Title


Science and Technology Indonesia
e-ISSN:2580-4391 p-ISSN:2580-4405
Vol. 8, No. 3, July 2023

Research Paper

Coprecipitation Synthesis and Antimicrobial Effect Study of Europium Doped Spinel
Manganese Ferrites Nanoparticles (MnEu0.1Fe1.9O4NPs)
Amina Chidouh1*, Tarek Tahraoui2, Badra Barhouchi3
1Laboratory of Chemistry, Physics and Materials Biology, Department of Natural Sciences, Higher Normal School of Technological Education, Skikda, 21000, Algeria2Mines Metallurgy Materials Laboratory L3M, National Higher School of Technology and Engineering - ENSTI Annaba, 23000, Algeria3Pharmaceutical Sciences Research Center (CRSP), Constantine, 25000, Algeria
*Corresponding author: amchidouh@gmail.com

AbstractDue to the high prevalence of micro-organisms resistant to conventional antimicrobials, the search for new antimicrobial drugs isunderway, with nanoparticles being one of the options. This study reports for the first time the use of the coprecipitation method tosynthesize europium (Eu) doped spinel manganese ferrites nanoparticles (MnEu0.1Fe1.9O4NPs). The purpose of this research is todetermine the antimicrobial activity of MnEu0.1Fe1.9O4NPs. MnEu0.1Fe1.9O4NPs were analyzed using Fourier Transform InfraredSpectroscopy (FTIR), X-Ray Diffraction (XRD), and Scanning Electron Microscopy (SEM) combined with Energy Dispersive X-RayAnalysis (EDX) to determine their structure, size, morphology and elemental compositions. The antimicrobial activity of synthesizednanoparticles was evaluated qualitatively using a diffusion disc on agar, followed by minimum inhibitory concentrations (MIC)determination. The findings show that all tested strains were adversely affected by the examined NPs, where E. coli exhibited thehighest sensitivity to NPs, followed by S. aureus. The NPs displayed a moderate level of anti-candida action. MnEu0.1Fe1.9O4NPscould be exploited in biomedical usages.
KeywordsCoprecipitation Method, Nanoparticles, Antibacterial Activity, Anti-candida Action, Ferrites Spinel

Received: 19 January 2023, Accepted: 20 June 2023
https://doi.org/10.26554/sti.2023.8.3.494-500

1. INTRODUCTION

Antibacterial resistance rapidly increases throughout numer-
ous bacterial species, becoming a significant clinical and public
health problem worldwide (Kadiyala et al., 2018) . Nanoscale
engineering of nanoparticles (NPs) and surfaces gives new ways
for antibacterial drugs that increase conventional organic chem-
istry methods (Bozon-Verduraz et al., 2009; Jiang et al., 2009).
Metal oxide NPs (MO-NPs) have shown promising results in
inhibiting bacterial growth and combating antibiotic resistance
(Djurišić et al., 2015; Horie et al., 2012). MO-NPs are also
desirable as antimicrobial medicaments because they are stable
and less toxic to human cells than organic NPs (Deravi et al.,
2007) . Their nanoscale and variable surface chemistry. Their
changeable surface chemistry and nanoscale permit MO-NPs
to cause toxicity of bacteria over different modes of action, like
proteolysis, enzyme inhibition, cell membrane lysis, oxidative
stress and lipid peroxidation (Djurišić et al., 2015; Coker et al.,
2012; Allahverdiyev et al., 2011).

Among the various MO-NPs, spinel ferrites (MFe2O4 such
as M=Mn2+, Fe2+, Co2+, Ni2+) have been widely used due to

their magnetic and electrical advantages (Reddy et al., 2012;
Kollu et al., 2015). Mn is of particular interest in the biomedical
field due to its different oxidation states. Mn2+ ions show the
highest stability compared to Mn3+ and Mn4+ ions (Kalaiselvan
et al., 2022) . A cubic spinel structure characterizes manganese
ferrite (MnFe2O4). These nanoparticles are necessary materials
based on their moderate magnetization, size, biocompatibility,
high sensitivity, good chemical stability and tunable toxicity in
biological systems (Al Zahrani et al., 2022; Kalaiselvan et al.,
2022).

These ferrites are highly important in biomedicine as mag-
netic carriers for bioseparation, protein immobilization and en-
zymes (Arulmurugan et al., 2005; Costa et al., 2003). MnFe2O4
nanostructures are promising for clinical cancer diagnosis and
therapy (Kalaiselvan et al., 2022) . The chemical formula AB2O4
generally represents the spinel structure, where A and B rep-
resent the tetrahedral (surrounded by 4 oxygen atoms) and
octahedral (surrounded by 6 oxygen atoms) sites (Kalaiselvan
et al., 2022) .

Different synthesizing methods, such as hydrothermal syn-
thesis, alcohol dehydration, and sol-gel and spray drying, have

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https://doi.org/10.26554/sti.2023.8.3.494-500


Chidouh et. al. Science and Technology Indonesia, 8 (2023) 494-500

been reported (Liu et al., 2018; Rahman and Ahmed, 2005).
However, these processes are not economically suitable for
large-scale production. The coprecipitation process enables
the production of ultrafine powders with chemically uniform
composition, orderly size and good reactivity (Bueno et al.,
2007; Verma and Chatterjee, 2006). The advantages of this
process are ease of processing, excellent production efficiency,
low energy loss and high-purity product (Bandekar et al., 2019) .
The main drawbacks of co-precipitation methods are poor crys-
tallinity and particle size dispersal, large agglomeration, and
the need for pH control (Houshiar et al., 2014; Zahraei et al.,
2015).

Recently, MO-NPs synthesized by rare earth (RE) ion dop-
ing have received considerable attention due to their field of
applications (Park et al., 2016; Prodi et al., 2015; Wang, 2008).
According to various studies (Bouzigues et al., 2011; Das and
Das, 2013; Yu et al., 2009; Zhuang et al., 2015), rare-earth
metals are essential in manufacturing modern technologies
such as mobile phones, computers, biomedical applications
and solar cells. Particular attention has been paid to rare-earth
cations, such as Eu3+, because of the resonance energy lev-
els and their suited spectroscopic properties. Therefore, eu-
ropium has been investigated for cancer treatment applications
and used in dyes for magnetic resonance applications (Jahani
et al., 2016) . Rare earth elements doped manganese ferrite has
many biological activities. Akhtar et al. (2019) concluded that
Mn0.5Zn0.5SmxEuxFe1.8−2xO4 (0.01≤×≤0.05) NPs have po-
tential probable anti-cancer and antibacterial abilities activities.
Mohafez et al. (2021) indicated that an acceptable antifun-
gal effect was observed in the presence of MnCe1.4Fe0.6O4
nanoparticles. Al Zahrani et al. (2022) reported that the higher
antibacterial activity of Ce3+ substituted MnFe2O4NCs was ob-
served. Therefore, to our knowledge, there are currently unde-
tailed studies about the synthesis and antimicrobial activity of
europium-doped manganese ferrite nanoparticles (MnEu0.1Fe
1.9O4NPs). To this end, this study investigates an original study
about the morphological, structural and antimicrobial activity
of MnEu0.1Fe1.9O4NPs. The findings that were achieved are
thoroughly described.

2. EXPERIMENTAL SECTION

2.1 Synthesis of Europium-doped Manganese Ferrite Nano
particles (MnEu0.1Fe1.9O4NPs)

The coprecipitation method was used to synthesize nanoparti-
cles of europium-doped manganese ferrite with the composi-
tion of MnEu0.1Fe1.9O4NPs. Manganese chloride tetrahy-
drate MnCl2.4H2O (Biochem, Chemopharma), europium
chloride EuCl3 (Sigma-Aldrich) and ferric chloride hexahy-
drate FeCl3.6H2O (Biochem, Chemopharma) were taken in
the molar ratio of 1:0.1:1.9 and dissolved in 100 mL of double-
distilled water with constant magnetic stirring for an hour.
Sodium hydroxide (2M) solution was added drop after drop
until the solution’s pH reached around 12. Instantaneously, the
orange solution turned dark brown. The solution was stirred
continuously in a water bath at 80°C. The precipitate was fil-

Figure 1. FTIR Spectrum of MnEu0.1Fe1.9O4 Nanoparticles

tered and repeatedly rinsed with deionized water and acetone,
then dehydrated at 80°C in the oven for 2 h. The sample was
annealed at 600°C in an electric muffle furnace for 6h (Ban-
dekar et al., 2019) . The co-precipitated ferrite was ground
using an agate pestle and mortar to obtain fine particles.

2.1.1 MnEu0.1Fe1.9O4NPs Characterization
Fourier Transform Infrared Spectroscopy (FTIR) analysis of
the synthesized nanoparticles was obtained with a SHIMADZU
FTIR-8000 spectrometer at room temperature utilizing KBr
pellets in the range of frequency of 4000-400 cm−1 with a
wavenumber resolution of 1 cm−1. The morphological and
elemental analysis was recorded using Scanning Electron Mi-
croscopy (SEM-EDX Quanta 250). X-ray Diffraction (XRD)
analysis has been recorded utilizing a Rigaku X-ray diffrac-
tometer with high-intensity Cu-K𝛼 radiation (_ = 1.54178
Å).

2.2 Microorganisms and Growth Conditions
This study utilized five classified bacteria ATCC: American
Type Culture Collection: Gram-positive bacteria: Staphylococ-
cus aureus 25923 and Enterococcus faecalis 29212, and Gram-
negative bacteria: Escherichia coli 25922, Klebsiella pneumoniae
700603 and Pseudomonas aeruginosa 25953. One clinical yeast,
Candida albicans, was also tested. The microorganisms were
graciously supplied from the culture collection of the Pharma-
ceutical Sciences Research Center, Algeria. Nutrient agar was
used as the growth media.

2.3 Antimicrobial Assays
The qualitative assessment of the synthesized nanoparticles
antimicrobial effect was performed using disk diffusion on agar
and minimum inhibitory concentration (MIC) (Vanden and
Vlirtinck, 1993) . The microbial strains were pre-grown at

© 2023 The Authors. Page 495 of 500



Chidouh et. al. Science and Technology Indonesia, 8 (2023) 494-500

Figure 2. SEM Micrographs with Average Particle Size
Distribution of MnEu0.1Fe1.9O4

Table 1. Elemental Composition of MnEu0.1Fe1.9O4 Nanopar-
ticles Obtained from EDX Analysis

Elements Weight% Atomic%

O K 47.89 77.28
EuL 5.08 0.86
MnK 15.48 7.27
FeK 31.55 14.59

37.0±0.1°C for 18 h on nutrient agar. Each strain was di-
luted in a saline solution at the McFarland scale 0.5, 1.5 ×
108 UFC/mL (equivalent to DO 0.08-0.1/_ = 625 nm). The
inoculum was streaked into agar plates using a sterile swab af-
ter the Mueller Hinton Agar (MHA) had solidified, and the
surface of the MHA was covered with a sterile filter disc (What-
man paper N°3) with a 6-mm diameter. A concentration of
2 mg/mL of NPs was prepared in methanol, and 50 `L of
the NPs was dropped onto the inserted discs. The plates have
been kept at 37°C for 18-24 h. Methanol was employed as a
negative control. Gentamicin (GEN) and ampicillin (AMP)
were used as positive controls. The effectiveness was deter-
mined by determining the zone’s diameter of microbial growth
inhibition (ZOI) atop the disc and recording the diameter with
a transparent ruler in millimeters. All tests were performed in
triplicate.

Figure 3. EDX Spectra and Elemental Composition for
MnEu0.1Fe1.9O4 Nanoparticles

Figure 4. XRD Pattern of MnEu0.1Fe1.9O4 Nanoparticles

MIC experiment was done in the 96-microplate using
the broth dilution method. 100 `L volume of the obtained
nanoparticle at different concentrations (16 to 0.5 mg/mL) was
prepared in wells previously inoculated with 50 `L of Mueller
Hinton Broth (MHB) broth medium. The freshly adjusted
inoculum to DO of 0.08 to 0.1 (50 `L) was added to the pre-
pared NPs solution and then incubated for 24 hours at 37°C.
Untreated bacteria with DMSO was used in the trial as the
negative control. The lowest concentration of the tested com-
pound that the microorganism does not demonstrate visible
growth was defined as the minimum inhibitory concentration
(MIC) (Rehman et al., 2019) .

3. RESULTS AND DISCUSSION

FTIR Spectroscopy is a valuable technique for structural anal-
ysis and cations redistribution between tetrahedral and octahe-
dral sites in spinel structures (Hakeem et al., 2016) . Figure 1
shows the FTIR spectrum of MnEu0.1Fe1.9O4NPs. The band
at 1425 cm−1 is for out-of-plane deformation vibration of the
C-H. A band characterizes adsorbed water at 3435 cm−1. The

© 2023 The Authors. Page 496 of 500



Chidouh et. al. Science and Technology Indonesia, 8 (2023) 494-500

bands at 2131 cm−1 and 1630 cm−1 are attributed to O-H
stretching and H-O-H bonded vibration modes, respectively.
The existence of the 576 cm−1 band is based on the vibration
stretching of octahedral and tetrahedral groups (Hakeem et al.,
2016) .

Figure 2 shows the Scanning Electron Microscopy (SEM)
micrographs of MnEu0.1Fe1.9O4 NPs. Micrographs were taken
at scales of 5, 10, 20 and 50 µm, it demontrates the formation
with uniform distribution of agglomerated nanoparticles (Devi
and Soibam, 2018) . Due to their high surface energy and mag-
netic properties, nanoparticles have the potential to aggregate
and develop into large assemblages (Sagadevan et al., 2018) .
The average nanoparticles size distribution was estimated in
the range of (30 nm-70 nm) using imageJ software .

Figure 3 displays the results of EDX measurments for the
elemental composition of the prepared sample. EDX spectra
show the purity, homogeneity and presence of Eu in the sam-
ple, indicating that Eu was successfully doped into the spinel
structure. As determined by the EDX analyses, the elemental
composition is listed in Table 1 with the atomic and weight
percentages.

X-ray diffraction measurements show that peaks of MnEu0.1
Fe1.9O4NPs correspond to peaks of a typical spinel structure
prepared by coprecipitation. Figure 4 displays the prepared
nanoparticles X-ray Diffraction pattern. The crystallite’s aver-
age size was in the range of 40.9 nm. Intense diffraction peaks
occur at 2\=18.15°, 29.78°, 34.99°, 42.63°, 52.70°, 56.18°,
61.73°, 64.68° corresponds to (111), (220), (311), (400), (422),
(511), (440), (531) usual planes of ferrite spinel arrangements
Fe2MnO4 that is characterized by face-centered cubic phase
belonging to space group of Fd3m (Hakeem et al., 2016; Mo-
hafez et al., 2021) according to the standard card JCPDS file
no. 100319 (Wang, 2008) . It is observed at about (20.24°,
33.25° and 40.90°) and (24.22°, 33.25°, 35.68°, 49.57° and
54.26°) the appearance of secondary phase peaks with an in-
significant amount which corresponds to the ortho ferrite (Eu-
FeO3) (01-074-1475) and hematite (Fe2O3) (01-089-0599)
phases respectively (Zubair et al., 2017) .

One of the crucial causes in the development of secondary
phases is attributed to the low solubility and electronic config-
uration of the rare earth element Eu3+ (Mohafez et al., 2021;
Shirsath et al., 2014). The Eu3+ ion with a bigger radius of
about (1.07 Å) prefers to occupy the octahedral sites of Fe3+,
whose ionic radius is 0.67 Å. Consequently, when the Eu3+ ions
substitution is high, this directs to the creation of secondary
phases as an impurity on the grain boundaries due to the dif-
fusion of RE-Eu3+ ions (Zubair et al., 2017) . The sample’s
average value of the peaks of XRD analysis is used to deter-
mine the crystallite size (D). The value of ‘D’ is calculated using
Scherrer’s formula Cullity (1978) as follows:

D = K𝛽 cos\ (1)

In which ‘D’ signifies the average crystallite size in nm, the
shape factor, ‘k’, has a value of 0.94, X-rays are employed, and
their wavelength is _ , \ is the Bragg’s diffraction angle, and 𝛽
is FWHM (Zubair et al., 2017) .

To the best of our knowledge, the antimicrobial action of
MnEu0.1Fe1.9O4NPs was investigated for the first time on both
yeast and Gram-positive/negative bacteria. Subsequently, the
results indicate that the synthesized MnEu0.1Fe1.9O4NPs had
exceptional antimicrobial activity against selected microbial
strains as shown in Table 2. Moreover, the checked NPs were
harmful to all the tested strains at the dose of 2 mg/mL and gave
moderate to strong activity, which indicated by the formation of
a clear zone. The inhibition zones (ZOI) were ranged from 12
to 24 mm. Among the tested bacterial strains, E. coli exhibited
the strongest sensitivity to NPs, followed by S. aureus, with ZOI
values of 24 ± 0.14 mm and 21 ± 0.24 mm, respectively. Fur-
thermore, the produced NPs also displayed an equally positive
effect against E. faecalis, K. pneumoniae and P. aeruginosa. Xiu
et al. (2012) included that inorganic nanoparticles are distin-
guished for their large surface-to-volume ratios and nanoscale
sizes, enhancing their response to pathogenic bacteria. Ac-
cording to Jesudoss et al. (2016) , the ferrite nanoparticles in-
volved significantly in combating infectious against both Gram-
positive and Gram-negative bacteria.

In addition, Candida albicans remains a potent yeast strain,
where the NPs showed a moderate anti-candida activity and pre-
sented the weakest inhibition zone (ZOI: 12 ± 0.24 mm). The
encouraging antibacterial potential of MnEu0.1Fe1.9O4NPs
nanoparticles was corroborated by the results of experiments
and compared with antibiotics, gentamicin and ampicillin, as a
positive control. Contrary to gentamicin which displayed a re-
markable antibacterial activity, ampicillin had no effect against
all tested bacteria. As revealed in Table 2, the MIC (minimum
inhibitory concentration) of MnEu0.1Fe1.9O4NPs was found
to be 1 mg/mL for E. coli, K. pneumoniae, P. aeruginosa and C.
albicans whereas 2 mg/mL MIC value was recorded for S. aureus
and E. faecalis. In agreement with our results, the synthesized
europium doped cerium dioxide (Eu3+CeO2) nanoparticles
exhibited outstanding antibacterial effectiveness against E. coli
and S. aureus with the following diameter of inhibition zone
(ZOI): 37 ± 0.3 mm and 18 ± 0.2 mm, respectively (Gnanam
et al., 2021) . Ashour et al. (2018) demonstrated that the fer-
rites had antibacterial activity against all pathogens tested. Zinc
cobalt ferrite nanoparticles (ZCFO) were the most effective,
with zones of inhibition of 13.0 mm against Bacillus subtilis and
Staphylococcus aureus.

However, Akhtar et al. (2019) demonstrated an improved
activity against E. coli compared with S. aureus of Mn0.5Zn0.5Sm
xEuxFe1.8−2xO4NPs. In contrast, Mohafez et al. (2021) con-
cluded that MnFe2O4 and MnCe1.4Fe0.6O4NP s did not show
antibacterial activity against infectious Gram-negative bacte-
ria: (Escherichia coli, Acinetobacter baumannii and Pseudomonas
aeruginosa), as well as infectious Gram-positive bacteria: (Bacil-
lus cereus, Staphylococcus epidermidis and Streptococcus pneumo-
niae) even at concentrations of 1024 `g mL−1. According to

© 2023 The Authors. Page 497 of 500



Chidouh et. al. Science and Technology Indonesia, 8 (2023) 494-500

Table 2. Antimicrobial Activity of MnEu0.1Fe1.9O4NPs Tested Against Various Microorganisms

Microbial Strains
Reference Number Zone of Inhibition ZOI (mm) MIC

(ATCC) MnEuFe2O4 Gentamicina Ampicillina Methanolb (mg/mL)

E. coli 25922 24± 0.14 20± 0.23 - - 1
S. aureus 25923 21± 0.24 23± 0.29 - - 2

K. pneumoniae 700603 15± 0.32 11± 0.30 - - 1
P. aeruginosa 25953 15± 0.15 15± 0.12 - - 1

E. faecalis 29212 15± 0.33 20± 0.29 - - 2
C. albicans Clinical 12± 0.24 / / - 1

ATCC: American Type Culture collection (a): Positive control (Antibiotics: Gentamicin GEN and Ampicillin AMP), (b): Negative control
(Methanol), (-): No effect.MIC: minimum inhibitory concentration of the tested NPs. The results were expressed as mean ± standard deviation.

Gheidari et al. (2020) , Pseudomonas aeruginosa and Staphylococ-
cus aureus were the most resistant bacteria against CoFe2O4
nanoparticles.

MnFe2O4 and MnCe1.4Fe0.6O4 nanoparticles were also
tested against pathogenic fungi Aspergillus fumigatus, Candida
albicans and Fusarium oxysporum. As a result, adequate antifun-
gal activities were detected in the presence of MnCe1.4Fe0.6O4
nanoparticles (Mohafez et al., 2021) . Al Zahrani et al. (2022)
showed that the increasing concentrations of MnFe2O4 and
Ce-doped MnFe2O4 nanocrystallites (NCs) eventually stop the
replication operation and cease the spread of Gram-negative/po
sitive bacterium when the MnCe0.3Fe1.7O4NCs had higher
activity than the other doped and undoped samples. Mean-
while, the bacterial status and inoculum size influenced the
effectiveness of antimicrobials as well as their applied concen-
trations (Li et al., 2017) . However, our findings showed that our
MnEu0.1Fe1.9O4NPs has a better antibacterial effect against
Gram-negative bacteria. It could be because this class of bacte-
ria has less rigid cell walls than Gram-positive bacteria, which
have a more complicated outer membrane (Jiang et al., 2020) .
Generally, their cell wall is thicker due to the peptidoglycan
layer absent in Gram-negative bacteria. This layer hinders
the penetration of metal ions into the cytoplasm, whereas in
Gram-negative bacteria, the NPs easily penetrate the cell mem-
brane and cause damage (Slavin et al., 2017) . The improved
antibacterial action might be associated with the exposure of
ions that have positive charges, like as Eu3+, Fe3+ and Mn2+,
which charge the bacteria membrane and affect membrane
wholeness, therefore, can decrease electrostatic interaction, al-
lowing membrane obstacles to be traversed, which disturbs the
respiratory chain that kills cells. Cell death is brought on by
breaching membrane barriers, which messes up the electron
transport chain (Al Zahrani et al., 2022) . Thus, the reported
antimicrobial property of MnEu0.1Fe1.9O4NPs in this research
will open up a new way to apply these nanoparticles as a new
source of antimicrobials, which can be further explored in the
biomedical sector.

4. CONCLUSION

Spinel europium-doped manganese ferrite nanoparticles (MnE
u0.1Fe1.9O4NPs) were synthesized using coprecipitation meth
od. By using XRD analysis, the sample’s spinel cubic struc-

ture was determined. The crystallite’s average size was in the
range of 40.9 nm. EDX spectra of the sample confirm the
successful doping of Eu into the spinel structure. The antimi-
crobial action of MnEu0.1Fe1.9O4NPs was investigated on both
yeast and Gram-positive/negative bacteria. The present study
showed that the synthesized MnEu0.1Fe1.9O4NPs had excep-
tional antimicrobial activity against selected microbial strains
and can be used in biomedical applications.

5. ACKNOWLEDGMENT

We are grateful to Professor Djekoun Abdelhamid, the Director
of Pharmaceutical Sciences Research Center (CRSP), Algeria
for his support. We respectfully acknowledge the contribution
made to this research by Dr. Boukhzar Skander, Research fel-
low at the Research Center in Industrial Technologies (CRTI),
Algeria.

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	INTRODUCTION
	EXPERIMENTAL SECTION
	Synthesis of Europium-doped Manganese Ferrite Nanoparticles (MnEu0.1Fe1.9O4NPs)
	MnEu0.1Fe1.9O4NPs Characterization

	Microorganisms and Growth Conditions
	Antimicrobial Assays

	RESULTS AND DISCUSSION
	CONCLUSION
	ACKNOWLEDGMENT