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

atikah.arifah93@gmail.com

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