Green Synthesis and High Efficacy Method for Reduced Graphene Oxide by Zataria Multiflora Extract

Journal of Environmental Treatment Techniques

2020, Volume 8, Issue 1, Pages: 488-496

J. Environ. Treat. Tech.

ISSN: 2309-1185

Journal web link: http://www.jett.dormaj.com

Green Synthesis and High Efficacy Method for

Reduced Graphene Oxide by Zataria Multiflora

Extract

Behrang Sabayan , Nooredin Goudarzian *, Mohammad Hossein Moslemin , Razieh

Mohebat ¹

Department of Chemistry, Yazd Branch, Islamic Azad University, Yazd, Iran

Chemistry Department, College of Sciences, Shiraz Branch, Islamic Azad University, Shiraz, Iran

Received: 14/12/2019

Accepted: 27/01/2020

Published: 20/02/2020

Abstract

There are many different methods for producing nanoparticles, but the use of plants due to the low cost and environmentally friendly

nature of nanoparticle synthesis is very much considered. The biological synthesis of reduced graphene oxide by extracts of Zataria

multiflora cultivar is reported in this work. In this study, Zataria multiflora extract was used as a reducing agent for the production of

nano-graphene oxide in high yield and avoids the use of toxic and environmentally hazardous reducing agents commonly used in the

reduction processes of GO. The reduced graphene oxide and optimization of reaction conditions were monitored by UV and analyzed

for determination of size and morphology of reduced graphene oxide with Transmission Electron Microscopy (TEM), Raman spectra,

Fourier Transform-infrared spectroscopy and X-ray diffraction devices (XRD) respectively.

Keyword: Green synthesis, Zataria multiflora, Reduced graphene oxide, Graphene oxide

hardness, and mechanical quality, high electrical and warm

Introduction¹

conductivity, adaptability and attraction. Along these lines,

numerous utilizations will be made, incorporating applications

in nanoelectronic, sun oriented cells, vitality stockpiling

gadgets, for example, batteries and supercapacitors. Graphene

and graphene oxide have high conductivity and are extremely

reasonable for use in circuits and electronic devices. While

graphene oxide has been introduced as the primary component

in all materials discussed thus far, nanocomposites of graphene

oxide use these sheets as a minor filler component fixed within

either a polymer or an inorganic matrix. Delivery their high

oxygen content, graphene oxide layers are the carbon analogs,

whose nanocomposites with polymers have been completely

investigated for a broad range of applications (6, 19). In

contrary, the huge surface to volume ratio of graphene oxide,

in addition to its high ability to dispersion in both water and

non-aqueous solvents and its broad spectrum of reactive

surface-bound functional groups, discourages aggregation,

make it easy for processing in solution and promoting gross

interaction between fillers and polymers. A diversity of

graphene oxide-based nanocomposites has been manufactured

as thin films, although they are usually reduced to graphene for

conductivity surveys.

Graphene and graphene oxide have enormous potential for

broad category of biomedical, catalytic and optical

applications, electronic, bioimaging, etc. due to their excellent

characteristics and their biocompatibility (1-6). Thus, there is a

growing need to develop the processes which are not harmful

to the environment of the graphene synthesis that does not use

toxic chemicals. Recent articles on the reduction of graphene

oxide discuss the substitution of natural reducing agents for

toxic hydrazine4–8and biologically reduction of graphene

oxide by microorganisms and leaves, seed or peels of a plant

extract have been suggested as possible eco-friendly

alternatives to chemical and physical methods (7-11).

The reduction of graphene oxide under specific conditions

(temperature and pressure) are costly and tedious and have

potential dangers for the earth, thus it’s required to have

minimal effort, non-dangerous and non-lethal strategies. One

of the strategies to produce reduced graphene oxide is the green

reduction, which is noticeable and has expanded nowdays (12-

5). Graphite oxide was first created in 1860, in view of known

techniques for Hummer and Hoffman. It is a blend of carbon,

hydrogen and oxygen molecules. Graphite oxide is extremely

hydrophilic and comprises a layered structure of graphene

oxide sheets. The most widely recognized technique for the

preparation of graphite oxide is to utilize at least one-

concentrated acid within the strong oxidizers. Brody utilized

this strategy in 1859 (16-18). Researchers trust that graphene

oxide is a standout amongst the toughest materials at any point

known, because of its C-C bond strengths. Graphene oxide has

specific properties, for example, nanometric size, high

The most common path toward bulk quantities of reduced

graphene Oxide starts with the conversion of graphite to

graphene Oxide [GO]. The first procedures for the synthesis of

GO were done many years ago by Brodie (20), Staudenmeier

(21) and Hummers et al. (16) and persist with only minor

changes. The degree of graphite oxidation, as quantified by the

carbon to oxygen atomic ratio, is dependent on the synthetic

Corresponding author: Nooredin Goudarzian, Chemistry

Department, College of Sciences, Shiraz Branch, Islamic Azad

University, Shiraz, Iran. E-mail: ngoudarzian@iaushiraz.ac.ir.

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2020, Volume 8, Issue 1, Pages: 488-496

method as well as the reaction procedure (22-24). The

Staudenmaier method 20 usually produces the most oxidized

GO, although the oxidation of graphite to GO breaks up the

sp2-hybridized unity of the stacked graphene layers, generating

errors that manifest as clear notches in the stack (18, 25) and

become more the distance between adjacent sheets (23, 24).

This increased spacing is different depending on the amount of

water interacted within the Stacked-sheet structure (25, 26) and

reduces sheets interaction, so helping the delamination of GO

to individual graphene oxide sheets by exposure to low-power

sonication through the water. At a little basic pH, negatively

charged, hydrophilic oxygenated functional groups on the

graphene oxide surface can settle down dispersions of these

layers in water-based media (18, 27). Within shortly of being

available in bulk quantities, graphene oxide, reduced graphene

oxide, and graphene have become highly practical, inexpensive

building blocks for the creation of several advanced

carbonaceous materials, with applications to be restricted only

by the imagination. For example, it is not difficult to imagine

the internalization of different additives into thin layer

composites of graphene and graphene oxide to create excellent

material compositions.

Thus, polymers have already been put together to graphene

papers and films, either in nanoparticle form (9, 28, 29), or by

in situ electro polymerization (1, 18, 22) for stabilizing

mechanically and to facilitate processing. The development of

ways to completely exfoliate graphite powder, in a wide range

of solvents (30, 31), and the without the presence of reducing

or stabilizing agents, will be essential to the generation of high-

quality graphene materials. Improved synthesis and processing

of graphene oxide and graphene being these highly versatile

materials can be achieved by more researchers and engineers,

so ensuring the quick development of many new materials with

amazing properties. For instance, monodisperse colloidal

solutions of single graphene sheets and few-layer graphene

piles have recently been separated from bulk graphene

nanosheet dispersions via density gradient ultracentrifugation

interest in the world as the most successful carbon allotrope for

the growth of next-generation carbon-based materials. On the

other hand, the number of papers that explain graphene and its

related materials (41, 42) has eminently exponentially from

2004 isolation of single graphene by use of mechanical

exfoliation3,42. Graphene can be made from different graphite

derivatives (43), bulk number of completely exfoliated

graphene-like sheets are commonly driven by the reduction of

graphene oxide distribution or powders. Graphene has attracted

significant interest in the scientific community has the most

promising carbon allotrope for the development of next-

generation carbon-based materials. Indeed, the number of peer-

reviewed articles that discuss graphene and its associated

materials (41, 42).

Zataria multiflora as a reducing agentZataria multiflora is a

kind of thyme. Zataria multiflora is the main type of local

Thymus in Iran. It is situated in Shiraz (Mount Sivand,

mountains neglecting the Maharlou Lake, between the

backwoods, among Jahrom and Mansurabad, near Tarom

Darab), Zataria multiflora is contained around 40% thymol, the

primary successful fixing with a specific end goal to restore

coriander and thyme because of the extravagance of this

substance is the best decision for biological recovery (43-45).

There are mixes, for example, bicyclic acid, propionic acid,

acrylic acid, unpredictable lalalum, and germabol. Mixes of the

plant incorporate lanthanide acetic acid derivation, coumarin,

flavonoids, and phytosterols. Studies have demonstrated that

the blend of green graphene oxide reduced by the utilization of

Zataria multiflorais remove is less known in the present

examination. The amalgamation of green-graphene oxide from

Zataria multiflora species was considered (43).

Materials and methods

.1 Preparation of Graphene oxide powder

Graphene oxide prepared by oxidation of regular graphite

powder by Modified Hummers techniqu (1, 16, 23). In the

ordinary strategy for preparation 5 gr graphite and 5 gr of

(1, 32) and used to make thin films that have lower sheet

NaNO

2 4

(sodium nitrate) were added to 200 ml of H SO (in Ice

was added to this

resistance than those made from bulk graphene dispersions.

The third frontier for later development hides in the production

of huge surface graphene thin films for use in electronic and

energy usage. However, to produce the highly conductive and

translucent monolayers necessary for these purposes, new

methods for large-area thin-film preparation must be mixed

with advances in the synthesis of large quantity, spitless

graphene sheets, as discussed in the next paragraph. As

graphene can also be a template for catalytic nanoparticles (33,

and water) and bit by bit 30 gr of KMnO

blend and amid this procedure, temperature kept on 30°C (since

the response is exothermic). The reaction mixture at that point

mixed for 18 hours at room temperature with the goal that thick

glue is shaped. The glue at that point filled 400 ml of distilled

water and following 20 minutes another 1 liter of distilled water

added to the blend. 30 ml of 30% H

the reaction mixture drop insightful to reduce the abundance

KMnO . The arrangement transformed into light dark-colored

in the wake of including H . The solution was expelled in a

2 2

O arrangement added to

5), having high procedures and turnover, the production of

catalytic thin films would afford large surface area

heterogeneous catalysts for utilization in fuel units. In another

application, graphene drop cast from a dispersion has recently

been utilized as a thin support film for TEM images to prepare

unmatched atomic resolution (36). With such decent

applications in one's head, the development of new graphene

oxide and graphene-based materials will lead to many future

developments in science and technology (1, 37). While

graphene particles can be detached from reduced slightly

dissolve of graphene oxide, thermal exfoliation prepares an

alternative way for its preparation directly from GO and

circumvents the usage solvents (1, 18, 25).

This method involves the speedily heating of GO, which

gets complete delaminated via the evolution of carbon dioxide

as surface hydroxyl and epoxy groups decompose (1, 23). With

its fully joined together, firm 2D structure, graphene has both

electrical and thermal ability to conduct (38, 39) and

mechanical possessions (40) that are superior to those of other

carbon allotropes. So, graphene has attracted significant

decanter and centrifuged and after that washed out with

distilled water to achieve the pH of 7 and afterward dried under

vacuum for 24 hours to frame GO powder (15-18).

.2 Preparation of Zataria multiflora leaf extracts

Leaves of the Zataria multiflora plant gathered from

different regions of Shiraz, Fars, Iran, washed independently

with water at that point dried for 7 days at room temperature45.

The plant leaf separates set up by the expansion of 10 g of

altogether washed and finely grounded leaves (utilizing a

residential blender) to 200 ml of deionized (DI) water in a 500

ml Erlenmeyer flask. The mixture boiled for 60 min before

sifted under encompassing conditions. The concentrates put

away at 4 °C and utilized inside seven days (37-46).

..3 Reduction of graphene oxide

In an ordinary methodology for the reduction of graphene

oxide, 50 mL of Zataria multiflora leaf extract added to 200 mL

of a homogeneous scattering of graphene oxide (1 mg/mL). In

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2020, Volume 8, Issue 1, Pages: 488-496

the wake of energetically shaking and sonicated for 20 minutes,

the mixture warmed in an oil shower at 98 °C under reflux for

concentration of the Zataria multiflora extracts expanded to

25%. Thus, 20% of Zataria multiflora extracts utilized for

4 hrs. The subsequent dark scattering centrifuged at 15,000

future tests. Figure 2 demonstrates the adjustment in

rpm for 60 min. The pellet then washed three times with DI

water to evacuate the undesirable materials. Finally, the

lessened graphene oxide (RGO) dried at 70 °C for 24 hrs (46,

absorbance at 600 nm for the reaction between 20% Zataria

multiflora extracts and 0.5 mg/mL of graphene oxide at three

distinct temperatures (25, 30, 45, 60, 75 and 96 °C). As the

temperature expanded from 25 to 96 °C, the level of reduction

likewise expanded of course (53). Both the reaction rate and

7).

values of final absorbance were much higher at 96 C than those

at 25, 30, 45, 60 were and 75 °C.

Results and Discussion

Optimization of reaction conditions for the reduction of

graphene oxide an unmistakable change in the shade of the

suspension on reduction can be an undeniable sign of the

reaction (48-50). The chemical reduction of the yellow-dark

colored colloidal suspension more often than not results in a

dark accelerate, which likely outcomes from an expansion in

the hydrophobicity of the graphene oxide sheets caused by a

decline in polar functionality on the surface of the sheets (49-

.1 Infrared spectra

Graphite powder utilized to make graphene oxide, which

acquired from Merck Inc. The aftereffects of the graphite

infrared spectroscopy test are watched, the graphite contains no

carbonyl and epoxide group, and the adsorption in the locale of

around 3422 cm-1 is identified with the hydroxyl group of the

atom water consumed by graphite (3, 13, 17) (Figure 3).

Graphene oxide arranged by changed Hummer strategy at

that point refined. The FT-IR spectroscopy of graphene oxide

demonstrated an absorption frequency of 1720 cm-1relative to

the tensile vibrations of the carbonyl group (C = O) and

additionally strong absorption in the 3421 cm-1 of the hydroxyl

group, (OH) 1384 cm group of the flexural vibrations of the

hydroxyl group (OH) and absorption in the 1031 cm-1 locale

of the epoxide tensile frequencies (Figure 4). Likewise, the

pinnacle contained in 1621 cm is identified with the functional

groups (C = C) staying on graphene plates, which have not

experienced any progressions amid the oxidation procedure (3,

1). Consequently, the reduction of graphene oxide on

introduction to Zataria multiflora extricates checked by an

adjustment in shading and an expansion in absorbance at 600

nm (optimized after scan) and furthermore it tends to be

observed by optical thickness estimation at 600 nm (4, 9, 53,

0). Serial dilution checked the straight connection between

absorbance at 600 nm (52) and the concentration of graphene

suspension. We examined the impact of Zataria multiflora

concentrates of various fixations on the reduction of graphene

oxide. Figure 1 demonstrates the adjustment in absorbance at

00 nm with time, for the response between various

concentrations of Zataria multiflora extracts (5, 10, 15, 20, 25

and 30 % v/v) and 0.5 mg/mL of the graphene oxide suspension

at 96 °C for 30 hrs.

1, 13, 17). . At that point, the concentrate of the leaf extract

of Zataria multiflora was added to arranged GO, refluxed then

sonicated, and at the last stage, the suspension was centrifuged

and the RGO dark shaded washed with water twice and went

away in the stove.

The level of reduction seen to increment with

concentration of the Zataria multiflora removes from 5% to

0%. No further increment in absorbance discovered when the

Zataria multiflora extracts 5% V/V

Zataria multiflora extracts 15% V/V

Zataria multiflora extracts 25% V/V

Zataria multiflora extracts 10% V/V

Zataria multiflora extracts 20% V/V

Time(hours)

Figure 1: Effect of the concentration of Zataria multiflora extracts on the reduction of graphene oxide (0.5 mg/mL) at 96 °C

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2020, Volume 8, Issue 1, Pages: 488-496

5 centigrade degree

0 centigrade degree

30 centigrade degree

75 centigrade degree

45 centigrade degree

96 centigrade degree

Time(hours)

Figure 2: Effect of reaction temperature on the reduction of graphene oxide (0.5 mg/mL) using 20 % v/v Zataria multiflora extracts

Figure 3: Infrared spectra of graphite

The examination of the FT-IR spectra in GO and RGO

demonstrates that in GO there is a tensile vibration of C = O in

the estimations of 3350 cm^-¹and 1100 cm^-¹(Figure 5), so it

tends to be presumed that amid the recovery procedure

(planning of RGO), the current oxygen species on the surface

of graphene sheets, the concentrate of Zataria multiflora plant

has been decreased and graphene oxide has been reduced to

graphene (3, 11–13, 21, 54).

cm-1 of 1720 cm , yet in RGO the ductile vibration of C = O

isn't so serious and moved to 1797 cm , and also, the power of

-1

the vibration retention of OH in 3421 cm and 1384 cm which

at RGO diminished fundamentally after reduction and achieved

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2020, Volume 8, Issue 1, Pages: 488-496

Figure 4: Infrared spectra of graphene oxide

Figure 5: Infra-red spectra of reduced graphene oxide

.2 XRD analysis

The X-ray beam diffraction test utilized to check the

structure of graphene oxide, graphene, and oxidized graphene.

As appeared in figure 7, X-rays beam diffraction (XRD) for

graphene oxide demonstrates a strong peak at 2θ = 10.5, which

shows the nearness of oxygen-containing functional groups in

void spaces between graphene oxide plates. As found in Figure

, the graphene X-ray beam diffraction example of this peak

has been exchanged to the range 2θ = 13 and speaks to the

graphene cover after the way toward recovering and emptying

water particles and oxygen bunches from the spaces between

the layers of graphene oxide (11, 12, 54).

Figure 6: XRD spectra of reduced graphene oxide and graphene oxide

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.3 Raman study

Raman scattering is a valuable device to portray graphite and

multilayer structure, after the reduction of GO, the 2D band is

extraordinary, which proposed about stacking of graphene

layers. As GO has diverse kinds of functional groups that may

avoid stacking of graphene layers yet after reduction because

of an abatement of such functional groups a couple of graphene

layers are stacking and framed multilayer RGO (54).

graphene materials as this dissipating emphatically relies upon

the electronic structure. Raman range of GO was observed to

be altogether changed after the reduction (Figures 7 and 8). In

the spectra of GO and RGO, two crucial vibration bands were

seen in the scope of 1250–1750 cm . The G vibration mode,

inferable from the first-order scattering of E2 g phonons by sp2

carbon of GO and RGO were discovered 1592 and 1587 cm-1

separately, while the D vibration band acquired from a

breathing method of j-point photons of A1g symmetry of GO

and RGO showed up at 1350 and 1345 cm-1 separately. .After

the reduction of GO, the force proportion of the D band to the

G band (ID/IG) was expanded essentially. As D band emerges

due to the sp2 carbon group, the higher power of D band

proposed the nearness of more detached graphene space in

RGO in contrast with GO and expulsion of oxygen moieties

from the former. Notably, the two-phonon (2D) Raman

dissipating of graphene-based materials is an important band to

separate the monolayer graphene from a twofold layer/multi-

layer graphene as it is profoundly discerning to stacking of

graphene layers. For the most part, a Lorentzian peak for the

3.4 Transmission Electron Microscopy (TEM) analysis

Figure 9 shows TEM images of the RGO sheet reduced

with Zataria multiflora leaf extract. The appearance of

transparent and silky sheets of RGO in TEM images verifies its

stability under a high-energy electron bar. The high-resolution

TEM images are utilized to get to the number of layers in

numerous areas. The edges of the suspended graphene films

tend to crease back, permitting the cross-sectional perspective

of the films. The collapse of a couple of layers at the edges of

the films shows up as a couple of dim lines, individually. The

arrangement of a few layers RGO is unmistakably obvious. It

has likewise been discovered that the forces of all the

diffraction spots are not equivalent and sharp enough.

Additionally, the diffraction spots are related to some uncertain

spots. Every one of these perceptions additionally supporting

the development of a few layers RGO (3, 13, 17, 21, 54).

D band of the multi-layer graphene sheets seen at 2700 and

900 cm-1 (11, 12, 54). This demonstrates RGO has a

Figure 7: Raman spectra of GO

Figure 8: Raman spectra of RGO

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2020, Volume 8, Issue 1, Pages: 488-496

Figure 9: TEM images at 200 nm magnifications, showing the formation of few layer of reduced graphene oxide

Conclusion

Ethical issue

We demonstrated that the studied phyto extracts have

Authors are aware of, and comply with, best practice in

publication ethics specifically with regard to authorship

(avoidance of guest authorship), dual submission, manipulation

of figures, competing interests and compliance with policies on

research ethics. Authors adhere to publication requirements

that submitted work is original and has not been published

elsewhere in any language.

tremendous potential to be used as reducing agents for the

reduction of GO with an environmentally benign synthetic

protocol. Graphene oxide was prepared from graphite powder

using a modified Hummers’ method followed by ultra-

sonication. The optimized reaction condition for the reduction

of graphene oxide was determined to be a reaction time of 20

hours and a temperature of 96 C using 20% v/v of Zataria

multiflora extracts. We showed that the considered phyto

extracts can possibly be utilized as diminishing operators for

the reduction of GO with an ecological amiable engineered

convention. Graphene oxide was set up from graphite powder

utilizing an altered Hummers' strategy trailed by ultra-

sonication. The upgraded response conditions for the reduction

of graphene oxide was resolved to be a reaction time of 20

hours and a temperature of 96 °C utilizing 20% v/v of Zataria

multiflora extracts. The most imperative favorable

circumstances of the phytochemicals are their wealth in nature,

cost-viability, and simple item segregation after reduction as

they extricated from non-palatable or squander plant items. The

estimations of particular capacitance, high electrical

conductivity, and high carbon to oxygen proportion of the

phyto extract RGO are adequate. In this way, this green strategy

can be utilized for vast scale generation of RGO. An eco-

accommodating and naturally considerate reduction framework

by utilizing Zataria multiflora removes as a biocatalyst for the

reduction of graphene oxide is portrayed. The reduction was

done in a watery medium at an alternate temperature. TEM

uncovers the development of few-layer of reduced graphene

oxide. FTIR and X-ray examination give proof to the end of

labile oxygen usefulness from the surface of GO. De-

oxygenation and the development of deformities in the RGOs

have been affirmed by Raman spectroscopy. The primary

points of interest of this system over the conventional synthetic

reduction are the cost-adequacy, ecologically agreeable

methodology, and basic item confinement process. This

ecologically amicable reduction of graphene oxide can possibly

be utilized in different fields, for example, biomedical

applications.

Competing interests

The authors declare that there is no conflict of interest that

would prejudice the impartiality of this scientific work.

Authors’ contribution

All authors of this study have a complete contribution for

data collection, data analyses and manuscript writing.

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