Investigation of Antioxidant for Polyvinylpyrrolidone-Silver ...xajzkjdx.cn/gallery/20-april2020.pdf · Investigation of Antioxidant for Polyvinylpyrrolidone-Silver Nanocomposite

Investigation of Antioxidant for Polyvinylpyrrolidone-Silver

Nanocomposite Using Pandanus atrocarpus Extract

Abdullah Hasan Jabbar 1*, Hayder Shkhair Obayes AL-Janabi 2, Maytham Qabel

Hamzah3, Salim Oudah Mezan 4, Alaa Nahad Tumah 5, Amira Saryati Binti Ameruddin 6,

Mohd Arif Agam 7*

1-7 Department of Physics and Chemistry, Faculty of Applied Sciences and Technology,

University Tun Hussain Onn Malaysia, Pagoh Campus, Jalan Pancor, 84600 Pancor, Johor,

Malaysia

1 Al-Hussein Teaching Hospital, Directorate of Al-Muthanna Health, Ministry of Health,

Republic of Iraq

2 Department of Biotechnology, Al-Qasim Green University, Iraq

3,4 Directorate of Education Al-Muthanna, Ministry of Education, Republic of Iraq.

Abstract

Nanocomposites are characterized as a multiphase material where one of the phases has a

dimension in the nanoscale. There has been huge enthusiasm for the commercialization of

nanocomposites for an assortment of uses including medicinal, electronic, and basic. The general

motivation behind this study was on the development of silver nanoparticles, due to the present

enthusiasm encompassing these metals due to their exceptional properties which are not quite the

same as the relating bulk material. A novel, simple, cost-effective, nontoxic, and environmentally

friendly technique was developed for synthesizing silver nanoparticle- (AgNP-)

Polyvinylpyrrolidone (PVP) nanocomposite using Pandanus atrocarpus aqueous extract. UV-

visible spectroscopic analysis was carried out to assess the formulation of AgNPs. The average

size of green AgNPs was about 40 nm. Images of spherical green nanoparticles were characterized

using field emission scanning electron microscopy (FESEM). The resultant green AgNPs were

added slowly to polymer (PVP) solution. The AgNPs encapsulated within the polymer chains were

characterized by X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR).

Journal of Xi'an University of Architecture & Technology

Volume XII, Issue IV, 2020

Issn No : 1006-7930

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Modification of thermal stabilities of AgNP/PVP nanocomposites was confirmed using UV- vis

(450nm). The green AgNP/PVP nanocomposites showed tested for antioxidant activity by DPPH

and ABTS. Both tests showed good results ABTS 76% and DPPH 73.10%. Thus, the key findings

of the work include the use of a safe and simple nanocomposite, which had marked antioxidant

activity.

Keywords: ABTS; DPPH; Nanocomposites; Polyvinylpyrrolidone; Polymers

Introduction

Preparation of noble metal nanoparticles by reducing metal salts has been extensively studied

(Jamkhande et al., 2019). To stabilize the dispersion of nanoparticles, it is necessary to use

protective agents, such as polymers, surfactants, and chelating agents (Ramanathan & Aqra, 2019).

The metal/polymer nanocomposite is being used as additives in polymer matrix and they are

excellent candidates for various applications (Boomi et al., 2014). In the past, Poly-N-vinyl-2-

pyrrolidone (PVP) a polymeric substance was used in many pharmaceutical applications as it is

biocompatibile with control rate of drug release so as to improve the in vivo pharmacokinetics

(Fahmi et al., 2009). PVP is one of the major additives to increase the solubility and dissolution

rate (i.e. restrain the drug crystallization in solution) of poorly water soluble drugs in

gastrointestinal pharmaceutical preparations and enhance the bioavailability of drugs Chadha et

al., 2006). Nowadays, the PVP polymer has been used as thin films, detoxicants and bio-active

compounds (Li et al., 2010).These nanoparticles have widely been studied with a view to

improving the quality of catalysts (Marson et al., 2018). All living organisms are suffered from the

damage caused by the free-radical oxygen species. Free-radical oxygen species damage cells by

attacking unsaturated fatty acids in the cell membrane. Fortunately, a protective enzyme,

superoxide dismutase, completely converts these free-radical oxygen species into two water

molecules and oxygen (Carneiro et al, 2018).

The enhanced biological properties of PVP doped PVP/AgNPs are to be explored. Therefore in

the present study, PVP- AgNPs have been synthesized using Pandanus atrocarpus extract (PAE).

Water was used as the environment benign reaction medium and plant metabolites as reducing and

capping agent, making the process greener one. Interaction between metal nanoparticles and PVP

is not much documented and properties that nanoparticles may acquire as a result of these



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interactions are even less reported. Antioxidants may be defined as any substance that when

present at low concentrations compared with those of an oxidizable substrate, significantly delays

or prevents oxidation of that substrate in a chain reaction (Moharram & Youssef, 2014).

Antioxidants have become a popular research topic because they cannot be generated by the human

body and hence have to be consumed in the diet. Many fruit and vegetables have been found to be

rich sources of antioxidants. Since a large portion of the human diet is based on fruit and

vegetables, it is important to understand the biological and biochemical interactions between these

dietary antioxidants and living systems.

Materials and Methods

Preparation of Pandanus atrocarpus aqueous extract

Pandanus atrocarpus (PA) plant was supplied by Ethno Resources Sdn. Bhd. (Selangor,

Malaysia). The plant sample was stored at 4 C until further use. The protocol described by Awad

et al. (2019) was used for the preparation of aqueous extract. In brief, 2 g of PE whole plant powder

was boiled in 100 mL distilled water. The filtered extract was concentrated to dryness in a hot air

oven at 40 ℃ for 48 hours to obtain a dark brown semisolid mass which was weighed and labelled

as PAE. The MCE was stored at 4 ℃ and used for further experiments.

Synthesis of PA-AgNPs

For the synthesis of the silver nanoparticles, the PAE (0.8) ml was added to the AgNO3 solution

and the volume was adjusted to 10 ml with de-ionized water. The final concentration of Ag+ was

1 × 10−3 M. The solution was stirred for 2 min. The reduction process Ag+ to Ag0 nanoparticles

was followed by the color change of the solution from yellow to brownish-yellow to deep brown

depending on parameters studied such as the extract concentration, temperature and pH. The

nanoparticles were prepared at different pH values, the pH of the solutions was adjusted using 0.1

N H3PO4 or 0.1 N NaOH solutions (Khalil et al., 2014).

Synthesis of PVP/Ag Nanocomposite

PVP-Ag NCs synthesized by the method of (Awad et al. 2015) with slightly modification. The

PVP dissolved in distilled water to prepare a 20 % stock solution. The synthesized PAE-AgNPs



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(Previous section) mixed with of PVP (40%) in different ratios (1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2

and 9:1) The resulting reaction mixture monitored for change in color visually as well as by

recording UV-vis spectra at regular intervals within the wavelength range of 300 to 700 nm. The

PVP-Ag nanocomposite, further characterized.

Characterization of PVP-AgNCs

The synthesis of PVP-AgNCs was confirmed by periodically scanning the absorption maxima of

aliquots (1.0 mL) of the reaction solution in1-cm path-length disposable cuvettes. The UV–

visspectrawere recorded every 30 minutes within the wavelength of 300 to 700 nm (Ahmed et al.,

2016). The possible biomolecules of PAE responsible for bioreduction of silver ions were

identified by recording the Fourier transform infrared (FTIR) spectrum of AgNPs using Perkin

Elmer Spectrum FTIR in the range of 4000–450 cm-1 at a resolution of 4 cm-1 (Ovais et al., 2016).

The powder AgNP sample was used for X-ray diffraction measurements performed on Shimadzu

XRD 6000 diffractometer operating at 40 kV in the region of 2θ from 30ᵒ to 80ᵒ at a speed of

0.02ᵒ/min (Ali et al., 2016). For size and shape analysis of PVP-AgNPs, the field emission

scanning electron microscope (FESEM) images were captured on JEOL JSM-7600F at an

accelerating voltage of 10 keV. The size analysis from FESEM images was performed on Image J

1.52g software (Mohseni et al., 2019).

Antioxidant Activity (ABTS free radical scavenging activity)

The ABTS free radical scavenging assay was performed by following the procedure as described

by Garcia-Leis et al. (2016). ABTS reagent was dissolved in water to a 7 mM concentration, the

ABTS stock solution was reacted with 2.45 mM potassium per sulfate (final concentration). The

ABTS•1 solution was diluted with ethanol in such a way that the absorbance was between 0.7-0.8.

The synthesized PVP-Ag NCs solution of different concentrations (100 µL, 200 µL, 300 µL, 400

µL and 500 µL, diluted to a total volume of 1 mL with distilled water) was mixed with 4 mL of

ABTS•1 solution in test tubes. The absorbance was measured at 734 nm exactly after 6 minutes

using UV-vis spectrophotometer (GENESYSTM 10S, Thermo Scientific, USA). ABTS•1 free

radical scavenging activity was expressed as percent inhibition (% I) calculated by the following

formula:



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% I = (C - S

C)×100

Where; C is the absorbance of control and S is the absorbance of sample.

All experiments were repeated thrice and the data were reported as means (±SD for triplicates).

Antioxidant Activity (DPPH free radical scavenging activity)

The DPPH free radical scavenging activity of CA-CrA was estimated according to the

procedure described by Rasheed et al. (2017), with slight modification. Briefly, 0.5 mL of 0.1 mM

DPPH (dissolved in ethanol) was mixed with 0.5 mL each of synthesized PVP-Ag NCs solution

of different concentrations (100 µL, 200 µL, 300 µL, 400 µL and 500 µL, diluted to a total volume

of 0.5 mL with distilled water, where needed) in test tubes. The absorbance was recorded at 517

nm by using UV-vis spectrophotometer using UV-vis spectrophotometer (GENESYSTM 10S,

Thermo Scientific, USA). DPPH free radical scavenging activity was expressed as percent

inhibition (% I) calculated by the following formula:

% I = (C - S

C)×100

Where; C is the absorbance of control and S is the absorbance of sample.

All experiments were repeated thrice and the data were reported as means (±SD for triplicates).

Results and Discussion

Uv-Vis (Ultraviolet–visible Spectroscopy)

The formation of PVP-AgNC was confirmed by UV–vis spectral study. The AgNPs was first

checked for change in color from colorless to brown and UV-Vis spectroscopy. The change in

color showed the formation of AgNPs because of the reduction of silver metal particles Ag+ to

nanoparticles Ag0. This color refers to the excitation of SPR. As shown in Figure 1, characteristic

SPR band for AgNPs and PVP-AgNCs was observed at around 432 nm and 420 nm respectively

(Edison & Sethuraman, 2017).



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Figure 1: UV-VIS absorbance spectra for various weight-of (wt %) AgNO3 and different concentrations

Fourier Transformed Infrared Spectrometry (FTIR)

The prominent peaks present in PAE are located at 3338, 2131, 1638, 1295, and 428 as shown in

Fig. 2. It may be inferred from that peak shifts after PVP-Ag NCs synthesis by varying the

concentration of PAE do not considerably differ from each other as all the five spectra very closely

overlap each other. In all of the PVP-Ag NCs samples, the same two peak shifts after Ag NCs

synthesis were observed as: 3338 to 3325/3326 cm-1 and 1638 to 1634/1635 cm-1, as recorded for

Ag NPs samples 1 to 5. The peak at 3338 belongs to the –OH group which, in this case, can be

attributed to the phenolic and flavonoid compounds of PAE. Second important stretching vibration

is recorded from 1638 to 1634/1635 cm-1 which, although may be considered as amide groupf of

proteins as well as benzene ring containing aromatic compounds, it may also be taken into account

for phenolic and flavonoid compounds as they exhibit strong vibration on this wavenumber

(Shivakumar et al., 2014; Prathna et al., 2011). In the context of all these stretching vibrations

found in the FTIR spectra of PVP-Ag NCs, it may be assumed that they have played a role in

fastening the rate of Ag NCs synthesis reaction leading to the nucleation along with stalling the

secondary growth of the nuclei.

0.00.20.40.60.81.01.21.41.61.82.0

300 400 500 600 700

Ab

sorb

ance

Wavelength (nm)

30 min 60 min 90 min 2 h 3 h



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Figure 2: Fourier Transformed Infrared Spectrometry (FTIR) spectrum of conductive film.

Field Emission Scanning Electron Microscopy (FESEM)

The particle size and morphology analysis was performed by scanning electron microscope (SEM).

The SEM micrographs of the AgNPs and PVP-AgNCs are shown at the magnification of 200,000×

in figure 3. The shape of the PVP-AgNCs is almost spherical and the smallest particle size is 27.34

nm. The function of PVP in the PVP-AgNCs nanocomposites is not only as a binder, but it also

prevents the process of agglomeration of Ag nanoparticles and limits the diameter of the

nanoparticles formed (Zhang et al., 2011; Flahaut et al., 2000). As for the morphology of particles

in nanocomposite, it is evident that the particles obtained in this study are mostly spherical in

shape.



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Figure 3. FESEM image of PVP/AgNCs

X-Ray Diffraction Analysis (XRD)

The XRD analysis of PVP/AgNC was carried out to confirm the crystalline nature of the

synthesized nanomaterials. Figure 4 show the XRD spectrum of PVP/AgNCs, respectively. Figure

4 show the XRD spectrum of MCE-AgNPs and PVP/AgNC, respectively. The X-ray

diffractograms, at 2θ from 20°-80°, clearly indicate the crystallinity of the tested samples as

indicated by the broadening of Bragg’s peaks. The presence of sharp peaks indicates the bioactive

compounds present on the surface of the NPs (Shabestarian et al., 2017). The Bragg’s reflection

values of 38°, 44°, 64° and 77°, obtained for MCE-AgNPs and PVP/AgNCs, corresponding to the

set of lattice planes, that is, (111), (200), (220) and (311) have been obtained at 2θ indicating the

formation of face centered cubic (fcc) crystalline structure of silver nanostructure. The peak related

to the lattice plane (111) is the most intense suggesting its most predominant orientation in addition

to the crystalline nature of the synthesized silver colloids. The XRD spectrum did not show any

other crystallographic peak confirming the high purity of synthesized Ag colloids. Similar results

have been reported recently for the synthesis of AgNPs-mediated nanocomposite (Awad et al.,

2015).



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Table 1. XRD Peak list of PVP/AgNCs

Pos. [°2Th.] Height [cts] FWHM Left [°2Th.] d-spacing [Å] Rel. Int. [%]

38.0589 4981.03 0.4428 2.36445 100.00

44.2253 1324.48 0.4723 2.04803 26.59

64.5287 1127.84 0.2066 1.44417 22.64

77.4782 1191.13 0.4723 1.23197 23.91

20 30 40 50 60 70 80

0

1000

2000

3000

4000

5000

6000

(311

)

(220

)

(200

)

(111

)

Inte

nsi

ty

2 Theta

PVP- AgNCs

Figure 4. XRD pattern of PVP/AgNCs

Antioxidant activity of PAE-AgNPs and PVP/AgNCs

Many research reports indicate that there is an intimate relationship between oxidative stress and

age-related neurodegenerative disorders. In this context, many studies have been performed to

examine the beneficial effects of antioxidants aiming at reducing or blocking the neuronal death

which is usually a pathophysiological consequence in such diseases (Ramassamy, 2006). As the

radical scavenging capacity of a compound is usually attributed to its antioxidant potential, two

antioxidant activity assays of as-synthesized PAE-AgNPs and PVP/AgNCs were performed for



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the evaluation of their antioxidant potential, that is, their capacity to inhibit ABTS and DPPH free

radicals.

ABTS Free Radical Scavenging Activity

The dose-response bar chart of ABTS scavenging assay of as-synthesized PAE and PVP/AgNC

are shown in Fig. 5. It can be clearly observed that all samples and PAE have good antioxidant

potential with effective ABTS radical inhibitory activity in a dose-dependent manner. At 500 µL

sample, the ABTS radical scavenging activity (% I) of PAE was recorded at 58.20 %, and

PVP/AgNC at 76.00 % (Table 2). The antioxidant activity of PVP/AgNC was higher than PAE.

However, the maximum ABTS radical scavenging activity was shown by ascorbic acid recorded

at 81.40 %.

This result is in conformity to the previous reports in which MNPs were reported to have

higher antioxidant activity as compared to the extract (Gomaa, 2017; He et al., 2017; Khan et al.,

2016). The antioxidant activity of ascorbic acid was higher than that of the other samples as shown

in Table 2.

Figure 5. ABTS free radical scavenging assay of PVP/AgNCs and PAE. Error bars represent 95%

confidence interval (n=3).

0.0

10.0

20.0

30.0

40.0

50.0

60.0

70.0

80.0

90.0

100 200 300 400 500

AB

TS %

I

Sample Concentration (µL)

PVP-AgNC

Extract

Ascorbic acid



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Table 2. ABTS free radical scavenging assay of PVP/AgNCs and PAE

Sample Concentration (µL) PA-AgNPs±SD PAE±SD Ascorbic acid±SD

100 21.4±1.35 12.0±1.19 41.7±1.25

200 34.6±1.36 21.8±1.26 49.9±0.94

300 48.4±1.54 34.9±1.20 60.9±1.10

400 59.0±1.13 48.9±1.73 71.2±1.40

500 64.2±1.95 58.7±0.85 81.9±1.85

DPPH Free Radical Scavenging Potential

The DPPH free radical scavenging potential of as-synthesized PVP/AgNCs and PAE show that

they have the hydrogen donating ability and hence they may serve as free radical inhibitors or

scavengers, acting possibly as potent antioxidants. The dose-response bar chart of DPPH free

radical scavenging assay of as-synthesize, PVP-AgNCs and PAE have been shown in Fig. 4.9. It

can be clearly observed that both the silver colloids and PAE have good antioxidant potential with

DPPH radical inhibitory activity in a dose-dependent manner. At 500 µL sample, the DPPH radical

scavenging activity (% I) of PVP/AgNCs recorded at 73.10 % and PAE at 58.70 % as shown in

Fig. 6. However, it is noteworthy that the antioxidant activity of PVP/ AgNCs is higher than CAP

that may be due to comparatively higher concentration of bioreductants adsorbed onto the surface

(Shabestarian et al., 2017). The antioxidant activity of ascorbic acid was significantly higher than

that of the PVP-AgNCs and PAE as shown in table 3.



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Figure 6. DPPH free radical scavenging assay of PVP/AgNCs and PAE. Error bars represent 95%

confidence interval (n=3).

Table 3. ABTS free radical scavenging assay of PVP/AgNCs and PAE

Concentration (µL) PVP/AgNCs±SD PAE±SD Ascorbic acid±SD

100 28.0±0.84 12.0±1.19 41.7±1.25

200 42.1±0.94 21.8±1.26 49.9±0.94

300 55.5±1.10 34.9±1.20 60.9±1.10

400 66.4±1.16 48.9±1.73 71.2±1.40

500 73.1±0.99 58.7±0.85 81.9±1.85

The electron transferring potential of a compound determines its reducing power and as such it

may be considered in terms of its antioxidant activity (Gooding et al., 2014). The presence of rich

polyphenols and flavonoids content, responsible for the biosynthesis of PAE-AgNPs, has been

reported for its antioxidant capacity (Gray et al., 2018). These antioxidant compounds of flavonoid

content might got adsorbed onto the surfaces of metallic NPs (Wang et al., 2009). Surface reactions

of the synthesized NPs, by virtue of the adsorbed antioxidant moieties onto the NPs surfaces, along

with a high surface area to volume ratio of the NPs, produce a tendency in the NPs to interact and

0.0

10.0

20.0

30.0

40.0

50.0

60.0

70.0

80.0

90.0

100 200 300 400 500

DP

PH

% I

Sample Concentration (µL)

PVP/AgNC

PAE

Ascorbic acid



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scavenge the free radicals (Dauthal & Mukhopadhyay, 2013). Nobel MNPs like AgNPs and GNPs

have been used under in vitro and in vivo conditions for the scavenging of free radicals (Suganthy

et al., 2017). It implies that the findings of this study signify the potential of PAE-AgNPs as

promising antioxidant moieties.

Conclusion

The introduced work demonstrated the fast greener synthesis of AgNPs utilizing TPandanus

atrocarpus and their composite with the PVP polymer. The technique here is nontoxic, ecologically

cordial, and straight forward and involves minimal effort and has no lethal chemicals. The

formation of greener AgNPs was determined by FESEM and UV-Vis spectroscopy, where UV-

vis recorded at 450 nm in the UV-Vis range. FESEM demonstrated the average size of the resulting

nanoparticles to be 40 nm. The nanocomposite was characterized utilizing FTIR spectroscopy and

XRD techniques. The nanocomposite films showed significant antioxidant activity both on ABTS

and DPPH assay. This guarantees a promising potential utilization of the nanocomposite as

antioxidant.

Acknowledgment

This work was funded by the Fundamental Research Grant awarded by the Ministry of Higher

Education (FRGS/1/2019/STG07/UTHM/02/5(FRGS K171).

Conflict of interest: The authors have no conflicts of interest to declare

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