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Journal of Environmental Chemical Engineering 9 (2021) 105290

Contents lists available at ScienceDirect

Journal of Environmental Chemical Engineering


journal homepage: www.elsevier.com/locate/jece

Green synthesis of a silver nanoparticle using Moringa oleifera seed and its
applications for antimicrobial and sun-light mediated photocatalytic
water detoxification
Hafiza Mahreen Mehwish a, Muhammad Shahid Riaz Rajoka a, Yongai Xiong a, Huiming Cai a,
Rana Muhammad Aadil b, Qaisar Mahmood c, Zhendan He a, d, *, Qinchang Zhu a, **
a
School of Pharmaceutical Science, Guangdong Key Laboratory for Genome Stability & Human Disease Prevention, Shenzhen Key Laboratory of Novel Natural Health
Care Products, Engineering Laboratory of Shenzhen Natural Small Molecule Innovative Drugs, Health Science Center, Shenzhen University, Shenzhen 518060, PR China
b
National Institute of Food Science and Technology, University of Agriculture, Faisalabad 38000, Pakistan
c
Department of Environmental Science, COMSAT University Islamabad, Pakistan
d
School of Pharmacy, Shenzhen Technology University, Shenzhen 518060, PR China

A R T I C L E I N F O A B S T R A C T

Editor: Dr. G. Palmisano Natural water resources are rapidly being polluted because of improper treatment of sewage from industrial
facilities; accordingly, there is an urgent need for environmentally friendly methods of treatment for this type of
Keywords: waste. Here, synthesis of cost effective, green silver nanoparticles (AgNPs) using Moringa oleifera seed (MOS) as a
Moringa oleifera seed reducing/capping agent and their applications in antimicrobial and photocatalytic oxidation for water treatment
Silver nanoparticles
are reported. The results revealed that the MOS-AgNPs were crystalline and spherical, with an average size of
Antimicrobial activity
4.0 nm. The MOS-AgNPs showed excellent antimicrobial activity against Gram positive (Staphylococcus aureus;
Photocatalytic activity
Water treatment 14.6 mm) and Gram negative (Escherichia coli; 30.6 mm, Salmonella enterica typhimurium (29 mm), and Pseudo­
monas aeruginosa; 22.8 mm) bacteria. Moreover, the MOS-AgNPs showed remarkable photocatalytic activity
toward organic dyes (methylene blue (> 81%), orange red (> 82%), and 4-nitrophenol (> 75%)) under sunlight
irradiation. In addition, > 80% of the toxic Pb metal ions were removed from the treated water by MOS-AgNPs.
Under similar conditions, the synthesized MOS-AgNPs retained their photocatalytic efficiency after 10 photo­
catalytic cycles. Overall, these findings indicate that MOS-AgNPs may be a powerful antimicrobial agent against
water borne pathogens as well as a promising and economic agent for use in the treatment of waste generated by
industrial dyeing processes.

1. Introduction easy, economically feasible, and environmentally friendly approach to


the removal of organic dyes from wastewater.
The consistent growth of the world’s population coupled with One major problem in water treatment is the removal of toxic pol­
industrialization and urbanization has led to increased demand for en­ lutants. Organic dyes from the textile industry are one of the major
ergy, chemicals, and materials [48]. Furthermore, discharge of highly pollutants that impact aquatic systems. These organic dyes are inevi­
undesirable waste into the environment is inevitably accompanied by tably released into environment, where they cause serious toxicity to
lower quality of environmental systems, especially aquatic reservoirs plants and animals [37]. Organic dye pollutants, including methylene
[53]. While the textile industry plays a vital role in the global economy, blue, Congo red and 4-nitrophenol, are non-degradable because of their
it also serves as one of the largest sources of environmental pollution [6]. complex structures (Fig. 1) and can therefore cause long-term pollution
Indeed, the wastewater released from the textile industry is widely re­ of aquatic systems [65]. Microbiological contaminants comprise another
ported to be of serious concern to the environment, especially to aquatic major pollutant in water, and serious health problems can occur when
and terrestrial habitats [22]. Therefore, there is an urgent need for an there is continuous transmission of bacterial and viral pathogens to

* Corresponding author at: School of Pharmacy, Shenzhen Technology University, Shenzhen, China.
** Corresponding author.
E-mail addresses: hezhendan@szu.edu.cn (Z. He), zhuqc@szu.edu.cn (Q. Zhu).

https://doi.org/10.1016/j.jece.2021.105290
Received 21 December 2020; Received in revised form 24 February 2021; Accepted 26 February 2021
Available online 1 March 2021
2213-3437/© 2021 Elsevier Ltd. All rights reserved.
H.M. Mehwish et al. Journal of Environmental Chemical Engineering 9 (2021) 105290

water. For example, water-borne diseases are mainly caused by the using Artemisia tournefortiana Rchb extract showed excellent photo­
consumption of microbial contaminated water or food that has been catalytic degradation of Coomassie brilliant dye under sunlight exposure
cross-contaminated with microbes during washing [25]. Therefore, within 60 min [5].
environmentally friendly practices are urgently needed for catalytic Moringa oleifera (MO) is a moderate sized tree cultivated throughout
degradation of organic pollutants and water disinfection. Recently, India and Myanmar. Almost all parts of this tree are edible, and it is
biogenic nanoparticles have been efficiently used to eradicate various commonly used as food in India [33]. Moringa oleifera plant extract is
organic dyes and bacterial pathogens from water [61]. rich in proteins, carbohydrates, phenol, vitamins, kaempferol, potas­
Nanotechnology is a rapidly growing scientific field that encom­ sium, calcium, and amino acids [60]. Recently, Moringa oleifera seed
passes a wide range of applications, from medicine to aeronautics, from (MOS) has attracted a lot of attention because of its contributions to
agricultural husbandry to molecular pharming [58]. Metal nanoparticles rural economies as well as its being an excellent source of oil and various
have unique size and shape related properties that make them of great other products. MOS is traditionally used to flocculate impurities in
interest for use in catalytic, sensing, optics, and antibacterial materials water, for which it has received scientific attention for approximately 40
[54]. Among metal nanoparticles, silver nanoparticles (AgNPs) have years [59]. Although MOS can be used as an excellent coagulant for
been shown to have high photocatalytic, antioxidant, and antimicrobial water purification, it does not show antibacterial activity. Therefore,
activities [21]. water treated with MOS is not suitable for drinking without further
AgNPs have been used as an antimicrobial agent against a wide range disinfection [36]. Another viable method to remove bacteria is use of
of Gram positive and negative bacteria [28]. Several bacterial pathogens metals with antimicrobial properties, such as silver. However, the major
generate antibiotics resistance genes because of the use of antibiotics MOS bioactive compounds and their role in the synthesis of MOS-AgNPs
and nanoparticles have been suggested as a substitute to traditional for wastewater treatment have not yet been investigated.
antibiotics because of their exceptional antimicrobial properties [24]. Therefore, the present study was conducted to investigate the green
Various chemical and physical methods used to synthesize AgNPs are synthesis of silver nanoparticles using MOS and their potential appli­
not cost effective. In addition, chemical methods used to synthesize cations in water treatment. The synthesized MOS-AgNPs were charac­
AgNPs often produce various toxic substances that pose ecological risks terized using FTIR, XRD, XPS, SEM, TEM, AFM, and TGA analysis and
[64]. Accordingly, it is necessary to develop alternative processes that further evaluated for their potential antimicrobial and photocatalytic
are safe for the environment and sustainable [19]. In recent years, activities. Finally, the MOS-AgNPs potential to remove heavy metals
synthesis of AgNPs using green chemistry to reduce the use and for­ such as Pb+2 from water was investigated.
mation of toxic substances using green chemistry has gained consider­
able attention [57]. 2. Materials and methods
In recent decades, various biomimetic processes have been evaluated
for their ability to synthesize AgNPs [13]. Although microbes are widely 2.1. Moringa oleifera seed (MOS) extract preparation
used to synthesize nanoparticles, they have some limitations, including
difficulty preserving microbial strains, difficulty maintaining aseptic The dried and cleaned seeds of Moringa oleifera (MO) were purchased
conditions and high culture media preparation costs. However, the use from Yunnan Siwu Biotechnology, Inc. (Kunming, Yunnan, China). The
of plants to synthesize nanoparticles has recently emerged as an alter­ cultivated variety was PKm2 and a voucher specimen (SZU-14CS9354)
native to avoid the limitations associated with generating nanoparticles was deposited in our laboratory. MOS (30 kg) was washed with distilled
using microbes. At present, various plants such as Laminaria japonica, water and refluxed with ethanol (Sigma Aldrich, China) for 2 h, twice,
Taxus baccata, Fritillaria flower, Trifolium resupinatum, Solanum melon­ after which the extracts were concentrated at reduced pressure and
gena, and Aloevera are used for green synthesis of AgNPs [46]. It was ethanol extract was obtained. Next, the ethanol extract was separated
recently shown that Gallnut extract mediated AgNPs composite con­ into petroleum ether (Sigma Aldrich) and alcohol (Sigma Aldrich) by
tained reduced nitrophenol (76.5%) relative to a control. Moreover, solid extraction. The alcohol portion was then added to the mixture to
synthesized AgNPs have shown excellent antimicrobial activity against collect the extract part, after which it was filtered using Whatman No. 1
E. coli and Staphylococcus aureus [11]. Additionally, AgNPs synthesized filter paper (Sigma Aldrich) and freeze dried using a rotary evaporator.

Fig. 1. Representative chemicals structure of dyes, (A); methylene blue, (B); 4-nitrophenol, and (C); congo red.

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H.M. Mehwish et al. Journal of Environmental Chemical Engineering 9 (2021) 105290

The dried MOS extract was stored at RT until use. 2.4.3. Bacterial viability assay
The bacterial viability of each tested strain was calculated by the
colony counting method [50], after which the percentage of viable mi­
2.2. Synthesis of MOS-AgNPs crobial cells was determined by comparing the controls with the treat­
ments using the following formula:
To optimize the amount of MOS required for the synthesis of AgNPs,
varying volumes (0–10 ml) of aqueous seed extract were added to the Bacterial viability (%) = (number of colonies on treated plate/number of col­
0.5 mM silver nitrate solution (final volume 100 ml) at pH 11.0. After onies on control plate) × 100 (1)
optimization, the MOS-AgNPs were synthesized by mixing 10 ml of MOS
extract (10 mg) and 90 ml of 0.5 mM silver nitrate (Sigma Aldrich)
while heating the reaction mixture (optimized pH 11) at 60 ◦ C and
stirring at 300 rpm using a magnetic stirrer. The reaction mixture was
2.5. Photocatalytic degradation of dyes
analyzed after 2 h using a UV–Vis spectrophotometer (LAMBDA, Per­
kinElmer, USA) to evaluate the standard peak (at 350–450 nm). At the
2.5.1. Photocatalytic activity toward Congo red and methylene blue
same time, variations in color were noted. After 2 h, the MOS-AgNPs
MOS-AgNPs potential for catalysis of the reduction of methylene
were purified via centrifugation at 8000 rpm for 20 min. The pellets
blue (MB) (Sigma Aldrich) and Congo red (CR) (Sigma Aldrich) under
were then suspended in distilled water and dialyzed (dialysis membrane,
sunlight (97,000 lx, 11:00 a.m. to 1:00 p.m.) was determined via
Beijing, Sigma) against distilled water three times to completely remove
UV–Vis spectroscopy (LAMBDA, PekinElmer, USA) as previously
the unbound compounds. Next, the reaction mixture was centrifuged to
described [65]. Finally, the organic dye degradation rate (k) was
obtained pellets, which were subsequently dried at 80 ◦ C and then
determined as follows:
stored at 4 ◦ C until use. Furthermore, the effects of temperature and
catalytic dose were optimized by maintaining the precursor solution of Organic dye degradation rate (k) = [(A0 − At)/A0] × 100 (2)
silver nitrate at various temperatures (25 ◦ C, 35 ◦ C, 45 ◦ C, 55 ◦ C, and
65 ◦ C) and varying catalyst quantities (0–2.0 mM) [18]. where k = the degradation rate, A0 = the original solution absor­
bance, and At = the final solution absorbance.
A wastewater sample was also used to authenticate the feasibility of
2.3. Characterization of MOS-AgNPs this method and determine the catalytic activity of MOS-AgNPs toward
MB (Sigma Aldrich) and CR (Sigma Aldrich). To accomplish this, a
SEM (JSM-7800F, Japan) was used to visualize the shape of bio­ wastewater sample was collected from a sewage treatment plant in
synthesized MOS-AgNPs. For transmission electron microscopy (JEM- Shenzhen, Guangdong province, China and mixed with organic dyes,
1230, Japan) analysis. A micrograph was obtained at 200 Kv and the size after which the catalytic activity of MOS-AgNPs toward organic dye was
and the diameter of MOS-AgNPs were determined by measuring at least determined as described above.
50 particles on the TEM micrograph using Image J. AFM (Dimension
ICON, Brooke, USA) is commonly used to investigate the height and 2.5.2. Photocatalytic activity toward 4-nitrophenol
roughness of nanoparticles [31]. FTIR spectroscopy (Nicolet 6700, The catalytic reduction of 4-nitrophenol by MOS-AgNPs was moni­
Nicolet, United State) analysis was performed in a range of tored using UV–Vis spectroscopy (LAMBDA, PekinElmer, USA) as pre­
4000–400 cm− 1 using KBr (Sigma Aldrich) pellets mixed with purified viously reported [38]. The absorption spectra were then recorded in the
powder of biosynthesized MOS-AgNPs at a resolution of 4 cm− 1 and a range of 300–700 nm via UV–Vis spectroscopy (LAMBDA, PerkinElmer)
scanning frequency of 32 [42]. X-ray diffraction (XRD) scanning (D8 at RT and the degradation rate was calculated using Eq. (2).
Advance, Bruker AXS, Germany) was conducted to analyze the crystal­
line nature of MOS-AgNPs within the range of 30–70 θ [56]. The thermal 2.5.3. Reusability of AgNPs
stability of MOS-AgNPs was studied using a thermo gravimetric analyzer AgNPs were added to the organic dye solutions at a final concen­
(TGA-Q50, American TA Instruments, USA) under nitrogen atmospheric tration of 1 mg/ml, after which the reaction mixtures were agitated at
pressure. RT for 60 min. Next, the AgNPs were separated via centrifugation and
washed with distilled water. The collected AgNPs were then oven dried
and reused for the next experimental cycle under the same conditions
2.4. Antimicrobial activity [1].

2.4.1. Agar well diffusion assay


The anti-bacterial potential of MOS-AgNPs was evaluated using an 2.6. Microbially contaminated wastewater disinfection
Agar well diffusion assay [4] against the Gram positive bacteria Staph­
ylococcus aureus ATCC29213 and the Gram negative bacteria Escherichia Wastewater samples (200 ml) were treated with MOS-AgNPs (pellet
coli ATCC 25922, Pseudomonas aeruginosa ATCC27853, and Salmonella on chitosan) by shaking at 120 rpm for 60 min. The wastewater samples
enterica serotype typhimurium ATCC14028. The diameters (mm) of the were subsequently evaluated by Escherichia coli quantification using
inhibition zones were measured in triplicate at the end of the incubation colony counting methods as previously described [16].
period.

2.4.2. Determination of minimum inhibitory concentration against each 2.7. Treatment of wastewater
pathogenic strain
The MIC of MOS-AgNPs against each pathogenic bacterial strain was For wastewater treatment, 50 mg of MOS-AgNPs was mixed with
determined as previously described [20], with slight modification. 200 ml of wastewater and then shaken for 6 h under sunlight (mid-day),
Briefly, 200 µl of MOS-AgNPs at each concentration (2.5–100 µg/ml) while untreated samples were used as a control. The pH, chemical ox­
was inoculated into 1.5 ml of nutrient broth supplemented with 100 µl ygen demand (COD), total suspended solids (TSS), total dissolved solids
of old cultures of pathogenic bacterial strains. Nutrient broth without (TDS), turbidity, and electrical conductivity (EC) were then determined
MOS-AgNPs was used as a control. After incubation at 30 ◦ C for 24 h, the using the standard methods for examination of wastewater as previously
MIC values were measured by calculating the OD at 630 nm using a described [40]. Additionally, the hardness, chloride, and sulfate con­
spectrophotometer (H1, BioTek, USA). tents of the wastewater were determined as described by Rump [52].

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H.M. Mehwish et al. Journal of Environmental Chemical Engineering 9 (2021) 105290

2.8. MOS-AgNPs Pb+2 removal efficiency in contaminated water was most intense and narrow, which indicates that this was the most
suitable pH for further experiments. However, increasing the amount of
Solutions containing various concentrations of Pb+2 (0.5–0.05 mg/ MOS extract resulted in a broadening absorption peak, which indicates
ml; Sigma Aldrich) were added to flasks containing water and treated that the MOS-AgNPs increased in size (data not shown). Based on these
with MOS-AgNPs (1 mg/ml) solution at RT for 30 min. A flask without results, a variety of MOS primary/secondary phytochemicals (gallic
MOS-AgNPs was used as a control. All experiments were performed in acid, flavonoids, and phenolic acids) act as stabilizing and capping
triplicate under RT and the percentage (%) reduction rate was calculated agents during the fabrication of MOS-AgNPs; hence, these MOS extract
as follows: phytochemicals were responsible for the Ag nanoparticle reduction. A
possible schematic representation of MOS-AgNPs formation is shown in
Reduction rate (%) = A/B × 100 (3)
Fig. 2C.
where A is the amount of the reduction and B is the original amount
that was reduced. 3.2.2. Morphological analysis of MOS-AgNPs
The FE-SEM image of the MOS-AgNPs revealed an even shape and
3. Results and discussion spherical surface morphology (Fig. 3A). Chemical analysis of MOS-
AgNPs using EDX analysis revealed the presence of Ag and other ele­
3.1. Synthesis of MOS-AgNPs ments covering the Ag aggregates (Fig. 3B). The presence of Ag was
because of the formation of MOS-AgNPs, while the other elements were
The optimized conditions for the biosynthesis of MOS-AgNPs were from the MOS extract. In the present study, the average size of MOS-
found to be 0.5 mM AgNO3 aqueous solution, 10 ml MOS extract AgNPs was 10.2 nm, which is in agreement with the results of a
(10 mg), pH 11, and 60 ◦ C in the dark. The color change of the resultant similar study conducted by Raj et al. [45], who found AgNPs synthesized
solution from yellow to reddish brown suggested the formation of MOS- via Enicostemma axillare (Lam.) leaf extract had an average size of
AgNPs had occurred (Fig. 2A). 18.12 nm.
The representative TEM image in Fig. 3C shows individual MOS-
3.2. Characterization of silver nanoparticles AgNPs as well as small aggregates. Most of the MOS-AgNPs were
spherical in shape with an average size of 4.0 nm (Fig. 3D). The MOS-
3.2.1. Ultraviolet–visible (UV–Vis) spectroscopy AgNPs were not in direct contact, even within aggregates, which in­
In the present study, UV–Vis absorption spectroscopy conducted dicates the stabilization of particles via MOS extract capping molecules.
after 2 h of reaction gave an absorbance at 421 nm, which indicates the Previous studies have reported different mechanisms of AgNPs biosyn­
formation of MOS-AgNPs (Fig. 2A). In addition, the surface plasmon thesis events that are related to oligopeptide catalysis, nitrate reductase,
resonance band of the MOS-AgNPs solution remained close to 421 nm, and NADH dependent reductase participating in nanoparticles that are
which further confirmed that silver nanoparticles in the aqueous phase spherical and triangular [39]. AFM probe analysis showed that the
were monodispersed. The stability of MOS-AgNPs could be a result of MOS-AgNPs varied in height and roughness (Fig. 3E), which is consis­
capping agents present in the MO, such as polysaccharides, proteins, and tent with the TEM and SEM observations.
phenolic compounds [32]. Furthermore, the UV–vis absorption spectra
taken after 2 h at different pH (11, 9, 7, and 5) revealed major differ­ 3.2.3. Fourier transform infrared (FT-IR) spectroscopy
ences in the spectra (Fig. 2B). Specifically, the absorption band at pH 11 Comparison of the FTIR spectrum of MOS-AgNPs with that of MOS

Fig. 2. UV–Vis absorption spectra taken after 2 h at different pH (A, B) and a schematic illustration of MOS-AgNPs synthesis (C).

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H.M. Mehwish et al. Journal of Environmental Chemical Engineering 9 (2021) 105290

Fig. 3. Morphology of MOS-AgNPs Scanning electron microscopy (A); EDX analysis (B); Transmission electron microscopy (C); MOS-AgNPs size distribution (D); and
Atomic force microscopy (E).

aqueous extract (Fig. 4A) revealed that noticeable peak shifts at MOS extract were also present in MOS-AgNPs with slight peak shifts,
3387–3392 cm− 1 (O‒H stretch) and 2927–2924 cm− 1 (C‒H stretch), which indicates the formation of MOS-AgNPs.
which were consistent with polyphenolic or flavonoid compounds and
alkanes, respectively, as well as stretching at 1651–1637.5 cm− 1 (‒ 3.2.4. X-ray diffraction (XRD) analysis
C˭C‒) and 1062–1058.9 cm− 1 (C‒O stretch), which is consistent with X-ray diffraction analysis revealed a peak at 37.8◦ (2θ), which in­
alkenes and primary alcohol, respectively, in both MOS-AgNPs and MOS dicates that the nanoparticles were composed of pure silver [63]. In
extract. These findings suggest that the functional groups present in addition, XRD analysis showed that the MOS-AgNPs were crystalline in

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H.M. Mehwish et al. Journal of Environmental Chemical Engineering 9 (2021) 105290

Fig. 4. FTIR (A), XRD (B), and thermogravimetric (C) analysis of MOS-AgNPs.

nature. There were also peaks at 42◦ (2θ), 65◦ (2θ), and 78◦ (2θ) con­ 3.2.5. Thermo-gravimetric analysis
forming to 200, 220, and 311 silver planes, respectively. Furthermore, The TGA results (Fig. 4C) showed that there were three major weight
the unassigned peaks in the XRD pattern might have been because of the loss ranges. The first weight loss (approximately 5.2%) range reflected
existence of bio-organic phases that occurred on the surface of the loss of absorbed water and volatile residues, as described previously
MOS-AgNPs (Fig. 4B). [51]. The second weight loss (approximately 11.4%), which occurred
between 200◦ and 420 ◦ C, was a result of the thermal degradation of

Fig. 5. Antibacterial activity measured by zone inhibition assay of MOS-AgNPs. The data is statistically significant at p < 0.001.

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H.M. Mehwish et al. Journal of Environmental Chemical Engineering 9 (2021) 105290

MOS phytochemical compounds found on the surface of MOS-AgNPs. 3.4. Photocatalytic degradation of organic dyes
Finally, transitions were observed at above 800 ◦ C, which is close to
the melting point of silver. 3.4.1. Photocatalytic degradation of Congo red and methylene blue
After treatment with MOS-AgNPs, there was a gradual decrease in
3.3. Antimicrobial activity color from deep blue for MB solution or red for CR solution to colorless,
which indicates the degradation of organic dyes. Moreover, an absorp­
The MOS-AgNPs exhibited significantly (p < 0.001) higher antimi­ tion peak was found at 660 nm for MB dye solution (Fig. 6A) and 490 nm
crobial activity than the MOS extract. The results showed that the MOS- for CR dye solution (Fig. 6D) during 75 min of exposure to solar light in
AgNPs (100 µg/ml) exerted the highest anti-bacterial activity against the presence of MOS-AgNPs. The reaction kinetics for the MB and CR
Gram negative bacteria. Specifically, E. coli strain ATCC25922 had an organic dyes were described by a pseudo first order reaction as shown in
inhibition zone of 28.6 mm, Pseudomonas aeruginosa strain ATCC25853 Fig. 6C and F. Overall, the results revealed the following linear corre­
had an inhibition zone of 22.8 mm, and Salmonella enterica typhimurium lations of ln (At/A0) vs time after 75 min: y = − 0.0475, x = − 0.2335
ATCC14028 had an inhibition zone of 29 mm, whereas the Gram posi­ R2 = 0.9514 for MB organic dye and y = − 0.0451, x = − 0.3874
tive organism Staphylococcus aureus strains ATCC29213 had an inhibi­ R2 = 0.9605 for CR organic dye.
tion zone of 15.6 mm (Fig. 5). This antimicrobial activity was higher The maximum MB degradation rate observed was 91.4% for waste­
than that reported for Moringa oleifera leaf extract mediated AgNPs [35] water and 82.4% for pure water, while the CR degradation rates for
against Gram negative and Gram-positive bacteria [2]. Previous studies wastewater and pure water were 86.2% and 82.7%, respectively (Fig. 6B
reported that Cannabis sativa leaf extract mediated AgNPs (1 mg/ml) and E). Furthermore, in the absence of MOS-AgNPs photocatalyst there
exhibited antimicrobial activity against E. coli and Staphylococcus aureus was no noticeable decrease in the MB and CR dye concentrations. In a
strains, forming zones of inhibition of 7.5 mm and 0.6 mm, respectively previous study, AgNPs photo-catalytically degraded MB (78%) under
[9]. sunlight irradiation after 90 min [14].

3.3.1. Minimum inhibitory concentration of each pathogenic strain 3.4.2. Photocatalytic degradation of 4-nitrophenol
The MOS-AgNPs showed significantly (p < 0.001) higher antimi­ The aqueous solution of 4-nitrophenol showed peak absorbance at
crobial activity with low MIC values against Gram negative bacteria 317 nm (Fig. 7A); however, a rapid shift to 400 nm was observed
(13 µg/ml for Escherichia coli ATCC25922, 18 µg/ml for Pseudomonas immediately after the addition of NaBH4 because of formation of
aeruginosa ATCC27853, and 9 µg/ml for Salmonella enterica typhimurium phenolate ion as an intermediate product. Additionally, reduction
ATCC14028) and Gram positive bacteria (35 µg/ml for Staphylococcus occurred after addition of MOS-AgNPs as indicated by the rapid decrease
aureus ATCC29213) than MOS extract, which was in accordance with in the phenolate peak (400 nm) with time (Fig. 7B). Overall, the results
the results of the agar well diffusion assay (Table S1). A previous study indicated that approximately 89% of the 4-nitrophenol was successfully
showed that AgNPs synthesized using leaf extract of Curcuma longa L. reduced into by-product within 75 min (Fig. 7C). As shown in Fig. 7D,
exhibited MIC values of 2.5 µl, 2.25 µl, and 2.5 µl against E. coli, Pseu­ the reaction kinetic was described by a pseudo first order reaction for 4-
domonas aeruginosa, and Staphylococcus aureus, respectively [30]. nitrophenol reduction. Additionally, the results showed a linear corre­
Another study reported that the mediated AgNPs exhibited MIC values lation of ln(At/A0) vs. time after 75 min for 4-nitrophenol reduction
of 235 µg and 120 µg against E. coli and Staphylococcus aureus, respec­ (R2 = 0.9614). These findings were consistent with the previous finding
tively [23]. As shown in Table 1, our synthesized MOS-AgNPs exhibited that Phaseolus vulgaris seed mediated AgNPs could be used as a catalyst
excellent antimicrobial activity relative to previously reported AgNPs. to degrade 4-nitrophenol (< 90%) successfully [48].
The degradation rates of the dyes (MB, 4-NP, and CR) also decreased
3.3.2. Bacterial viability assay as their initial concentrations increased. This was because of over­
The antibacterial activity of MOS-AgNPs determined based on the crowding of organic dye molecules on the catalyst surface under high
colony forming unit assays showed that MOS-AgNPs significantly concentration conditions. Accordingly, increasing the dose of catalyst
(p < 0.001) inhibited the growth of all tested pathogenic microbes enhanced the photocatalytic degradation of organic dyes. Furthermore,
(Table 2) relative to MOS extract. Among the tested pathogens, Gram the organic dye degradation rates increased as the pH increased from 5.0
negative bacteria were more susceptible than Gram positive bacteria, as to 9.0, above which they decreased. Therefore, in the present study, pH
indicated by inhibition rates of 98.2 ± 0.9%, 89.1 ± 1.0% and 92 ± 1.5 9.0 was used as the optimal pH to obtain the maximum degradation of
for Escherichia coli ATCC25922, Pseudomonas aeruginosa ATCC27853 organic dyes. A possible comparsion of dyes degrdation rate (Table 3)
and Salmonella enterica typhimurium ATCC14028, respectively, and schematic representation of the photocatalytic activity of MOS-
compared to 78.7 ± 1.3% for Staphylococcus aureus ATCC29213 after AgNPs during degradation of organic dyes is presented in Fig. S2.
24 h of exposure of 200 µl of MOS-AgNPs (100 µg/ml). Fig. S1 shows a
simple illustration of a possible antibacterial mechanism of MOS-AgNPs.
3.5. Recyclability of MOS-AgNPs

Although the above results confirm that the MOS-AgNPs have

Table 1
Comparison of antimicrobial activity of various AgNPs with prepared MOS-AgNPs.
Plant and its parts Nanoparticle Size (nm) Test pathogens Zone of inhibition (mm) References

Withania somnifera leaves AgNPs 5–30 S. aureus 11 [49]


Zingiber officinale rhizome AgNPs 10–20 S. aureus 6.5 [62]
Chitosan/Alginate AgNPs 10 E. coli N/A [43]
Cannabis sativa leaf AgNPs 26–29 Bacillus subtilis 22.8 [9]
Curcuma longa L. AgNPs 15–40 P. aeruginosa 15 [30]
Moringa oleifera seed AgNPs 4 E. coli 28.6 Present study
Moringa oleifera seed AgNPs 4 P. aeruginosa 22.8 Present study
Moringa oleifera seed AgNPs 4 S. aureus 13.5 Present study
Moringa oleifera seed AgNPs 4 Salmonella enterica typhimurium 29 Present study

*N/A = Not applicable.

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H.M. Mehwish et al. Journal of Environmental Chemical Engineering 9 (2021) 105290

Table 2
Percentage (%) inhibition of tested bacterial strains treated with Moringa oleifera extract and synthesized AgNPs (n = 3 ± SDa).
Samples Incubation time (h) Inhibition of tested bacterial strains (%)

S. aureus ATCC29213 E. coli ATCC25922 P. aeruginosa ATCC27853 Salmonella typhimurium

Extract 12 N/A 2.5 ± 0.5a N/A N/A


24 N/A 3.0 ± 0.8a N/A N/A
AgNPs 12 59.2 ± 0.7a 75.3 ± 0.9a 69.9 ± 0.7a 65 ± 0.8a
24 78.7 ± 1.3a 98.2 ± 0.9a 89.3 ± 1.0a 92 ± 1.5a
a
SD = Standard deviation, N/A = No activity, AgNPs = Synthesized silver nanoparticles.

Fig. 6. Photocatalytic activity of MOS-AgNPs Degradation rate of MB by solar light (A–C) and Degradation rate of CR by solar light (D–F).

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H.M. Mehwish et al. Journal of Environmental Chemical Engineering 9 (2021) 105290

Fig. 7. Absorption spectra of 4-nitrophenol and 4-NP + NaBH4 in aqueous medium (A); absorption spectra for reduction of 4-NP + NaBH4 in aqueous medium in
presence of AgNPs (B); degradation (%) of 4-nitrophenol by AgNPs (C); and reaction kinetic for reduction of 4-NP + NaBH4 in aqueous medium in presence of MOS-
AgNPs (D).

treatment with MOS-AgNPs for 4-NP, which indicates high stability;


Table 3
however, slight reductions were observed for both MB and CR.
Comparison of catalytic activity of various AgNPs with prepared MOS-AgNPs.
Catalysts Pollutants Degradation Time pH References 3.6. Microbial contaminated wastewater disinfection
(%) (min)

AgNPs Rose Bengal 92 100 N/ [8] Control wastewater without MOS-AgNPs showed initial contamina­
A
tion by E. coli and Salmonella typhimurium at approximately 55 cfu/ml.
AgNPs Acid Orange- 40 150 N/ [27]
8 dye A After treatment with MOS-AgNPs for 1 h, the concentration decreased
Ag/ZnO AY99 80 80 6 [15] by 87.4% for E. coli and 95.6% for Salmonella typhimurium, which in­
NPs dicates that they have excellent potential for use in wastewater disin­
MOS- Methylene 91 75 9 Present fection. These results were consistent with or better than previously
AgNPs blue study
MOS- Orange red 86 75 9 Present
reported results that showed microbial contaminated wastewater after
AgNPs study treated with AgNPs for 90 min a reduction of 98.5% occurred in initial
MOS- 4-NP 89 75 9 Present bacterial concentration [29] (Table 3).
AgNPs study

*N/A = Not applicable. 3.7. Wastewater treatment with MOS-AgNPs

excellent photocatalytic activity, it is important to study their recycla­ As shown in Table 4, the control wastewater sample showed higher
bility. Therefore, the MOS-AgNPs were separated and reapplied to MB, pH, turbidity, TDS, TSS, EC, COD, hardness, sulfate and chloride levels
CR, and 4-NP degradation to determine their recycling efficiency. As than the MOS-AgNPs treated wastewater sample. The results of the
shown in Fig. 8, no significant loss was observed after 10 cycles of present study revealed that wastewater treatment with MOS-AgNPs
significantly reduced pH (10.1–7.9), EC values (11.8–3.7 mg/l), TSS

9
H.M. Mehwish et al. Journal of Environmental Chemical Engineering 9 (2021) 105290

Fig. 8. Reusability of MOS-AgNPs for reduction of 4-NP, MB, and CR.

values (2508–645 mg/l), COD values (865–252 mg/l), sulfate content nanoparticles. This finding is similar to the results of a recent study
(675–211 mg/l) and chloride content (2517–1061 mg/l). The waste­ involving green synthesis of silver nanoparticles using Moringa oleifera
water sample treated with ferric chloride showed decreased pH, EC, TSS, flowers that showed the maximum absorption to be 429 nm [7].
COD, chloride and sulfate values relative to the control group [3]. The size and shape of MOS-AgNPs was measured by SEM and TEM
Furthermore, MgO nanoparticle treated wastewater showed signifi­ analysis, respectively. The results confirmed that MOS-AgNPs were
cantly COD and total organic carbon values as compared with the con­ spherical in shape with an average size of 4.0 nm. The mechanism of
trol group [10]. MOS-AgNPs was confirmed by FTIR analysis, which showed slight shifts
in absorption peaks indicating the formation of nanoparticles. Taken
3.8. MOS-AgNPs Pb+2 removal efficiency in contaminated water together, these findings indicate that polysaccharides, phenols, proteins,
and amides are the main groups responsible for reduction, capping and
AgNPs have also been used to treat water contaminated with Zn+2, stabilizing MOS-AgNPs [47].
Cd , Ag+, Pb+2, and Hg+2 [65]. In the present study, MOS-AgNPs were
+2 In present study, the antimicrobial activity of synthesized MOS-
evaluated for the ability to remove Pb+2 from water. The results showed AgNPs against pathogenic bacterial species (Gram negative E. coli
that precipitate was formed within 30 min of mixing MOS-AgNPs with strain ATCC25922, Pseudomonas aeruginosa strain ATCC25853, and
Pb+2 solutions, and Pb+2 concentrations in supernatant significantly Gram positive Staphylococcus aureus strain ATCC29213) was explored.
decreased. In short, our results demonstrated that MOS-AgNPs promoted The results showed that MOS-AgNPs exhibited better antimicrobial ac­
the removal of Pb+2 from water (Fig. 9). tivity against Gram negative bacteria than Gram positive bacteria. A
recent study revealed that AgNPs synthesized from Moringa oleifera
4. Discussion leaves exhibited no antimicrobial activity against K. pneumoniae [44].
Furthermore, it has been reported that AgNPs produced using Moringa
In the present study, a green and efficient method for biosynthesis of oleifera seeds have the potential to control the dengue vector Aedes
silver nanoparticles was developed using Moringa oleifera seed extract. aegypti [55]. The binding of AgNPs to bacterial cell membranes depends
The developed method has numerous significant advantages over con­ on the available surface area to volume ratio of AgNPs. Because smaller
ventional methods. Specifically, the synthesis of AgNPs using Moringa AgNPs have a larger surface area, they provide a higher bactericidal
oleifera seed extract does not produce any toxic substances [34]. Addi­ effect [41]. The MOS-AgNPs developed in the present study were found
tionally, there was an increase in color intensity when the incubation to have an average size of 4.0 nm, which resulted in their having a
time was increased from 0 to 2 h from yellow to black because of surface relatively high antimicrobial activity against pathogenic microbes. It is
plasmon resonance phenomena [12], after which no further color also possible that MOS-AgNPs penetrate the bacteria cells and then
changes were observed. These findings indicate the stability of bio­ interact with macromolecules within the cells [26].
synthesized MOS-AgNPs, as previously described [60], suggesting the In the present study, the catalytic activity of synthesized MOS-AgNPs
availability of various plant molecules that reduce silver ions. In this applied to sunlight mediated photocatalytic degradation of organic dyes
study, the MOS-AgNPs surface plasmon resonance band remained close was also explored. The results indicated that the synthesized MOS-
to 421 nm, which further confirmed the formation and stability of AgNPs effectively catalyzed the decolorization of organic dyes in the

10
H.M. Mehwish et al. Journal of Environmental Chemical Engineering 9 (2021) 105290

1061 ± 0.5a
T
Chloride (mg/l)

2517 ± 0.6a
C

211 ± 0.5a
Sulphate (mg/l)

675 ± 0.5a
C

252 ± 1.1a

Fig. 9. Pb+2 removal reduction rate of MOS-AgNPs from polluted water.


T

presence of sunlight. This degradation process takes place on the surface


COD (mg/l)

of MOS-AgNPs, which act as a catalyst. The average particle sized


865 ± 0.6a

measured by TEM was 9.4 nm, so smaller size MOS-AgNPs have greater
surface area; hence, the greater surface area of MOS-AgNPs offers more
C

available active sites where the degradation of organic dyes (MB and
CR) takes place via electron transfer (Fig. S1). Since, the prepared MOS-
3.7 ± 1.6a

AgNPs has half-filled 5s orbital. Upon sunlight exposure, the electron


gets excited and moves from the valance band of silver to an empty
T

conduction band, resulting in the formation of electron hole pairs, which


were considered good oxidizing and reducing agents. Upon reacting
11.8 ± 1.3a
EC (dS/m)

with oxygen, the excited electron produces free radicals at the surface of
the catalyst (MOS-AgNPs) active site, which leads to the degradation of
C

organic dyes [17]. However, trapping of the electron by Ag is only


temporary because of its stable 4d10 configuration; thus, it can easily
645 ± 0.6a

transfer the trapped electron to reacting species that can participate in


the reaction, thereby enhancing the photocatalytic activity.
T

5. Conclusions
2508 ± 0.5a
TSS (mg/l)

Treatment of industrial wastewater, especially that produced by the


textile dying process, is a large environmental challenge. Silver nano­
C

SD = Standard deviation; C = control sample and T = AgNPs treated sample.

materials have the potential for use in the treatment of industrial


1143 ± 1.1a

wastewater because of their activities in catalytic degradation of organic


Various parameter comparisons of control and AgNPs treated wastewater.

dyes and microbes. In the present study, stable silver nanoparticles were
produced using MOS via a simple, economic, and green synthesis route.
T

Specifically, AgNPs with an average size of 9.4 nm were synthesized


using MOS and silver nitrate solution via reduction of Ag+ ion and sta­
3540 ± 1.3a
TDS (mg/l)

bilization of silver nanoparticles by MOS secondary metabolites. The


MOS phytochemicals served as reductants and capping agents for the
C

silver nanoparticles. The generated MOS-AgNPs exhibited good anti­


bacterial activity against both Gram positive and negative bacteria
41 ± 0.9a

commonly found in wastewater. Moreover, the MOS-AgNPs showed


excellent photocatalytic activity toward the reduction of Congo red, 4-
Turbidity (NTU)

nitrophenol and methylene blue organic dyes because of their smaller


size and dispersity. Furthermore, MOS-AgNPs displayed excellent po­
85 ± 0.5a

tential for Pb removal from polluted water. The current findings suggest
that MOS-AgNPs have great potential for use in water treatment without
C

producing toxic or hazardous chemicals. However, further studies at the


7.9 ± 0.6

molecular level and industrial scales are needed to verify its actual
applicability. Additionally, the use of clean water and wastewater to
prepare working solutions and using a control sample in each experi­
T

ment and its comparison with experimental samples will strengthen the
Parameters

10.1 ± 1.5a

findings presented in the present study.


Table 4

pH

11
H.M. Mehwish et al. Journal of Environmental Chemical Engineering 9 (2021) 105290

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Declaration of Competing Interest
photocatalytic activity of ZnO nanoparticles by silver doping for the degradation of
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