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Neem Leaf Extract ResearchPublication Seminar
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Presented on The 2nd
International Conference on Pharmaceutical Nanotechnology/Nanomedicine 2015, 6th
June 2015
ANTIBACTERIAL CREAM FORMULATION FROM NEEM LEAF EXTRACT
(Azadirachta indica A. Juss) FOR ACNE TREATMENT Atikah Arifah
1, M.F. Arifin
1, Lisa T J
1, Aris H S
1, Winda D J
1
Faculty of Pharmacy Pancasila University Jakarta1
ABSTRACT
Neem leaf contains antibacterial compounds such as flavonoids, alkaloids, triterpenoids, and tannins
and can be used in the treatment of acne. In this study, neem leaves are formulated as o/w creams
to increase its convenience when applied to the skin. Neem leaves were extracted with 96% ethanol
and the Minimum Inhibitory Concentration (MIC) against Staphylococcus aureus was determined
using dilution method. Based on MIC values, creams are made in 3 formula with variety of extract
concentration. The cream preparations were evaluated in physical, chemical and microbiological
aspects including organoleptic, homogenity, emulsion type, spreadability, globule size, viscosity and
rheology, centrifugation, pH, and microbial contamination test. Accelerated stability test is also
conducted at a temperature of 40°C and sampled at week 0, 2, and 4. The antiacne effectivity of the
cream preparations were tested in vitro by inhibition zone observation and in vivo by Region of
Interest (ROI) method, Accelerated stability test’s data were analyzed using non-parametric Kruskal
Wallis method to draw conclusions. The results showed that neem extract has MIC values of 4%.
Accelerated stability test showed that the cream remains stable after storage for 4 weeks at 40°C.
Neem leaf extract creams showed inhibition zone of 8.95 to 14.45 mm. In vivo testing showed that
the test subjects’ acnes are healed after using the cream for 7 days. It can be concluded that neem
leaf extract may be formulated into a stable cream preparations in physics, chemistry, and
microbiology aspects and effective as an antibacterial for acne.
Keywords: Neem, o/w cream, acne, Staphylococcus aureus
INTRODUCTION
Acne is a skin disease caused by the
Staphylococcus aureus bacteria and generally
treated with topical antibiotics, which can
lead to resistance problems, so that nowadays
herbal actives based acne treatment are often
developed. Neem is a plant that grows well
and may easily cultivated in Indonesia. Neem
leaves contain alkaloids, flavonoids,
triterpenoids, and tannins which is effective
as an antibacterial and anti-inflammatory, so
that neem leaves juice is often used in
traditional acne treatment. This treatment is
considered impractical because the juice
should be fresh and can not be stored for a
long time. To overcome that problems, in this
research, the neem leaves were extracted and
formulated into oil-in-water (o/w) creams.
The cream preparations were evaluated in
physical, chemical, microbiological, and
effectivity aspects.
INSTRUMENTS
Instruments used in this research are: Rotary
vacuum evaporator, stirrer (Hsiangtai), oven
(Memmert), refrigerator, spreadability tester
kit, Brookfield viscometer, microscope
(Olympus), micrometer, vernier calipers, pH
meter (Hanna, HI-2211), centrifuge (Kokusan,
H-103N), incubator (Memmert), waterbath
(Memmert, W-600), colony counter (Galaxy-
320), laboratory glassware (Pyrex).
MATERIALS
Neem leaves, ethanol 96%, stearic acid, cethyl
alcohol, glyceril monostearic, triethanolamine,
isopropyl myristate, propylene glycol, methyl
paraben, propyl paraben, sodium
metabisulfite, citric acid, aquadest, nutrient
agar, bacteria pepton broth, potato dextrose
Agar, trypticase soy broth (TSB), vogel
johnson agar, cetrimide agar, chromogenic
agar, Staphylococcus aureus, Sudan III,
Methylene Blue, antibacterial cream (contains
Gentamisin).
Presented on The 2nd
International Conference on Pharmaceutical Nanotechnology/Nanomedicine 2015
June 2015
RESEARCH PRINCIPAL
The neem leaves were cut into small pieces,
dried, extracted by maceration with 96%
ethanol, thickened with a rotary vacuum
evaporator and the Minimum Inhibitory
Concentration (MIC) against Staphylococcus
aureus were determined using dilution
method. Based on MIC values, cream
made in 3 formula with variety of extract
concentration.
The cream preparations were evaluated in
physical, chemical and microbiological aspects
including organoleptic, homogenity, emulsion
type, spreadability, globule size, viscosity and
rheology, centrifugation, pH, and microbial
contamination test. Accelerated stability test
is also conducted at a temperature of 40°C
and sampled at week 0, 2, and 4. The antiacne
effectivity of the cream preparations were
tested in vitro by inhibition zone obs
and in vivo by Region of Interest
method, Accelerated stability test’s data were
analyzed using non-parametric Kruskal Wallis
method to draw conclusions.
RESULT AND DISCUSSION
The cream preparations pictures and
evaluation results could be seen below:
Figure 1. Neem leaf extract cream preparations
Figure 2. Inhibition zone results
0
5
10
15
20
25
Bla
ng
ko
Ko
ntr
ol +
Ko
ntr
ol -
Ek
stra
k 4
%
Ek
stra
k 8
%
Ek
stra
k …
F1
DD
H (
mm
)
International Conference on Pharmaceutical Nanotechnology/Nanomedicine 2015
cut into small pieces,
dried, extracted by maceration with 96%
ethanol, thickened with a rotary vacuum
evaporator and the Minimum Inhibitory
Staphylococcus
were determined using dilution
Based on MIC values, creams are
made in 3 formula with variety of extract
The cream preparations were evaluated in
physical, chemical and microbiological aspects
including organoleptic, homogenity, emulsion
type, spreadability, globule size, viscosity and
centrifugation, pH, and microbial
test. Accelerated stability test
is also conducted at a temperature of 40°C
and 4. The antiacne
effectivity of the cream preparations were
by inhibition zone observation
Region of Interest (ROI)
method, Accelerated stability test’s data were
parametric Kruskal Wallis
The cream preparations pictures and
seen below:
Figure 1. Neem leaf extract cream preparations
results
Figure 3. Accelerated stability test results
Accelerated stability test for cream
preparation’s result showed that all formulas
remain stable after 4 weeks
40°C. The organoleptic test showed that the
creams are acceptable for skin application,
and all formulas are homogenous. The
cream’s viscosity increased while the
spreadability decreased following the increase
of extract concentration used in cream
formulas. Cream type is o/w so that the
creams are easy to washed with water and
doesn’t feel oily on the skin.
rheology is plastic thixotropic, which
characterized by a high viscosity at low stress,
but decreased when stress is applied and
starts flowing when the yield value is
exceeded. The cream’s pH varies between
5,99-7,01. In vitro test showed that the
creams have an inhibition zone of 8,94
mm and in vivo test showed that subject’s
acnes are improved as the redness and size of
acnes are reduced.
CONCLUSION
The ethanol extract of neem
formulated into a cream
physics, chemistry and microbiology
and effective as an antibacterial
F1
F2
F3
International Conference on Pharmaceutical Nanotechnology/Nanomedicine 2015, 6th
Figure 3. Accelerated stability test results (week 4)
Accelerated stability test for cream
preparation’s result showed that all formulas
remain stable after 4 weeks of storage at
40°C. The organoleptic test showed that the
creams are acceptable for skin application,
and all formulas are homogenous. The
s viscosity increased while the
spreadability decreased following the increase
of extract concentration used in cream
formulas. Cream type is o/w so that the
creams are easy to washed with water and
doesn’t feel oily on the skin. The cream’s
astic thixotropic, which
characterized by a high viscosity at low stress,
but decreased when stress is applied and
starts flowing when the yield value is
ed. The cream’s pH varies between
7,01. In vitro test showed that the
tion zone of 8,94-14,45
mm and in vivo test showed that subject’s
acnes are improved as the redness and size of
of neem leaves can be
cream which is stable at
microbiology aspects
as an antibacterial for acne
Presented on The 2nd
International Conference on Pharmaceutical Nanotechnology/Nanomedicine 2015, 6th
June 2015
REFERENCES
1. Soegihardjo CJ. Mimba (Azadirachta indica A. Juss, suku Meliaceae), Tanaman Multi Manfaat yang Dapat Menanggulangi Persoalan Rakyat Indonesia. SIGMA. 2007; 10 (1): 83 – 102.
2. Debjit B, Chiranjib L, Jitender Y, K.K Tripathi, K.P. Sampath K. Herbal Remedies of Azadirachta indica and its Medicinal Application. J Chem Pharm. Res. 2010; 2(1): 62 – 72.
3. Nestri H, M.Widyo W, Riskha KM. Identifikasi dan Uji Aktivitas Antibakteri Fraksi Teraktif Daun Mimba (Azadirachta indica A. Juss). Hibah bersaing. Surakarta: Universitas Sebelas Maret. 2009.
4. Hanni M. Aktivitas Antibakteri Ekstrak Daun Mimba (Azadirachta indica) pada Staphylococcus aureus (skripsi). Malang: Akademi Putra Indonesia; 2012. p. 1 – 5.
Thank you for: Faculty of Pharmacy, University of Pancasila, Jagakarsa, South Jakarta