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Antioxidant activity and chemical constituents of some Indonesian fruit peels

Article  in  Medicinal Plants - International Journal of Phytomedicines and Related Industries · March 2014
DOI: 10.5958/j.0975-6892.6.1.006

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 Antioxidant activity and chemical constituents of some Indonesian fruit peels 1
Research Article

Antioxidant activity and chemical constituents of some

Indonesian fruit peels
Muhtadi1, Haryoto1, Tanti Azizah Sujono1, Peni Indrayudha1, Andi Suhendi1 and Khong Heng Yen 2
1Faculty of Pharmacy, Muhammadiyah University of Surakarta, Jl. Achmad Yani Tromol Pos 1,

Pabelan Kartasura, Surakarta 57102, Indonesia

2School of Chemistry nd Environtmental Studies, Faculty of Applied Sciences, University Teknologi MARA,

94300 Kota Samarahan, Sarawak, Malaysia

ABSTRACT

The ethanolic extracts and their fractions of some Indonesian fruit peels were screened for their free radical
scavenging properties using vitamin-E as standard antioxidant. Free radical scavenging activity was evaluated
using 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical. The strongest antioxidant activity was shown by the
Rambutan (Nephelium lappaceum Linn.), followed by Kelengkeng (Euphoria longan Lour. Steud) and Durian
(Durio zibethinus Murr.) with IC50 values of 7.74, 11.85, and 28.83 µg/mL, respectively comparable to vitamin-
E (IC50 = 8.48 ± 0.1 µg/mL). The ethyl acetate fraction of these fruit peels extracts demonstrated the most active
antioxidant activity compared with hexane, chloroform and methanol fractions. The chemical constituents of
Rambutan and Durian were successfully isolated using vacuum liquid chromatography and radial chromatography
technique, and these structures were characterizated based on the proton (1H) and carbon (13C) NMR spectra.
The isolated compounds were ethyl gallate (1) from Rambutan (Nephelium lappaceum Linn.) rind, 4,4-dimethyl-
poriferasta-18(19)-en-3-ol (2) and 3α-E-ferulyloxy-lup-20(29)-en-28-oic acid (3) from Durian (Durio zibethinus
Mur r.) peels. The results showed that prospective antioxidant contents in Rambutan’s fruit peel extract was higher
than Kelengkeng and Durian fruit peel extracts. [Medicinal Plants 2014; 6(1) : 00-00]

Keywords : Antioxidant activity, fruit peel, Rambutan (Nephelium lappaceum Linn.), Kelengkeng (Euphoria
longan Lour. Steud), Durian (Durio zibethinus Murr.)

INTRODUCTION these fruits are used as the valued materials, instead

Indonesia is one of the country in southeast Asia which
is rich with tropical fruits, besides Malaysia, Myanmar
and Thailand (Chomchalow et al., 2008; UN ESCAP,
2007). Rambutan ( Nephelium lappaceum Linn),
Kelengkeng (Euphoria longan Lour. Steud) and Durian
(Durio zibethinus Murr.) are the main fruits commonly
found in Indonesia. Unfortunately, not all the parts of
Corresponding author : Muhtadi
e-mail : muhtadi@ums.ac.id
Received : ; Accepted :
doi: 10.5958/j.0975-6892.5.2.009
their peels are still regarded as waste. Previous chemical
studies showed that these fruit peels exhibit bioactive
compounds. For instance, the Mangosteen peel was
proven rich in a-mangostin and xanthone derivatives,
in which antioxidant potential are identif ied (Jung et
al., 2006; Jose et al, 2008). Furthermore, our previous
study has also shown that the extract of Euphoria
longan peels contains a potent substance as antioxidant
and antibacterial activity (Muhtadi et al., 2013).
The present study has been undertaken to assess the
antioxidant activity and chemical constituents of three
Indonesian fruit peels, viz., Rambutan (Nephelium
lappaceum Linn), Kelengkeng (Euphoria longan Lour.
Steud) and Durian (Durio zibethinus Murr.).

Medicinal Plants, 6(1) March 2014

2 Muhtadi et al.
MATERIAL AND METHODS
Plant Material
The fruit peel samples of Nephelium lappaceum ,
Euphoria longan and Durio zibethinus were collected
from local area of Surakarta, Central of Java, Indonesia.
Herbarium voucher specimens were prepared and
deposited in the Herbarium of Pharmacy Biology at
Faculty of Phar macy, Muhammadiyah University of
Surakarta, Indonesia.
Spectral Studies
Infrared spectra were measured using the universal
attenuated total reflection (UATR) technique on a
Perkin-Elmer 100 series. NMR spectra were obtained
using JEOL 300MHz FT NMR spectrometer using
tetramethylsilane (TMS) as internal standard. Ultra
violet spectra were recorded in acetone on a Shimadzu
UV-160°, UV-Visible Recording Spectrophotometer.
Extraction and isolation
The samples were air-dried and ground to powder and
extracted with ethanol for 3 X 24 hours at room
temperature. The extracts were fractionated with the
par tition method, using n-hexane, chlorofor m, ethyl
acetate, and methanol-water solvents. The extracts and
their fractions were dried under reduced pressure and
stored at 4ºC until used. The ethanol extract of Durian
peels (350.0 mg) was fractionated and purif ied using a
stepwise gradient system (SiO 2, hexane/chloroform,
started 3.5:6.5) on radial chromatography technique to
give 23 fractions. Fractions 7-10 were combined and
characterized 3α-E-ferulyloxy-lup-20(29)-en-28-oic
acid (3) (6 mg), and from fractions 17-19 were combined
and characterized 4,4-dimethyl-poriferasta-18(19)-en-3-
ol (2) (2 mg). On other hand, the ethanol extract of
Ramb utan peels (422.0 mg) over a silica radial
chromatography (chloroform/hexane, chloroform/ethyl
acetate and chloroform/methanol gradient) provided 22
fractions. Fractions 14-19 were combined and
characterized as ethyl gallate (1) (30 mg).
The details of fractions having characterstics of
ethyl gallate, 4,4-dimethyl-poriferasta-18(19)-en-3-ol
and 3α-E- ferulyloxy-lup-20(29)-en-28-oic acid are
listed below:
The Ethyl gallate (1)
White amor phous powder UV λ max 215, 273 nm
(methanol); 1H-NMR (300 MHz; aceton-d 6): δ 7.13 (s,
2H, H-2/6), 4.26 (q, J = 7.1 Hz, 2H, H-8), 1.32 (t, J =
7.1 Hz, 8H, H-9); 13C NMR (75 MHz; acetone): δ 165.8
(C-7), 145.16 (C-5), 145.11 (C-3), 137.8 (C-4), 121.2
(C-1), 108.8 (C-2/C-6), 60.0 (C-8), 13.7 (C-9).
4,4-dimethyl-poriferasta-18(19)-en-3-ol (2)
White amor phous powder UV λ max 215, 273 nm
(methanol); 1H-NMR (300 MHz; aceton-d6): δ 5.24 (m,
1H, H-19), 1.14 (s, 3H, H-28), 1.00 (s, 3H, H-25), 0.97
(d, J = 7.5 Hz, 3H, H-29), 0.96 (s, 3H, H-24), 0.91 (t, J
= 7.5 Hz, 3H, H-23), 0.85 (s, 3H, C-27), 0.79 (s, 3H, H-
26). 13C NMR (75 MHz; acetone): δ 135.9 (C-18),
126.2 (C-19), 78.6 (C-3), 57.0 (C-5), 56.2 (C-21), 53.9
(C-9),48.5 (C-14),46.8 (C-13),42.9 (C-10), 39.9 (C-17),
39.8 (C-8), 39.5 (C-20), 39.3 (C-4), 37.6 (C-12), 34.0
(C-1), 31.4 (C-11),29.2 (C-15), 28.8 (C-2), 28.7(C-25),
28.1 (C-16), 25.0 (C-7), 24.1 (C-22), 24.0 (C-28), 21.5
(C-29), 19.1 (C-6), 17.7 (C-27), 17.5 (C-23), 16.4 (C-
24), 15.9 (C-26).
3α-E-ferulyloxy-lup-20(29)-en-28-oic acid (3)
White amor phous powder UV λ max 215, 273 nm
(methanol); 1H-NMR (300 MHz; aceton-d 6): δ 8.23 (s,
1H), 7.60 (d, J = 15.8 Hz, 1H. H-2’), 7.34 (d, J = 1.8 Hz,
1H, H-9’), 7.18 (dd, J = 8.2, 1.9 Hz, 1H, H-5’), 6.88 (d,
J = 8.2 Hz, 1H, H-8’), 6.71 (d, J = 15.8 Hz, 1H, H-3’),
4.89 (m, 1H, H-3), 4.71 (s, 1H, H-29a), 4.59 (s, 1H, H-
29b), 3.92 (s, 3H, OCH3), 1.70 (s, 3H, H-30), 1.28 (s,
3H, H-27), 1.01 (s, 3H, H-26), 0.95 (s, 3H, H-23), 0.85
(s, 3H, H-24), 0.75 (s, 3H, H-25). 13C NMR (75 MHz;
acetone): δ 184.5 (C-28), 177.6 (C-1’), 151.6 (C-20/C-
7’), 150.1 (C-6’), 141.4 (C-3’), 128.1 (C-4’), 123.8 (C-
9’), 122.2 (C-8’), 116.3 (C-2’), 111.6 (C-5’), 110.0 (C-
29), 78.6 (OCH3), 56.8 (C-17), 56.3 (C-9), 51.4 (C-19),
49.9 (C-18), 47.9 (C-18), 43.2 (C-14), 41.5 (C-8), 39.60
(C-4), 39.56 (C-1), 39.0 (C-13), 38.0 (C-10), 37.5 (C-
22), 35.2 (C-16), 32.8 (C-15), 31.3 (C-21), 30.4 (C-2),
28.6 (C-23), 28.2 (C-12), 26.4 (C-7), 21.7 (C-11), 19.5
(C-30), 19.1 (C-6), 16.62 (C-24), 16.53 (C-25), 16.1 (C-
26), 15.0 (C-27).
Antioxidant Activity
The extracts and their fractions were evaluated for the
antioxidant activity using the 1,1-diphenyl-2-
picrylhydrazyl (DPPH) as described by Loo et al.,
(2008), with minor modif ications. Each sample of
stock solution (1.0 mg/mL) was diluted to a f inal
concentration of 1000, 500, 250, 125, 62.5, 31.3, 15.6,
and 7.8 µg/mL. Then, a total of 3.8 mL of 50 µM DPPH
methanolic solution (1 mg/50 mL) was added to 0.2
mL of each sample solution and allowed to react at
room temperature for 30 min. The absobance of the
Medicinal Plants, 6(1) March 2014
 Antioxidant activity and chemical constituents of some Indonesian fruit peels 3

mixtures was measured at 517 nm. A control was
prepared without sample or standard and measured
immediately at 0 min. The precent inhibition (I %) of
DPPH radical were calculated as follows:
Ablank – Asample
I % = × 100
A blank
where Ablank is the absorbance value of the control
reaction (containing all reagents except the test sample)
and Asample is the absorbance value of the test samples.
The sample concentration provide 50% inhibition (IC50)
was calculated by plotting inhibition percentages
against concentrations of the sample. All tests were
carried out in triplicate and IC50 value were reported as
means ± SD of triplicates.
RESULTS AND DISCUSSION
Antioxidant activity
The ethanolic extract and their fractions of fruit preels
of Rambutan, Kelengkeng and Durian were evaluated
for their antioxidant properties by DPPH radical
scavenging (Table 1). It has been reported that free
Table 1. Antioxidant activity of some Indonesian fruit
peelsa
Sample DPPH IC 50
(mg/mL)
radical scavenging activity is greatly influenced by
the phenolic component of samples (Cheung et al.,
2003). In the present study, ethanolic extract and their
fractions of Rambutan fruit peels demonstrated more
activity than vitamin-E and other extract and fractions.
This indicated abundance of phenolic compounds and
prospective antioxidant ingredients in the Rambutan
fruit peels compared to Durian and Kelengkeng. All of
the ethyl acetate fractions were more active than the
crude extract and their fraction, which showed that the
phenolic compounds were readily soluble in ethyl
acetate solvent.
The isolated chemical constituents
One chemical constituent namely ethyl gallate (1) had
been isolated from Rambutan and two compounds from
Durian, namely 4,4-dimethyl-poriferasta-18(19)-en-3-ol
(2) and 3α-E-ferulyloxy-lup-20(29)-en-28-oic acid (3).
These compounds (Fig. 1) were elucidated based on
their proton and carbon NMR spectra, and the structure
of the ethyl gallate (1) was conf irmed with reference
spectral data (Mahajan and Pai, 2010), 4,4-dimethyl-
poriferasta-18(19)-en-3-ol (2) with the Ersoz’s NMR
spectral data (Ersoz et al., 2002) and 3α-E-ferulyloxy-
lup-20(29)-en-28-oic acid (3) with Khumar and Sharma’s
spectral data (Khumar and Sharma, 2006).
The ethyl gallate (1) has been reported to exhibit
good antioxidant properties (Romero et al., 2010; Kubo
et al., 2010; Bonacorsi et al., 2011), antiviral (Juliana,
2006), and induces apoptosis of HL-60 Cells (Kim et

Ethanol extract of Kelengkeng fruit peels 11.85 ± 0.96

Hexane fraction of Kelengkeng fruit peels 374.81 ± 9.73
Ethyl acetate fraction of Kelengkeng fruit 9.23 ± 0.15 HO

peels O
HO
Methanol-water fraction of Kelengkeng 42.89 ± 1.91 OCH2CH3 HO
fruit peels HO

Ethanol extract of Durian fruit peels 28.83 ± 0.48 Ethyl gallate (1) 4,4-dimethyl-poriferasta-18(19)-en-3-ol (2)
Chloroform fraction of Durian fruit peels 32.81 ± 2.64
Ethyl acetate fraction of Durian fruit peels 14.91 ± 3.23
Ethanol extract of Rambutan fruit peels 7.74 ± 0.76
Chloroform fraction of Rambutan fruit 6.64 ± 1.14 O

peels O OH

Ethyl acetate fraction of Rambutan fruit 4.29 ± 0.72 O

peels
HO
Methanol-water fraction of Rambutan fruit 8.34 ± 0.59
OCH3
peels
Vitamin E as standard 8.48 ± 0.77 3-E-ferulyloxy-lup-20(29)-en-28-oic acid (3)

aData represent meand ± standard deviation of three independent Fig. 1. The chemical structure of the isolated compounds from
experiment. Rambutan and Durian fruit peels

Medicinal Plants, 6(1) March 2014

 4 Muhtadi et al.
al., 2012). In addition, Shabana et al. (2013) also
reported that the sterol glycosides from fruit peels of
Solanum melongena L. have a promising anticancer
activity against hepatocellular carcinoma. The sterol
glycosides has similar skeleton as that of 4, 4-dimethyl-
poriferasta-18(19)-en-3-ol (2) and therefore, compound
2 is expected to have cytotoxic effect against cancer
cell lines whereas 3α-E-ferulyloxy-lup-20(29)-en-28-oic
acid (3), is one of the alkyl ferulate derivative which
is expected to be antioxidative (Kikuzaki et al., 2002).
CONCLUSION
The bioassay studies showed that the peels of Rambutan
(Nephelium lappaceum Linn), Kelengkeng (Euphoria
longan Lour. Steud) and Durian (Durio zibethinus Murr.)
exhibited very potent anti-oxidative properties.
Rambutan ( Nephelium lappaceum Linn) peels
demonstrated the highest antioxidant acitivity,
comparable with vitamin-E. In addition, the ethyl
gallate was succesfully isolated from Rambutan fruit
peels extract whereas 4,4-dimethyl-poriferasta-18(19)-
en-3-ol and 3α-E-ferulyloxy-lup-20(29)-en-28-oic acid
were isolated from Durian fruit peels extract.
ACKNOWLEDGEMENT
The authors wish to thank Muhammadiyah University
of Surakarta and Ministry of Education and Cultural
Republic, Indonesia for Excellent Research Grant
Scheme for f inancial support. The authors also wish to
thank the staff of Phar macy Biology Laborator y,
Universitas Muhammadiyah Surakarta for their
assistance in the determination of the samples and
Universiti Teknologi MARA Sarawak for laboratory
facilities.
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