218. http://doi.org/10.17503/agrivita.v39i2.810
Accredited: SK No. 60/E/KPT/2016
215

Heriad Daud Salusu et al.: Phytochemical Screening and Antioxidant Activity of Selekop................................................

This plant is considered of possessing a Preparation of Fruit Extract

minor economic value, wildly grown in the forest The extraction method used was cold
and uncultivated (Milow, Malek, Edo, & Ong, extraction with etanol as a solvent. Cold extraction
2014). Although the leaves of this plant have was done by dissolving the powder in the ratio of
traditionally been known to be beneficial, the fruit powder : solvent (1:10). The extraction was done
is still unidentified and it requires further study by immersing each sample with etanol. The sample
about its benefits and contents especially bioactive was then shaken using a shaker for 48 hours at
components that have the potential for further room temperature, then filtered to separate the
development. extract from fruit material. Furthermore, by using a
All bioactive food compounds are obtained rotary evaporator, the extract was evaporated at a
from the plants; they are called the phytochemicals. temperature of 30-40 0C, to obtain a crude extract.
Most of these compounds are active molecules
Phytochemical Screening
and named as antioxidants. The antioxidants
The secondary metabolites from each of
can diminish reactive oxygen, nitrogen, and free
the extracts were analyzed using phytocemical
radicals. The reactive species cause many chronic
screening according to standard procedures such
diseases (Carlsen et al., 2010). Food from plants like
as flavonoids, tannins, alkaloids, triterpenoids/
vegetables, fruits, and whole grains, had abundant
steroids, and saponins analysis. For the tests for
of bioactive compounds and give contribution
flavonoids (Kokate, 2001), a few drops of dilute
to human health than basic nutrition to combat
sodium hydroxide (NaOH 1 %) were added into 1
diseases. Zhang et al. (2015) suggested that more
ml of sample extract. The emergence of the clear
studies need to be done to identify the bio actives
yellow color in the extract solution, which turned
and mechanism of antioxidant phytochemicals
colorless after the addition of dilute acid (HCl 1 %),
actions. Thus, it is expected that more antioxidant
indicated that flavonoids were present. Meanwhile,
phytochemicals in food as well as in medicinal
for the tests for tannins (Kokate, 2001), 10 ml of
plants will be explored not only for its positive but
the extract solution was added to a 1 % solution of
also potential undesirable effects to human.
lead acetate. The tannins were tested positive if the
Therefore, this research was carried out to
yellow precipitate was formed. Furthermore, the
determine the antioxidant activity and phytochemical
tests for alkaloids (Kokate, 2001), 2 ml of HCl was
in Selekop fruit extracts contained in the flesh, seed
added to 5 ml of the extract solution, then combined
and pericarp in order to collect data and information
with 1 ml of Dragendorff solution. The color of the
to enhance the benefits of this fruit in the future.
solution became orange or red indicating that
MATERIALS AND METHODS the extract contained alkaloids. While the tests
for triterpenoids and steroids (Harborne, 1987),
The research was conducted for two months
Anhydride acetic acid was put about 10 drops and
(November-December 2015) in the Laboratory
2 drops of concentrated sulfuric acid were put into
of Wood Properties and Product Analysis, State
sequentially to 1 ml of sample test that had been
Agricultural Polytechnic Samarinda.
dissolved in acetone. The test sample was shaken
Materials and left for a few minutes. If it turned to red and
The tools used were a rotary vacuum purple, the test would be declared positive for
evaporator, spectrophotometer, and other glass triterpenoids and if it turned green and blue, this
tools, while the material was fresh Selekop fruit would indicate a positive test for the presence of
separated into three different parts (i.e., flesh, seed steroids. Moreover, for the tests for saponins
and pericarp). In addition, a number of chemical (Harborne, 1987), 10 ml the hot water was put into
materials namely etanol, acetone, aquades, solvent 1 ml of sample test which had been dissolved in
of DPPH (1,1-diphenyl-2-picrylhydrazyl), dimethyl acetone. The solution was cooled and shaken for
sulfoxide (DMSO), acetic acid anhydride, sulfuric 10 seconds. The formation of a steady froth with
acid), Dragendorff reagent, Liebermann-Burchard a height of 1-10 cm would take approximately 10
regent, Molisch reagent, Sodium hydroxide, minutes and remained even after the addition of
hydrochloric acid, lead acetate, and ascorbic acid 1 drop of 2N HCl, which indicated that the tested
were used in the analysis process. extracts contained saponins.
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Heriad Daud Salusu et al.: Phytochemical Screening and Antioxidant Activity of Selekop................................................

Antioxidants Activities Meanwhile, flavonoids were detected in

The testing of antioxidant activity used a all parts of the fruit because these compounds
spectrophotometer at room temperature (25 0C) with belonged to the natural polyphenol group which can
a wavelength of 514 nm and a solution of DPPH be found in many fruits, vegetables, nuts, seeds,
(1,1-diphenyl-2-picrylhydrazyl) acted as a free radical, flowers, leaves, barks, etc.
and ascorbic acid as a positive control. The amount Furthermore, flavonoids belong to one group
of antioxidant activity was characterized by the IC50 of aromatic compounds including polyphenols and
(inhibitory concentration), that is a concentration of antioxidants and the abundant numbers in nature,
the sample solution indicated a potential antioxidant so they are more widely used than other compounds
that was needed to reduce DPPH by 50 % (Arung, as antioxidants (Waluyo & Pasaribu, 2013).
Kusuma, Christy, Shimizu, & Kondo, 2009). Tannins are usually found in almost all green
The activity of the antioxidant from the sample plants, and more in barks. The test results show
extract was determined based on the relative all three parts (flesh, seed and percarp) of the fruit
inhibition percentage compared to the control using contain tannins. Taste astringent on some fruits
the following equation: including fruit Selekop indicates the presence of
A DPPH  A sample tannins. The more tannin content, the greater the
Relative Inhibition Activity (%)   100% antioxidant activity presents due to the fact that
A DPPH
tannins contain polyphenolic compounds that have
Where ADPPH represents the absorption from free radical catcher activities (Malangngi, Sangi, &
DPPH and Asample the absorption from the sample. Paendong, 2012).
The result was then integrated into the regression Table 1. Phytochemical in extracts of flesh, seed
equation with extract concentration as the abscissa and pericarp of Selekop
(x-axis) and the percentage of antioxidant inhibition
as the ordinate (y-axis). The value of IC50 was Parts of Selekop fruit
Phytochemicals
Flesh Seed Pericarp
calculated when the value of inhibition percentage Flavonoids + + +
reached 50 % using the equation y = ax + b. Alkaloids - + +
Saponins + + +
RESULTS AND DISCUSSION
Tannins + + +
In this study, ethanol was used for extraction Triterpenoids - + +
because it is an alcohol derivative that would easily Steroids - - -
dissolve the groups of secondary metabolites, both Remarks: + = present; - = absent
polar and non-polar. Furthermore, ethanol would not Alkaloid content in fruits were found only in the
hydrolyze compounds; be volatile at a low boiling pericarp and seed but not in the flesh. Alkaloids are
point, and good for thermolabile compounds. As one of the secondary metabolites found in plants,
stated by Harborne (1987), ethanol was good to be which can be found in leaves, branches, seed, and
used for preliminary extraction. bark, (Aksara, Musa, & Alio, 2013). Novelli, Lorena,
Phytochemical testing was conducted to & Antonella (2014) reported that alkaloid contained
determine the content of secondary metabolites in Ficus benjamina L. crude extract, had a strong
contained in the each of the extract in order to find correlation with its antioxidant.
out the potential utilization by testing the active Similar to alkaloids content, triterpenoid
compounds such as flavonoids, tannins, alkaloids, was not found in the flesh but only in the seed and
triterpenoids, steroids, and saponins. pericarp. Harborne (1987), stated that triterpenoids
Phytochemical contents in the crude extract compounds generally found in resin and sap,
of flesh, seed and pericarp from Selekop fruit are classified as essential oils, were volatile and fragrant.
shown in Table 1. The data shows that the extract Steroids were not detected in all of the extracts.
from flesh only contains flavonoids, tannins, and Generall, steroids are found in leaf photosynthesis
saponins, while the extracts from the seed and the and are rarely found in fruits. Steroids in plants work
pericarp contain almost all components except for to increase the rate of plant cell renewal, retard
steroids which were not found in all three parts of leaves aging (senescence), hamper leaves drop,
the fruit extract. and stimulate cell renewal in plant shoots.
217

Heriad Daud Salusu et al.: Phytochemical Screening and Antioxidant Activity of Selekop................................................

Saponins were detected in all parts of the fruit. antioxidant activity expressed by Ahmed et al. (2014)
This confirms the statement of Fahrunnida & Pratiwi in his study of Calamus tenuis Roxb suggested a
(2015), which reported that the highest amount of correlation between antioxidant activity and the total
saponin was found on the fruit rather than on the flavonoid content. Regarding with phytochemicals,
leaves and leaf stalks in the case of starfruit. the peel and seed were higher than flesh and useful
Result of the analysis of the antioxidant activity as reducing agents and radical scavengers. These
based on IC50 values and compared with ascorbic antioxidants activities had a close relationship with
acid as a positive control is shown in Table 2. its total phenols and flavonoids (Bakar, Karim, &
Based on IC50 values antioxidant of sample Perisamy, 2015).
is presented in Table 2, as well as ascorbic acid as
CONCLUSION AND SUGGESTION
positive control.
This study showed that extract of pericarp,
Table 2. IC50 value of extract flesh, seed and
flesh and seed from Selekop fruit can serve as
pericarp on Selekop fruit potential source of natural antioxidants. Out of the
Samples IC50 (ppm) three extracts, seed in particular, prospective to be
Flesh 122.51 developed as a source of natural antioxidants and
Seed 63.30 has the higher medicinal benefit. Further studies
Pericarp 53.21 are required for the isolation and characterizing
Ascorbic acid 3.06 of antioxidant components as well as the active
phytochemicals responsible their biological
The strength of the antioxidant activity was activities.
determined based on IC50 value, referring to Jun et
al. (2003) where the value of < 50 ppm is considered ACKNOWLEDGEMENT
as strong, 50-100 ppm as active, 100-250 ppm as This research support in part from grant of
moderate, 250-500 ppm as weak, and > 500 ppm Newton Fund Institutional Links (ID 172719105).
as inactive.
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