Chemical Papers (2022) 76:7313–7325

https://doi.org/10.1007/s11696-022-02392-w

ORIGINAL PAPER

Green synthesis and characterization of silver nanoparticles

using Morinda lucida leaf extract and evaluation of its antioxidant
and antimicrobial activity
Ayomide H. Labulo1 · Oyinade A. David2 · Augustine D. Terna3

Received: 18 May 2022 / Accepted: 22 July 2022 / Published online: 16 August 2022
© Institute of Chemistry, Slovak Academy of Sciences 2022

Abstract
This study emphasizes the production of eco-friendly silver nanoparticles from a medicinal plant extract of Morinda lucida
(M. lucida) and investigated its antioxidant and antimicrobial activity. Phytochemical screening of M. lucida (ML) leave
extract was carried out and observed to contain some fundamental phyto-reducing agents such as reducing sugar, proteins,
and alkaloids. The green synthesized AgNPs (ML-AgNPs) were characterized by UV–vis spectroscopy, Fourier transform
infrared spectroscopy (FTIR), transmission emission microscopy (TEM), scanning electron microscopy (SEM), X-ray dif-
fraction (XRD), and Energy dispersive X-ray analysis (EDX). Thermo gravimetric analysis (TGA) was performed on the
synthesized ML-capped AgNPs to determine the thermal stability and the formation of the green synthesized AgNPs. The
formation of AgNPs was confirmed by the UV–vis absorption spectra, which showed an absorption band at 420 nm. The
morphology of ML extract-mediated AgNPs was mostly spherical and rough-edged crystallite nanostructures, with an aver-
age particle size of 11 nm. The FTIR analyses revealed distinctive functional groups which were directly involved in the
synthesis and stability of AgNPs. The crystallite size was 8.79 nm, with four intense peaks at 2θ angles of 38°, 44°, 64°,
and 77°. At an energy level of 3.4 keV, a significant signal was observed indicating the production of thermally stable and
pure crystallite AgNPs. The antioxidant property of green synthesized ML-AgNPs was determined to be 40% higher than
that of crude M. lucida leaf extract. The ability of green synthesized ML-AgNPs to scavenge free radicals also increased in
the order of ­OH− < NO < ­H2O2. The ML-AgNPs have strong activities with a maximum against P. vulgaris and a minimum
with E. faecalis.

Keywords Morinda lucida · Green synthesis · Silver nanoparticles · Antioxidant · Antimicrobial activity

Introduction (Naikoo et al. 2021), agricultural production (Ndlovu et al.

2020), engineering (Dhanapal 2012), and catalysis (Wu et al.
Nanoscience is a fascinating branch of science concerned 2020; Li et al. 2021; Xu et al. 2021). Nanoparticles (NPs)
with the use and development of structures and materials are small particles with a diameter of 1–100 nm that can be
with nanometer-scale dimensions. Nanotechnology advance- made in a variety of ways. When compared to bulk materi-
ments have benefited scientists in their search for a novel als, NPs are articles with different physical, thermal, and
approach to manufacturing nanomaterials for antimicrobials chemical properties (Khan et al. 2019). NPs can be syn-
thesized using various methods, but some involved the use
of hazardous and poisonous substances that provide a high
* Ayomide H. Labulo risk of toxicity (Khan et al. 2019). Green synthesis is one of
ayomide.labulo@science.fulafia.edu.ng
the most environmentally friendly and low-cost approaches
1
Department of Chemistry, Federal University of Lafia, Lafia, with high reproducibility and yield (Hano and Abbasi 2022).
Nasarawa State, Nigeria Apart from the use of bacteria, fungi, and algae, aqueous
2
Department of Plant Science and Biotechnology, Federal extracts of leaves have been found to reduce silver salt and
University Oye-Ekiti, Oye‑Ekiti, Ekiti State, Nigeria instantly capped the nanosized silver to reduce agglomera-
3
Department of Chemistry, Federal University of Technology, tion (Ansar et al. 2020). Metal nanoparticles, such as silver
PMB 1526, Owerri, Imo State, Nigeria nanoparticles (AgNPs), are appealing due to their diverse

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7314 Chemical Papers (2022) 76:7313–7325

applications in pharmaceuticals (Contreras 2012), including Materials and methods

the treatment of acne and dermatitis, as well as post-surgical
infection (Contreras 2012). Silver, among other metals, has Collection of plant materials and chemical reagents
been proven to be more hazardous to microbes while being
less toxic to mammalian cells (Verma and Stellacci 2010). The medicinal plants used for this study were collected in
AgNPs have found widespread application in the design of February 2020 around Ibadan metropolis, Oyo state Nigeria
multi-resistant medications (Ojemaye et al. 2021) because Latitude 7.3775° N, and Longitude 3.9470° E. The herbal
of their unusual stability, diverse geometries, and ease of plant was identified as M. lucida (Brimstone) (Fig. 1) also
release of silver to impede the growth of bacterial cells known as Oruwo in the Yoruba language and was brought to
by creating reactive hydrogen peroxide. Water treatment the laboratory for preparation. The plant leaves were identi-
(Zhao et al. 2021), agriculture (Khan et al. 2021), biomedi- fied as Morinda lucida at the herbarium of the Department
cal (Akintelu et al. 2021), catalysis (Ajitha et al. 2021), and of Botany and Plant Science, Federal University of Lafia.
sensing (Jiang et al. 2022) are only a few other applications Silver nitrate ­(AgNO3 99.80%) was supplied by Sigma-
of AgNPs that have been reported. In this study, AgNPs Aldrich Chemicals (South Africa).
were synthesized from the extract of the brimstone tree (M.
lucida), a medium-sized tree with crooked, short branches. Preparation of M. lucida leaf aqueous extract
M. lucida plant can be commonly found in Senegal, Nige-
ria, Sudan, Angola, and Zambia (Agyare et al. 2013). It is Plant leaves were thoroughly cleaned with distilled water
sometimes planted around villages, e.g., in Benin (Tran et al. before being dried in the open air. After the leaves had com-
2013). In Nigeria, the stem is popular as a chewing stick pletely dried, they were pulverized with a porcelain mortar
(Agyare et al. 2013). The root, bark, and leaves decoctions and pestle, and the samples were stored in well-labeled air-
and infusions are used as remedies against malaria, diabetes, tight plastic containers (Labulo et al. 2015). 10 g of aqueous
stomach aches, ulcer, leprosy, and different type of fever leaf extracts were weighed and boiled in 600 mL of deion-
(Abbiw 1990). M. lucida has been found to contain high ized water at 100 °C for 20–30 min. The extract was then
content of alkaloids, flavonoids, anthraquinones, and phy- filtered through Whatman 185 m filter paper, yielding a light
tochemicals which are good anti-carcinogenic, anti-inflam- brown colored filtrate (Fig. 2). The extract was kept in the
matory, and antioxidants (Feng et al. 2018). M. lucida was refrigerator between 4 and 10 °C for 10–15 days and utilized
investigated for the first time in this study for the synthesis of for the synthesis of AgNPs.
AgNPs. The synthesized ML-AgNPs were characterized and
we investigated the total antioxidant potency of the aqueous Phytochemicals screening
extract of M. lucida and the green synthesized ML-AgNPs.
Phytochemical screening was carried out using the plant
extracts to determine the presence of the following com-
pounds: phenols and tannins, saponins, triterpenes, flavo-
noids, alkaloids, and steroids according to the procedure
outlined by Hedge et al. (2010).

Fig. 1  Images of the leaves of

M. lucida

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Chemical Papers (2022) 76:7313–7325 7315

Fig. 2  a M. lucida plant extract

b M. lucida plant extract and
silver nitrate after 2 h c M.
lucida-silver nanoparticles (ML-
AgNPs) after 24 h of incubation

Test for phenols and tannins Test for alkaloids

A few drops of 3% F ­ eCl3 were added to a 1 mL solution The alkaloid test was carried out by adding 5 mL of HCl to
of the extract and shaken a little. The presence of phenolic the extract solution. The solution was agitated, filtered, and
compounds and tannins was detected by the appearance of kept for further analysis. Meyer’s test was done afterward
a deep blue coloration formed. by adding 2 mL of the filtrate to 5 mL of Meyer’s reagent.
The formation of a yellow precipitate indicates the presence
of alkaloids.
Test for saponins

A portion of the extract solution for both plants was put Test for steroids
in two different clean test tubes and shaken vigorously,
then left for a few minutes. The presence of saponins was 2 mL of the extract solution was mixed with 1 mL of acetic
detected by the formation of a stable froth in the solution. anhydride and heated for a few minutes then cooled. Few
drops of conc. ­H2SO4 was added by sliding it down the side
of the test tube containing the solution. The presence of ster-
Test for triterpenes oids was indicated by the appearance of a blue coloration.

1 mL of chloroform was added to the extract solutions and

reacted with 1 mL of conc. ­H2SO4 by carefully sliding it Synthesis of silver nanoparticles from plant extracts
down the walls of the test tubes containing the solutions.
The presence of Triterpenes was confirmed by the forma- The synthesis of silver (Ag) nanoparticles was carried out
tion of red coloration in the solution. according to the method described by Dada et al. (2015). In
a typical experiment, 10 mL of the aqueous leaf extract was
measured and placed into a clean 250 mL beaker, where
Test for flavonoids it was reacted with 40 mL of 1 mM ­AgNO3 at room tem-
perature (Fig. 3). The synthesized mixture was allowed to
Lead acetate solution was added to 1 mL of the extract stand for 24 h before being separated by centrifugation at
solutions and the presence of flavonoids was confirmed 4000 rpm for 30 min. The transparent liquid was decanted,
by the formation of a yellow precipitate in the solution. and the settling layer (i.e., nanoparticles) was dried in the

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Fig. 3  Proposed schematic

diagram of the reduction
mechanism of silver nitrate by
the aqueous leaf extract of M.
lucida

oven at 30 °C and stored in a 5 mL plastic sample vial with Anti‑oxidant property

the appropriate labeling. The NPs formed were labeled as
the M. lucida-silver nanoparticles (ML-AgNPs). Cupric reducing antioxidant capacity (CUPRAC) assay

The CUPRAC assay method was utilized with slight adjust-

Characterization ments to evaluate the cupric ions (­ Cu2+) reduction ability of
the extracts, as described by Gulcin et al. (2004). In a test
The Biochrom Libra PCB 1500 UV–vis spectrophotometer tube, 0.25 mL ­CuCl2 solution (0.01 M), 0.25 mL ethanolic
was used to measure the absorbance of the green-produced neocuproine solution (7.5 × ­10–3 M), and 0.25 mL acetate
AgNPs. The absorbance of silver nanoparticles dispersed buffer (1 M) were mixed altogether, then 0.25 mL extracts
in a quartz cuvette with a 1 cm optical path was measured were mixed. With distilled water, the total reaction volume
by removing a small aliquot from the reaction mixture. The was set at 2 mL, and the solution was thoroughly mixed.
intrinsic characteristics of the formed AgNPs were investi- The tubes were sealed and maintained at room temperature
gated using energy dispersive X-ray (EDX), X-ray diffraction for 30 min before being tested for absorbance at 450 nm.
(XRD), scanning electron microscopy (SEM), transmission Increased absorbance indicates greater reduction potential,
electron microscopy (TEM), Fourier-transform infrared which is represented as Trolox equivalent (TEAC) when
spectroscopy (FTIR), and thermogravimetric analysis Trolox is used as the standard.
(TGA). The approximate sizes and shapes of AgNPs were
examined using SEM (TESCAN Vega TS 5136LM SEM Ferrous ion‑chelating ability assay The ferrous ion chelat-
typically at 20 kV at a working distance of 20 nm) and TEM ing (FIC) assay was performed using Singh and Rajini
by dropping an aqueous solution containing the silver nano- method (Singh and Rajini 2004), with significant modifica-
particles onto the carbon-coated grids and drying under an tions. 2 mM F­ eCl2·4H2O and 5 mM ferrozine solutions were
infrared lamp. Micrographs were obtained using a Philips diluted 20 times. In a nutshell, an aliquot (1 mL) of various
Morgagni (M-268) operating at 80 kV. XRD analysis was extract strengths was combined with 1 mL ­FeCl2·0.4H2O.
done by model D8 ADVANCE apparatus (Bruker, USA), The reaction was started by adding ferrozine after 5 min of
and FTIR analysis was performed using a SHIMADZU incubation (1 mL). The mixture was vigorously shaken, and
FTIR model IR8400s spectrophotometer, which provided the absorbance of the solution was measured spectrophoto-
information on functional groups and biomolecules present metrically at 562 nm after another 10 min of incubation.
on the surface of AgNPs during their formation. Thermo- The percentage inhibition of ferrozine—Fe2+ complex
gravimetric analyses for thermal stability of the AgNPs were formation was calculated by using the formula:
performed using TA instrument Q series™ thermal ana- [( )/ ]
lyzer DSC/TGA (Q600). XRD pattern was recorded using % Chelating effect = Acontrol − Asample Acontrol × 100
a graphite monochromatized high-density Cu Kα radiation (1)
(λ = 0.15406 Å) by Rigaku/Dmax RB.

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Chemical Papers (2022) 76:7313–7325 7317

where, Acontrol = absorbance of the control sample (the con- et al. 1982), as reported by Marcocci et al. (Marcocci et al.
trol contains ­FeCl2 and ferrozine, complex formation mol- 1994). Nitric oxide is produced when sodium nitroprus-
ecules) and Asample = absorbance of tested samples. side in an aqueous solution at physiological pH reacts with
oxygen to form nitrite ions, which may be quantified using
the Griess reaction. The reaction mixture, which included
Reductive potential 0.1 mL of oil extract at various concentrations (10, 5, 2.5,
1.25, 0.625, 0.3125 mg/mL) and 0.9 mL of sodium nitro-
The reductive potential of the extract was determined accord- prusside (2.5 mM) in phosphate buffer saline, was incu-
ing to the method of Oyaizu (1986) and as described by Gul- bated for 150 min under illumination. Following incuba-
cin et al. (2004). The extracts (1 mL) were mixed with 1 mL of tion, 0.5 mL of 1% sulphanilamide in 5% phosphoric acid
phosphate buffer (pH 6.6, 0.2 M) and refluxed before adding was added and incubated in the dark for 10 min, followed
2.5 mL of 1% potassium ferricyanide. The reaction mixture by 0.5 mL 0.1% NED (N-1-naphthyl ethylenediamine dihy-
was then incubated for 20 min in a water bath at 50 °C. The drochloride) (Marcocci et al. 1994).
mixture was then treated with 2.5 mL of 10% trichloroacetic
acid and centrifuged for 10 min. The mixture was vortexed
Scavenging of hydrogen peroxide ­(H2O2)
after adding 2.5 mL (100 L) of supernatant, 2.5 mL (distilled
water), and 0.5 mL (20 L) of 0.1% ferric chloride. At 700 nm,
In phosphate-buffered (PBS) saline, a solution of hydrogen
the absorbance was measured. The reaction mixture’s higher
peroxide (20 mM) was prepared at pH 7.4. 1 mL extracts or
absorbance suggests a stronger reductive potential.
standards in methanol were added to 2 mL hydrogen perox-
The reductive potential was expressed as a reductive
ide solutions in PBS at various concentrations. After 10 min,
potential index (REI) as shown below;
the absorbance was measured at 230 nm in comparison with
REI = Absorbance of extract a blank solution containing extracts in PBS without hydro-
(2) gen peroxide (Jayaprakasha et al. 2004).
∕Absorbance of 10 𝜇g/mL vitamin C standard

Antibacterial activity
−
Assay of hydroxyl radical scavenging activity ­(OH )
Antimicrobial sensitivity
According to the method of Halliwell et al. (1987), the
hydroxyl radical scavenging activity was assessed by evalu- The antibacterial potency of ML-AgNPs and plant extract
ating the competition between deoxyribose and the fractions was assessed against Citrobacter, E. coli, P. vulgaris, S.
for hydroxyl radicals generated from the ­Fe3+/ascorbate/ typhi, V. cholerae, and E. faecalis using the disk diffusion
EDTA/H2O2 system. Reaction mixture of 1.0 mL reagent method (Green et al. 1982). The prepared Whatman filter
composed of 3.0 mM deoxyribose, 0.1 mM EDTA, 2 mM paper No.1 discs were soaked in ML-AgNPs and plant
­H2O2, 0.1 mM l-Ascorbic acid, 0.1 mM F ­ eCl3·6H2O in extract solutions and air-dried in a sterile environment. Agar
10 mM phosphate buffer, pH 7.4 were prepared and various plates were prepared by swabbing the bacteria cultures uni-
fraction concentrations (50–350 g/mL) were added to the formly unto individual plates. The plates were divided into
reaction mixture. After 1 h of incubation at 37 °C, 1.0 mL four and placed upon the earlier prepared discs, then plates
of 1% (w/v) TBA (in 0.25 N HCl) and 1.0 mL of 10% (w/v) were incubated at 37 °C for 1–2 days and the zone of inhi-
TCA were added to the reaction solutions. The reaction bition was measured by an ordinary scale (Khanom et al.
mixtures were heated in a boiling water bath for 20 min at 2018).
100 °C, and the pink chromogen (malondialdehyde-(TBA)
adduct) was extracted into 1.0 mL of butan-1-ol, with the
absorbance measured at 532 nm against a blank solution.
The percentage inhibition was calculated using the Results and discussion
expression:
Phytochemical compounds in the plant extract
(3)
( )/
Percentage % inhibition = Abs(control) − Abs(sample) Abs(control) × 100
Table 1 summarizes the phytochemical components found
in the plant extract. Aqueous leaves extract of M. lucida
Inhibition of nitric oxide radical (NO) contained alkaloids, flavonoids, tannins, terpenoids, anth-
raquinones, phlobatanin, carbohydrates, saponins, pro-
The extracts were tested for their ability to inhibit nitric teins, amino acids, and phytosterols while steroids, cardiac
oxide radical activity using the Green et al. method (Green

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Table 1  Qualitative analysis Phytochemicals Pres- UV–visible spectroscopy

of the aqueous extracts of M. ence/
lucida leaf Absence The reduction in A ­ g+ to A­ g0 was confirmed by a color
Alkaloids + change from light brown to a deep brown, indicating the for-
Steroids − mation of nanosilver (Fig. 2a–c). This was further confirmed
Flavonoids + using UV–vis spectrophotometer. The surface plasmon reso-
Tannins + nance (SPR) was observed for ML-AgNPs, with a band peak
Terpenoids + at 420 nm absorption maxima (Fig. 4a). A yield of 56%
Anthraquinone + was calculated after centrifuging at 6000 rpm. According
Phlobatanins + to Aref and Salem (2020), silver nanoparticles produced
Cardiac glycosides − from Cinnamomum camphora had a wavelength of 420 nm.
Reducing sugars − Hamelian et al. (2018) also found a strong and single peak
Carbohydrates + in the UV–vis spectrum of colloidal AgNPs synthesized
Saponins + from Thymus Kotschyanus extract. In a study conducted by
Coumarin − Aldosary and Abd El-Rahman (2019), an absorbance peak
Protein + of 427–437 nm was identified for C. sinensis extract-AgNPs.
Phenols − Phenols, steroids, flavonoids, tannins, steroids, and saponins
Amino acids + are among the metabolites found in M. lucida aqueous leaf
Phytosterols + extract that can reduce A­ g+ ions. The presence of flavonoids
and protein in M. lucida aqueous extract can be linked to the
+ present, − absent plant extracts’ ability to serve as a stabilizing and capping
agent.

glycosides, reducing sugars, coumarin, and phenols were Total phenolic compounds
absent. According to Gutorova et al. (2021), the presence of
both alkaloids and flavonoids aids in the reduction in silver The secondary metabolites present in green plants such as
ions to generate AgNPs. Flavonoids were also found in green phenolic compounds are essential for the reduction in metal
synthesized nanoparticles made from corn silk aqueous ions into their corresponding nanosized metals (Konappa
extract, according to Li et al. (2020). Previous phytochemi- et al. 2021). The concentration of the metabolites can influ-
cal screening on M. lucida revealed that phenols, flavonoids, ence the yield, morphology, and particle size of the NPs. In
and reducing sugars are the major components of plants that Fig. 4b, it was observed that the total phenolic compounds in
have reducing power (Medina-Cruz et al. 2020). Therefore, the extract and the AgNPs increase, with increased concen-
as a result of the presence of both proteins and alkaloids in tration. The leaf extract shows a phenolic content of about
the M. lucida extract used in this study, the green-produced 18 µg/mL at the highest concentration of 160 µg/mL, while
AgNPs should have high antioxidant potency. AgNPs showed a total phenolic content of 10 µg/mL at a

Fig. 4  a UV–visible spectra of green synthesized ML-AgNPs and b concentration of total phenolic compounds

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Fig. 5  a FTIR spectra of M. lucida plant extract and ML-AgNPs b XRD spectra of ML-AgNPs

similar concentration. This confirms that not all the metab- alcohol, phenol, esters, ethers, aldehydes, alkanes, and
olites and the antioxidant compounds are used up for the proteins, which were involved in the synthesis and stabi-
reduction in the Ag salt, but also act as the capping agent lization of AgNPs. Comparing the two spectra, C–O was
(Shoor and Lodise 2006). observed in the ML-AgNPs spectra (Fig. 5a), implying
that the system's stabilization was caused by the carbonyl
FTIR analysis group of the reducing sugars adhering to the silver (Ag)
(Singh et al. 2013).
The functional groups involved in capping and stabilizing
the Ag nanoparticles were identified using FTIR between
4000 and 400 ­cm−1. Figure 5a shows the spectra of the XRD
aqueous extract of M. lucida and ML-AgNPs. The aqueous
extract of M. lucida showed characteristic peaks at 3307, The XRD pattern of ML-AgNPs is shown in Fig. 5b. XRD
2890, and 1649 ­cm−1 which could be attributed to hydroxyl pattern reveals four intense peaks at 38°, 44°, 64°, and
group O–H stretching (Labulo et al. 2015), C–H stretching 77°, which correspond to the face cubic centre (fcc) 111,
vibrations of methyl, methylene, or methoxy groups, and 200, 220, and 311 planes of AgNPs, respectively. The
C–O stretching in the carbonyl group (Shoor and Lodise results are consistent with those of Rahimi-nasrabadi et al.
2006), and the N–H group of amines (Kumar et al. 2019), (2014). The other peaks in the spectra could be attributed
respectively. The broadband peak at 3307 ­cm−1 observed in to metabolite capping the AgNPs.
the ML-AgNPs spectra could be attributed to the overlap- The size of the AgNPs was calculated by the
ping stretching mode of N–H and O–H functional groups Debye–Scherrer equation as follows:
(Kumar et al. 2019).
k𝜆
The C–O band at 1045 ­c m −1 may be assigned to the D= (4)
𝛽 cos 𝜃
polyols (i.e., flavones, terpenoids, and carbohydrates)
present in the plant extract. The various assignments are where D is the crystallite size of the AgNPs, λ is the wave-
consistent with those published in the literature for related length of the X-ray source (1.54056 Å), β is full width at
compounds (Medina-Cruz et al. 2020; Aref and Salem half maximum (FWHM) of the diffraction peak in radian,
2020; Gutorova et al. 2021). These organic compounds k is the Scherrer constant that varies from 0.9 to 1, and θ
were identified as stabilizing groups and capping agents is the Bragg angle in radian (Raja et al. 2017). The XRD
that contributed to the silver ions reduction to metallic pattern analysis of ML-AgNPs gave a crystallite size of the
silver. FTIR measurements indicated peaks corresponding nanoparticles was 8.79 nm.
to different functional groups such as carboxylic acids,

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Fig. 6  a TEM and b SEM

images of ML-AgNPs

TEM 3.4 keV, with weak signals from N, O, C, Si, P, and Cl. The
pure crystalline character of the ML-AgNPs was completely
TEM is a powerful tool in determining the morphologies, composed of silver, as indicated by the predominant emis-
shapes, and sizes of metal NPs. The TEM image of ML- sion energy at 3.4 keV. The findings of Matin et al. (Matin
AgNPs shows the polydispersed-capped NPs (Fig. 6a). The et al. 2017) on silver nanoparticles synthesized from Pega-
obtained NPs are spherical with particle average particle num harmala plant extract via the green approach are con-
size of 11 nm (Fig. 7a) similar to the findings of Zangeneh sistent with the findings in this work. Palani et al. (2015)
et al. (2019). It could be observed that the NPs are evenly reported a similar result utilizing synthetic melanoidin bac-
distributed without agglomeration with transparent capped terial extracts and a 3.3 keV energy level for green manu-
edges. This could be a result of the presence of phytosterol factured AgNPs.
and amino acids from the M. lucida aqueous extract. The
SEM image (Fig. 6b) shows a flake-like surface morphology Thermo‑gravimetric analysis (TGA)
of the ML-AgNPs.
The TGA plot of ML-AgNPs is shown in Fig. 8. TGA was
EDX carried out to determine the thermal stability and presence of
oxygen functionality on the capped green synthesized ML-
Figure 7b shows the elemental composition of ML-AgNPs. AgNPs (Nityanada and Kaushik 2012). TGA thermogram of
The synthesized nanosilver showed a strong signal at the AgNPs showed three identical and continuous weight loss

Fig. 7  a Particle size distribution b EDX spectra of ML-AgNPs

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larger surface-area-to-volume ratio of ML-AgNPs, which

enhances their reactivity towards radicals, can be attributed
to the increased antioxidant activity of ML-AgNPs. Further-
more, the antioxidant mechanism of the plant-mediated silver
nanoparticles, ML-AgNPs, can be attributed to the preferential
sorption of extract components on the surface of the silver
nanoparticles, AgNPs (Patil Shriniwas et al. 2017; Khandel
et al. 2018). The result obtained is similar to the antioxidant
property of L. betulina-capped AgNPs (Sytu and Camacho
2018). Previous studies have shown that the level of antioxi-
dant capacity is dependent on the plant species and the botani-
cal family (Ranjitham et al. 2013).
Fig. 8  TGA and DTA thermograms of ML-AgNPs ML-AgNPs inhibited 75–90% hydrogen peroxide from
concentrations ranged from 0.25 to 1.0 mg/l while M. lucida
of stages (Fig. 10). The first decomposition stage occurs from extract inhibited 50–68% ­H2O2. The ML-AgNPs had no sig-
room temperature to 160 °C with 1.96% weight loss indicating nificant inhibitory effect on Hydroxyl ions ­(OH−) whereas,
the removal of water containing impurities in the ML-AgNPs. 0.25–1.0 mg/l. ML-AgNPs scavenged 50–60% Nitric oxide
The second stage was observed between the temperatures (NO) compared to the M. lucida extract. Thus the inhibi-
160–559 °C with 4.39% weight loss, attributed to the decom- tory factor of ML-NPs increased along with concentration.
position of the capping organic compounds (such as carbonyl As a result, the average inhibitory capacity ­(IC50) of ML-
and the reducing sugar) agents acting as the core–shell. This AgNPs improved as concentrations increased. The ability of
result is in agreement with the result obtained from FTIR spec- ML-AgNPs to scavenge free radicals increased in the order
tra (Fig. 5a). The third and final step is attributed to the decom- ­OH− < NO < ­H2O2 (Table 3). The ­IC50 values required to
position of ML-AgNPs at elevated temperatures between 560 scavenge NO, OH–, and ­H2O2 were lower in ML-AgNPs
and 800 °C with a weight loss of 2.17%. The percentage purity compared to the M. lucida extract, implying that only small
of the green synthesized ML-AgNPs is estimated to be 91.48% amounts of ML-AgNPs are required to scavenge the free rad-
pure silver. For a one-pot crystalline AgNPs synthesis, Nity- icals, further confirming better antioxidant properties than
ananda and Kaushik (2012) reported a similar range of TGA. M. lucida extract (Fig. 9). The increased antioxidant activity
of ML-AgNPs relative to the raw extract is intriguing since
Antioxidant property antioxidants protect humans from oxidative stress-related
illnesses through chemoprotection (Ulewicz-Magulska and
Antioxidant, metal chelating, and free radical scavenging Wesolowski 2019).
abilities of M. lucida extract and ML‑AgNPs The metal chelating ability of EDTA, ML-AgNPs,
and M. lucida extract ranged from 23–92, 19–65, and
The approaches used to measure antioxidant capacity 2.0–47%, respectively (Fig. 10), and the results revealed
include an electron transfer mechanism (Flieger et al. 2021),
which includes a copper antioxidant capacity reduction test
Table 3  IC50 values of M. lucida and ML-AgNPs on free radicals
(CUPRAC). ML-AgNPs had a total antioxidant capacity that
was 40% higher than the crude M. lucida extract (Table 2). IC50
The CUPRAC and reducing potential of the ML-AgNPs was NO OH− H2O2
two folds greater than the M. lucida extract (Table 2). The
antioxidant property of green synthesized AgNPs as a result ML-AgNPs 0.52 ± 0.09b 1.68 ± 0.18b 0.14 ± 0.0005b
of the plant extract being a natural antioxidant material, hence, M. lucida extract 1.26 ± 0.025a 1.93 ± 0.09a 0.21 ± 0.004a
acted as a reducing and stabilizing agent in the green syn- Values with identical alphabets a and b are not significantly different
thesis, resulting in the surface modification of AgNPs. The at DMRT (p < 0.05)

Table 2  Antioxidant potential TAC​ CUPRAC​ Reductive potential

of M. lucida and ML-AgNPs (AAE/g sample) (mg TE/g sample)

ML-AgNPs 229.89 ± 0.01a 65.62 ± 1.07a 2.07 ± 0.02a

M. lucida extract 158.49 ± 0.001b 28.63 ± 1.03b 0.90 ± 0.005b

Values with identical alphabets a and b are not significantly different at DMRT (p < 0.05)

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Fig. 10  Metal chelating abilities of M. lucida and ML-AgNPs

Fig. 9  Free radical scavenging potential of M. lucida extract and ML-
AgNPs
adequate antioxidant properties. Ponmani et al. (2020),
obtained comparable results to those reported in this
investigation.
that ML-AgNPs had a stronger chelating impact than
the crude plant extract and EDTA. In aqueous extracts
of M. lucida, AgNPs of M. lucida, and EDTA, regres- Anti‑microbial activity
sion coefficient (R 2 ) values of 0.8592, 0.9488, and
0.8267 were observed, indicating that ML-AgNPs are Anti‑microbial screening
capable of dispersing in solutions with high surface
area to volume ratio while providing active sites for The antimicrobial potency of both M. lucida extract and
the interaction between the metal ions and the chelate ML-AgNPs was examined against E. coli, S. typhi, V.
ML-AgNPs. The metal chelating action intensified cholera, P. vulgaris, E. faecalis, and Citrobacter using the
as the concentration of ML-AgNPs increased. When disc diffusion method (Fig. 11). The results showed the
evaluated using two-way ANOVA, all of the values extent of susceptibility of the examined organisms. The
were found to be significant at p < 0.05, as shown in organisms showed varied zone of inhibition against the M.
Table 3. Overall, the findings show that green-produced lucida extract and ML-AgNPs. The ML-AgNPs showed
ML-AgNPs exhibit significant metal chelating activity the maximum zone of inhibition of 43 nm against the P.
toward metal ions (Fig. 10). The green synthesis of sil- vulgaris compared to 11 nm obtained for M. lucida extract
ver nanoparticles with S. wightii resulted in AgNPs with (Table 4). The minimum inhibition of 16 and 9 nm was

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Fig. 11  Antimicrobial screening of a E. coli b S. typhi c V. cholerae d P. vulgaris e E. faecalis f Citrobacter

Table 4  The antibacterial potency and zone of inhibition of M. lucida Conclusions

extract and ML-AgNPs against bacteria
S/N Bacteria Zone of inhibition (nm) In this study, ML-AgNPs were synthesized using the eco-
M. lucida extract ML-AgNPs
friendly greener approach. The aqueous extract of the M.
lucida was employed not only as the reducing but also as a
1 E. coli 10 ± 0.95 26 ± 0.10 capping agent and antioxidant. The color change, UV–vis
2 P. vulgaris 11 ± 0.42 43 ± 0.64 adsorption and EDX confirm the reduction in ­Ag+ to ­Ag0.
3 S. typhi 13 ± 1.05 31 ± 0.59 The TEM showed that the ML-AgNPs’ average particle size
4 V. cholerae 8 ± 1.53 29 ± 0.68 was 11 nm and was spherical. The FTIR results are in agree-
5 E. faecalis 10 ± 3.01 18 ± 1.12 ment with the total phenolic compound analysis. The ML-
6 Citrobacter 9 ± 1.48 16 ± 6.23 AgNPs are stable and have good antioxidant and antimicro-
bial activities. The green synthesized ML-AgNPs approach
is quick, cost-effective, environmentally benign, non-toxic,
obtained for ML-AgNPs and M. lucida extract, respec- and suited for large-scale manufacturing. However, more
tively, against Citrobacter. Additionally, the high suscep- research is needed to show various biological properties
tibility of ML-AgNPs against these organisms indicated (such as antifungal, antidiabetic, anti-inflammatory, and
that the synthesized ML-AgNPs could be a valuable can- cytotoxic potential) as well as their mechanism of action.
didate for the treatment of infectious diseases especially
as it has been reported that AgNPs have proved effective Acknowledgements The authors acknowledge the contribution of
against SARS-CoV-2 or the new variant COVID-19 that Nwodo Anastasia and the entire members of the Nanotechnology
Research Group of the Federal University of Lafia, Nigeria.
have claimed over 1.8 million lives (Almanza-Reyes 2021).
Thus, exploiting the use of readily available green plants
Declarations
for the synthesis of nanoparticles could create wealth and
proffer solutions to problems ravaging mankind in the Conflict of interest The authors declare no conflict of interest.
health sector.

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7324 Chemical Papers (2022) 76:7313–7325

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