Vol 3, Issue 1, 2021 (1-8)
http://journal.unpad.ac.id/idjp
*Corresponding author,
e-mail: anis.yohana.chaerunisaa@unpad.ac.id (A. Y. Chaerunnisa)
https://doi.org/10.24198/idjp.v3i1.30946
© 2021 A. Y. Chaerunnisa et al
Peel Off Gel Mask Containing Green Tea Leaf Extract (Camellia Sinesis L) with Antioxidant
Activity
Chaerunisaa, A. Y.*, Khuzaimah, Z., Surachman, E.
Faculty of Pharmacy, Padjadjaran University, Jatinangor, Jl. Raya Jatinangor Km 21.5 Sumedang,
Indonesia
Received: 2 Jan 2021, Revised: 12 Feb 2021, Accepted: 23 Feb 2021, Published: 27 Feb 2021
ABSTRACT
Peel off mask has been very popular and preferred due to its ease in application. Gel base
has been preferred for its cold and moist sensation during application. When herbal
ingredients such as extract is introduced into gel formulation, the stability then will become
an issue. The aim of the study was to formulate the peel-off mask gels containing green tea
leaves extract (Camellia sinensis L.) which effective, stable, and safe to be used. The
antioxidant study was conducted by using Diphenylhydrazylpicryl (DPPH) method. The
peel-off mask gel formula were Hidroxypropyl Methylcelulosa (HPMC) as gel base at
0.5% concentration and plasticizer Polyvinyl Alcohol (PVA) at concentration of 12%.
Evaluation of peel-off mask gel included observation on pH, viscosity, dispersing power,
and drying time during the 90 days of storage at room temperature. The IC50 values of the
green tea (Camellia sinensis L.) extract was 5.511 ppm. The concentration of green tea
leaves extract in the preparation were 1x IC50 (F1); 2x IC50 (F2) and 3x IC50 (F3). The
result showed that all preparations were stable in the aspect of consistency, color, and odor.
Viscosity, drying time and dispersing power remained unchanged after 90 days of storage.
The pH value changed, but still in the range of acceptable pH of topical preparations. The
results of antioxidant activity study showed that formula F3 (peel-off mask gels with 3 x
IC50 of green tea extracts) had the highest antioxidant activity.
Keywords: antioxidant, Camellia sinensis L., peel off gel masks, green tea
1. Introduction
Aging skin can be inhibited by using cosmetics
that contain antioxidants. Antioxidants can
reduce free radicals to be less reactive
molecules, thus it can avoid and reduce
oxidative damage. The use of antioxidants in
anti-aging skin care is essential to prevent
further skin damage1.
Antioxidants are grouped into two categories
based on the source, which is an antioxidant
that is naturally and synthetically derived2. The
concerns about possible side effects that are not
known from synthetic antioxidants cause
natural antioxidant potential alternatives to be
developed2. One of the potent plant as a
powerful antioxidant is green tea leaves
(Camellia sinensis L.). Polyphenol compounds
are very important compounds in tea, especially
in catechins class. The antioxidant activity of
tea is related to the presence of catechins.
Epigallocatechin gallate (EGCG) and
epicatechin gallate (ECG) are types of major
catechins in green tea. This compound has a
A. Y. Chaerunnisa et al / Indo J Pharm 3 (2021) 1-8
2
strong activity to prevent free radicals such as
peroxynitrite, superoxide, hydrogen peroxide3.
With its high antioxidant content, green tea
leaves can be used as one of the raw materials
of cosmetic preparation. The selected cosmetic
preparation is dosage of gel peel-off masks.
Peel-off gel mask is a mask that is a practical
one, because after the mask is dry, it can be
immediately removed without the need to
rinse4. The benefits of gel mask peel-off can
remove impurities and dead skin cells to keep
skin clean and fresh. This mask can also restore
the freshness and softness of the skin, even with
regular use it can reduce slight wrinkles on the
skin5,6.
Based on the background, it is necessary to
study the formulation of green tea leaves
antioxidant extracts in gel peel-off mask
(Camellia sinensis L.). The antioxidant activity
of green tea will be measured before and after
preparation is made into a mask. The stability
of the preparation which includes physical and
microbiological stability will also be observed
within 90 days of storage.
2. Materials and Methods
2.1 Instrument
Instrument used included Digital scales
(Mettler Toledo), water bath (Memmert),
parchment paper, spatula, evaporating basin,
stir bar, a mechanical stirrer, a pH meter
(Hanna), Rion Viscometer, incubator (Sakura,
IF-4), UV spectrometer (Specord 200 ), and
glassware commonly used in Pharmaceutics,
Microbiology, Phytochemistry and Natural
Products Laboratory.
2.2 Plant material
Dried Green tea leaves (Camellia sinensis L.)
were obtained from Plantation Manoko,
Lembang, Indonesia and determinated in the
Department of Biology Faculty of Mathematics
and natural Science, Padjadjaran University.
2.3 Chemical Material
Diphenylhydrazylpicryl, Tri aethanol amine
(PT.Quadrant), Glycerin (PT. Brataco),
Eutanol G (PT. BASF), 10% ammonia,
chloroform, 2N HCl, Mayer reagent,
Dragendorff reagent, Lieberman-Burchard
reagent, Nessler reagent, distilled water, FeCl3
%, 1% gelatin solution has been used as
recieved. Mg powder, ether, methanol,
vanillin10% solution, concentrated H2SO4 5%
KOH, α-naphthol solution of 5%, 1%
Ninhydrin solution, vitamin C, Mueller Hinton
Agar (MHA), Sabouraud Dextrose Agar
(SDA), Gelatin (PT. Brataco), nipagin (PT.
Quadrant), Nipasol (PT. Brataco), Carbopol
940 (PT. Brataco), triethanolamine (PT.
Quadrant), polyvinyl alcohol (PT. Brataco),
HPMC (PT. Brataco), Propilenglikol (PT.
Brataco) and distilled water were used as
received.
2.3 Experimental Methods
Extraction of Simplisia
2 kg of dried green tea leaves powder were
macerated by using 96% ethanol for 3x24 hours
at room temperature. The ethanol extract was
then evaporated using a rotary evaporator under
vacuum pressure at a temperature of
approximately 35˚C - 40˚C and kept at 4oC
until used. Examination on parameters of the
green tea leaves extract included water content,
water and ethanol soluble extract content.
Phytochemical screening
Phytochemical screening was conducted to
determine the secondary metabolite content of
alkaloids, tannins, polyphenols, flavonoids,
steroids, triterpenoids, quinones, saponins,
monoterpenoid, and sesquierpenoid in
simplisia drugs and ethanol extracts.
Antioxidant Activity
Investigation on antioxidant activity of the
extract was conducted by using DPPH reagent
and measured with UV-Visible
spectrophotometric method. A total of 1 mL
DPPH solution was added into a solution of
A. Y. Chaerunnisa et al / Indo J Pharm 3 (2021) 1-8
3
sample at particular concentration. Vitamin C
was used as a standard at concentration of 4, 2,
1 and 0.5ppm. The absorbance of each sample
were measured at a maximum wavelength of
DPPH which is 517nm. Percentage of
inhibition was calculated as follow:
% inhibition = [1- (A test / A control)] x 100%
In which:
A test = average absorption of DPPH solution
in the sample
A control = Absorption of DPPH solution in
ethanol
% inhibition = Percentage inhibition of free
radical
Optimation of Base for peel off gel mask
Table 1. The Base Formula of Gel Masks
Material Formula
F01 F02 F03
PVA (%) 12 12 12
HPMC (%) 0,5 0,5 0,5
Gliserin (%) 12 - 12
PPG (%) - 10 10
TEA (%) 2 2 2
Nipagin (%) 0,2 0,2 0,2
Nipasol (%) 0,05 0,05 0,05
Aquadest Ad 100 Ad 100 Ad 100
F01 = Formula with humectant glycerin
F02 = Formula with humectants propilenglikol
F03 = Formula with humectant glycerin and
propilenglikol
The PVA was dispersed in hot water (mixt. 1),
HPMC was dispersed in hot water (mixt. 2),
nipagin and nipasol were dissolved in ethanol
(mixt. 3). Mixt. 2 and 3 was mixed, stirred and
was added to a mixt. 1 and stirred by using a
mechanical stirrer. Triatanolamin was added to
form a clear mass gel. Next, then glycerine
(F01), or propilenglikol (F02) or glycerin and
propilenglikol (F03) was added and mixed
vigorously until the gel become clear and
homogeneous. The organoleptic investigation,
pH, viscosity, dispersion power and drying
time were examined for 14 days of
investigation.
Formulation of Peel Off Gel Mask with Green
Tea Leaves Extract
Gel mask were formulated using the optimized
gel using different humectant (F01, F02 and
F03). To the optimum formula (F02), green tea
leaves extract were added as much as 1, 3, and
5 times of the IC50 value. Extract were
dissolved into ethanol and added to the gels,
then stirred until homogeneous. Peel-off gel
mask were prepared as follows: gel mask
without green tea leaves extract as standar base
of gel mask (F0): gel mask with tea leaves
extract containing 1 x IC50 (F1): 3 x IC50 (F2)
and 5 x IC50 of green tea extracts (F3).
Physical Quality of gels
Observation quality gel peel-off mask
preparation was conducted by observing the
changes in organoleptic, pH, viscosity,
dispersion power, and drying time during the
90 days of storage.
Drying Time Test
The test was performed by applying the peel off
gel mask to the back part of the hand and was
observed until the gels dried, and formed easy-
to-peel-off layer. The test was conducted over
period of 90 days of storage.
Microbial Contamination of Gel Mask
Microbial contamination examing were
conducted on the first day and the 90th day of
storage. This examination was conducted by
using the media Muller Hinton Agar (MHA) as
a medium for bacteria and Soburo Dextrosa
Agar (SDA) as a medium for fungi.
Antioxidant Activity of gel masks
Antioxidant activity of the gel maks containing
extract were determined by using DPPH
method as mentioned earlier on the section of
antioxidant activity of extract. Gel masks
A. Y. Chaerunnisa et al / Indo J Pharm 3 (2021) 1-8
4
formula F1, F2 and F3 were diluted with water
into concentration of 16, 8, 4, 2, and 1 ppm. The
absorbance sample were measured bu Uv
visible spectrophotometry, and campared with
exract and Vitamin C as standard.
Irritation Test
Irritation test was conducted on 10 volunteers
by using a closed patch test (Patch Test). The
peel-off gel mask with the highest
concentration of extract were applied on the
back part of the right hands of volunteers
(measuring 4 cm2) while that without extract
was on the left hand. The film were left
attached for 10 hours, then the reaction were
marked.
Data Analysis
Data observation were made in the form of a
table and then analyzed with a statistical way
analysis of variance (ANOVA) with the help of
software Statistical Product and Service
Solutions (SPSS) version 16.0.
3. Results and Discussion
3.1 Extraction and Phytochemical
Screening
Results obtained by extraction constant
viscous extract 538.362g giving the yield value
of 30.03%. Table 2. Showed the result of
Phytochemical Screening. It can be concluded
that the extract contain flavonoid, alkaloid and
many other biological compound which
responsible to its activity as antioidant.
The results of the moisture content
determination of green tea leaves extract was
0.83%. This result was in the range of the
quality, which should not be more than 10%, in
order to avoid microbial growth especially
fungus on the extracts.
The results of the water soluble content from
green tea leaves extract was 28.5%, while that
which ethanol soluble was 56.5%. These results
showed that green tea leaves extract is more
soluble in ethanol than water.
Table 2. Phytochemical screening of the
extract
Secondary metabolites
Result
Alkaloids:
a. Reagents Dragendroff
+
flavonoids:
a. Reagents
concentrated HCl + Mg
+
b. 2N H
2
SO4 reagent
+
c. Reagents NaOH 10%
+
Polyphenols
+
Tannins
+
Saponin
-
Steroids and
Triterpenoid
+
Monoterpenes and
Sesquiterpenoids
-
Quinone
+
(+) = Detected (-) = Not detected
3.2 Antioxidant Activity of Green Tea
Leaves Extract
The results showed that Green tea leaves
extract has IC50 value as much as 5.51ppm.
IC50 is the concentration that can inhibit 50%
of free radicals. Based on comparison study
with vitamin C as standard, the extract have
0.558 times of the activity compared with
vitamin C (IC50 of 3.075ppm) as shown in
Table 3 and 4. IC50 was later then used as the
reference in determining the concentration of
the extract to be prepared in gel masks.
Table 3. IC50 Value and % Inhibition of
Green Tea Leaves Extract
C
(ppm)
Absorbance
Average
%
Inhibition
IC50
(ppm)
0
0.832
-
2
0.651
21.755
4
0.496
40.385
5.511
6
0.369
55.649
8
0.276
66.827
3.3 Formulation of Peel Off Gel Mask
Based on the result of optimation process on the
gel base formula, to which drying time, F02
was the one which will be used as a basis for
further experiment in preparation gel mask due
A. Y. Chaerunnisa et al / Indo J Pharm 3 (2021) 1-8
5
to its short drying time. This conclusion also
been supported by physical observation on the
formula made in optimation process. After 2
weeks of storage of the three formulas were
investigated for its elasticity in order to be
unbreakable when it is exfoliated (peel-off).
Table 4. IC50 Value and % Inhibition of
Vitamin C
C
(ppm)
Absorbance
Average
%
Inhibition
IC50
(ppm)
0
0.807
-
0.25
0.758
6.071
1
0.657
18.587
3.075
2
0.539
33.209
4
0.289
64.188
C = concentration of sample
Further formulations of antioxidant green tea
leaves extract gel mask were performed by
using F0 base with addition of the extract at
concentration of 0.0005; 0.0015; and 0.0025%.
3.4 Physical observation results of the gel
masks
Formula of gel mask with various
concentrations of green tea leaf extract showed
the consistency, smell, and color which
unchanged for 90 days of storage, indicating
that no decomposition of materials or excipient
of gel mask interact and causing changes in
consistency, smell, and color.
3.5 pH of gel masks
The study were performed to investigate the
effect of different concentration of extract on
pH of gel formulation. Formula gel masks with
various concentration of green tea extract tend
to have acidic pH which between 5-5.5, that
probaly due to the green tea leaves extract
which has a pH of 4.4. During the storage
period, the pH of each formula gel mask
changed but still in the area of pH for topical
preparation (Figure 2). The change in pH can
be due to chemical reaction in the gel or by the
effect of extract itself which contain many
biochemical constituent 7,8,9.
Figure 2. Changes in the pH curve Gel Mask
with different concentration of Green Tea
Leaves Extract for 90 Days of Storage
3.6 Viscosity of gel masks
The study on viscosity of all gel mask formula
were conducted to evaluate the effect of
different concentration of extract on viscosity
of gel formulations. The results showed that the
viscosity of gel containing different
concentration of extract did not significantly
change during 90 days of storage (Figure 3).
Accordingly, viscosity of the gel were all in the
range of required viscosity10,11. The
exception was found on gel formula without the
addition of extract to which it revealed the
increase after 28 days of storage. This
phenomena can be explained by the possibility
of water evaporation from the gels.
3.7 Drying Time
Based on the result of drying time test during
optimation, gel formulation with propylen
glycol (F02) was that with short drying time.
Further investigation were conducted using
propylen glycol as humecta, with different
concentration of extract as active ingredient.
The results of investigation on drying time of
gel mask showed that the drying time were in
the range of drying mask products in the
market, which lied between 10-20 minutes12.
The drying time between the three of the
formula was not significantly different due to
small difference of extract concentration as
active substance in the formula (Figure 4).
A. Y. Chaerunnisa et al / Indo J Pharm 3 (2021) 1-8
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Figure 3. Changes in Viscosity Curve Gel Mask
with Green Tea Leaves Extract for 90 Days of
Storage
Figure 4. Drying Time of gel masks with
Green Tea leaves extract during 90 days of
storage
3.8 Dispersion power of Gel masks
The purpose of dispersion power test was to
determine the ability of gel to easily being
spread on the surface of the skin without
significant pressure. From the data, it can be
concluded that the dispersion power of all gel
masks formulation were in the range of 5-7cm
and revealed no significant difference during
90 days of storage. The test results of the
dispersion power can be seen in Figure 5.
Dispersion power were highly influenced by
gel strength thus the type of gel base were very
influencing13, 14, 15.
Figure 5. Curve Changes in Dispersion Power
with Green Tea Extract Leaves for 90 Days of
Storage
3.9 Determination of Antioxidant activity of
gel masks
With regard to the IC50 values, a
compound is defined to be the strong
antioxidant when the IC50 value is less than
50ppm. Strong antioxidant has the IC50 value
of 50-100ppm, moderate antioxidant has IC50
value between 100-150 ppm IC50, and is is
undermined when IC50 value is 150-200 ppm
or over 200. Antioxidant activity of three gel
masks formulas showed the IC50 below 20
which revealed strong antioxidant activity.
Compared to the IC50 of extract (found to be
5.511 ppm) the antioxidant activity of extract
were decreased which due to dilution effect of
excipients. Accordingly, an increase in IC50
value, which revealed the decrease in
antioxidant activity can be seen after 90 days of
storage (Figure 6). It is noticable that the
decrease was much lower when the extract was
used at higher concentration (Figure 6).
Decrease in antioxidant activity was may due to
the influence of the interaction between the
extract with excipient of gel mask used in the
formula16. Mechanism of reaction between the
extract and excipients need to be further
investigated. Somehow, this problem can be
overcomed by increasing the concentration of
the extract as active substance.
A. Y. Chaerunnisa et al / Indo J Pharm 3 (2021) 1-8
7
3.10 Microbial contamination
Investigation on microbial contamination was
conducted as the base of preparation were water
based hidrogel polymer which higly in risk to
be contaminated by microbial growth, therefore
the microbial testing is obligatory for those
formulations. Based on the results, microbial
contamination carried out on the first day of
preparations were zero, and so did after day 90
of storage. It can be concluded that
antimicrobial used as the preservative in the
formulation is effective either to maintain the
microbial growth or to preserve the
formulations.
Figure 6. Results of Measurement of
Antioxidant Activity Preparations Gel Mask
with Green Tea Leaf Extract on Day 1and Day
90
F1 = gel mask with tea leaves extract 1 x IC50
F2 = gel mask with tea leaves extract 3 x IC50
F3 = gel mask with tea leaves extract 5 x IC50
3.11 Skin irritation study
Gel masks with or without addition of tea
leaves extract did not cause any irritation to the
20 volunteers, both primary and secondary
irritation. The results marked as no occurrence
of burnt, redness, and swelling in volunteer
skin, and can be concluded that either the
material or extract used in gel masks is safe to
be used.
4. Conclusion
The antioxidant activity of green tea leaves
extract was 0.558 times compared with vitamin
C as vitamin C has IC50 of 3.075 ppm while
that of green tea leaves extract was IC50 5.511
ppm. Formula of antioxidant gel mask used
green tea leaf extract concentration as much as:
1x IC50(F1); 2x IC 50 (F2) and 3 x IC 50 (F3).
All formulation were generally stable including
pH, drying time, viscosity and dispersion
power after 90 days of storage. Antioxidant
activity of gel masks were defined as the strong
antioxidant due to its IC50 which less than 50
ppm.
Conflict Of Interest
None
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