RESEARCH ARTICLE | MAY 24 2023

Structural and antifungal behaviours of Co0.6Fe2.4O4

nanoparticles covered by Syzygium polyanthum extract 
Trio Erik Setyawan; Ahmad Taufiq  ; Sunaryono Sunaryono; ... et. al

AIP Conference Proceedings 2748, 020055 (2023)

https://doi.org/10.1063/5.0138546

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 Structural and Antifungal Behaviours of Co0.6Fe2.4O4
Nanoparticles Covered by Syzygium polyanthum Extract
Trio Erik Setyawan, Ahmad Taufiqa), Sunaryono Sunaryono, Arif Hidayat, and
Markus Diantoro

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Department of Physics, Faculty of Mathematics and Natural Sciences, Universitas Negeri Malang,
Jl. Semarang 5 Malang, 65145 Indonesia
a)
Corresponding author: ahmad.taufiq.fmipa@um.ac.id

Abstract. In this research, we produced Co0.6Fe2.4O4 nanoparticles covered by natural surfactants from Syzygium
polyanthum extract. The results of XRD analysis showed that the Co0.6Fe2.4O4 nanoparticles were successfully synthesized,
which was indicated by the presence of the spinel phase without other phases. The Co0.6Fe2.4O4 nanoparticles had a particle
size of 12.1 nm. SEM image indicated that the morphology of the nanoparticles was spherical and agglomerated with an
average particle size value of 34.2 nm. Furthermore, the FTIR spectrum showed the vibrational peaks of C=C and C-H at
the wavenumbers of 1633 cm−1 and 2881 cm−1, indicating the presence of the surfactant Syzygium polyanthum. At the
wavenumbers of 500₋600 cm−1, the vibrational peaks of Fe-O and Co-O were overlapped, which indicated that the Co2+
ion has succeeded in replacing the position of the Fe3+ ion. Our data analysis results also captured that the Co0.6Fe2.4O4
nanoparticles using natural surfactants from Syzygium polyanthum are potential candidates as high-performance antifungal
agents.

INTRODUCTION
In the last decades, microbial contamination has contributed to the increasing number of illnesses and deaths
worldwide. It is also reported that nearly half of the population in developing countries is exposed to microbial
contamination [1]. The microbial contamination caused by the fungus C. albicans has increased significantly [2].
Uncontrolled growth of C. albicans can cause various types of diseases [3]. Therefore, various attempts have been
conducted to overcome the growth of C. albicans, one of which is the use of antibiotics. However, most microbes are
capable of developing resistance to antibiotic treatment [4]. Therefore, it is central to design antimicrobial agents to
reduce the level of microbial contamination.
Recently, the use of nanomaterials as antimicrobial agents has become an important way in recent years. Based on
the previous work, Fe3O4-based nanomaterials have been intensively carried out and have become one of the promising
approaches as antimicrobial agents. In general, Fe3O4 nanoparticles have superparamagnetic properties, a large surface
area [5], and are biocompatible [6]. Recent research also portrays that Fe3O4 nanoparticles were believed to have
excellent antibacterial activity [7] and antimicrobial performance against C. albicans [3]. On the other hand, Fe3O4
entails limitations such as easy oxidation, agglomeration, and low stability [8], so that further innovation is needed to
improve Fe3O4 performance. This innovation can be achieved by administering cobalt doping. Cobalt is able to
increase stability and minimize air oxidation in magnetite [9]. Previous research reported that cobalt was able to show
antimicrobial activity against dermatophyte and phytopathogenic fungi [7]. In addition, cobalt, in the form of Co2+
ions, has been shown to be able to penetrate bacterial cell walls [10]. The combination of magnetite and cobalt material
is believed to increase the effectiveness of the antifungal. In a previous study, Naik et al. found that the CoxFe3-xO4
nanoparticles had antifungal activity [11].
In addition, further innovations are also necessary to improve the biocompatibility properties of the CoxFe3-xO4,
such as in the form of Co0.6Fe2.4O4 nanoparticles. This innovation can be achieved by administering the surfactant
prepared from Syzygium polyanthum. Syzygium polyanthum has been used as a traditional medicine that has efficiency

Proceedings of 2nd International Conference on Condensed Matters and Advanced Materials (IC2MAM)
AIP Conf. Proc. 2748, 020055-1–020055-6; https://doi.org/10.1063/5.0138546
Published by AIP Publishing. 978-0-7354-4493-5/$30.00

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against diarrhea, diabetes, and inflammation [12]. Syzygium polyanthum was also believed to have antifungal activity
against Aspergillus, Eurotium, Penicillium [13] and antibacterial activity against S. aureus [14], S. mutans [15].
Syzygium polyanthum is also useful as a reducing agent of organic and inorganic compounds [16]. Based on that note,
Syzygium polyanthum has great potentials as a natural surfactant as well as improving the antibacterial activity of
nanoparticles. Interestingly, up to now, there are no reports in fabrication of magnetic nanoparticles using Syzygium
polyanthum as surfactant agent as well as in fabrication Co0.6Fe2.4O4 nnaoparticles. Thus, in this case, the use of
Syzygium polyanthum as a surfactant is predicted to be able to improve biocompatibility properties and prevent
Co0.6Fe2.4O4 agglomeration. Overall, it is believed that the antifungal performance of the Co0.6Fe2.4O4 nanoparticles
with the Syzygium polyanthum surfactant is supferior to the Fe3O4 nanoparticles and the individual Syzygium
polyanthum surfactant.

EXPERIMENTAL METHOD

Synthesis of Co0.6Fe2.4O4 nanoparticles

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The Fe3O4 obtained from iron sand was reacted with HCl to get a solution of FeCl3 and FeCl2. Then, the solution
was filtered and separated from the sand residue using 60 mesh filter paper and its solution was taken. Furthermore,
CoCl2 powder was dissolved in distilled water to be mixed with the prepared FeCl2 and FeCl3 solution through the
titration process. The solution was stirred with a magnetic stirrer hotplate for 30 min. The resulting solution was
reacted with Syzygium polyanthum extract and titrated with NH4OH solution. After the titration process was complete,
a precipitate formed Co0.6Fe2.4O4 particles. The precipitate was then washed with distilled water to normal pH and
dried at 100 °C for one h.

Characterization
Several instrumentations were used to characterize the Co0.6Fe2.4O4 nanoparticles. X-ray diffraction (XRD) was
used to determine the crystal structure, phase, crystal size, and lattice parameters. The XRD used in this study was
PANalytical type X'Pert Pro, with a Cu-Kα radiation beam of 1.5406 Å. To determine the functional groups in the
sample, FTIR characterization was carried out. The FTIR used in this research was the Shimadzu brand with a
spectrum range of wavenumbers (4000 ₋ 400 cm-1). To determine the morphology and elemental content of
Co0.6Fe2.4O4 nanoparticles, SEM-EDX characterization type FEI (Inspect S50) with a magnification of up to 200,000
times and a high voltage of 25 kV was carried out. To determine the inhibition zone value of the Co0.6Fe2.4O4
nanoparticles against C. albicans, a disc diffusion test was performed.

RESULTS AND DISCUSSION

Fig. 1 showcases the results of the XRD characterization analysis of the Co0.6Fe2.4O4 nanoparticles. The diffraction
peaks indicate that the Co0.6Fe2.4O4 nanoparticles have been formed at an angle of 2θ = 30.1°; 35.5°; 43.1°; 53.5°;
57.1°; 62.7°; 74.1° by associated to crystal plane of (2 2 0), (3 1 1), (4 0 0), (4 2 2), (5 1 1), (4 4 0), and (5 3 3). From
the refinement results, it can be seen that the Co0.6Fe2.4O4 nanoparticles have a diffraction pattern that matches the data
model (AMCSD 0002402). The lattice parameter value of Co0.6Fe2.4O4 nanoparticles is 8.375 Å with the space group
of F d-3 m (α = β = γ = 90°). These results correspond to the previous research conducted by Gahrouei et al., which
indicated 8.40 Å of the lattice parameters [17]. In the spinel ferrite structure, the Co2+ ion substitution tends to replace
the Fe3+ ion in the tetrahedral position [18]. Thus, it causes stretching of the crystal axis and increases the distance of
the crystal plane, which is influenced by the Co2+ ionic radius (74 pm) which is greater than those of Fe3+ ionic radius
(64 pm) [18]. The detail of crystal structure can be found in Fig. 2.
It is believed that the presence of Co2+ ion in the tetrahedral position tends to reduce the fraction of Fe3+ ion so that
the particle size enlarges. In the previous research conducted by Wahda and Mohamed, it has been confirmed that the
lattice parameters and crystallite size tend to increase monotonically with the increase in the molar fraction of Co2+
[19]. The crystallite size formed from the Co0.6Fe2.4O4 nanoparticles is 12.1 nm. The result of the particle size is smaller
when compared to the particle size in the previous work prepared by using seaweed surfactant (Sargassum muticum),
which is 17 nm [20]. Giving Syzygium polyanthum surfactant is believed to reduce the crystallinity of the Co0.6Fe2.4O4
nanoparticles and leads to a decrease in the crystallite size.

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 100

(311)
80

Intensity (arb.u)
60

40
(220) (440)
(400) (511)
20

0
10 20 30 40 50 60 70 80

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2-Theta (deg.)
FIGURE 1. X-ray Diffraction Pattern of Co0.6Fe2.4O4 Nanoparticles

FIGURE 2. Ferrite Spinel Crystal Structure of Co0.6Fe2.4O4 Nanoparticles

The functional groups of Co0.6Fe2.4O4 nanoparticles were analyzed using FTIR, as shown in Fig. 3.
Characterization of FTIR produces vibrations between ionic bonds and forms a peak at a specific wavenumber. Based
on the FTIR characterization, there were Fe-O vibrational peaks at the wavenumbers of 400₋600 cm−1 [21]. The peak
vibration at the wavenumber of 412 cm−1 occurs due to the stretching of the Fe-O ion bond at the octahedral site, while
the peak at the wavenumber of 542 cm−1 is caused by stretching the Fe-O ion bond at the tetrahedral site. There is an
overlap between the Fe-O and Co-O ionic bonds at the tetrahedral site so that the Co peak cannot be detected. [22].
Subsequently, there is a vibration peak of the double bond C=C at the wavenumber of 1633 cm−1 which indicates the
content of phenolic compounds in the surfactant Syzygium polyanthum [23].

C=C
% Transmittance (arb. u.)

C-H

CO2
C-H

Fe-O

-OH Fe-O
4000 3600 3200 2800 2400 2000 1600 1200 800 400
Wavenumber (cm-1)

FIGURE 3. FTIR Spectrum of Co0.6Fe2.4O4 Nanoparticles

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 There are vibrational peaks of C-H and C-H sp2 at wavenumbers of 1361 cm−1 [24] and 2881 cm−1 [25] which
indicate the content of aromatic compounds in the surfactant Syzygium polyanthum. In addition, the peak vibration at
the wavenumber of 2351 cm−1 indicates the formation of CO2 molecular bonds in the air [26]. Furthermore, there is
an OH functional group bond at the peak of the vibration of 3439 cm−1. The presence of OH functional groups and
aromatic compounds indicates that the phenolic group compounds in the surfactant Syzygium polyanthum can perform
as antioxidants [25].

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FIGURE 4. SEM Image and Size Distribution of Co0.6Fe2.4O4 Nanoparticles

SEM characterization was carried out to generate information about the particle size distribution and morphology
of the sample. SEM informs the agglomeration rate of a material. The results of SEM characterization and sample
particle size distribution are shown in Fig. 4. The synthesized Co0.6Fe2.4O4 nanoparticles are spherical and have
undergone agglomeration with uniform size distribution. Theoretically, the agglomeration of Co0.6Fe2.4O4
nanoparticles is caused by the influence of magnetic interactions between the nanoparticles, which act as
ferromagnetic materials [27]. Agglomeration can also be affected by the increase in kinetic energy caused by collisions
between particles [28]. As a result, the nanoparticles tend to be sturdy to overcome the barrier between them and
agglomerate into large particles. The Co0.6Fe2.4O4 nanoparticles have an average particle size distribution of 34.24 nm.
The results of the average particle size distribution are nearly close to the results of previous studies conducted by
Jouyandeh et al., which indicates the size of 35 nm [29].

FIGURE 5. Inhibition Zone Diameter of Co0.6Fe2.4O4 Nanoparticles

In this research, the antifungal activity of the Co0.6Fe2.4O4 nanoparticles sample were tested using the agar diffusion
method by observing the diameter of the inhibition zone against Candida albicans [30]. This method was chosen as it
is easy and accurate in measuring the inhibitory area formed by the penetration of the active compound. From the test,
a clear zone was formed around the sinkhole. The results of testing the antifungal activity of Co0.6Fe2.4O4 nanoparticles
against C. albicans are shown in Fig. 5. Based on the measurement results, it appears that the Co0.6Fe2.4O4
nanoparticles have a fairly high inhibition zone diameter, which is 7.2 mm. This indicates that the Co0.6Fe2.4O4
nanoparticles are effective in their use as an antifungal agent against C. albicans.

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 The antifungal activity is closely related to the presence of reactive oxygen species such as hydroxyl peroxide,
hydroxyl radicals, superoxide radical anions, and hydroxyl ions contained in the sample [31]. The addition of
surfactant Syzygium polyanthum is believed to be able to increase the effectiveness of the inhibition zone in C.
albicans. Polyphenol compounds contained in the surfactant Syzygium polyanthum are able to damage the membrane
of fungal and bacterial cells [32]. On the other hand, the surfactant Syzygium polyanthum has biocompatibility agents
and leads to an increase in reactive oxygen species in the sample [33].

CONCLUSION
In this research, the Co0.6Fe2.4O4 nanoparticles were successfully synthesized using the coprecipitation method,
which is characterized by no other peaks in the diffraction peaks of magnetite nanoparticles with Fd-3m space group
and the form of a cubic spinel crystal structure. Based on the XRD analysis, it is observed that the particle size is 12.12
nm. The FTIR characterization results of Co0.6Fe2.4O4 nanoparticles showed vibrational peaks of C=C and C-H at the
wavenumbers of 1633 cm−1 and 2881 cm−1 which indicated the presence of bay leaf surfactant. There was overlapping
at the peak of Fe-O and Co-O vibrations at the wavenumber 5of 42 cm-1. SEM analysis results showed that the

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Co0.6Fe2.4O4 nanoparticles were spherical and agglomerated with an average particle size of 34.24 nm. The antifungal
test results against C. albicans indicated that the Co0.6Fe2.4O4 nanoparticles had a superior antifungal performance with
an inhibition zone diameter of 7.2 mm.

ACKNOWLEDGMENTS
This reserach was supported by the PNBP Universitas Negeri Malang research grant for AT.

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