microwave mediated green synthesis of copper nanoparticles using aqueous extract...

Microwave Mediated Green Synthesis Of Copper Nanoparticles

Using Aqueous Extract Of Piper Nigrum Seeds And Particles

Characterisation

N. Gandhi1 D. Sirisha2 * and Smita Asthana3 1. Research Scholar, Centre for Environment and Climate Change, School of Environmental Sciences,

Jawaharlal Nehru Institute of Advanced Studies, Hyderabad, India.

2. Head, Centre for Environment and Climate Change, School of Environmental Sciences,

Jawaharlal Nehru Institute of Advanced Studies, Hyderabad, India.

3. Reader in Chemistry, Department of Chemistry,

St. Ann’s College for Women-Mehedipatnam, Hyderabad, Telangana, India.

Corresponding Author: Dr. D. Sirisha, (Head, Centre for Environment and Climate Change, School of Environmental

Science, Jawaharlal Nehru Institute of Advanced Studies (JNIAS), Hyderabad, Telangana.) & Department of Chemistry,

St. Ann’s College for Women, Hyderabad, Telangan. Email: [email protected], [email protected]

Abstract: Green synthesis of nanoparticles using biological molecules derived from the plant sources in the form of

extracts are exhibiting superiority over chemical and physical methods. Development of green nanotechnology is

generating interest of researchers towards eco-friendly green synthesis of nanoparticles. In present study, green synthesis

of stable copper nanoparticles were done using Piper nigrum seed extract. The seed extract prepared by using deionised

water was mixed with 0.01M of copper sulphate (CuSO4.5H2O) solution and an alternative energy source microwave

irradiation used to get nanoparticles in a short incubation period. There is a change in colour (Blue to Green) observed and

that is indicating the formation of copper nanoparticles. The plant based molecules present in the seed extract have highly

controlled assembly, which is suitable for synthesis of nanoparticles. These green synthesised copper nanoparticles were

characterised with the help of Uv-visible spectrophotometer, X-ray diffraction analysis (XRD). It was observed that the

Piper nigrum seed extract can reduce copper ions to copper nanoparticles within 2 to 5 minutes of reaction time. Thus this

method can be used for rapid and eco friendly green synthesis of suitable copper nanoparticles.

Key words: Copper nanoparticles, Green Synthesis, Uv-visible spectrophotometer, X-ray diffraction analysis (XRD),

Piper nigrum seed.

INTRODUCTION

Nanoparticles preparation and study about nanoparticles are importance in the research. The characters of metal

nanoparticles like optical, electrical, magnetic and catalytic are depending on their size, shape and morphology. Several

methods are available which have been extensively used to produce nano sized copper (Figure-1). The micro emulsion

method (Nasser & Husein, 2007), sub merged nanoparticles synthesis system (Kao et al, 2007), flame based aerosol

method (Chiang et al, 2012), sono chemical (Vijaya kumar et al, 2001) hydrothermal (Zhang et al, 2006) and solid state

techniques (Wang et al, 2004) the use of toxic chemicals are subjects of most important concern.

IAETSD JOURNAL FOR ADVANCED RESEARCH IN APPLIED SCIENCES

VOLUME 5, ISSUE 2, FEB/2018

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http://iaetsdjaras.org/859

Figure-1: Various methods of Synthesis of Copper nanoparticles

The use of toxic chemicals for the synthesis of nanoparticles limit their applications. Therefore, development of

clean, biocompatible non toxic and eco friendly method for nanoparticles synthesis deserves merit. The interest in this

field has shifted toward ‘green’ chemistry and bio-processor approach. These approaches focus on utilization of

environmental-friendly, cost-effective and biocompatible reducing agents for synthesis of copper nanoparticles. Here, in

this work we report microwave assisted green synthesis of Copper nanoparticles, reducing the copper ions by the aqueous

extract of Piper nigrum seed, characterized by Uv-Vis spectroscopy, XRD, TGA and DTA.

MATERIALS & METHOD

Materials

Piper nigrum seed (Figure- 2 A) for the biosynthesis of the copper nanoparticles was procured from the local

supermarket. Copper sulphate used in the study is of analytical grade.

Preparation of Sample Extract

5 gm of Piper nigrum seeds were accurately weighed thoroughly washed under running tap water followed by

washing it with double deionised water to remove surface impurities. They were crushed using a blender and finely

macerated. After homogenization 100ml of double deionised water was added and heated over a water bath maintained at

80°C for 15 minutes. The extract obtained was filtered through muslin cloth and then through Whatmann no: 1 Filter

paper (pore size 25µm) and used immediately for the biosynthesis of copper nanoparticles.



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Figure-2 (A): Classification and general information about Piper nigrum

Preparation of Copper Nanoparticles:

For the preparation of 100 ml copper nanoparticles 80 ml of 1mM CuSO4.5H2O solution mixed with 20 ml of

Piper nigrum seed extract and incubated for 3 hours at room temperature. Another set of same experiment conducted with

supply of microwave irradiation (900 waats for 2 minutes) as alternate energy sources. The change in colour indicating the

formation of copper nanoparticles (Figure-2 B).

Piper nigrum seed weighing 5 g were thoroughly washed and dried and crushed into 100 ml sterile distilled water.

Filtered through Whatt man No.1 filter paper (pore size 45 μm) followed by further filtered through 0.22μm sized filters

0.001M Copper Sulphate (CuSO4) was prepared in 1000 ml of distilled water

20 ml of Piper nigrum seed extract was added into 80 ml of aqueous solution of 0.001M Copper Sulphate (CuSO4)

Incubation in microwave oven at 750 watts for 5 minutes

Copper (Cu) nanoparticles formation



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Figure-2 (B): Green synthesis of Copper nanoparticles A) CuSO4 solution B) Piper nigrum seed extract and C)

CuNP solution

Characterisation of Copper Nanoparticles:

The microwave mediated and green synthesized copper nanoparticles were characterized by Uv-

spectrophotometer, XRD, TGA and DTA respectively.

Advantages of using Plant extracts:

1. The generation of nanoparticles utilizing the chemical techniques has been raising worry among the environmentalists as

they have an antagonistic effect on their biology, henceforth the utilization of plant extracts for the arrangement of

nanoparticles is being favored due its salubrious nature towards the earth. Indeed, even in the business it delivers

considerably less harmful waste.

2. The plants supplement both the lessening and also balancing out specialists for the nanoparticles which generally must

be remotely included different strategies.

3. The synthetic strategy is being demonstrated less financially gainful when contrasted with the plant technique as the

support cost is significantly less and the waste transfer requires less exertion among different variables.

4. This technique is shockingly better than utilizing the natural strategy as the support of entire plant framework is

substantially less than a culture of microscopic organisms which needs a horde of marvels to be dealt with.

5. Late investigations have demonstrated that the helpful impacts of plants, from which the nanoparticles are being

determined, can likewise be permeated upon the particles subsequently giving us idealize vehicles to the restorative

materials to follow up on the site of activity and also taking out the need to falsely build up a medication for that specific

affliction

RESULTS & DISCUSSION

Uv-Visible Spectrophotometer

UV-Visible spectroscopy refers to the absorption spectroscopy in the ultraviolet visible spectral region. It uses

light in the visible region and adjacent near infrared (NIR) ranges. In this region of electromagnetic spectrum, molecules

undergo electronic transitions nanoparticles have certain optical properties such as size, shape, concentration,

agglomeration state and refractive index which can be identified by UV-visible spectrometer. Nanoparticles made from

certain metals strongly interact with certain wavelength of light and their unique optical properties leads a phenomena

known as surface Plasmon resonance. In the present study the UV visible spectra for copper nanoparticles synthesized

from Piper nigrum seed extract at different time intervals and different temperatures were represented in Fig-3, Fig-4 and

Fig-5 respectively. The Plasmon resonance produced the peaks at about 300 to 310 nm for copper nanoparticles

synthesized from Piper nigrum seeds extract.



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X-Ray Diffraction Studies

Powder X-ray Diffraction (XRD) is one of the primary techniques used by mineralogists and solid state chemists

to examine the physico-chemical make-up of unknown materials. X-ray diffraction is one of the most important

characterization tools used in solid state chemistry and materials science. XRD is an easy tool to determine the size and the

shape of the unit cell for any compound. Powder Diffraction Methods is useful for Qualitative analysis (Phase

Identification), Quantitative analysis (Lattice parameter determination & Phase fraction analysis) etc. Diffraction pattern

gives information on translational symmetry - size and shape of the unit cell from Peak Positions and information on

electron density inside the unit cell, namely where the atoms are located from Peak Intensities. It also gives information on

deviations from a perfect particle, if size is less than roughly 100 – 200nm, extended defects and micro strain from Peak

Shapes & Widths.

Peak Indexing

Indexing is the process of determining the unit cell dimensions from the peak positions. It is the first step in

diffraction pattern analysis. To index a powder diffraction pattern it is necessary to assign Miller Indices (h k l) to each

peak. Unfortunately it is not just the simple reverse of calculating peak positions from the unit cell dimensions and

wavelength (Cullity 1978).

Figure-6: XRD showing peak indices and 2 θ positions



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XRD analysis of the prepared sample of Copper nanoparticles was done by a Goniometer. Data was taken for the 2θ

range of 20 to 80 degrees with a step of 0.02 degree. Indexing process of powder diffraction pattern was done and Miller

Indices (h k l) to each peak was assigned in first step. Diffractogram of the entire data is in Fig.6.

Table-1: Simple Peak Indexing

Peak position, 2θ 1000 X Sin2 θ 1000XSin2 θ/46 Remark

42.6 132 2.86 12 + 12 + 12 = 3

51.8 188 4.08 22 + 02 + 02 = 4

72.4 359 7.80 22 + 22 + 02 = 8

Indexing has been done in two different methods and data are in Table.1 & Table.2. In table.1, one need to find a

dividing constant and the values in the 3rd column becomes integers (approximately). Here, the constant is 56 (=

188−132). Moreover, the high intense peak for FCC materials is generally (1 1 1) reflection, which is observed in the

sample.

Table-2: Peak indexing from d-spacing

2θ d 1000/d2 (1000/d2)77.32

42.6 2.121 235 3.03

51.4 1.776 282 4.01

73.6 1.286 389 5.03

Table.3: Experimental and standard diffraction angles of Cu specimen

Experimental diffraction angle (2θ in degrees) Standard diffraction angle (2θ in degrees)

JCPDS Copper:04-0836

42.6 43.297

51.4 50.433

73.6 74.130

Three peaks at 2θ values of 42.640, 51.400, and 73.620 deg corresponding to (111), (200), and (220) planes of

copper were observed and compared with the standard powder diffraction card of JCPDS, copper file No. 04–0836.

Table.4 shows the experimentally obtained X-ray diffraction angle and the standard diffraction angle of Cu specimen. The

XRD study confirms / indicates that the resultant particles are (FCC) Copper Nanoparticles.

Particle Size Calculation

From this study, considering the peak at degrees, average particle size has been estimated by using Debye-Scherrer

formula (Nat et al, 2008; Das et al, 2009; Nath et al, 2007; Hall et al, 2000)

Where K is instrument constant (0.9), λ is wavelength of X ray diffraction (0.1541 nm) ‘β’ is FWHM (full width at half

maximum), ‘θ’ is the diffraction angle and ‘D’ is particle diameter size.

1). 2θ = 42.6

2). 2θ = 51.4

3). 2θ = 73.6



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The calculated results were concluding that the microwave mediated green synthesized copper nanoparticles by

using Piper nigrum seeds extract is less than 40 nm in size.

Calculation of d-Spacing

The value of d (the interplanar spacing between the atoms) is calculated using Bragg’s Law: 2dsinθ = nλ

Wavelength λ = 1.5418 Å for Cu

1). 2θ = 42.6

θ = 21.3

2). 2θ = 51.4

θ = 25.7

3). 2θ = 73.6

θ = 36.8

TGA/DTA Analysis

TGA measures the amount of weight change of material either as a function of temperature or isothermally as a

function of temperature in an inert atmosphere of nitrogen. TGA, measured weight loss curve gives information on

changes in sample composition, thermal stability, and kinetic parameters for chemical reactions in the sample. TGA gives

the following characteristics.

1) Thermal stability

2) Material purity

3) Determination of humidity. Is also examines corrosion studies, gasification process and kinetic process. The Figure-7,

TGA curve shows a loss of 50% indicating the loss of water and other volatile components which were used as stabilising

agents. Two exothermic peaks were attributed in this TGA curve. The first weight loss peak was related to the removal of

water from the surface at 250oc and it is due to removal of organic molecules. At higher temperatures no significant

weight loss was observed, there by supporting the crystallinty with high purity. These observations indicate that

phytochemicals present in the Piper nigrum seeds extract serve the dual purpose of reducing the copper salt to nano

dimensional copper particles and it is serving as capping agent.

DTA curve is shown in Figure-8, for green synthesized copper nanoparticle. In this curve two different peaks

were observed and those were representing the exothermic reactions. 1st peak is observed 260oc indicating loss of water

and 2nd peak is observed at 504oc. The initial weight loss from 13.0 mg to 6.5 mg is observed which exactly 50% is. The

area enclosed in the peaks will give the enthalpy of the reaction which is equal to.

Figure-7: TG (%)/DTA spectrum of Copper Nanoparticles



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Figure-8: TG (mg)/DTA spectrum of Copper Nanoparticles

Mechanism of formation CuNPs

The utilization of plant and plant extracts in nanoparticle amalgamation is viewed as profitable over microbial

based framework since it decreases the intricate procedure of keeping up cell cultures. The molecule estimate development

can likewise be controlled by changing combination conditions like pH, reductant focus, temperature, blending proportion

of the reactants and so forth. The plant based combination can be done either extracellularly or intracellularly. Intracellular

blend happens inside the plant though the extracellular union happens in vitro. The examinations uncover that

extracellular blend utilizing plant separates has been viewed as better when contrasted with intracellular amalgamation

(Makarov et al, 2014) on the grounds that it takes out the extraction and decontamination strategies. Biosynthesis of

CuNPs by plant extracts, for example, Eclipta prostrate (Chung et al, 2017), Plantago asiatica leaf extract (Nasrollahzadeh et al,

2017), Ocimum sanctum (Kulkarni & Kulkarni 2013; Bhasker et al, 2014), Camellia sinensis leaf extract (Mohindru & Garg

2017), Gloriosa superba leaf extract (Powar et al, 2016), Carica papaya (Suresh et al, 2014), Mangolia kobus (Lee et al, 2013),

Syzygium aromaticum (Subhankari et al, 2013), Artabotrys odoratissimus (Umesh et al, 2014), Capparis zeylanica (Renganthanan et

al, 2014), Vitis vinifera (Subbaiya et al, 2014), Nerium oleander (Gopinath et al, 2014), Datura metel (Makwana et al, 2014),

Pisidium guajava (Kote et al, 2014), Cinnamum (Koizhaiganova et al, 2014), Aloe vera (Javad et al, 2015), Citrus medica

(Mahendra et al, 2015), Eucalyptus (Pramod et al, 2015), Eupatorium glandulousm (Subbaiya et al, 2015), Cassia fistula (Valli et al,

2015), Phyllanthus embilica (Caroling et al, 2015), Guava (Caroling et al, 2015), Cassia Auriculata (Valli et al, 2016), Ocimum

tenuiflorum (Vemila et al, 2016) and so on have been accounted for. Till date, part of papers has been distributed around

there which portrays the system and part of dynamic biomolecules in blend (Parashav et al, 2011). These examinations

recommended that nearness of phytochemicals in plant separates are the key segment in diminishment and adjustment of

copper particles (Parashav et al, 2011). The phytochemicals which are in charge of lessening are terpenoids, flavonoids,

ketones, aldehydes, amides, and carboxylic acids. The water dissolvable metabolites, for example, flavones, natural acids,

and quinones are exclusively in charge of the bioreduction particles. A few analysts have revealed that a keto-enol progress

of anthraquinone is in charge of arrangement of CuNPs. It has been likewise watched that mesophytes contain three sorts

of benzoquinones: cyperoquinone, dietchequinone, and remirin which may be in charge of lessening of particles and

development of CuNPs. One of the biomolecule which significantly partake is terpenoids. Terpenoids are otherwise called

isoprene, a normally happening natural mixes in plants, they contain five-carbon isoprene units. It has been investigated by

a few specialists that Geranium leaf remove contain terpenoids, which go about as significant player in biosynthesis of

AgNPs (Shanker et al, 2003). Similar results were found with Cinnamomum zeylanicum (cinnamon) removes contains

eugenol which may be in charge of the decrease silver nitrate to AgNPs (Satish kumar et al, 2009).



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Based on previous investigations and information available in the literature, it have been recommended that the

deprotonation of the hydroxyl particle of eugenol prompt arrangement of reverberation balanced out structures which can

additionally oxidized, by reducting metal particles into its nano extend (Satish kumar et al, 2009). Another significant class

of plant metabolite is flavonoids. Falvonoids are gathering of polyphenolic mixes containing 15 carbon particles and are

water dissolvable. Flavonoids can be grouped into: isoflavonoids, bioflavonoids and neoflavonoids, which can go about as

chelating and diminishing operators for metal particles. The practical gathering present in flavonoids are exclusively in

charge of nanoparticle arrangement. The change of flavonoids from the enol to the keto may prompt decrease of metal

particles to frame nanoparticles (Makarov et al, 2014). Ahmad et al. reported that Ocimum basilicum(sweet basil) extract

contains of flavonoids, eugenol and polyphenols that assume enter part in the development of AgNPs from silver particles

by tautomerization of enol to keto shape (Ahmad et al, 2010). A few investigations have been demonstrates that flavonoids

can go about as chelating operators for instance quercetin is a flavonoid which can chelate at three positions including the

carbonyl and hydroxyls at theC3 and C5 positions and the catechol aggregate at the C3' and C4' site (Makarov et al, 2014).

These aides in understanding that flavonoids are associated with start of nanoparticle arrangement (nucleation) and further

accumulation, notwithstanding the bioreduction organize. Glucose a straight monosaccharides having free aldehydic

gathering can specifically go about as decreasing specialists while fructose which contains keto-gathering can go about as

cell reinforcements if tautomeric changes happens from ketone to an aldehyde (Makarov et al, 2014). It has been accounted

for that when glucose was utilized as a diminishing specialist the nanoparticles with various morphologies were watched

while with fructose just monodispersed nanoparticles were watched. It has been hypothesized that aldehydic gathering of

sugar get oxidized into a carboxylic gathering by means of the nucleophilic expansion of OH-, which eventually prompt

decrease of metal particles and blend of nanoparticles (Makarov et al, 2014).

There are three principle stages which incorporated into the plant interceded amalgamation. Beginning stage

otherwise called enactment stage amid which metal particles get lessened and the decreased metal iotas get nucleated;

another is the development stage in which unconstrained accumulation of little contiguous nanoparticles jumps out at

frame particles of a bigger measurement, which are thermodynamically more steady; last stage is the end stage which

decides the last state of the nanoparticles (Makarov et al, 2014; Si & Mandal 2007). Increment in the development stage,

prompt total of nanoparticles into nanotubes, nanorods and nanotriangles and so on (Makarov et al, 2014).. In the end

stage, nanoparticles experience conformational change which is thermodynamically steady, which affirms the part of plant

concentrate to settle metal nanoparticles.

CONCLUSION

The present examination speaks to a clean, non-dangerous and in addition eco-accommodating technique for

combining CuNPs. The topping around every molecule gives normal substance condition framed by the bio-natural

compound present in the Piper nigrum seed extract, which might be mainly in charge of the particles to wind up plainly

balanced out. This method gives us a straightforward and proficient route for the combination of nanoparticles with

tunable optical properties administered by molecule measure. From the of nanotechnology perspective, this is a critical

improvement for incorporating CuNPs financially. All in all, this green science approach toward the combination of

CuNPs has a few focal points viz, simple process by which this might be scaled up, financial suitability, and so forth.

ACKNOWLEDGEMENTS

The authors are grateful to Dr. Shilpa Chakra, Head of the Department, Centre for Nano Science and

Technology, Institute of Science and Technology, Jawaharlal Nehru Technological University Hyderabad, Kukatpally,

Telanagana for providing instruments for the characterisation of green synthesized Copper Nanoparticles.



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