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International Conference on Green Energy and Environment 2019 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 353 (2019) 012038 doi:10.1088/1755-1315/353/1/012038

Antibacterial Effectiveness Of Formulations Nanoemulsion

Cratoxylum Glaucum Korth. Extract

S Enggiwanto, N Riyani, Y Pratama, O Roanisca and R G Mahardikaa

Departement of Chemistry, Universitas Bangka Belitung

Jl. Kampus Peradaban, Merawang, Bangka, 33172
a
E-mail: [email protected]

Abstract. Cratoxylum glaucum Korth. is a members of the family Hypericaceae which are
spread in Indonesia, one of which is found in the forest of the Bangka Belitung Islands.
Cratoxylum glaucum Korth is often used by the people of Bangka Belitung as a flavoring
ingredient from ancient times until now it is still consumed and utilized by people around Bangka
Belitung. One source of natural antioxidants is the top of the crust. The effort of increasing its
activity is very strong, so in this study we try to develop extraction of Cratoxylum glaucum made
in the form of nanoemulsion. This study aims to determine the comparison of Cratoxylum
glaucum extract in nanoemulsion preparations with extracts as antibacterial. Nanoemulsion is
made with the composition of Virgin Coconut Oil (VCO) as an oil phase, tween 80 as surfactant
and aquades as a water phase with a ratio of 2.5: 10: 25. The nanoemulsion extract of Cratoxylum
glaucum have a size 422.9 nm and PI value 0.493. The antibacterial activity of nanoemulsion
formulation has inhibitory growth power in moderate categorized Staphylococcus aureus
bacteria, whereas in the extract of Cratoxylum glaucum it has the inhibitory growth of active
Staphylococcus aureus bacteria.

1. Introduction

Cratoxylum glaucum are one of the members of the family Hypericaceae which are spread in Indonesia,
one of which is found in the forest of the Bangka Belitung Islands. Cratoxylum glaucum Korth. plants
are known by the people of Bangka Belitung as endemic plants, usually given the name “Pucuk Idat”.
The species Cratoxylum glaucum is often used by the people of Bangka Belitung as a flavoring
ingredient from ancient times, until now it is still consumed and utilized by people around Bangka
Belitung .
Cratoxylum glaucum are widely used such as the use of the roots, stems, leaves and bark as traditional
medicine to facilitate breastfeeding, tighten the skin, treat diarrhea, anticancer, antibacterial, antiviral
and other diseases. The major secondary metabolites of Cratoxylum glaucum Korth. include
anthraquinone and xanthones. The major metabolites of Cratoylum glaucum Korth. has similarities to
the major metabolites of mangosteen (Garcinia mangostana) which have bioactivity as an antioxidant
[1].
The antioxidant activity of Cratoxylum glaucum Korth. ethyl acetate extract showed very strong
activity with an IC50 value of 32.212 µg / mL [2]. The effort of increasing its activity is very strong, so
Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution
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Published under licence by IOP Publishing Ltd 1
International Conference on Green Energy and Environment 2019 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 353 (2019) 012038 doi:10.1088/1755-1315/353/1/012038

in this study we try to develop extraction of Cratoxylum glaucum Korth. to made in the form of
nanoemulsion. The use of nanoemulsion in the drug delivery system is called Self-Nanoemulsifying
Drug Delivery System (SNEDDS). SNEDDS is a corporation of oil, surfactants, cosurfactants and active
substances when mixed with water to form oil / water type nanoemulsions [3]. Nanoemulsion is a new
drug delivery system having a size range between 50 - 1000 nm [4]. The term nanoemulsi is said to be
a stable, clear preparation of two insoluble liquids, such as oil and water, stabilized by the interface film
of surfactant molecules [4].
The advantage of nanoemulsion is to have a good surface area compared to macroemulsion so that it
is more effective as a carrier system. The small size of nanoemulsion particles can penetrate easily
through the skin layer and increase skin penetration in the drug inserted [5]. In addition nanoemulsi
preparations are used as an alternative to make small droplet sizes can facilitate penetration and
absorption across bacterial membranes, with this measure nanoemulsion has a clear and transparent
physical form and increases the effectiveness of the Cratoxylum glaucum Korth as antibacterial.
Therefore, in this study, the formulation of the Cratoxylum glaucum Korth. acetone extract of
nanoemulsion which has high stability and small droplet size as an antibacterial effectiveness against
Staphylococcus aureus.

2. Research methods

2.1 Sample Preparation

Leaves of Cratoxylum glaucum Korth. are obtained from Kimak Village, Pemali District, Bangka
Regency. Then the sample is dried under the sun. The dried sample is ground into dry powder.
Furthermore, sifted and macerated.

2.2 Extraction
Dry powder leaf of Cratoxylum glaucum Korth. was taken 50 grams macerated with acetone solvents as
much as 500 mL for 3 x 24 hours. After that the obtained filtrate was concentrated with rotary evaporator
vacum until the concentrated extract of Aseton was obtained.

2.3 Preparation of Nanoemulsion Acetone Extract Chratoxylum glaucum Korth.

Chratoxylum glaucum Korth. extract was made by varying the addition of acetone extracts by 0.01, 0.02,
0.03, 0.04, 0.05, 0.075, 0.1, 0.25, 0.5, 0.75 g which is suspended with an emulsifier. Each extract will
be suspended in virgin coconut oil, with magnetic stirring. Then added with tween 80, added with
distilled water and stirring using homogenezer at a speed of 8,000 for 30 minutes. Then measurements
were made of physical stability and percent transmittance [6]. Furthermore, the product nanoemulsion
product was analyzed by particle size analyzer (PSA).

2.4 Antibacterial Activity Testing

Antibacterial testing was carried out by the inhibitory zone method. 50 μL were taken from each test
solution with a concentration of 20%, 50% and 80% as well as a positive control solution and a negative
control. Then put into a hole that has been formed. After that, it was incubated for 24 hours at 37 oC.
Measurements are made by looking at the inhibition zone formed.

3. Result and Discaussion

3.1 Physical Stability of Nanoemultion

One of the physical stability tests was carried out by measuring nanoemulsion transmittance. The percent
transmittance test obtained showed that the more extracts used, the smaller the yield value of
transmittance was obtained because the extracts added with more concentrations would produce more
concentrated colors or the mixture of oil and surfactant would be incorporated by extract. Good
nanoemulsion has a clear or clear visual appearance with a high percent transmittance value. the addition

2
International Conference on Green Energy and Environment 2019 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 353 (2019) 012038 doi:10.1088/1755-1315/353/1/012038

of Cratoxylum glaucum depends on the solubility in the system used [6],[7]. PH testing decreased in the
concentration range of 0.075 - 0.1 to be more acidic. this shows that the more extracts, the smaller or
acidic pH values will be obtained [8].

Table 1. Physical Stability Test Results of Nanoemulsion Cratoxylum glaucum

Concentration Transmittance
Formula pH Density
of Extract (gr) (%)
E1 0.01 6 69.2 0.99138
E2 0.02 6 58.07 0.98990
E3 0.03 6 44.06 0.98778
E4 0.04 6 43.56 0.98909
E5 0.05 6 37.26 0.98915
E6 0.075 6 25.17 0.98830
E7 0.1 5 10.68 0.99204
E8 0.25 5 1.29 0.99009
E9 0.5 5 0.043 0.99122
E10 0.75 5 0.014 0.98958

3.2. Particle Size Analysis of Nanoemulsion Formulations

Nanoemulsion measurement using PSA was done three times. The results of the analysis of the size of
nanoemulsion extract of Cratoxylum glaucum extract are shown in Figure 1. The particle size analyzer
results showed that nano size was formed from formulation E1 which consisted of 0.01 gr extract with
the ratio of Virgin Coconut Oil (VCO) as an oil phase, tween 80 and aquades 2.5: 10: 25. From the
results of the PSA it shows that the size formed by the size of nanoemulsion <10 nm normalized intensity
distribution 12-20%.

Figure 1. Graph of particle size analyzer results (normalized intensity distribution) of formulation
nanoemultion Cratoxylum glaucum Korth.

The results of droplet size and the value of the polydispersity index (PI) shown in Table 2. PI which
can help provide information about the stability and uniformity of nanoemulsion sizes. The results

3
International Conference on Green Energy and Environment 2019 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 353 (2019) 012038 doi:10.1088/1755-1315/353/1/012038

showed that the droplets size of Cratoxylum glaucum extract nanoemulsion was 422.9 nm. The
nanoemulsion size ranges from 50-1000 nm [4]. So, the results of the droplet size obtained in this study
met the nanoemulsion preparation criteria. Furthermore, the results of the PI value indicate the
homogeneity or stability of nanoemulsion. The smaller the value of PI, the size of the droplet will be
more uniform and homogeneous [9]. The Good PI value ranged from 0.01 - 0.7. The results of the PI
value of this study amounted to 0.493. So that the Cratoxylum glaucum extraction nanoemulsion
formulation is included in the general nanoemulsion criteria.

Table 2. Analysis of nanoemulsion of Cratoxylum glaucum extract using Particle Size Analyzer
E1 Diameter (nm) PI
Average 422.9 0.493

3.3. Antibacterial Activiy

Based on the results of Antibacterial testing obtained showed that the results of the thick extract of
Cratoxylum glaucum had a greater inhibitory value than in the form of nanoemulsion for testing
Stapylococcus aureus bacteria. The extract of Cratoxylum glaucum which is used in testing is
completely pure or without any comparison of mixtures whereas in the form of nanoemulsion the extract
ratio is different. The level of inhibition of bacterial growth was categorized as <5 mm weak, medium
5-10 mm, 10-19 active and 20 mm very active [10] (Table 3). Size of nanoemulsions particularly
effective delivery systems for Cratoxylum glaucum extract to increase antibacterial efficacy. This is
because the small size of the nanoemulsion makes it easy for the extract to interact with the surface of
the cell membrane [11].

Table 3. Atibacterial activity of Staphylococcus aureus

No Extract Concentration % Inhibition (mm)

1. Extract 20% 14.025
2. Extract 50% 15.275
3. Extract 80% 15.215
4. E1 20% 9.57
5. E1 50% 7.865

4. Conclusion
Based on the research carried out, a formulation of nanoemulsion extract of Cratoxylum glaucum have
a size 422.9 nm and PI value 0.493. Composition of formulation nanoemulsion in this study consists of
0.01 g (2%) extract with the ratio of VCO: tween 80: aquades 2.5: 10: 25. The antibacterial activity of
nanoemulsion formulation has inhibitory growth power in moderate categorized Staphylococcus aureus
bacteria, whereas in the extract of Cratoxylum glaucum it has the inhibitory growth of active
Staphylococcus aureus bacteria.

References
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International Conference on Green Energy and Environment 2019 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 353 (2019) 012038 doi:10.1088/1755-1315/353/1/012038

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Acknowledgments
This research partially funded by Directorate General of Higher Education (DIKTI) Ministry of
Education and Research, Republic of Indonesia through Program Kreatifitas Mahasiswa Penelitian
Eksakta (PKMPE) grant 2018 and USAID through the SHERA program - Centre for Development of
Sustainable Region (CDSR). In year 2017-2021 CDSR is led by Center for Energy Studies - UGM.