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Online - 2455-3891
Print - 0974-2441Vol 9, Suppl. 3, 2016
ANTIOXIDANT ACTIVITY OF COMBINATION BANANA PEEL (MUSA PARADISIACA) AND
WATERMELON RIND (CITRULLUS VULGARIS) EXTRACT IN LOTION DOSAGE FORM
NATALIA ALAMSYAH1*, RATNA DJAMIL2, DENI RAHMAT3
1Faculty of Pharmacy, Pancasila University, Jakarta 12640, Indonesia. 2Departement of Biological Pharmacy, Faculty of Pharmacy,
Pancasila University, Jakarta 12640, Indonesia. 3Departement of Pharmaceutical Technology, Faculty of Pharmacy, Pancasila University,
Jakarta 12640, Indonesia. Email: natalia_alamsyah@yahoo.com
Received: 28 August 2016, Revised and Accepted: 14 September 2016
ABSTRACT
Objective: Test the antioxidant activity of banana peel extract, watermelon rind extract, and combinations, also in lotion dosage form.
Methods: The antioxidant activity of each extract and their combinations were tested with 2,2-diphenyl-1-picrylhydrazil method and formulated into
a lotion dosage forms. Evaluation of the lotion dosage forms, including organoleptic, homogeneity, pH, viscosity and rheology, microbiology, acute
dermal irritation test, as well as the antioxidant activity test.
Results: Antioxidant activity test on the extract showed inhibitory concentration 50% (IC50) values of banana peel is 64.03 part per million (ppm)
and watermelon rind extract of 300.12 ppm. Combination of banana peel and watermelon rind extract with three different concentrations, IC50:IC50,
inhibitory concentration at 75% (IC75):inhibitory concentration at 25% (IC25), IC25:IC75 resulted the best antioxidant activity is a combination of banana
peel, and watermelon rind extract IC75:IC25 with IC50 value of 177.56 ppm. The antioxidant activity test in lotion dosage form showed IC50 values of
lotion base is 853.16 ppm, lotion banana peel extract is 472.50 ppm, lotion watermelon rind extract is 496.71 ppm, and the combination of banana
peel and watermelon rind extract are 300.04 ppm.
Conclusion: Lotion dosage form formula that meets the requirements of quality, efficacy, and safety with the best antioxidant activity is lotion dosage
form combination of banana peel and watermelon rind extract formula (IC75:IC25).
Keywords: Antioxidant activity, Banana peel (Musa paradisiaca), Watermelon rind (Citrullus vulgaris), Combination of banana peel and watermelon
rind extract, Lotion of antioxidant.
INTRODUCTION
Damages from free radicals at the cellular level cause cell
inflammation, increasing the risk of damage from sunlight radiation,
and related development of skin cancer. Antioxidants work by
preventing onset of disorder at the cellular level. The first body
protection that most simple and flexible is on the outside of the body,
that cause more appearances of antioxidant products on the market
as a cosmetic preparation. But currently, a large part of preparations
that available using chemical antioxidants. The development of
natural antioxidants is still continuing to be studied because it is
difficult to maintain the antioxidant activity of material and its
stability [1-3].
Waste of yellow banana peels is rich in flavonoids and another
phenolic in addition to carbohydrates, cellulose, minerals such
as potassium and sodium. Flavonoid and phenolic are bioactive
compounds that useful as antioxidant [4-6]. According to Yunovan,
the antioxidant activity of “Kepok” banana peel extract in ethanol
has inhibitory concentration 50% (IC50) values of 156.67 part
per million (ppm) [7]. White layer of watermelon rind that is not
consumed contains citrulline. Citrulline is one of the antioxidants
that are beneficial for skin health [8]. According to Damayanti
et al. research, the antioxidant activity juice white layer of round
watermelon rind has IC50 values 214.369 ppm and oval watermelon
rind is 376.266 ppm [9]. Traditionally, watermelon that rich in water
content and other fruits that rich in antioxidants such as banana can
be used for skin detoxifying, providing essential nutrients to skin by
moisturizing, brightening and protecting the outer layer of skin [10].
The objective of this research was to study antioxidant activity of
banana peel, watermelon rind, and the combinations, also in lotion
dosage form using antioxidant testing 2,2-diphenyl-1-picrylhydrazil
(DPPH) to see the stability of extract antioxidant activity when
formulated into lotion dosage form.
METHODS
Chemicals and equipment
Three months old “Kepok” banana peels from Jakarta - Indonesia,
white layer of three months old watermelon rinds from Central
Java - Indonesia, ethanol 70%, glyceryl monostearate (Msnchem),
cera alba (Xiamen Fengstone), tween 80 (Kao Corporation), glycerin
(P&G Chemicals), paraffin liquid (Tudapetrol), nipagin (Ueno Fine
Chemicals), nipasol (Ueno Fine Chemicals), parfume, water, methanol
(Ajax Finechem), DPPH (Sigma-Aldrich), plate count agar (BD), potato
dextrose agar (BD), semi-micro scales, blender (Philips), rotary
evaporator (Buchi), freezedryer (Telstar), ultra violet–visible (UV-Vis)
spectrophotometer (Shimadzu 1240), viscometer (Brookfield type RV),
pH meter (Mettler Toledo).
Manufacture of extract
Banana peel extract
Banana skin was macerated using ethanol 70% ratio 1:2 using a shaker
for 24 h at room temperature, repeated 3 times and filtered. The filtrate
was concentrated by rotary evaporator.
Watermelon rind extract
White layer of watermelon rind is refined using blender and filtered.
The filtrate was powdered by freeze dryer.
Quality inspection of extract
Examinations involve in non-specific parameters are organoleptic,
drying shrinkage, specific gravity, water content, total ash and acid
Research Article
© 2016 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.2016.v9s3.14926
Asian J Pharm Clin Res, Vol 9, Suppl. 3, 2016, 300-304
Alamsyah et al.
301
insoluble ash, solvent residues, pesticide residues, heavy metal
contaminations, and microbial contaminations (total plate count and
total yeast mold). As well as specific parameters are phytochemical
screening (alkaloids, saponins, tannins, phenolics, flavonoids,
triterpenoids, steroids, glycosides, essential oils, quinones,
coumarin) and total flavonoid content by spectrophotometric
method [11,12].
Antioxidant activity test by DPPH method
Antioxidant activity was tested as a release of the stable DPPH
radical. In the radical form, DPPH maximum absorption at λ 515 nm.
The mixture reaction (5 ml) contain 1 ml of 0.4 mM DPPH (15.8 mg
DPPH in 100 ml of methanol) and various concentrations of test
solution dissolved in methanol. After incubation 30 minutes at 37°C
in the dark room, absorbance was measured at λ 515 nm using UV-
Vis spectrophotometer. Lower absorbance indicates higher restriction
of free radical scavenging (% inhibition). Determination of IC50 value
using linear regression between concentration and percent inhibition
by replacing the percent inhibition to 50 [13-18].
% Inhibition=(Abscontrol−Abssample)/Abscontrol×100%
Prepare a series of test solution with a concentration of 12.5; 25; 37.5;
50; and 62.5 mg/ml of banana peel extract, watermelon rind extract,
combination banana peel, and watermelon rind extract.
Manufacture of antioxidant lotion dosage form
Cera alba that has been melted is poured into beaker glass containing
paraffin liquid at the top of a hotplate temperature 40-60°C and stirrer
at speed around 700 rpm. Then, add glyceryl monostearate, tween 80,
glycerin, extracts which have been mixed with nipagin and nipasol,
water, and perfume as fragrances (Table 1) [19].
Evaluation of lotion dosage form
Organoleptic includes examination of color, odor, and sense when an
application on the skin.
Homogeneity by applying the preparation on glass object, drops of
methylene blue, and observed under a microscope.
pH tested using a pH meter.
Viscosity and rheology tested using a Brookfield viscometer type RV,
spindle number 4 with three different speed (2.5, 4, and 5).
Microbiology tested of total plate count and total yeast mold with
sample concentration 0.5 ml.
Acute dermal irritation test using albino rabbit (weight around 2 kg)
with test dose 0.5 g. Response rating on 1, 24, 48, and 72 h after the
opening of the patch. Ethical clearance approval from Airlangga
University, Surabaya – Indonesia.
Antioxidant activity test of preparation using DPPH method with test
solution concentration 20, 40, 60, 80, and 100 μg/ml.
RESULTS
Table 2 shows examination results of extract. A study of antioxidant
activity of banana peel extract, watermelon rind extract, and their
combinations are shown in Table 3. The combination of banana peel
and watermelon rind extract with three different combinations based
on the comparison IC value of its single extract, which combination of
banana peel:watermelon rind extract with a ratio of IC50:IC50, inhibitory
concentration 75% (IC75):Inhibitory concentration 25% (IC25), and
IC25:IC75. Banana peel extract, watermelon rind extract, and the best
combination of banana peel and watermelon rind extract (combination
of banana peel:watermelon extract [75:25]) are formulated into lotion
dosage form. Evaluation of lotion dosage form include organoleptic
(Fig. 1 and Table 4), homogeneity (Table 5), pH (Table 6), viscosity
(Table 7) and rheology (Fig. 2), microbiology (Table 8), acute dermal
irritation (Table 9), also antioxidant activity test of preparation
(Table 10).
DISCUSSION
The result from organoleptic and homogeneity test between lotion
formulas showed that there was no significant difference which
is white-like milk color, typically weak odor, soft and non-sticky
feel when applied to the skin, and the preparation is homogeneous
with emulsion type O/W (oil in water). The soft feeling in lotion
shows that glycerin as humectant and paraffin liquid as emollient
is function properly. Non-sticky feeling is because the base of lotion
is O/W (oil in water) with a composition ratio of water: oil around
80:20. pH of lotion between 5.0 and 5.5 which are in range pH of
skin acid mantle (4.5-6.5) so expectantly the lotion will not damage
the skin acid mantle layer. Test results show the viscosity of formula
lotion F1> F3> F2> F4. Formula F1 (base of lotion) resulted in the
most viscous lotion. Extract of banana peel and watermelon rind
are hydrophilic which causing an attractive force to water molecules
with a base of lotion, so the viscosity becomes more aqueous.
Rheology of lotion is plastic. Characteristic chart of plastic rheology
Table 1: Formulation of antioxidant lotion dosage form
Formula materials Composition (%)
F1 F2 F3 F4
Banana peel extract - 12.81 mg - 13.86 mg
Watermelon rind extract - 60.02 mg 21.65 mg
Glyceryl monostearate 2.8 2.8 2.8 2.8
Cera alba 7.5 7.5 7.5 7.5
Tween 80 4.7 4.7 4.7 4.7
Glycerin 10 10 10 10
Paraffin liquid 10 10 10 10
Nipagin 0.15 0.15 0.15 0.15
Nipasol 0.15 0.15 0.15 0.15
Parfume Qs qs qs qs
Water ad (ml) 100 100 100 100
Fig. 1: Antioxidant lotion dosage form. F1: Formula 1 is a control,
base of lotion without addition of extract. F2: Formula 2 is a
lotion banana peel extract. F3: Formula 3 is a lotion watermelon
rind extract. F4: Formula 4 is a lotion combination of banana
peel:watermelon rind extract ratio inhibitory concentration at
25%:inhibitory concentration at 75%
Fig. 2: Rheology of lotion
Asian J Pharm Clin Res, Vol 9, Suppl. 3, 2016, 300-304
Alamsyah et al.
302
Table 2: Quality inspection of extract
No Type of testing Results Method
Banana peel Watermelon rind
1 Organoleptic Organoleptic
Shape Viscous liquid Viscous liquid
Color Dark brown Yellow
Odor Latex aromatic Watermelon aromatic
2 Drying shrinkage 39.66% 39.91% Oven 105°C
3 Specific gravity 1.07 g/ml 1.12 g/ml Pycnometer
4 Water content 19.96% 33.30% Karl-Fisher
5 Total ash 2.92% 2.25% Gravimetry
6 Acid insoluble ash 0.08% 0.51% Gravimetry
7 Solvent residues Not detected Not detected GC
8 Pesticide residues LC MS/MS
Organophosphates Not detected Not detected
9 Heavy metal contaminations ICP-OES
Pb, Cd, Sn, HG, As Not detected Not detected
10 Microbial contaminations Total count
Total plate count <10 cfu/ml <10 cfu/ml
Total yeast mold <10 cfu/ml <10 cfu/ml
11 Total flavonoid 0.129% 0.133% Spectrophotometric
12 Phytochemical screening Qualitative
Alkaloids + +
Saponins + +
Tannins + -
Phenolics + -
Flavonoids + +
Triterpenoids + +
Steroids - -
Glycosides - -
Essential oils - -
Quinones - -
Coumarin - -
GC: Gas chromatography, LC-MS/MS: Liquid chromatography-mass spectrometry/mass spectrometry, ICP-OES: Inductively coupled plasma-optical emission
spectrometry, cfu: Colony forming unit
Table 3: Antioxidant activity test of extract
Material IC50 (ppm)
Banana peel extract 64.03±2.78
Watermelon rind extract 300.12±15.32
Combination of banana peel: watermelon rind extract (IC50:IC50) 226.82±18.60
Combination of banana peel: watermelon rind extract (IC75:IC25) 177.56±10.20
Combination of banana peel: watermelon rind extract (IC25:IC75) 292.52±2.91
IC75: Inhibitory concentration at 75%, IC25: Inhibitory concentration at 25%, IC50: Inhibitory concentration at 50%, ppm: Part per million
Table 4: Organoleptic of lotion
Formula F1 F2 F3 F4
Color White-like milk White-like milk White-like milk White-like milk
Odor Typically weak Typically weak Typically weak Typically weak
Sense Not sticky Not sticky Not sticky Not sticky
Table 5: Homogeneity of lotion
Formula F1 F2 F3 F4
Homogeneity Homogeneous Homogeneous Homogeneous Homogeneous
Emulsion type O/W O/W O/W O/W
Table 6: pH of lotion
Formula F1 F2 F3 F4
pH 5.26±0.06 5.27±0.03 5.29±0.04 5.27±0.08
Asian J Pharm Clin Res, Vol 9, Suppl. 3, 2016, 300-304
Alamsyah et al.
303
is consistency curve for plastic material not passing through the
point (0,0) but intersects the shearing stress axis (or will be cut
off if the straight part of the curve is extrapolated to the axis) at a
point known as yield value. The preparation will not flow until the
shearing stress is achieved by yield value. This plastic rheology is
characteristic of cera alba that used in the base of lotion, when idle
and low temperatures, these materials tend to be a flocculation form.
Microbiology test shows the growth of microorganisms <10 cfu/ml,
so qualify according to “A Guidance Document on Microbiological
Control of Cosmetic Products” that limit of microorganisms (bacteria,
yeasts, and molds) in leave-on product should not more than 100
cfu/ml in 0.5 ml of product. This shows that nipagin and nipasol used
in lotion worked effectively to keep the lotion from microorganism
growth. Acute dermal irritation test shows the primary irritation
index is zero, means no indication of erythema and edema, it can be
categorized as very mild irritation response.
The best antioxidant activity of extract is banana peel extract with
IC50 value 64.03 ppm. The results of antioxidant activity test of lotion,
formula F1 is base of lotion do not have antioxidant activity so in
conclusion that the base of lotion is not affecting antioxidant activity of
extract in lotion dosage form. Antioxidant activity of formula F2, F3, and
F4 decreased if compare with antioxidant activity of extract. Formula
F2 using banana peel extract has decreased approximately 7.4. Formula
F3 using watermelon rind extract and formula F4 is a combination
of banana peel and watermelon rind extract have decreased
approximately 1.7. Watermelon rind extract and combinations extract
when formulated into a lotion dosage form are decreased smaller than
banana peel extract. It can be concluded that the antioxidant activity
of banana peel extract can be protected by watermelon rind extract of
decomposition, so the combination of both extracts shows the highest
antioxidant activity in lotion dosage form. The antioxidant activity of
extract can not guarantee the stability of antioxidant activity when
formulation into a dosage form.
CONCLUSION
Banana peel extract with the highest antioxidant activity when
formulated into lotion has significant decrease of antioxidant
activity, but when combined with watermelon rind extract has better
antioxidant activity.
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Table 7: Viscosity of lotion
Formula F1 F2 F3 F4
Viscosity (cps) 23,765.00±1,192 19,931.76±490 20,753.33±1,276 19,235.00±1,437
cps: Centi poise
Table 8: Microbiology of lotion
Formula F1 F2 F3 F4
TPC (cfu/ml) <10 <10 <10 <10
TYM (cfu/ml) <10 <10 <10 <10
cfu: Colony forming unit, TPC: Total plate count, TYM: Total yeast mold
Table 9: Acute dermal irritation test of lotion
Formula F1 F2 F3 F4
Primary irritation index 0 0 0 0
Table 10: Antioxidant activity test of lotion
Formula F1 F2 F3 F4
IC50 (ppm) 853.16±12.99 472.50±28.48 496.71±39.88 300.04±5.51
IC50: Inhibitory concentration at 50%, ppm: Part per million
Asian J Pharm Clin Res, Vol 9, Suppl. 3, 2016, 300-304
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