Antioxidant Activity of Melon Fruit (Cucumis melo L.

‘GMP’) Ethanolic Extract

Muhammad Zulfikar1, Faiza Senja Widya1, Wiko Arif Wibowo2, Budi Setiadi
Daryono2 and Slamet Widiyanto1, a)
1
Laboratory of animal physiology, Faculty of Biology, Universitas Gadjah Mada, Jalan Teknika Selatan, Sekip
Utara, 55281 Yogyakarta, Indonesia.
2
Laboratory of Genetic and Breeding, Faculty of Biology, Universitas Gadjah Mada, Jalan Teknika Selatan, Sekip
Utara, 55281 Yogyakarta, Indonesia.
a)
Corresponding Author: slametbio@ugm.ac.id

Abstract : Melon (Cucumis melo L.) cv. Gama Melon Parfum (GMP) is a new melon cultivar made by cross breeding
between female ♀NO-3 (Natsumo Omoide) dan ♂MR-5 (Miyamauri) parents has a distinctive scent at ripening stage.
During the ripening stages there are chemical changes of the fruit including antioxidant properties. The aims of this
research were to study the changes of antioxidant activity and antioxidant compound during ripening stages of melon cv.
Gama Melon Parfum (GMP). Melon with three ripening stages (10 DAA, 20 DAA and 30 DAA) were harvested and
analyzed their antioxidant activity, ascorbic acid, and flavonoid. The methods were used flavonoid quantitative analysis
by extraction and spectrophotometer, antioxidant activity was quantified by free- radical 2,2-diphenyl-1-picrylhydrazyl
(DPPH) method and ascorbic acid content was determined by iodometric method. The results showed that antioxidant
activity, ascorbic acid, and flavonoid increased during ripening. Content of antioxidant activity 10 DAA (27,29 %), 20
DAA (48,56 %) and 30 DAA (73,78 %), content of ascorbic acid 10 DAA (19,53 mg/100g), 20 DAA (23,33 mg/100 g)
and 30 DAA (27,19 mg/100g), and flavonoid content 10 DAA (70,62 μg/g), 20 DAA (73,14 μg/g) and 30 DAA (81,87
μg/g). Antioxidant activity positively correlated with ascorbic acid and flavonoid content.

INTRODUCTION
The fruits are a source of antioxidant compounds, such as phenolics, vitamins, carotenoids and minerals, which
contribute to their chemopreventive potential [1]. All these substances are related to lower the risk for the
development of health problems such as certain types of cancer, cardiovascular diseases, type 2 diabetes and obesity.
Melon cv. Gama Melon Parfum (GMP), a new cultivar of melon, has been developed in the Laboratory of
Genetic and Breeding, Faculty of Biology, Universitas Gadjah Mada (UGM), Indonesia. Melon cv. Gama Melon
Parfum (GMP) has a phenotypic character stand out is that it has a very fragrant fruit aroma like perfume but tastes
bitter, so it is not suitable for consumption. Cultivar Gama Melon This perfume is the result of breeding between the
melon breeders ♀NO3 (Natsumo Omoide) and ♂MR5 (Miyamauri) [2].
Volatile compounds and antioxidant compounds in melons will form and develop during fruit development [3].
It has been reported that the levels of high bioactive compounds of fruits are strongly influenced by ripening stage
[4, 5]. The study of changes in bioactive compounds during ripening of fruit has great relevance both to human
health and commercial purposes. Those study provides valuable information to understand the alteration of those
compounds and to evaluate the best harvesting time to get the highest antioxidant potential. Harvesting time is
essential to get a high-quality fruit with good nutrition potential. As a new melon cultivar, the study of antioxidant
potential in this fruit is still rare. The study that focused on antioxidant activity alteration during ripening stages of
melon cv. Gama Melon Parfum (GMP) has never been done before. Therefore, the aims of this research were to
study the changes of antioxidant activity and antioxidant compound during ripening stages of melon cv. Gama
Melon Parfum (GMP).

The 6th International Conference on Biological Science ICBS 2019

AIP Conf. Proc. 2260, 040029-1–040029-5; https://doi.org/10.1063/5.0015748
Published by AIP Publishing. 978-0-7354-2020-5/$30.00

040029-1
 MATERIALS AND METHODS

Plant Materials and Chemicals

Melon cv. Gama Melon Parfum (GMP) was cultivated in Agricultural Training, Research and Development
Station of Universitas Gadjah Mada, Desa Kalitirto, Berbah Sub-District, Sleman Regency of the Special Region of
Yogyakarta. Melon with three ripening stages (10 DAA, 20 DAA dan 30 DAA).
Etanol 96%, aluminium trichloride (AlCl3), amilum, yodium, aquades, n-Butanol, quercetin, Sodium nitrite,
aluminium chloride, sodium hydroxide, rutin, acetone, 1,1-diphenyl-2-picrylhydrazyl (DPPH).

Extract Preparation
Dried melon flesh were pulverised into fine powder using a stainless steel blender and mixed with methanol at
the ratio of 1:10 (w/v). Then these mixtures were manually swirled for 15 min and left in a sonicator for 60 min.
After that, the mixture was incubated in an incubator shaker at 40 °C with 140 rpm for 48 hours and the mixture was
filtered through a whatman filter paper no 1 and the filtrate obtained was evaporated, concentrated at room
temperature and stored at 4 °C. The extract obtained after double extraction with solvent was dar k yellowish brown
in color.
Ascorbic Acid Content (AAC)
Determination of vitamin C levels using the iodine titration method. Melon meat is crushed in a blender of 300 g.
Melon flesh that has been crushed is taken 30 g and put in a 100 mL measuring flask and add aquades. Filter with
centrifuges to adjust the filtrate. 10 mL of filtrate is taken with a pipette and put in 125 mL erlenmeyer, add 2 mL of
1 % starch solution (soluble starch) and 20 mL of distilled water, then titrate with 0.01 N standard iodine.

Total Flavonoid Content (TFC)

The total flavonoid content (TFC) of melon cv. Gama Melon Parfum (GMP) extracts was determined as [6].
Briefly, 0.5 mL of diluted sample was mixed with 2 mL of deionized water followed by the addition of 150 μL of
5% (w/v) sodium nitrite solution. After 6 min, 150 μL of 10% (w/v) aluminum chloride was added and the mixture
was incubated for another 6 min. Then, 2 mL of 4% (w/v) sodium hydroxide was added and the solution was
immediately made up to 5 mL with deionised water, mixed thoroughly and placed in the dark at room temperature
for 15 min. The absorption of the solution was measured at 510 nm against a reagent blank. Rutin was used as a
standard and the TFC was expressed as mg of Rutin Equivalents (RE) per g of bitter melon powder on a dry basis
(mg RE/g).

Statistical Analysis
Results were expressed as mean values with standard deviations (mean ± SD). Significant differences between
mean values in the different experiments were tested using the one-way analysis of variance (ANOVA) and the
Bonferroni post-hoc test using a 5% significance level (P < 0.05). All statistical analysis was performed using SPSS
23.

Radical Scavenging Activity (DPPH)

This assay was carried out according to Zegin et al [7] with some modifications in which increase in absorbance
due to reduction of the DPPH radical determined spectrophotometrically at 515 nm for 60 min based on preliminary
experiments. In this method, a total of 0.1 mg/mL of the extracts and synthetic antioxidants (catechin and BHT) in
ethanol were added into 3 mL of ethanol solution of DPPH. The mixture was vortex for 20 s at room temperature.
The blank sample (A blank) absorbance which contains 0.1 mL ethanol instead of extracts was also carried out.
Absorbance measurements were done immediately in a 10 mm quartz cell from 1 min up to 60 min with 10 min
intervals using a UV-visible spectrophotometer. The percentage of DPPH radical-scavenging activity was calculated
using the following equation:

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 Percentage Radical Scavenging Activity = Abs.blank - Abs.sample x 100%
Abs.blank

Statistical Analysis
The effects of ripening stages on the antioxidant properties of melon cv. Gama Melon Parfum (GMP) were
analyzed by oneway analysis of variance (ANOVA). Tukey test was carried out to identify significant differences
between ripening stages. Mean values with p < 0.05 were considered statistically significant. For analysis of
correlation between antioxidant capacity and antioxidant compounds, the regression analysis was carried out. All
statistical analysis were performed using SPSS 23.

RESULT AND DISCUSSION

This research uses Melon cv. Gama Melon Parfum (GMP) with three stages of maturation which are 10DAA,
20DAA and 30 DAA.
Vitamin C, also known as ascorbic acid (AA), is considered as an enzymatic cofactor. It plays a key role as an
essential compound for plant tissues due to its considerable antioxidant role [8]. Vitamin C is one of the most potent
antioxidant vitamins required for growth, healthy body tissue and wound repair and for an efficient immune system.

Ascorbic acid content (AAC)

40
a
30
(mg/100g)

b
c
20
10
0
DAA 10 DAA 20 DAA 30
Ripening stage (DAA)

FIGURE 1. Ascorbic acid content (AAC) melon cv. Gama Melon Parfum (GMP).

The results show that ascorbic acid content increased during ripening stages as shown in Fig.1. Ascorbic acid
reached the highest value at ripening stage DAA 30 (27.19 mg/100g). It showed that ascorbic acid was synthesized
during ripening of melon cv. Gama Melon Parfum (GMP). These results are agreement with several studies reported
in tomato. The increase in ascorbic acid content with ripening has been attributed to the increase in lipid
peroxidation considering that fruit ripening which is an oxidative phenomenon that requires turnover of active
oxygen species [9]. Under this condition, antioxidant compounds including ascorbic acid usually increased.
Flavonoids are the most common group of polyphenolic compounds that are found ubiquitously in plants, which
usually are very effective antioxidants. They are important in the plants for normal growth, development and
defense against infection and injury. These plant secondary metabolites also show anti-allergic, anti-inflammatory,
antimicrobial and anticancer activities [10]. Flavonoids are a class of secondary plant metabolites with significant
antioxidant and chelating properties [11].

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 Total flavonoid content (TFC)
100 a
c b
80
60

(μg/g)
40
20
0
DAA 10 DAA 20 DAA 30
Ripening stage (DAA)

FIGURE 2. Total flavonoid content (TFC) melon cv. Gama Melon Parfum (GMP).

Total flavonoid content increased during ripening stages as shown in Fig.2. Flavonoid reached the highest value
at ripening stage DAA 30 (81.87 μg/g). It showed that flavonoid was synthesized during ripening of melon cv.
Gama Melon Parfum (GMP), this indicates that the flavonoids are the major phenolic compounds present in
cantaloupe plant. Variation in flavonoid content in fruits is strongly influenced by extrinsic factors such as fruit type
and growth, season, climate, degree of ripeness, food preparation and processing [12]. A study done by Zuhair et al
[13] reported that total flavonoid content increases significantly during the ripening of papaya through different
maturity stages.
Radical scavenging activity (DPPH) increased during ripening stages as shown in Fig.3. DPPH reached the
highest value at ripening stage DAA 30 (73.78 %RSA). In general, the antioxidant activity of melon cv. Gama
Melon Perfume (GMP) increases during ripening, due to an increase in acid ascorbic and total flavonoid content. It
is known that flavonoid compounds are responsible for scavenging free radicals. However, antioxidant activity
depends not only on the flavonoids themselves, but also on them other compounds such as ascorbic acid, phenols,
and carotenoids [14]. Because of this, antioxidant activity in fruit cannot be obtained only with flavonoid content,
but also in the combined action of antioxidants present in fruits.

Radical scavenging activity (DPPH)

100
a
80
b
% RSA

60
c
40
20
0
DAA 10 DAA 20 DAA 30
Ripening stage (DAA)

FIGURE 3. Radical scavenging activity (DPPH) melon cv. Gama Melon Parfum (GMP).

There is a relationship between antioxidant activity and antioxidant content of cv melons. Gama Melon Perfume
(GMP). Antioxidant activity (DPPH test) is positively correlated with total flavonoids, and ascorbic acid content.
The results are in accordance with several studies published in bitter melon [15] and guava [16].

CONCLUSIONS
Melon cv. Gama Melon Perfume (GMP) provides a variety of dietary antioxidants such as flavonoids and
ascorbic acid. All stages of melon cv maturation. Gama Melon Perfume (GMP) shows an increase in free radicals.
Melon cv. Gama Melon Perfume (GMP) in stage DAA 30 showed the highest antioxidant activity and was also so
with total flavonoid content and ascorbic acid.

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 ACKNOWLEDGMENT
This study was funded by a grant from Rekognisi Tugas Akhir (RTA) 2019. The authors thank Prof. Dr. Budi S.
Daryono, M.Agr.Sc. as providing and growing Melon cv. Gama Melon Parfum (GMP).

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