Research Article

Characterization and Assessment of Antibacterial Potential of Garlic Based Silver Nanoparticles

Mehwish Saleem1* and Amina Atiq2

1Lahore College for Women University, Lahore, Pakistan; 2Islamia College Cooper Road, Lahore, Pakistan.

Received | September 25, 2022; Accepted | October 31, 2022; Published | November 26, 2022

*Correspondence | Mehwish Saleem, Lahore College for Women University, Lahore, Pakistan; Email: [email protected]

Citation | Saleem, M., and A. Atiq. 2022. Characterization and assessment of antibacterial potential of garlic based silver nanoparticles. Journal of Innovative Sciences, 8(2): 250-255.

DOI | https://dx.doi.org/10.17582/journal.jis/2022/8.2.250.255

Keywords | Nanotechnology, Garlic, Silver nanoparticle, Antibiotic resistance, Pathogens

Copyright: 2022 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

1. Introduction

A well-known medicinal herb known for its therapeutic and insecticidal capabilities is garlic (Allium sativum) (Ankri et al., 1997; Chen et al., 2018). Additionally, it has a lot of phytochemicals that shield people from illnesses (Kaur et al., 2019). Similarly, silver is well known for its bactericidal and inhibitive properties (Shankar et al., 2004). Antimicrobial agent resistance in bacteria has become a major global issue, increasing mortality in most cases (Ashour et al., 2015). As a result, it has become essential to identify novel antibiotic alternatives (Azam et al., 2012).

The wide range of uses for nanometals can be attributed to their high surface/ volume ratio (Ashour et al., 2015) which makes them superior to conventional antibiotics (Wang et al., 2017). Nanoparticles have additional antioxidant and antibacterial capabilities (Pasupuleti et al., 2008). Plant extracts and phytochemicals may function as both stabilizing and reducing agents during the formation of nanoparticles (Akhtar et al., 2013; Mittal et al., 2013). In recent decades, one of the most widely studied materials has been silver nanoparticles (Saravanan et al., 2018). Additionally, they have minimal costs, little cytotoxicity, and little immunological reaction (Samuel et al., 2020).

Garlic’s properties include actions that are antibacterial, antioxidant, immunomodulatory, anti-cancer, anti-inflammatory, hypoglycemic, and cardiovascular (Arshad et al., 2020). Recently, drug delivery utilizing nanoparticles has advanced quickly (Farrag et al., 2019; Girish et al., 2019), and the potential biological uses of nanoparticles derived from garlic have become more apparent. Furthermore, a wide variety of bacterial pathogens are not as commonly used as silver-based nanoparticles (Gu et al., 2013). As a result, there has been a lot of interest in the biosynthesis of silver nanoparticles, which is particularly intriguing for plants and could be a significant alternative in the treatment and control of infection by human diseases (Wang et al., 2017). Due to their outstanding qualities that make them suitable for various applications, many types of garlic extracts and their separated bio actives are used as an ingredient in the biogenesis of nanoparticles (Hussein et al., 2017; Kim et al., 2007; Jakobsen et al., 2012).

2. Materials and Methods

The garlic was purchased at a local market in Lahore. Garlic aqueous extracts were made using the maceration method. Garlic ethanolic extract was also made by dissolving 10g of sample in 100 mL of ethanol. For 24 hours, the suspension was shaken on a flask shaker at 280 rpm (Abbas et al., 2022). 0.01g, 0.025g and 0.05g aqueous and ethanol extracts were mixed with 100 mL of distilled water to make 0.01%, 0.025%, and 0.05% extract solutions, respectively (Table 1, Figure 1). The extracts were then mixed with silver nitrate to make silver nitrate nanoparticles (Figure 2). Furthermore, streptomycin was used as a positive control as a standard antibiotic, and a simple silver nitrate solution was used as a negative control to compare the antimicrobial results (Tahir et al., 2020).

2.1 Characterization

The size of the silver nanoparticles conjugated with garlic was determined using the BT-90 nano laser particle size analyzer. The 4 cm cuvette was briefly filled with nanoparticle solution to determine particle size. Triplicate runs of freshly manufactured samples were performed, and the findings were considered as average measurements (Salem et al., 2018). The diagnostic peaks of garlic and garlic coupled with silver were examined using UV-17000. To determine the particle nature of the AgNPs, an XRD analysis (X-ray diffractometer) was executed. The chemical formation of prepared NPs in LUMS was evaluated using (Fourier transform infrared) analysis (Tahir et al., 2020).

 

Table 1: Groups of garlic silver nanoparticles for antibacterial activity.

S. No.	Groups
1	0.01M Silver Nitrate Solution
2	Antibiotic Streptomycin
3	0.01% Aqueous Extract + 0.01M Silver Nitrate
4	0.025% Aqueous Extract + 0.01M Silver Nitrate
5	0.05% Aqueous Extract + 0.01M Silver Nitrate
6	0.01% Ethanolic Extract + 0.01M Silver Nitrate
7	0.025% Ethanolic Extract + 0.01M Silver Nitrate
8	0.05% Ethanolic Extract + 0.01M Silver Nitrate

 

 

 

2.2 Culture media preparation

To prepare a culture media for bacterial growth, 7.4g nutrient agar and 2.1g agarose gel were mixed in 250ml of distilled water in a flask that was placed on a hot plate for 10-15 minutes. For sterilization, the culture media was autoclaved for 1 hour. The sterilized culture media was poured into Petri dishes that were placed in laminar air flow to solidify. After solidification the Petri plates were incubated in incubator (A. Polok-Kowalska, S. Kowalski Typ; CLN 32 STD S/N; CN32SF 140474) at 37º C for 24 hours (Fahmy and Mamdouh, 2018).

2.3 Antimicrobial assay

The agar well diffusion method was used to assess the antibacterial activity of garlic extract attached NPs against E. coli, S. aureus, B. subtilis, and K. pneumoniae (He et al., 2017). Petri plates containing solidified culture media were transferred to a laminar air flow chamber, and each Petri plate was divided into four wells. Then, to observe antibacterial activity, each well was labelled with a name. The antibiotic streptomycin was poured into the first well in a volume of 10: l (as a positive control). The second well was filled with 60µl of silver solution (negative control). The remaining two wells were pored with 60µl of ethanolic and hot water extracts containing similar concentrations of silver-based nanoparticles. In three separate Petri plates, a 0.01 M solution of 0.01%, 0.025%, and 0.05% garlic aqueous extract and ethanolic extract based nanoparticle solution was poured (He et al., 2017).

3. Results and Discussion

3.1 UV-visible spectrophotometer

UV-visible (UV-Vis) spectrophotometry is the most crucial and simple method for verifying the creation of nanoparticles. Using a Shimadzu (Kyoto, Japan) UV-Vis spectrophotometer model 1800 with a wavelength between 190 and 800 nm. The absorbance spectra of numerous substances were captured. Garlic AgNPs suspension is a deep dark brown color. As a result, it is possible to forecast particle size and stability using absorbance peaks. The absorbance maximum of smaller Ag NPs is around 400 nm, and it rises with particle size before disappearing once the particle size reaches the nanoscale (Figure 3). Our research revealed that green produced Ag NPs had their highest absorbance at 408 nm. Ag NPs made by the green method have a limited size distribution, with smaller particles predominating, as shown by a narrow absorption peak at 408 nm.

3.2 Antibacterial activity

The negative control’s growth inhibition zone against bacterial strains (E. coli, S. aureus, K. pneumoniae and B. subtilis) was 5mm, 5mm, 12mm, and 9 mm, respectively. The growth inhibition zone of aqueous garlic extract against E. coli was 10mm, 10mm, and 13mm at 0.01%, 0.025%, and 0.05%, respectively. 9mm, 10mm and 12mm at 0.01%, 0.025%, and 0.05%. The growth inhibition zone of aqueous garlic extract against K. pneumoniae was 10mm, 9mm and 11mm at 0.01%, 0.025%, and 0.05%, respectively, and 10mm, 8mm, and 10mm at 0.01%, 0.025%, and 0.05% (Figure 5).

 

 

The growth inhibition zone of aqueous garlic extract against B. subtilis was 9mm, 9mm, and 10mm at 0.01%, 0.025%, and 0.05%, respectively, and 11mm, 13mm, and 9mm at 0.01%, 0.025%, and 0.05%. When compared to other bacterial strains, G-AgNPs had a greater effect on S. aureus. The zone of inhibition of

 

Table 2: Zone of inhibition of experimental trial against for different strains of bacteria by using agar well diffusion method.

	E. coli	K. pneumoniae	S. aureus	B. subtilis
Positive control	14mm	11mm	16mm	12mm
Negative control	5mm	5mm	12mm	9mm
0.01% AGE + 0.01M AgNO3	10mm	10mm	19mm	9mm
0.025% AGE+ 0.01M AgNO3	10mm	9mm	17mm	9mm
0.05% AGE + 0.01M AgNO3	13mm	11mm	18mm	10mm
0.01% EGE + 0.01M AgNO3	9mm	10mm	19mm	11mm
0.025% EGE+ 0.01M AgNO3	10mm	8mm	17mm	13mm
0.05% EGE+ 0.01M AgNO3	12mm	10mm	15mm	9mm

 

aqueous garlic extracts against S. aureus was 19mm, 17mm, and 18mm at 0.01%, 0.025%, and 0.05%, respectively, and 19mm, 17mm, and 15mm at 0.01%, 0.025%, and 0.05%, respectively (Table 2, Figure 4).

 

 

The current study confirms that antibacterial property of ethanolic extract is far better that aqueous extract. Several studies have been published that support our findings that an ethanolic (30%) extract of fermented black garlic has higher antibacterial activity against 11 bacterial strains than an aqueous extract that causes oral diseases. This extract inhibited the growth of more than 90% of salivary bacteria in both short and long incubations (Vlachojannis et al., 2018). Water extract of garlic Toluene extract has been reported to reduce the mortality of Caenorhabditis elegans from P. aeruginosa infections and clean the lungs of P. aeruginosa infected mice by modulating inflammation (Bapat et al., 2018).

Furthermore, garlic extracts are said to be effective against both gram-positive and gram-negative bacteria, such as B. subtilis, S. aureus, E. coli and K. pneumoniae (Gabriel et al., 2022). According to our findings, green-produced Ag NPs had the highest absorbance at 408 nm. The green approach produces Ag NPs with a restricted size distribution, with smaller particles predominating, as evidenced by a narrow absorption peak at 408 nm. According to this paper, nano-particles have a brown color (Shafea et al., 2021). The absorbance spectrum of the silver nanoparticles prepared in the reaction mixture was obtained using UV-Vis analysis, with the highest peak occurring at about 433 nm (Vijayakumar et al., 2019).

Conclusions and Recommendations

According to considerable studies, organosulfur compounds from garlic have potent antibacterial properties against a variety of bacteria, including MDR strains. The effectiveness of garlic extracts against Gram-positive and Gram-negative bacteria such E. coli, K. pneumoniae, B. subtilis and S. aureus has also been reported. Garlic and its components have been studied extensively for their antibacterial properties, but more recent gaps in our knowledge must be filled before we can use them as antibacterial agents in clinical settings.

Novelty Statement

Addition of garlic based silver nanoparticles proved an alternative to replace antibiotics, hence proved an effective step to combat diseases.

Author’s Contribution

Mehwish Saleem designed the study, wrote and edit the paper, paid publication fee. Amina Atiq performed the experimental work.

Conflict of interest

The authors have declared no conflict of interest.

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