Formulation and Evaluation of Platelet Rich Plasma (PRP) Lotion
Preparations with Asiaticoside as an Antioxidant
Yanni D. Mardhiani*1, Nita Selifiana1, Endah Arna Ningsih1, Taofik Rusdiana2
1Research Group of Pharmaceutics, Bhakti Kencana University, Bandung city, West Java,
Indonesia;
2Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy,
Universitas Padjadjaran, Sumedang, Indonesia
Submitted: 19/12/2023, Revised: 17/01/2024, Accepted: 28/01/2024, Published: 24/01/2025
ABSTRACT
Currently, the use of PRP requires a person to visit a skin care clinic by doing a
series of treatments directly. Therefore, in order to facilitate the use of PRP,
asiaticoside lotion base products containing antioxidants can be used as a PRP
product base. asiaticoside antioxidants serve to stabilize PRP when added to the
base. This research uses Platelet-Rich Plasma (PRP) and Asiaticosida as active
substances, Fancor® Uni-Embase as emollient and emulgator, DMDM hydantoin
as preservative, and distilled water as carrier. The stages in this research include
preparation of tools and materials, lotion base optimization, lotion preparation
formulation, addition of active substances, lotion preparation evaluation, physical
stability test, and antioxidant activity test. The results of the PRP lotion stability
test with asiaticoside for 10 days showed significant results (p<0.05) which means
that for 10 days there is a tendency to be unstable. Antioxidant activity test results
obtained IC50 results on Asiaticosida of 57.63 µg/mL, on PRP of 196.1 µg/mL, on
the base with the addition of Asiaticosida of 77.19 µg/mL, on F0 of 166.90 µg/mL,
on F1 of 96.727 µg/mL, on F2 of 88.395 µg/mL, and on F3 of 82.017 µg/mL.
Key words: Lotion, PRP, asiaticoside, Antioxidant
Vol. 5, Issue 2, 2023 (405-421)
http://journal.unpad.ac.id/IdJP
*Corresponding author,
e-mail: yanni.dhiani@bku.ac.id (Y. D. Mardiani )
https://doi.org/10.24198/idjp.v5i2.51883
© 2023 Y. D. Mardiani et al
406
1. Introduction
At this time many people use
various kinds of cosmetic preparations to
care for the skin of both women and men.
One of the cosmetic preparations for skin
care is lotion. According to Indonesian
Pharmacopeia III defines lotion as a
liquid preparation used as an external
drug in the form of a suspension or
dispersion. Lotions can be solid in the
form of a fine powder with a suitable
suspending agent or an emulsion of the
type seen in oil in water with a suitable
surfactant (Ministry of Health RI, 1979).
Indonesia is a tropical country
with various natural resources, including
plants, some of which have been utilized
both as cosmetics and as a means of
preventing or treating diseases. One of the
herbal plants, Centella asiatica, has a
strong antioxidant effect. In Centella
asiatica, there are many components of
triterpenoid compounds and asiaticosides
which are the main compounds that have
antioxidant activity (Zainol et al in
(Saputri & Damayanthi, 2015). Used as
herbs, parts of Centella asiatica
containing asiaticosides are found in
stems (15.9%), leaves (82.6%), and roots
(1.5%), so they are used as herbs
(Zulkarnaen et al., 2015).
Currently, the use of PRP requires
a person to visit a beauty clinic by doing
a series of treatments directly. PRP
treatment requires the use of needles to
insert PRP into the skin. Therefore, in
order to facilitate the use of PRP,
researchers are interested in making a
lotion base containing asiaticoside as an
antioxidant that can be used as a PRP
product base, it is hoped that this
antioxidant can stabilize PRP when added
to the preparation product. PRP (platelet-
rich plasma) is a blood component of
autologous biologically active products
enriched with a number of growth factors,
cytokines, and other plasma proteins (Lin
et al., 2020). PRP contains a high
concentration of platelets with a variety
of bioactive proteins, suggesting that PRP
can accelerate the process of tissue
regeneration and healing (Lee et al.,
2020). PRP has many platelets consisting
of growth factors and other elements,
which greatly help the regenerative
healing process. PRP works by destroying
platelet components that contain growth
factors. PRP contains seven growth
factors, including VEGF, EGF, FGF, IGF-
1, PDGF, TGF β-1, and HGF (Taniguchi
et al., 2019). PDGF plays a role in tissue
remodeling and promotes the production
of other growth factors. Growth factors in
PRP can induce β-cell regeneration and
increase β-cell mass by stimulating β-cell
neogenesis and ductal cell differentiation
into β-cells, as detected by increased c-
peptide levels (Younis, 2019). PRP has
experienced development and is
increasingly advanced, many benefits of
PRP include in terms of medical action,
dermatology, and in terms of beauty. PRP
can rejuvenate the skin and treat burns,
chronic ulcers, diabetic ulcers, and hair
loss (Satriyo et al, in Younis, 2019).
2. Method
This study is a laboratory
experimental study conducted from
March to June 2023. Platelet-Rich Plasma
(PRP) obtained from volunteers and
asiaticoside employed as active
substance, Fancor® Uni-Embase used as
emollient and emulgator, DMDM
hydantoin used as preservative, and
distilled water used as carrier. There are 4
formulas in this study, F0 with the
addition of 2.5% PRP concentration
without asiaticoside, F1 with the addition
of 2.5% PRP concentration and 1%
asiaticoside, F2 with the addition of 2.5%
PRP concentration and 2% asiaticoside,
Y. D. Mardiani et al / Indo J Pharm 5 (2023) 405-421
407
and F3 with the addition of 2.5% PRP
concentration and 3% asiaticoside.
Evaluation of lotion preparation includes
organoleptic test, spreadability test,
viscosity test, emulsion type test,
homogeneity test, pH test, accelerated
stability test, and antioxidant activity test
(Uv-Vis spectrophotometry). The data
obtained are presented in the form of
tables and graphs and the evaluation
results will be processed using IBM SPSS
software.
3. Result
3.1 Optimization Results
The purpose of this optimization is to find
out the right formula to determine the
lotion base formula that meets the
requirements of the literature.
Y. D. Mardiani et al / Indo J Pharm 5 (2023) 405-421
Table 1. Lotion Base Formula
Material name F1 F2 F3
Fancor® Uni
-Embase 5 7.5 10
DMDM Hydantoin 5 5 5
Aquadest
Ad 100 Ad 100 Ad 100
Fancor® Uni-Embase was mixed with hot
water at 70oC - 80oC and stirred until
homogeneous, then waited until cool.
After cooling, DMDM hydantoin was
added. Then put into the lotion container.
After optimization, the best emulgator
concentration was obtained to make
lotion base, so PRP lotion with
asiaticoside was formulated by making
four different formulas, namely F0 with
the addition of 2.5% PRP concentration
without asiaticoside, F1 with the addition
of 2.5% PRP concentration and 1%
asiaticoside, F2 with the addition of 2.5%
PRP concentration and 2% asiaticoside,
and F3 with the addition of 2.5% PRP
concentration and 3% asiaticoside.
Table 2: PRP lotion formula with asiaticoside
Composition (%) F0 F1 F2 F3
PRP 2,5 2,5 2,5 2,5
Asiaticoside -1 2 3
Fancor® Uni
-Embase 55 5 5
DMDM Hydantoin
0,6 0,6 0,6 0,6
Buffer pH 7
0,5 0,5 0,5 0,5
Triethanolamine (TEA)
ad pH 7.4
qs qs qs qs
Distilled water
ad 100
ad100
ad 100
ad100
Fancor® Uni-Embase was mixed with hot
water and stirred until homogeneous, then
waited to cool. After cooling, DMDM
hydantoin, TEA, and Buffer were added.
Then the pH was checked, after which
asiaticoside and PRP were added and day
10.
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3.2 Characterization
Lotion Base Reproducibility Test
Organoleptic Reproducibility Test
Table 3. Reproducibility Test on Organoleptic Characterization
Reproducibility Test for Homogeneity
Table 4. Reproducibility Test for Homogeneity
Reproducibility Test on Emulsion Type
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In the pH test, the test was repeated three
times, based on Figure 1, the average pH
obtained in batch 1 was 7.44, batch 2 was
7.42, and batch 3 was 7.43. The value
obtained is in accordance with the lotion
pH requirements of 4.5-8 (SNI 16-4399-
1996).
Reproducibility Test on Viscosity
In viscosity testing using a Brookfield LV
manual viscometer with a speed of 60
rpm and using spindle no 64. Viscosity
testing was repeated three times, based on
the diagram above, the average viscosity
value obtained in batch 1 was 3,500,
batch 2 was 3,583, and batch 3 was 3,583.
The results obtained meet the lotion
viscosity requirements of 2,000-50,000
cPs (SNI 16-4399-1996).
Reproducibility Test on Spreadability
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Figure 3: Reproducibility Test of Scatterability
In the spreadability test, the test was
repeated three times, based on Figure 3
above, the average spreadability results
obtained in batch 1 were 6.43, batch 2
was 6.45, and batch 3 was 6.35. The
results obtained are in accordance with
the lotion spreadability requirements of 5-
7 cm (Garg, 2002).
Stability Test
Organoleptic Stability Test
Organoleptic test is done by observing
changes using human senses including
odor, consistency, and color (Mardikasari,
et al., 2017).
Table 6: Stability test results on organoleptic.
Stability Test for Homogeneity
Table 7. Results of Stability Test for Homogeneity
Description: F0 = Lotion Formulation with PRP 2.5%
F1 = Lotion Formulation with PRP 2.5% and 1% asiaticoside
F2 = Lotion Formulation with PRP 2.5% and asiaticoside 2%
F3 = Lotion formulation with PRP 2.5% and asiaticoside 3%
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Description: F0 = Lotion Formulation with PRP 2.5%
F1 = Lotion Formulation with PRP 2.5% and asiaticoside 1%
F2 = Lotion Formulation with PRP 2.5% and asiaticoside 2%
F3 = Lotion formulation with 2.5% PRP and 3% asiaticoside
In the pH test, 3 repetitions were carried
out to ensure accuracy. Based on Figure
4, the pH test for 10 days obtained results
ranging from 6.83 to 7.59, in the four
formulas the more days the average pH
value drops but is still within the range of
pH requirements. The value obtained is in
accordance with the lotion pH
requirements of 4.5-8 (SNI 16-4399-
1996).
Stability Test on Viscosity
Image 5. Diagram of Stability Test Results on Viscosity
Description: F0 = Lotion Formulation with PRP 2.5%
F1 = Lotion Formulation with PRP 2.5% and 1% asiaticoside
F2 = Lotion Formulation with PRP 2.5% and asiaticoside 2%
F3 = Lotion formulation with PRP 2.5% and asiaticoside 3%
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Viscosity testing was carried out 3 times
to ensure the accuracy of the
measurements. Based on Figure 5.10, the
viscosity test for 10 days obtained
viscosity results ranging from 2,050-
4,117 cPs. During the 10 days there is an
increase and decrease in viscosity value
but the value obtained is still included in
the lotion viscosity requirements of
2,000-50,000 cPs (SNI 16-4399-1996).
Stability Test on Spreadability
Figure 6: Diagram of Stability Test Results for Spreadability
Description: F0 = Lotion Formulation with PRP 2.5%
F1 = Lotion Formulation with PRP 2.5% and 1% asiaticoside
F2 = Lotion Formulation with PRP 2.5% and asiaticoside 2%
F3 = Lotion formulation with PRP 2.5% and asiaticoside 3%
The spreadability test was conducted 3
times to ensure the accuracy of the
measurement. Based on Figure 6, the
spreadability test for 10 days obtained
results ranging from 5.7-6.82 cm, in the
four formulas on average the longer
stored, the more the spreadability area
increases, but there is also a decrease in
between storage times. However, the
values obtained are still in accordance
with the lotion spreadability requirements
of 5-7 cm (Garg, 2002).
Stability Test on Emulsion Type
Table 8. Results of Stability Test on Emulsion Type
Description: F0 = Lotion Formulation with PRP 2.5%
F1 = Lotion Formulation with PRP 2.5% and 1% asiaticoside
F2 = Lotion Formulation with PRP 2.5% and asiaticoside 2%
F3 = Lotion formulation with PRP 2.5% and asiaticoside 3%
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Antioxidant Activity Test
Description: F0 = Lotion Formulation with PRP 2.5%
F1 = Lotion Formulation with PRP 2.5% and 1% asiaticoside
F2 = Lotion Formulation with PRP 2.5% and asiaticoside 2%
F3 = Lotion formulation with PRP 2.5% and asiaticoside 3%
Based on Figure 7, the results of
antioxidant activity test conducted on
asiaticoside obtained IC50 value of 57.63
µg/mL, on PRP of 196.1 µg/mL, on the
base with the addition of asiaticoside of
77.19 µg/mL, on F0 obtained a value of
166.90 µg/mL, on F1 obtained a value of
96.727 µg/mL, on F2 obtained IC50 value
of 88.395 µg/mL, and on F3 obtained
IC50 of 82.017 µg/mL. PRP has weak
antioxidant, asiaticoside has strong
antioxidant, base with added asiaticoside
has strong antioxidant, F0 has weak
antioxidant, F1 has strong antioxidant, F2
has strong antioxidant, and F3 has strong
antioxidant.
4. Discussion
4.1. Reproducibility Test of Lotion
Base
Reproducibility refers to the ability to
reproduce the results of an analysis or
experiment with the same procedures,
data, and methods as originally used, thus
obtaining maximum results. The
reproducibility test used an optimized
base, formula B1 with a concentration of
5% Fancor® Uni-Embase with the
addition of 2% asiaticoside. In this test,
there were 3 batches of repetition with
manufacture on different days. Tests
carried out include organoleptic test, pH
test, homogeneity test, viscosity test,
spreadability test, and emulsion type test.
4.2 Reproducibility Test on
Organoleptic, Homogeneity, and
Emulsion Type
The results of the organoleptic test were
yellowish white in color, characteristic
odor of asiaticoside, and slightly thick.
While the homogeneity test is
homogeneous and the emulsion type test
is oil in water (M/A). In the three batches
produced the same results of the
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organoleptic test were yellowish white in
color, characteristic odor of asiaticoside,
and slightly thick. While the homogeneity
test is homogeneous and the emulsion
type test is oil in water (M/A). In the
three batches produced the same results
there were no changes, so it can be said
that the organoleptic test, emulsion type
test, and homogeneity test of lotion
preparations made have good
reproducibility.
4.3. Reproducibility Test for pH
In the pH test, the test was
repeated three times, based on the
diagram above, the average pH result
obtained in batch 1 was 7.44, batch 2 was
7.42, and batch 3 was 7.43. When the pH
test results were obtained, the data were
statistically evaluated with SPSS to
determine whether the formula repeated
as many as three batches had differences
between formulas. The pH data analyzed
resulted in normal data (p>0.05) and
homogeneous data of 0.653 (p>0.05). So
that data processing is continued with the
One Way Anova test, getting the result of
0.269 (p>0.05) which can be said that
there is no significant difference in pH
value. So it can be interpreted that the
preparations made in terms of pH have
good reproducibility.
4.4. Reproducibility Test on Viscosity
In the viscosity test, the test was
repeated three times, based on the
diagram above, the average viscosity
value obtained in batch 1 was 3,500,
batch 2 was 3,583, and batch 3 was 3,583.
The viscosity test data obtained were then
statistically analyzed using SPSS to
determine whether there were differences
in the formulas that were repeated as
many as 3 batches. The viscosity data
analyzed produced normal data, namely
(p>0.05) and homogeneous data, namely
0.653 (p>0.05). So that data processing is
continued with the One Way Anova test,
getting the result of 0.302 (p>0.05) which
can be said that there is no significant
difference. So it can be interpreted that
the preparations made have good
reproducibility.
4.5. Reproducibility Test for
Spreadability
In the spreadability test, the test was
repeated three times, based on the
diagram above, the average spreadability
results obtained in batch 1 were 6.43,
batch 2 was 6.45, and batch 3 was 6.35.
Based on the results of the spreadability
test obtained, it was then statistically
analyzed using SPSS to determine
whether there were differences in the
formulas that were repeated as many as 3
batches. The analyzed spreadability data
resulted in normal data, namely (p>0.05)
and homogeneous data, namely (p>0.05).
So that data processing is continued with
the One Way Anova test, getting the result
of 0.173 (p>0.05) which can be said that
there is no significant difference. So it
can be interpreted that the preparations
made have good reproducibility.
4.6. Stability Test of PRP Lotion with
Asiaticoside
At this stage, physical stability
testing of PRP lotion preparations with
asiaticoside was carried out at cold
temperature storage (4oC) for 10 days, as
for the day points tested, namely day 1,
day 4, day 7, and day 10. This test was
conducted on the four lotion formulations
with the addition of 2.5% PRP
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in each formula. There were three
variations of asiaticoside concentration
used, namely F1 by 1%, F2 by 2%, and
F3 by 3%, while F0 was not given the
addition of asiaticoside. Stability testing
was conducted to determine the physical
quality of lotion preparations using PRP
with asiaticoside as antioxidant, including
organoleptic test, viscosity test,
homogeneity test, spreadability test,
emulsion type test, and pH test.
4.7. Organoleptic Stability Test
Organoleptic observation of PRP lotion
preparations with asiaticoside to
determine the physical characteristics of
the preparations made including odor,
color, and shape (Mardikasari, et al.,
2017). The results of organoleptic testing
can be seen in table 5.9. The results of
organoleptic testing of PRP lotion
preparations with asiaticoside after
storage for 10 days at 4oC include day 1,
day 4, day 7, and day 10 have relatively
the same organoleptic. In the four
formulas made, the odor in F0 is odorless,
while F1, F2, and F3 are typical of
asiaticoside, this is because in F1, F2, and
F3 there is the addition of asiaticoside.
The shape produced from the four
formulas is rather thick. Meanwhile, the
color produced in F0 is white, this is
because the lotion base is already white
and there is no addition of asiaticoside to
F0, this also indicates that PRP does not
affect the color. In F1, F2, and F3 colors
have a yellowish white color, this is due
to the addition of asiaticoside to the lotion
preparation. The observation results of
the four formulas made did not show too
significant changes in terms of changes in
odor, color, and shape so that it can be
said that the preparation is stable in terms
of organoleptic.
4.8. Stability Test for Homogeneity
Homogeneity testing by checking the
presence or absence of clumped granules
in lotion preparations, the homogeneity
test aims to determine the mixing of
ingredients for lotion preparations. The
homogeneity of the preparation can be
influenced by the mixing procedure used
during the preparation (Noer & Sundari,
2016). The homogeneity of the emulsion
system can be influenced by the mixing
method and equipment used during the
emulsion manufacturing process (Noer
and Sundari, 2016). After 10 days of
testing at the points of day 1, day 4, day
7, and day 10 there was no significant
change. PRP lotion preparations with
asiaticoside showed homogeneous results
characterized by the absence of clumped
granules in the lotion.
preparation. In terms of the homogeneity
test, it shows that the PRP lotion
preparation with asiaticoside is not
damaged because of the homogeneous
preparation results.
4.9. Stability Test Against pH
The efficiency and stability of the drug,
as well as its safety not to irritate the skin,
are all verified through pH testing. pH in
preparations is related to the effectiveness
of preservatives, the stability of active
substances, and the state of the skin
(Fajriyah in Wulanawati et al., 2019).
The pH test for 10 days obtained results
ranged from 6.83-7.59, in the four
formulas the more days the average pH
value decreased but was still within the
pH requirement range.
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The value obtained is in accordance with
the lotion pH requirements of 4.5-8 (SNI
16-4399-1996). Changes in pH indicate a
lack of stability of the preparation which
can cause the preparation to be damaged
during storage. Differences in the pH
value of the preparation during storage
can be caused by decomposing media
such as storage temperature which can
increase the acid or base content of a
product (M.M et al., 2018). Changes in
the pH of the preparation can also be
caused by differences in the concentration
of the active substance, with a higher
concentration of extract, the pH of the
preparation will decrease (Sugiharto &
Dr. Cikra Ikhda Nur Hamida Safitri,
2020).
The data obtained were then processed
using SPSS version 20, the initial stage is
the requirements test including the
normality test (Shapiro Wilk) and the
homogeneity test (Levene's test). In the
normality test and homogeneity test, the
sig value was obtained. > 0.05 which
means that the data is normally
distributed and the data variance is
homogeneous. So that data processing is
continued with One Way Anova with the
results obtained sig value. <0.05 which
means that the average storage time of
each formula is significantly different. So
that data processing is continued using
the Post Hoc Test. Post Hoc Test results in
the pH F0 test day 1 with day 4, day 7,
and day 10; day 4 with day 1, day 7, and
day 10; day 7 with day 1 and day 4; day
10 with day-1 and day 4 there is a
significant difference between storage
times. In the pH test F1 day 1 with day 4,
day 7, and day 10; day 4 with day 1, day
7, and day 10; day 7 with day 1, day 10
and day 4; day 10 with day- 1, day 7 and
day 4 there is a significant difference
between storage times. In F2 day 1 with
day 4, day 7, and day 10; day 4 with day
1, day 7, and day 10; day 7 with day 1,
day 10 and day 4; day 10 with day 1, day
7 and day 4 there is a significant
difference between storage times. In the
pH test of F3 day 1 with day 7 and day
10; day 4 with day 7 and day 10; day 7
with day 1, day 10 and day 4; day 10 with
day 1, day 7 and day 4 there is a
significant difference between storage
times. So it can be interpreted that the
four formulas are not very stable in cold
temperature storage (4oC) for 10 days.
This is reinforced by the instability of the
pH value which is decreasing.
4.10. Stability Test for Spreadability
The purpose of the spreadability test is to
evaluate the spreading properties of PRP
lotion formulations containing
asiaticosides when applied to the skin to
ensure user comfort. The spreadability of
the lotion preparation decreases when the
same amount of pressure is applied in
each test, and the relationship between
spreadability and viscosity is inverse. The
ability to spread is inversely correlated
with the viscosity value where the low
viscosity value increases the spreadability
and vice versa, the high viscosity value
decreases the spreadability (Arisanty, et
al., 2020).
The spreadability test for 10 days
obtained results ranging from 5.7-6.82
cm, in the four formulas on average the
longer it is stored, the more the
spreadability increases, but there is also a
decrease between storage times.
However, the value obtained is still in
accordance with the lotion spreadability
requirements of 5-7 cm (Garg, 2002).
This is in accordance with the
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lotion spreadability requirements of 5-7
cm (Garg, 2002). This is in line with the
viscosity value that the longer stored the
more liquid or down, so that the
spreadability of the lotion is inversely
proportional to the viscosity value
(Oktaviasari & Zulkarnain, 2017). Some
factors that affect changes in the
spreadability value are temperature,
storage, and excipients used. The longer
the storage of the preparation, the lower
the binding power of the thickener
(Armadany et al., 2019). Changes in
spreadability can also be influenced by
less stable PRP.
The data obtained were then processed
using SPSS version 20, the initial stage
was the requirements test including the
normality test (Shapiro Wilk) and the
homogeneity test (Levene's test). In the
normality test and homogeneity test, the
sig value was obtained. > 0.05 which
means that the data is normally
distributed and the data variance is
homogeneous. So that data processing is
continued with One Way Anova with the
results obtained sig value. <0.05 which
means that the average storage time of
each formula is significantly different. So
that data processing is continued using
the Post Hoc Test. Post Hoc Test results
on the spreadability test F0 day 1 with
day 7 and day 10; day 4 with day 10; day
7 with day 1; day 10 with day 1 and day 4
there are significant differences between
storage times. In the F1 spreadability test
day 1 with day 10; day 10 with day 1
there is a significant difference between
storage times. In the spreadability test F2
day 1 with day 4, day 7, and day 10; day
4 with day 1; day 7 with day 1 and day
10; day 10 with day-1, day 7 and day 4
there is a significant difference between
storage times. In the spreadability test F3
day 1 with day 7 and day 10; day 4 with
day 7 and day 10; day 7 with day 1 and
day 4; day 10 with day 1 and day 4 there
was a significant difference between
storage times. So it can be interpreted that
the four formulas are not very stable in
cold temperature storage (4oC) for 10
days because in the One Way Anova test
there are significant differences. This is
reinforced by the instability of the
spreadability area that changes during
storage.
4.11. Viscosity Stability Test
The purpose of viscosity testing is to
evaluate the viscosity of the lotion
product made. Preparations with too high
viscosity will be more difficult to extract
from the container and more difficult to
flow. The packaging process will also be
affected by excessive viscosity because
high viscosity produces significant
resistance when putting the preparation
into the container. In addition, too low
viscosity is undesirable as it will cause
dripping when applied (Martin, et al.,
1993). The viscosity test used a
Brookfield LV manual viscometer with a
speed of 60 rpm and used spindle no 64.
The viscosity test for 10 days obtained
viscosity results ranging from 2,050-
4,117 cPs. During the 10 days there was
an increase and decrease in viscosity
values but the values obtained were still
included in the lotion viscosity
requirements of 2,000-50,000 cPs (SNI
16-4399-1996). Changes in viscosity can
be caused by an increase or decrease in
particle diameter size which causes the
surface area to get smaller or larger
(Mardikasari et al., 2020).
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Y. D. Mardiani et al / Indo J Pharm 5 (2023) 405-421
Changes in viscosity can also be
influenced by PRP active ingredients that
are less stable in the preparation.
The data obtained were then processed
using SPSS version 20, the initial stage is
the requirements test including the
normality test (Shapiro Wilk) and the
homogeneity test (Levene's test). In the
normality test and homogeneity test, the
sig value was obtained. > 0.05 which
means that the data is normally
distributed and the data variance is
homogeneous. So that data processing is
continued with One Way Anova with the
results obtained sig value. <0.05 which
means that the average storage time of
each formula is significantly different. So
that data processing is continued using
the Post Hoc Test. The Post Hoc Test
results in the F0 viscosity test day 1 with
day 4, day 7, and day 10; day 4 with day
1, day 7, and day 10; day 7 with day 1,
day 10 and day 4; day 10 with day-1, day
7 and day 0to-4 there is a significant
difference between storage times. In the
F1 viscosity test day 1 with day 7 and day
10; day 4 with day 10; day 7 with day 1
and day 10; day 10 with day- 1, day 7 and
day 4 there is a significant difference
between storage times. In the F2 viscosity
test day 1 with day 7 and day 10; day 4
with day 10; day 7 with day 1; day 10
with day 1 and day 4 there is a significant
difference between storage times. In the
viscosity test of F3 day 1 with day 7 and
day 10; day 4 with day 7 and day 10; day
7 with day 1, day 10 and day 4; day 10
with day-1, day 7 and day 4 there is a
significant difference between storage
times. So it can be interpreted that the
four formulas are not very stable in cold
temperature storage (4oC) for 10 days
because in the One Way Anova test there
are significant differences.
4.12. Stability Test Against Emulsion
Types
Emulsion type testing is carried out to
determine whether the preparation made
is an oil-in-water (O/W) or water-in-oil
(O/W) emulsion type using the dilution
method. The dilution method is carried
out by diluting 1 gram of lotion
preparation into 10 ml of distilled water
(Megantara, et al., 2017).
The emulsion type test of the four
formulas, namely the oil-in-water (O/W)
emulsion type, is characterized by the
mixing of the lotion preparation with
water. The use of emulsifiers which tend
to be more soluble in water causes this
type of emulsion preparation, and can
also be caused by water being used as a
solvent (Rowe, et al., 2009). In the
emulsion type test, the four formulas after
being stored for 10 days at cold
temperature (4oC) showed the same
results from day 1, day 4, day 7, and day
10. This shows that the PRP lotion
preparation with asiaticoside does not
experience a change in emulsion type
during storage time.
4.13. Antioxidant Activity Test
In this study, quantitative antioxidant
activity was tested using the DPPH (1,1-
diphenyl-2-picrylhydrazyl) technique
against a base containing asiaticoside,
PRP, asiaticoside, F1 (2.5% PRP and 1%
asiaticoside), F2 (2, 5% PRP and 2%
asiaticoside), and F3 (2.5% PRP and 3%
asiaticoside). When performing
antioxidant assays, the DPPH approach
offers the advantage of being fast, easy,
and requiring less sample and reagents.
419
Y. D. Mardiani et al / Indo J Pharm 5 (2023) 405-421
The DPPH wavelength obtained in this
test was λ 519 nm in methanol solvent in
accordance with the literature which
states that the maximum wavelength for
DPPH is in the range 515-520 nm
(Molyneux P, 2004). The next absorbance
measurement will be carried out at this
wavelength. Measurement of antioxidant
activity in samples was carried out by
reacting the sample and DPPH solution
(1:1) in a dark place, covered with
aluminum foil and tightly closed, to avoid
the decomposition of the DPPH solution
because it is easily oxidized. Vitamin C is
used as a reference because it has electron
donating groups on the C2 and C3 atoms.
Vitamin C can absorb free radicals
because of the presence of this group. The
IC50 value for vitamin C is 8.147 g/mL,
which shows that vitamin C is a strong
antioxidant. Antioxidant activity was
calculated by the IC50 value (Inhibition
Concentration 50%) using the linear
regression equation (y=bx+a).
The results of antioxidant activity tests
carried out on asiaticoside obtained an
IC50 value of 57.63 µg/mL, on PRP it
was 196.1 µg/mL, on a basis with the
addition of asiaticoside it was 77.19
µg/mL, on F0 a value of 166.90 was
obtained. µg/mL, at F1 the value was
96.727 µg/mL, at F2 the IC50 value was
88.395 µg/mL, and at F3 the IC50 was
82.017 µg/mL. PRP has a weak
antioxidant, asiaticoside has a strong
antioxidant, base with added asiaticoside
has a strong antioxidant, F0 has a weak
antioxidant, F1 has a strong antioxidant,
F2 has a strong antioxidant, and F3 has a
strong antioxidant.
5. Conclusion
Based on the results of the
research that has been carried out, it can
be concluded that: Formula F0 with a
concentration of 2.5% PRP, F1 with a
concentration of 2.5% PRP and 1%
asiaticoside, F2 with a concentration of
2.5% PRP and 2% asiaticoside, and F3
with a concentration of 2.5% PRP and 3
% asiaticoside can be formulated into
lotion preparations. The four PRP lotion
formulas with asiaticoside as an
antioxidant during 10 days of storage at
cold temperatures (4oC) tend to be less
stable because there are significant
differences in the spreadability test,
viscosity test and pH test.
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