Biosynthesis of iron oxide nanoparticles via a composite of Psidium guavaja-Moringa oleifera and their antibacterial and photocatalytic study

Journal of Photochemistry & Photobiology, B: Biology 199 (2019) 111601

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Biosynthesis of iron oxide nanoparticles via a composite of Psidium guavajaMoringa oleifera and their antibacterial and photocatalytic study

T

Ngozi Madubuonua, Samson O. Aisidaa,b, , Awais Alic, Ishaq Ahmadb,d,e,f, Ting-kai Zhaod,e, ⁎ S. Bothag, M. Maazaf,h, Fabian I. Ezemaa,f,h,i, ⁎

a

Department of Physics and Astronomy, University of Nigeria Nsukka, Nigeria National Centre for Physics, Quaid-i-Azam University campus, Islamabad 44000, Pakistan c Center for Micro and Nano Devices, Department of Physics, COMSATS University Islamabad, Pakistan d NPU-NCP Joint International Research Center on Advanced Nanomaterials and Defects Engineering, Northwestern Polytechnical University, Xi'an 710072, China e School of Materials Science & Engineering, Northwestern Polytechnical University, Xi'an 710072, China f Nanosciences African Network (NANOAFNET), iThemba LABS-National Research, South Africa g Microscopy Unit, University of Western Cape, South Africa h UNESCO-UNISA Africa Chair in Nanosciences/Nanotechnology, College of Graduate Studies, University of South Africa (UNISA), Muckleneuk Ridge, P.O. Box 392, Pretoria, South Africa i Department of Physics, Faculty of Natural and Applied Sciences, Coal City University, Enugu, Nigeria b

ARTICLE INFO

ABSTRACT

Keywords: Nanoparticles Nanomaterials Biosynthesis Antibacterial Photocatalytic

Human pathogenic diseases are on the rampage in the list of debilitating diseases globally. The endless quest to salvage this menace through various therapies via innocuous agents is essential to overcome these drug-resistant pathogens. This study engaged a benign, facile, biocompatible, cost-effective and eco-friendly approach to synthesized iron oxide nanoparticles (FeO-NPs) via a composite of Psidium guavaja-Moringa oleifera (PMC) leaf extract to address six most debilitating bacterial strain in vitro as an antibacterial agent. Physicochemical analysis of PMC formed nanoparticles (PMC_NPs) was effectuated through Fourier Transform Infrared Spectroscopy (FT-IR), UV–Visible Spectroscopy, X-ray Diffraction Spectroscopy (XRD), Transmission Electron Microscopy (TEM), and Vibrating Sample Magnetometer (VSM). The PMC_NPs inhibited the growth of six human pathogens with higher activity at lower concentrations. It is noteworthy from our observations that, the bacterial strains show functional susceptibility to the PMC_NPs at lower concentrations compared to the orthodox antibacterial drugs. Photocatalytic degradation was observed with a decrease in the absorbance of Methylene blue dyes with the help of PMC_NPs apropos irradiation time under visible light irradiation. Consequently, PMC_NPs serve as an enhanced substitute for the orthodox antibacterial drugs in therapeutic biomedical field sequel to its pharmacodynamics against the bacterial strains at lower concentrations and also serves as a good component for water purification.

1. Introduction Magnetic nanoparticles (MNPs) in the current nanotechnology applications have been given curious attention due to its sundry applications in the biomedical field as a potential therapeutic and theranostic tools. MNPs has received a cutting edge applications in the biomedical, bioengineering and nanomedicines via targeted drug delivery for cancer therapy [1–3], tissue repair synergy [4], magnetic fluid hyperthermia applications [5–7], contrast agents in magnetic resonance imaging (MRI) [8–12] and antimicrobial/antibacterial agent [13–17].

⁎

Iron oxide (FeO) as one of the functional MNPs exists majorly as magnetite (Fe3O4), hematite (α-Fe2O3) or maghemite (γ-Fe2O3). This unique quality and diversity have drawn massive attention to Iron oxide nanoparticles (FeO-NPs) for sundry applications such as catalysis, food coatings, cosmetics, antimicrobial and antibacterial agents [18]. These various applications are emplaced due to their diversified properties such as nanometric size, high magnetic permeability, cost-effectiveness, surface modification, good chemical stabilities, facile synthesis, colloidal stability and dispersion in aqueous media [19–21]. In recent years, FeO oxide are has been explored to address some of these debilitating pathogens [22–27]. The antibacterial applications of FeO-NPs

Corresponding authors at: Department of Physics and Astronomy, University of Nigeria Nsukka, Nigeria. E-mail addresses: [email protected] (S.O. Aisida), [email protected] (F.I. Ezema).

https://doi.org/10.1016/j.jphotobiol.2019.111601 Received 24 June 2019; Received in revised form 23 July 2019; Accepted 20 August 2019 Available online 21 August 2019 1011-1344/ © 2019 Elsevier B.V. All rights reserved.

Journal of Photochemistry & Photobiology, B: Biology 199 (2019) 111601

N. Madubuonu, et al.

are due to the noxiousness of the compound to the bacterial strains through the interface in the thiol group located in the respiratory base of the bacterial cells [28,29]. To optimized the various applications of FeO-NPs, effort has been entrained into the synthesis procedures through chemical and physical methods such as sol-gel method [30], microwave-assisted [31,32], Micro-emulsion [33], hydrothermal [34], Sono-electrochemical synthesis [35], Electrochemical method [36,37], Laser irradiation [38], Solvothermal method [39], Tollens process [40,41]. As delineated in these reports, the formed nanoparticles are embroiled with various toxic chemicals that are not biocompatible as a reducing and stabilizing agent [42]. The current applications of FeONPs in the nanotechnology industries chiefly in vivo biomedical and bioengineering applications required a nanoparticle that is biocompatible and environmental innocuous and benign. Biosynthesis/biogenic method serves as a better substitute to chemical and physical methods. The biosynthesis method involved the use of innocuous materials such as an extract from a plant that is biocompatible and benign to serve as a potential stabilizing and reducing agent. Plant extract engendered nanoparticles are facile, cost-effective and easily scaled up. The metabolites such as flavonoid, alkaloid, phenols, proteins, terpenoids, tannin and carbohydrate inherent in the plants extract enhanced the biosynthesis fabrication of the nanoparticles. The different shapes and sizes of the nanoparticles are based on the extract intrinsic compositions [43–45]. PG and MO are both wonderful plants owing to the presence of metabolites intrinsic in their leaves such as phenolic, flavonoids, isoquercitrin, astragalin, anthocyanins, proanthocyanidins, cinnamates, glucosinolates and isothiocyanates iron, potassium, Vitamin C, alkaloids, terpenes, quinines, tannins, saponins, and proteins which can serve both as an effective metal reducing agent and as capping agents [46,47]. In this work, biosynthesis of FeO-NPs via the composite of Psidium guavaja-Moringa oleifera aqueous leaves extract emplaced with metabolites such as flavonoid, alkaloid, phenols, proteins, terpenoids, tannin intrinsic in Psidium guavaja and Moringa oleifera was synthesized for the first time with the propitious potential to control six human most drugresistant bacterial strains as well as an agent of purification in the degradation of Methylene blue dyes. FeO-NPs were emplaced through the biosynthesis method and their antibacterial potency against drug resistance drugs was evaluated alongside with the photocatalytic efficacy.

release of the phytochemicals. The homogenized solution was allowed to cool at room temperature followed by filtration of extract using nylon mesh and then by mill pored filter paper. The PMC extract obtained was stored at 4 °C and used for the biogenic synthesis. The same procedures were observed for the extraction of P and M respectively. 2.3. Biosynthesis of PMC Capped Iron Nanoparticles (PMC-NPs) 0.5 M of Iron (III) chloride (FeCl3) were liquified in 100 mL of DW and stirred at 800 rpm for 1 h under room temperature for a complete dissolution of FeCl3. 20 mL PMC extract was introduced gently to the solution of FeCl3 in a volume ratio of 8:2 (V/V) of the FeCl3 and the PMC extract under vigorous stirring. A color change was observed after 2 min from orange to dark brown, an affirmation of the formation of PMC-NPs. The stirring continues for another 1 h after which the obtained homogeneous solution was transferred to a hot air oven at 100 °C for 24 h. The dark brown solid obtained was pulverized in a ceramic mortar with a pestle and washed four times with DW and re-dry under vacuum oven at 60 °C for 4 h before characterization. The above mechanisms were also observed for the production of P and M nanoparticles, respectively. 2.4. Antimicrobial Assay of PMC Formed Nanoparticles An adopted procedure of Agar Well-Diffusion Method (AWDM) was enthralled to analyze the antibacterial activity of P-NPs, M-NPs and PMC-NPs against the selected Gram-positive and Gram-negative drug resistance bacteria strains [48–50]. A quantity of Mueller Hinton Agar (MHA) was measured and liquefied in a measured volume of DW followed by autoclaved for 30 min. The autoclaved mixture was disgorged in each of the Petri plates (2/3 in volume), this is allowed to jell for 1 h. Each of the freshly cultured isolates was smeared on the surfaces of the MHA plates. The concentration of P-NPs, M-NPs and PMC-NPs ranges from 0.625 μg/mL, 1.25 μg/mL, 2.5 μg/mL, 5.0 μg/mL, and 10 μg/ mL were soused in a hole of about 8 mm in diameter created in each plate using a sterile cork borer. The plates were incubated at room temperature at (37 °C) for 24 h for proper diffusion of the doses on the surfaces of the plates. Afterwards, the diameters of the different levels of inhibition Zones were measured and recorded in millimeters (mm) units. The inhibition zone of the formulated nanoparticles were compared with eleven standard antibiotic drugs namely: Chloramphenicol (CRO-30 μg), Ampiciox (APX−30 μg), Pefloxacin (PEF−10 μg), Spectrin (CN−30 μg), Erythromycin (E−10 μg), Amoxicillin (AMX−30 μg), Septrin (SXT−30 μg), Ciprofloxacin (CPX−10 μg), Mixofloxacin (MXF−10 μg), Gentamycin (CH−10 μg) and Ofloxaxin (OFX−30 μg) as the positive control as well as the DW as the negative control.

2. Experimental Details 2.1. Materials and Reagents Iron (III) chloride hexahydrate 99.9%, Muller-Hinton agar, Methylene blue (MB) and Muller-Hinton broth (Analytical reagents from Sigma Aldrich product) were used as acquired in their purity grade. Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa, Staphylococcus aureus, Shigella, and Pasteurella multocida were obtained from Safety Molecular pathology laboratory (SMPL) in Enugu, Nigeria. Psidium guajava and Moringa oleifera leaves were collected from the earth within the University campus. All the homogenization process and washing of glassware were done using distilled water (DW).

2.5. Hemolytic Assay of PMC Formed Nanoparticles The toxicity of PMC-NPs was verified through the hemolytic assay following the nitty-gritty of Andrej et al. [51]. 5 mL normal saline solution in a tube containing 0.1 mL diluted blood together with a reasonable quantity of PMC was centrifuged for 2 min and incubated at 35 °C for 1 h. Also, 0.2 mL and 0.1 mL of diluted blood were homogenized in 4 mL and 5 mL sodium carbonate and normal saline solution in a centrifuge test tube, respectively. The test tubes were incubator at 35 °C for 1 h and later centrifuged at 4000 rpm for 5 min. The mixture with the sodium carbonate solution acts as the positive control by obliterating the red blood cell. While the dilution with standard saline solution without any noticeable destruction of red blood cell acts as the negative control. The optical density (OD) of PMC-NPs, positive and negative control samples, as shown in Table 1 was calculated at 545 nm from 1 ml supernatant [50,52]. Generally, it has been asserted through various reports that the tolerable limit of hemolysis for biomedical applications must not be higher than 5% [53]. Hence, the percentage of

2.2. Preparation of Psidium Guajava-Moringa Oleifera Composite (PMC) Extract Fresh leaves of Psidium guajava (P) and Moringa oleifera (M) were collected from the earth and washed firstly through running tap water. The leaves were re-washed again three times thoroughly with DW to take away unnecessary particles. This is followed by air-dried for 72 h at room temperature (32 °C) maximum. The dried leaves were blended using Hitachi electronic blender to form a powder. Then 10 g of PMC powdered leaves were liquefied in a flat bottom conical flask containing 100 mL of DW and stirred continuously for 1 h at 80 °C to enhance the 2

0.0487 ± 0.00367 0.680 0.026

3.2% 100 0

*

diluted blood with PMC-NPs was calculated to be 3.2%.

OD of (PMC PMCNPs) OD (+ control) Hemolysis (%) = × 100% OD ( + control) OD ( control)

* (1)

t

× 100%

*

±-Fe2O3 ³ -Fe2O3

.*

*

PMC-NPs

*

** M-NPs

* *

**

**

*

30

40

50

60

70

Fig. 1. XRD analysis of P, M and PMC formed nanoparticles.

The degradation of the MB was performed using PMC-NPs. A quantity of PMC-NPs was weighed and added to a known volume of an aqueous solution of MB for the photocatalytic degradation processes under the sunlight for 1 h. The degradation in the solution was observed at an interval of 30 min in UV–Vis absorption spectroscopy. The degradation percentage of MB solution was estimated using Eq. (1). 0

. * .

*

P-NPs

2.6. Photocatalytic Degradation of PMC Formed Nanoparticles

0

.*

*

. .

20

=

*

*

*

(214) (300)

FeNPs Positive control Negative control

*

(024)

Hemolysis (%)

(113)

Optical density

(012)

Nanoparticle

(104) (110)

*

(018)

Table 1 Hemolytic activities of PMC-NPs.

(116)

Journal of Photochemistry & Photobiology, B: Biology 199 (2019) 111601

N. Madubuonu, et al.

PMC formed nanoparticles. (XRD )

=

K Cos

(3)

where Ω(XRD) is the crystallite size (nm), K is a constant with value (0.9), the X-rays wavelength (λ = 0.15406 nm), ϵ is the full width at half maximum (FWHM) intensity measured in radians and θ is the Bragg diffraction angle of the plane [54]. The crystallite size are in the range of 40–90 nm.

(2)

where Ω is the degradation percentage (%), δ0 and δt are the initial absorbance and the absorbance at a chosen time interval of MB expressed in mg/L.

3.2. TEM, SAED and EDX Analysis of P, M and PMC Formed Nanoparticles The TEM analysis as shown in Fig. 2 revealed a spherical morphology for the P formed nanoparticles (Fig. 2ai), non-uniformed rodlike shape for the M formed nanoparticles (Fig. 2bi) and spherical-rodlike clusters of different shapes for PMC-NPs (Fig. 2ci) with their respective size distributions as shown in (Fig. 2iii of a, b & c). The pattern of the morphologies was determined by the SAED (Fig. 2 ii of a, b & c) with several sharp rings affirmed the polycrystalline structure of the formulated P-NPs, M-NPs and PMC-NPs. The formed rings are in tandem with the diffraction peaks from planes of a face-centered cubic (fcc) in XRD analysis. The EDX analysis (Fig. 2 iv of a, b & c) shows the elemental compositions of P-NPs, M-NPs, and PMC-NPs with the profound signal at 8.5 KeV showing the presence of iron coupled with the weak signals for Oxygen in the composite precursor.

2.7. Characterization Techniques The samples P, M and PMC formed nanoparticles were examined by X-ray diffraction (XRD) analysis using powder X-ray diffractometer Shimadzu-7000 with Cu-Kα radiation (λ = 1.5406 Å and a = 4.08620 Å) at room temperature in the continuous scanning mode of 2θ = 15o - 80°. The morphologies were examined by a high-resolution scanning electron microscope (Auriga Zeiss HRSEM). A High-resolution transmission electron microscopy (HRTEM) using a Tecnai F20 HRTEM operated at 200 kV was used by dropping a diluted amount of FeO-NRs to the TEM grid to determine the particle size through averaging measurement by randomly selections of some particles in the ambit of the TEM image. The elemental compositions were determined by the Energy Dispersive X-ray Spectrometer (EDS) attached to the HRTEM. The surface functional groups were determined by the Fourier Transform Infrared (FTIR) Spectrophotometer (PerkinElmer FT-IR spectra 1650, version 10.03.02) model with 256 scans and 6 cm−1 resolution in the range of 4000–500 cm−1. The absorbance of the samples was determined by the UV–visible spectrometer. The magnetizations against the magnetic field (M-H) curves were recorded by the Vibrating Sample Magnetometer (VSM) Lake Shore 4700 model at room temperature measured with a maximum magnetic field of ± 15 KOe.

3.3. FTIR Analysis of P, M and PMC Formed Nanoparticles Fig. 3 presents the FTIR spectra analysis of P, M, and PMC formed nanoparticles with various vibrational bands of biomolecules observed in the range of 500–4000 cm−1. The biomolecules observed in the functional groups are responsible for the reduction of iron in the formation of PMC-NPs. The band observed in this range 3034–3366 cm−1 shows the stretching vibration associated with OeH group in polysaccharide, proteins or polyphenols of water molecules bound in FeO surface [50,55,56]. The band between 1527 and 1669 cm−1 gives the stretching vibration adduced to carbonyl (C]O) group. Bending vibration of OeH, CeN (of aromatic amines) and CeOeC functional groups were observed in the region between 1338 and 1449 cm−1, and 1019–1123 cm-1, respectively. The observed vibration bands below 600 cm−1 gives birth to the stretching mode of the FeeO bond, an evidence of the successful immobilization of P, M and PMC on the surface of the formed nanoparticles [57–59].

3. Results and Discussion 3.1. XRD Analysis of P, M and PMC Formed Nanoparticles The XRD pattern of P, M and PMC formed nanoparticles (Fig. 1) showed the major diffraction peaks at 2 theta values of 22.9 (012), 34.5 (104), 35.5 (110), 42.1 (113), 50.9 (024), 55.9 (116), 58.8 (018), 62.9 (214), and 64.6 (300) degrees. This shows that the synthesized samples are well crystallized with peaks corresponding to the hexagonal hematite structure (JCPDS File No. 33-0664) and lattice parameter of a = 5.03560 Å and c = 13.7489 Å. Debye-Scherer's formula (Eq. (1)) was actuated to calculate the crystallite size. The average crystallite size of 75 ± 7.0, 76 ± 2.0, and 82 ± 7.0 nm was obtained for P, M and

3.4. UV Analysis of P, M and PMC Formed Nanoparticles The UV–Visible absorption spectra analysis of P, M, and PMC formed nanoparticles (Fig. 4) gives a color change from dark brown to 3

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Fig. 2. TEM, SAED, size distribution and EDX analysis of nanoparticles formed by (a) P (b) M and (c) PMC.

4

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N. Madubuonu, et al.

a c

(a) (b) (c)

b -4000

-2000

0

2000

4000

Magnetic Field (Oe) Fig. 5. VSM analysis of nanoparticles formed by P (a), M (b) and PMC (c). Table 2 The magnetic properties of P-NPs, M-Nps and PMC-NPs. Sample

Magnetic properties

P-NPs M-NPs PMC-NPs

Ms

Mc

Mr

(emu/g)

(Oe)

(emu/g)

5.56 2.82 5.87

94.3 33.2 20.92

7.48 2.42 2.42

Mr/Ms

1.345 0.858 0.412

Ms- Saturation magnetization, Mc- Coercivity, Mr- Remanence magnetization. Table 3 Inhibition Zone of nanoparticles formed by PMC against gram negative and gram positive bacteria. Dose μg/ml

DW PMC_1 PMC_2 PMC_3 PMC_4 PMC_5 CPX OFX PEF AMX CN MXF SXT CH CRO E APX

Fig. 3. FT-IR spectra of nanoparticles formed by (i) P, M, and PMC (ii) the expanded inset.

3.0 C (315)

2.5 2.0

a (310) b (314)

1.5 1.0 0.5

Inhibition zone (mm) E. coli

S. typhi

S. aureus

Shigella

P. m

P. a

0 0 0 0 0 20 0 0 0 0 0 0 0 0 0 0 0

0 0 0 0 0 24 0 0 0 0 0 0 0 0 0 0 0

0 0 0 0 0 26 20 0 0 0 0 22 0 0 0 12 0

0 0 0 0 0 24 22 0 22 8 16 20 4 18 0 0 0

0 0 0 0 0 18 0 0 0 0 0 20 10 14 0 0 0

0 0 0 0 0 16 0 0 0 0 0 22 12 6 0 0 0

PMC_1 = (0.625 μg/mL), PMC_2 = (1.25 μg/mL), PMC_3 = (2.5 μg/mL), PMC_4 = (5.0 μg/mL), PMC_5 = (10 μg/mL).

0.0

to the Surface Plasmon Resonance (SPR) peak of sample P, M and PMC formed nanoparticles occurs at 310, 314, and 315 nm respectively. The absorption bands of FeO nanoparticles have been reported within the range of 280–400 nm by many researchers [60,61]. The nanoparticles formed from PMC shows the highest SPR which may be adduced to the bio-conjugate of the phytochemicals found in P and M.

Wavelength (nm) Fig. 4. UV–Vis spectra of nanoparticles formed by P (a), M (b) and PMC (c).

light orange in aqueous solution. This is due to the reduction of ferric to ferrous by the reactive functional moieties present in the extract. The color change is as a result of the excitation in the UV–vis spectrum depending on the size of the particle [57]. The absorption band ascribed 5

Journal of Photochemistry & Photobiology, B: Biology 199 (2019) 111601

N. Madubuonu, et al.

morphology and particle interactions are synthesis dependent, invariable, the saturation magnetization is also synthesis-dependent [57]. Hence the smaller saturation value of P-NPs, M-NPs and PMC-NPs is adduced to the small particle size which may be ascribed to the disordered layers in the increased surface spin [63]. 3.6. Antibacterial Analysis of PMC Formed Nanoparticles The AWDM confirmed the antibacterial activities of P, M and PMC formed nanoparticles against the bacterial strains. The inhibition zones of PMC against common standard antibiotic drugs are summarized in Table 3. The nanoparticles formed by PMC at 10 μg/mL inhibit the growth of E. coli and S. aureus. It also shows strong activities against all the organisms when compared with the standard drugs. Antibacterial activities of PMC formed nanoparticles was found to increase with higher concentrations, (i.e the inhibition zone increases with higher concentrations) hence, the PMC formed nanoparticles is concentration dependent [64]. Owing to the sensitivity of PMC formed nanoparticles against the bacterial strains, its shows good and efficient antibacterial activities against the bacterial pathogens (Figs. 6–8). The performance of PMC against P and M formulated nanoparticles was also investigated as shown in Table 4 and Fig. 9. It was clear from Fig. 9 that the PMC composite formed nanoparticles inhibits the bacterial strains more than the nanoparticles formed by P and M. Furthermore, a confirmatory test at lower concentration of 2.5 μg/mL and 5.0 μg/mL of PMC formulated nanoparticles being the most auspicious was conducted on two most common drug resistance bacterial strains E. coli and S. aureus as shown in Table 5. It was obvious from Fig. 9 that the activities increases with the concentration of PMC formulated nanoparticles. Hence, PMC formulated nanoparticles stand a better chance as a potential substitute for the conventional antibacterial drugs sequel to its activities at lower concentrations.

Fig. 6. The assay of the minimum inhibition of PMC formed nanoparticles against bacterial strains.

P-NPs

M-NPs

PMC-NPs

25 20 15

3.7. Photocatalytic Analysis of PMC Formed Nanoparticles

10

Photocatalytic degradation was observed with a decrease in the absorbance of MB dyes in the presence of formulated PMC-NPs concerning irradiation time under visible light irradiation (as shown in Fig. 10). The degradation of PMC-NPs was identified by a color change in the Methylene blue (MB). The color of PMC-NPs was initially blue, which changed into light blue after 30 min and then light green after 1 h incubation with PMC-NPs under sunlight exposure. This color change from blue to light green after addition of PMC-NPs to MB can be adduced to the interaction of phytochemicals coated PMC-NPs with MB. We also observed that, with the increase in the exposure time to the sunlight, the degradation percentage of MB increased as calculated by Eq. (1) with a decrease in the intensity given by the UV–absorption spectra as shown in the inset of Fig. 10. This is as a result of the presence of surface hydroxyl groups that facilitates the trapping of photoinduced electrons and holes, thereby enhancing the photocatalytic degradation process. Studies showed that the photocatalytic activity of iron oxide NPs is morphology, crystallite size, and preparation method dependent [65–69].

5

a P.

m P.

Sh ig el la

S. Ty ph i

E. C

ol i

0

Fig. 7. The assay of the minimum inhibition of PMC formed nanoparticles against bacterial strains.

3.5. VSM Analysis of P, M and PMC Formed Nanoparticles The vibrating sample magnetometer (VSM) (Fig. 5) was used to measure the magnetic properties of sample a, b, and c by measuring the magnetization versus the applied magnetic field (M-H) curve at room temperature in the magnetic field range of ± 5 ΚΟe. The M-H curve revealed a superparamagnetic behavior. The calculated values for the saturation magnetization (Ms), coercivity (Hc) and remanence magnetization (Mr) are summarized in Table 2. This shows that the synthesized nanoparticles has strong magnetic properties and can be used for biomedical applications. The variation in the magnetic properties of the formed nanosized particles may be influence by the method of synthesis, surface and lattice spin and the magnetocrystalline anisotropy [57]. The magnetization of NPs depends strongly on the particle size, hence, the smaller the particle, the smaller the saturation magnetization compared to the bulk value [62]. Thus, since the particle size,

4. Conclusion A facile and eco-friendly method using an aqueous extract from the composites of P and M as an effective reducing agent was presented in this work. The nanoparticles formed from P and M were compared with the PMC formed nanoparticles through various characterization techniques. The XRD showed a single phase crystalline hexagonal hematite structure for the P-NPs, M-NPs, and PMC-NPs. The FTIR spectra gave

6

Journal of Photochemistry & Photobiology, B: Biology 199 (2019) 111601

N. Madubuonu, et al.

Fig. 8. (i) Inhibition zone of (i) PMC-NPs against E. coli and S. aureus (ii) positive control. Table 4 Comparison of the inhibition zone of P, M and PMC formed nanoparticles against the bacterial strains. Dose μg/ml

P-NPs_5 M-NPs _5 PMC-NPs _5

Table 5 Confirmatory test of PMC-NPs against two of the most resistive bacteria. Dose μg/ml

Zone of inhibition (mm) E. coli

S. typhi

S. aureus

Shigella

P.m

P.a

8.4 14.2 20.1

10.3 16.1 24.2

12.3 18.4 26.2

18.4 18.2 24.4

16.4 16.5 18.2

16.2 14.2 16.3

DW PMC-NPs PMC-NPs PMC-NPs PMC-NPs PMC-NPs

_1 _2 _3 _4 _5

Zone of inhibition (mm) E. coli

S. aureus

0 0 0 10.2 20.3 25.2

0 0 0 15.1 20.4 30.3

PMC_3 = (2.5 μg/mL), PMC_4 = (5.0 μg/mL), (Concentration = 3 g/7 ml in 3:10 dilution)

PMC_5 = (10

μg/mL)

the different functional groups responsible for the formation of the PNPs, M-NPs, and PMC-NPs. The UV–visible confirmed the formation of the nanoparticles through a visible color change from orange to dark brown. The TEM analysis of the PMC-NPs revealed a cluster of spherical-rod like morphologies. P-NPs, M-NPs, and PMC-NPs exhibited superparamagnetic behavior. The PMC-NPs inhibited the growth of Staphylococcus aureus, Escherichia coli, Shigella, Pseudomonas aeruginosa, Salmonella typhi and Pasteurella multocida with higher activity at lower concentration when compared to P-NPs and M-NPs. It is noteworthy from our observations that, the bacterial strains show strong and effective susceptibility to the PMC_NPS at lower concentrations compared to the orthodox antibacterial drugs. Photocatalytic degradation was observed with a decrease in the absorbance of MB dyes in the presence of formulated PMC-NPs apropos irradiation time under visible light irradiation. Consequently, the formulated PMC-NPs serve as an enhanced substitute for the orthodox antibacterial drugs in therapeutic biomedical field sequel to its pharmacodynamics against the pathogens and good components for water purification.

Fig. 9. A confirmatory test of PMC-NPs against E. coli and S. aureus at lower concentrations (3 g/7 ml). 7

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PMC-NPs (0 mins) PMC-NPs (30 mins) PMC-NPs (60 mins)

0.2 t=0

t = 30

t = 60

0.1

0.0

-0.1

-0.2 400

600

800

Wavelength (nm) Fig. 10. Photocatalytic degradation pattern of MB in the application of PMCNPs.

Declaration of Competing Interest The authors declare that they have no conflicts of interest Acknowledgments Samson O. Aisida acknowledges the NCP-TWAS Postdoc Fellowship award (NCP-CAAD/TWAS_Fellow8408). FIE (90407830) acknowledges UNISA for VRSP Fellowship award; he also acknowledges the grant by TETFUND under contract number TETF/DESS/UNN/NSUKKA/STI/VOL.I/B4.33. Also, we thank Engr. Emeka Okwuosa for the sponsorship of 2014, 2016 and 2018 nanoconferences/workshops. References [1] A. Aliosmanoglu, I. Basaran, Nanotechnology in cancer treatment, J. Nanomed Biotherapaeutic Dis. 21 (2012) 1–3. [2] M. Nadeem, M. Ahmad, M.S. Akhtar, A. Shaari, S. Riaz, Magnetic properties of polyvinyl alcohol and Doxorubicine loaded Iron oxide nanoparticles for anticancer drug delivery applications, PLoS One 11 (2016) e0158084. [3] J. Park, N.R. Kadasala, S.A. Abouelmagd, M.A. Castanares, D.S. Collins, A. Wei, Y. Yeo, Polymer-Iron oxide composite nanoparticles for EPR-independent drug delivery, Biomaterials 101 (2016) 285–295. [4] Z.-P. Ofra, M. Shlomo, S. Abraham, Application of iron oxide anoparticles in neuronal tissue engineering, Neural Regen. Res. 10 (2015) 189–191. [5] H. Choi, J. Frangioni, Nanoparticles for biomedical imaging, fundamentals of clinical translation, Mol. Imaging 9 (2010) 291–310. [6] A. Jordan, R. Scholz, P. Wust, H. Fähling, R. Felix, Magnetic fluid hyperthermia (MFH): cancer treatment with AC magnetic field induced excitation of biocompabiocompatible superparamagnetic nanoparticles, J. Magn. Magn. Mater. 201 (1999) 413–419. [7] I. Sato, M. Umemura, K. Mitsudo, M. Kioi, H. Nakashima, T. Iwai, X. Feng, K. Oda, A. Miyajima, A. Makino, M. Iwai, T. Fujita, U. Yokoyama, S. Okumura, M. Sato, P. Tiberto, Hysteresis losses and specific absorption rate measurements in magnetic nanoparticles for hyperthermia applications, Biochim. Biophys. Acta-Gen. Subj. 1861 (2017) 1545–1558. [8] J. Huang, L. Wang, X. Zhong, Y. Li, L. Yang, H. Mao, Facile non-hydrothermal synthesis of oligosaccharide coated Sub-5 nm magnetic Iron oxide nanoparticles with dual MRI contrast enhancement effects, J. Mater. Chem. B 2 (2014) 5344–5351. [9] Y. Li, C.H. Li, D.R. Talham, One-step synthesis of gradient gadolinium Ironhexacyanoferrate nanoparticles: a new particle design easily combining MRI contrast and photothermal therapy, Nanoscale 7 (2015) 5209–5216. [10] B. Gleich, J. Weizenecker, Tomographic imaging using the nonlinear response of magnetic particles, Nature 435 (2005) 1214–1217. [11] R. Duschka, H. Wojtczyk, N. Panagiotopoulos, J. Haegele, G. Bringout, T.M. Buzug, J. Barkhausen, F.M. Vogt, Safety measurements for heating of instruments for cardiovascular interventions in magnetic particle imaging (MPI) – first experiences, J. Healthc. Eng. 5 (2014) 79–93. [12] K. Kluchova, R. Zboril, J. Tucek, M. Pecova, L. Zajoncova, I. Safarik, M. Mashlan, I. Markova, D. Jancik, M. Sebela, H. Bartonkova, V. Bellesi, P. Novak, D. Petridis, Superparamagnetic maghemite nanoparticles from solid-state synthesis e their,

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