In the present work, Gamma Aluminum oxide (γ-Al2O3) is prepared from wastes (Pepsi Cans). These cans have a structure that contains more than 98 percent aluminum oxide, and they were effectively manufactured to generate nano-sized gamma alumina under mild parameters. Characterization of the prepared nanoparticles are characterizes using different techniques as X-ray diffraction (XRD), High resolution transmission electron microscope (HRTEM), Field Emission Scanning Electron Microscope (FESEM), Attenuated total Reflection-Fourier Transform Infrared (ATR-FTIR), Ultraviolet-Visible (UV-Vis) spectrophotometer and Thermogravimetric analysis (TGA). Also, antifungal activity of γ Al2O3 was applied to widespread pathogenic fungi: Aspergillus flavus, Fusarium oxysporum and Alternaria sp..at different nanoparticles concentrations to determine the maximum fungicidal concentration of the tested nanoparticles on the fungal strains. All characterization confirmed the formation of γ Al2O3 with nearly 2-3 nm and TGA data explains it is thermal stability. By investigating the fungicidal concentration, Data showed the antifungal activity of the selected nanoparticles shows concentration dependent manner based on the fungal strain sensitivity. The maximum antifungal activity of aluminum nanoparticles detected at concentration (3, 6 and 50mg/100ml) with inhibition percentage of (94.2, 95 and 93.3 %) for A flavus, F.oxsporum andAlternaria sp., respectively.
Research Article
Structural, Morphological, Optical, Thermal and Fungicidal Effects of Gamma Aluminum Nanoparticles Synthesized from Pepsi Cans
https://doi.org/10.21203/rs.3.rs-1127052/v1
This work is licensed under a CC BY 4.0 License
Version 1
posted 08 Dec, 2021
You are reading this latest preprint version
In today's world, nanoscience has become a popular field of study. Nanoparticles (NPs), both natural and man-made, have been entering the environment and industrial field [1]. For the past few decades, a variety of nanoparticles have been generated using different processes and used in pharmaceutical, feed and food, in addition to develop environmental technologies such as water treatment, the recognition of persistent contaminants [2, 3]. The field of materials science and engineering is becoming increasingly focused on improving the sustainability of the methods used to make nanoparticles, which is driving research into alternate nanoparticle inputs as well as the use of green synthesis techniques [4–7].
According to the United Nations Environment Program, approximately 11.2 billion tons of wastes are formed each year, posing a significant source of environmental pollution and negative health consequences, especially in low income countries, where more than 90% of waste is dumped or burned openly [8]. Closed-loop treatment of engineered materials on a wide scale, however, faces substantial technical, economic, environmental, and social hurdles. Many plastic and metal recycling procedures, for example, require the extraction and purification of hazardous compounds, posing new health concerns to humans and the environment [9]. Furthermore, the economic worth of the end-products is closely tied to the efficiency of recycling and repurposing initiatives. As a result, employing "clean manufacturing" technologies to produce "value-added" commodities from waste materials appears to be a promising synergy for accomplishing both circularity and sustainability goals [10].
Aluminum foundry tailing (AFT) is an aluminum oxide-rich hazardous solid waste generated during the aluminum metal industry [11, 12]. Nano-sized alumina has better catalytic and adsorption characteristics than micro-sized alumina. Gamma alumina is a porous substance distinguished by its high specific surface area and high reactivity among the many forms of alumina. Gamma alumina in nanoscale will have excellent chemical, mechanical, low modification temperature and thermal stability [13–16]. Rare research papers studied the antifungal effect of nano aluminum oxide as antifungal but many papers studied their antibacterial effect [17, 18].
In this paper, structural, optical and thermal characterization of gamma aluminum oxide prepared from wastes (Pepsi cans) are studied to be used for antifungal application on Aspergillus flavus, Fusarium oxysporumand Alternaria sp.
2.1. Synthesis of γ- Al2O3
Gamma-alumina was Synthesized using cans metals (collected from wastes), the solvents used were NaOH (5N), HCl (5N), and deionized water. Cut the Pepsi cans into small pieces. Slowly adding of these pieces into 5N NaOH to avoid any risks during the dissolving of aluminum, after the complete dissolved of cans, making filtration by Whatman™ 42 filter paper to separate the supernatant. Then adding the 5N HCl drop wise to the solution till the precipitate was obtained. Washed the precipitate three times using centrifuge at 5000 rbm/15min for elimination salts. Finally, the precipitate was dried at 70°C overnight and calcinated at 350°C/3h.
2.2. Measurements Techniques
X-ray diffraction (XRD) was analysed by using PANalytical X’Pert Pro with Cu-Kα as a target with secondary monochromator Holland radiation, the tube was operated at 45 kV and wavelength 0.1540 nm). High – Resolution Transmission Electron Microscope (HRTEM) was measured by JEM-2100F electron microscope with accelerated voltage 200 kV. Field Emission Scanning Electron Microscope (FESEM) with EDX detector was measured using SEM Model Quanta 250 FEG. ATR-FTIR spectral data was performed using Vertex 80 Bruker (Germany) with resolution 4 cm−1 within range 4000-400 cm−1. Optical Properties were performed using Jasco V-630 spectrophotometer in the range 200-1000 nm. Thermogravimetric analysis (TGA) was performed with an SDT Q600 V20.9 Build 20 thermal analyzer from ambient temperature to 1000°C with heating rate of 10°C/min.
2.3. Screening of aluminum nanoparticles’ antifungal activity
γ-Aluminum nanoparticles solution was prepared in concentration (2mg/3ml DDH2O) and tested for antifungal activity against Aspergillus flavus, Fusarium oxysporumand Alternaria sp. 7mm fungal disk of each isolate was cut and inoculated in 10mldistilled sterilized H2O then vortexed vigorously till obtain Fungal spore suspensions of 105-106. 1ml of the fungal spore suspension was inoculated in sterilized petri dishes, covered with warm PDA sterilized medium, and mixed homogenously then left till solidification. By mean of sterilize tip end, 7mm agar well was made and inoculated with 50ul (33.3 ug/50ul) of the prepared nanoparticles solution. The PDA plates were left in the fridge for 30 minutes to allow the solution diffusion, and then incubated at 30c for 5 day. The antifungal activity was detected after the incubation period by measuring the inhibition zone around the well by ruler in mm.
2.4. Determination of the fungicidal concentration:
Actively growing fungal discs of 7mm size in diameter were transferred to 100ml sterilized PDB medium. Different concentrations of Al-nanoparticles (B) were prepared according to the isolated fungal strain as follows (based on the screening experiment results): For A. flavus a concentration gradient of (0, 0.5, 1, 2 and 3 mg/100ml), while for Fusarium sp. (0, 0.5, 2, 4, 5 and 6mg/100ml) and Alternaria sp. (0, 5, 10, 20, 30 and 50 mg/100ml) to be incubated separately in the PDB culture medium altogether with the fungal discs. The cultures were allowed to grow on shaker incubator 180rpm at 30oC for 5 days under ambient light condition. the fungal mycelia were harvested by filtration through desiccated, pre- weighted Whatman filter paper No.1 and dried overnight at 70oC then the fungal mycelial weights were recorded two following days till a constant volume reached.
4.1. Structural Characterization
Figure 1 illustrates XRD of the preparation of gamma Aluminum using Pepsi cans treated base medium. For Figure 1 shows the crystalline diffraction peaks at 2q = 31.7°, 45.2°, 56.4°and 66.5° with tetragonal structure which agreed with JCPDS card 98-009-9836 which corresponded to boehmite-derived γ-alumina.
The crystal size of gamma Aluminum is calculated using scherrerr equation [19]
Where D is the crystal size, λ is the wavelength of X-ray, β (in radian) is the full width at half maximum (FWHM) of the diffraction peak and θ is the Bragg’s diffraction angle (in degree) of the peak maximum. The average crystal size of gamma Aluminum is equal 32.7 nm.
The dislocation density, S, is a calculation of the extent of defects and vacancies in a crystal that can be calculated using the formula from the crystallite size (D) and found to be equal 1.89x10-4 nm-2 [20].
Figure 2 shows HRTEM image of γ- Al2O3. As seen γ- Al2O3 shows interconnected rod-like particles porous structure with average particle size 2-3 nm.
Figure 3 represent Pepsi cans treated in basic medium, the surface is seen to be very rough not smooth with pointed edges. For EDX analysis as seen confirmed the presence of Al and Oxygen and there are also some element like Na and Cl which present during the washing process.
Figure 4 represents ATR-FTIR of γ-Al2O3. Bands at 3393 cm-1and 1638 cm-1 are assigned to the O-H stretching and bending vibration, respectively of moisture. Bands at 732 cm-1, 542 cm-1 and 481 cm-1 are corresponded to the Al-O vibrations [21, 22].
4.2. Optical Properties
The absorption spectra and the band gap energy of γ-Al2O3 are illustrated in Figure 5. Strong absorption peaks at 235 nm may be seen in the spectra because of the electron that excited from the valence band to the conduction band [23].
Tauc relation is used for calculating band gap energy of γ-Al2O3 [24]:
Where Eg is the band gap energy, b is constant and a is the absorption coefficient and can be determined using the below relation [25]:
Where A and d are the optical absorbance and the thickness of the prepared sample, respectively. The calculated band gap energy is calculated by extrapolating the linear part of the curve with x- axis which represents photon energy as seen in Figure 5 and it is equal 3.5 eV.
4.3. Thermogravimetric Analysis
Thermal behavior of γ- Al2O3 NPs is investigated by TG analysis throughout a temperature range from room temperature up to 1000 °C as seen in Figure 6. At around 150 °C, the thermal degradation shows around 10 % weight loss, showing the presence of strong interactions within the alumina structure. When the temperature approaches 1000 °C, the total weight loss is only 32%, demonstrating that γ- Al2O3 NPs has excellent thermal stability which is an important attribute for future industrial applications such as high-temperature catalysis and biomaterial manufacturing.
4.4. Evaluation of the antifungal activity
Figure 7 and Table 1 represent the effect of aluminum nanoparticles on the selected fungal strains growth inhibition in mm. all the fungal strains show variable sensitivity behavior against γ-Al2O3 NPs. The most affected strain with the maximum inhibitory activity was A. flavus followed by F. oxysporum with inhibition zone 9 and 7mm respectively. Alternaria sp. shows the less sensitivity action exposing to the selected nanoparticles with 1.5 mm inhibition zone.
4.5. Determination of the optimum fungicidal concentration against the fungal species.
4.5.1. Aspergillus flavus
According to A. Flavus, it can be seen from Table 2 and Figure 8 that, the maximum inhibitory concentration activity was detected in 3mg/100ml with inhibition percentage 94.2%.
4.5.2. Fusariumoxysporum
As seen in Figure 9 and Table 3 that the maximum inhibitory concentration was detected in ratio of 6mg/100ml with inhibition percentage (95%).
The research data output is in correspondence to Suryavanshi et al., 2017 [26] study who evaluated the antifungal activity of aluminum nanoparticles against different food born pathogenic fungi included fusarium oxysporum and Aspergillus flavus. The data showed that aluminum nanoparticles exhibited antifungal activity against F. oxysporum and A. flavus with MIC concentration of 250 and 150µg/ml respectively.
Moreover, Shenashen et al., 2017 [27] detected the antifungal activity of mesoporous aluminum nanoparticles against Fusarium Oxysporum and found that the highest antifungal activity was recorded at concentration 400mg/L with maximum inhibition percentage of 78.57% after growth on PDA plates in comparison to the control treatment.
4.5.3. Alternaria sp.
For Alternaria sp. It can be posed from Figure 10 and Table 4 that, the highest growth inhibitory concentration of γ-Al2O3 NPs was detected at 50mg/100ml value with growth inhibition percentage of 93.3%.
The fungicidal effect of aluminum nanoparticles may be explained by multiple cellular site destruction: either due to its direct linkage to protein and enzymes molecules or DNA contact which causes mutation and adversely affect cellular replication. The antifungal activity may also attribute to linkage to the free hydrogen bond on the cellular membrane which alters the lipopolysaccharide layer configurations that eventually lead to cell membrane damage and cell lysis reported by Capeletti et al., 2014 [28].
Gamma Aluminum oxide was successfully prepared from Pepsi Cans as a waste material and this preparation of nanoparticles is one of most successive method to recycling waste to be one of the green technology to preserve environment from pollution. XRD, HRTEM, ATR-FTIR, SEM-EDX, UV-Vis and TGA analysis are used to confirm and provide information about prepared nanoparticles to be used as antifungal material against three type of toxic fungus (producing mycotoxins) as A flavus, F.oxsporum andAlternaria sp.. γ-Al2O3 NPs structure confirmed from XRD and ATR-FTIR. HRTEM provide that the size of nanoparticles was approximately ranges from 2 to 3 nm. SEM images shows that the prepared nanoparticles surface has very rough not smooth with pointed edges and TGA affirmed the thermal stability of γ-Al2O3. The highest antifungal activity of aluminum nanoparticles was identified at concentrations of (3, 6 and 50 mg/100 ml) for A flavus, F. oxsporum, and Alternaria sp., respectively, with inhibition percentages of (94.2, 95, and 93.3).
Acknowledgment
This study was financially supported by National Research Centre of Egypt through internal research project no. 12020216.
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Table 1: Values of inhibition in mm of the selected fungal strains
|
Strain ID |
Inhibition zone/mm |
|
(A. Flavus) |
9.0 |
|
(Alternaria sp.) |
1.5 |
|
(Fusarium oxysporum) |
7.0 |
Table 2: Shows the fungal strain's response to γ-Al2O3 NPs concentration gradients for A. flavus.
|
|
A. flavus |
|
|
DW |
Inhibition % |
|
|
0 |
1.7 |
|
|
0.5 |
0.15 |
90.3 |
|
1 |
0.13 |
91.6 |
|
2 |
0.11 |
92.9 |
|
3 |
0.09 |
94.2 |
Table 3: shows the fungal strain's response to γ-Al2O3 NPs concentration gradients for Fusarium oxysporum
|
|
Fusarium oxysporum
|
|
|
DW |
Inhibition % |
|
|
Control |
0.8 |
|
|
0.5 |
0.2 |
75 |
|
2 |
0.18 |
77.5 |
|
4 |
0.13 |
83.75 |
|
5 |
0.11 |
86.25 |
Table 4: shows the fungal strain's response to γ-Al2O3 NPs concentration gradients for Alternaria sp.
|
|
Alternaria sp. |
|
|
DW |
Inhibition % |
|
|
Control |
0.9 |
|
|
5 |
0.2 |
77.78 |
|
10 |
0.13 |
85.56 |
|
20 |
0.07 |
92.2 |
|
30 |
0.07 |
92.2 |
|
50 |
0.06 |
93.3 |