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* Corresponding author: Ganesan, VCentre for Research and Postgraduate studies in Botany, Ayya Nadar Janaki Ammal College, Sivakasi, Tamilnadu, India

ISSN: 0976-3031

RESEARCH ARTICLEGREEN SYNTHESIS OF SILVER NANOPARTICLES USING LEAVES OF

MURRAYA PANICULATA (L.)JACK

*Ganesan, V., Arunkumar, C., Nima, P and Astalakshmi, A

Centre for Research and Postgraduate studies in Botany, Ayya Nadar Janaki Ammal College, Sivakasi, Tamilnadu, India

ARTICLE INFO ABSTRACT

The present study deals with the synthesis of silver nanoparticles using the leaf broth of

Murraya paniculata (Family: Rutaceae). The leaf broth was prepared and resuspended inaqueous solution of silver nitrate and it is known as reaction medium. This reaction medium

was kept in an incubator cum shaker (Orbitek) with 250rpm at 27

0

C for 24 hours to reducethe silver nitrate in to silver nanoparticles. The colour change in reaction medium from pale

yellow to dark brown was observed during the incubation period. It indicates the formationof silver nanoparticles and they were characterized by UV-Visible Spectroscopy, FourierTransform Infra-Red (FT-IR) Spectroscopy and X-ray diffraction (XRD) analyses. It isevident that the synthesized silver nanoparticles are capped by biomolecules which areresponsible for reduction of silver ions. Energy Dispersive X-Ray (EDX) analysis and

Scanning Electron Microscopy (SEM) confirmed the formation of silver nanoparticles. Thistype of phyto-mediated synthesis appears to be cost effective, eco-friendly and easyalternative green synthesis to conventional, physical and chemical methods of silver

nanoparticles synthesis.

INTRODUCTION

The tremendous developments in the field of nanotechnologymake the emergence of more new technologies to synthesize thenanoparticles of particular shape and size (Shivshankar et al.,

2004). Silver is a known metal that came into use before Neolithicreduction, even Greeks used it for cooking and to keep watersafer. There are several approaches used for the synthesis of silver

nanoparticles such as Chemical reduction (Santosh et al.,1999),phytochemical (Bera et al., 2010), Reverse micelles (Lim etal.,2004), Thermal decomposistion (Plante et al.,2010), Radiation

assisted (Cheng et al., 2011), Electrochemical (Hirsch et al.,2005), Sonochemical (Kurotchenkov et al., 2006), Microwaveassisted method (Nadagouda et al., 2011) and recently via green

chemistry method (Raveendran et al.,2003). The physical (Xu etal., 2008) and chemical processes (Wang et al., 2005) are thegeneral methods for the fabrication of nanoparticles but these are

not environmental friendly, where the chemicals used are toxicand flammable. To overcome this, green synthesis of silvernanoparticles using environmentally benign materials like plant

extracts (Parashar et al.,2009), fungi (Saifuddin et al., 2009a),bacteria (Bhansa et al., 2006) and enzymes (Willneret al., 2007)is a single step, time reducing, lowcost offers numerous benefits

of eco-friendliness and compatibility for pharmaceutical and otherbiomedical applications (Bar et al., 2009a). There are severalplants have been reported for the plant mediated biosynthesis ofsilver nanoparticles such as Phyllanthus amarus (Annamalai et al.

2011), Diopyros kaki (Vyom et al., 2009)andCapsicum annum(Harekrishna et al., 2009). Hence the present study is aimed to

synthesize and characterize the silver nanoparticles using the leafextract ofMurraya paniculata.

MATERIALS AND METHODS

Biological Synthesis of silver nanoparticles

All the reagents used in the present study were obtained fromHimedia Laboratories Pvt Ltd (Mumbai, India). Murrayapaniculata (Figure 1)belongs toRutaceaewere collected from the

Botanical garden of Ayya Nadar Janaki Ammal College, Sivakasi,Tamilnadu, India. The collected leaf samples were throughlywashed with tap water followed by distilled water to remove the

surface contaminants and dried for 48 hours under shade. Thedried leaves were taken and ground to make fine powder usingmixer and the sieved using 20 mesh sieve to get uniform size

range. 10g the sieved leaf powder was added to 100ml of distilledwater and boiled at 70 C for 10 minutes to prepare the leaf broth

(Sathishkumaret al., 2009). 10 ml freshly prepared leaf broth wasresuspended in 190ml of aqueous solution of silver nitrate and thismixture is used as reaction medium (Muyabi et al., 2012).

This medium was kept in an Incubator cum shaker (Orbitek-Model) with 250 rpm at 27C for 24 hours. From this reaction

medium a small aliquot of the sample was used to characterize thepresence of silver nanoparticles synthesized during the abovereaction. The characterization was performed through the

following analyses: UV- Visible spectroscopy (UV-Vis), FourierTransform Infrared spectroscopy (FT-IR), X-ray Diffraction(XRD) analysis, Scanning Electron Microscopy (SEM),Energy Dispersive X-Ray Analysis (EDX) and Transmission

Electron Microscopy (TEM). The quantity of Silver nanoparticlessynthesized by leaf tissue was estimated using Atomic Absorption

Spectroscopy (AAS) in accordance with the following equation(Dias et al., 2002) q= [(C0 C) / X] where: q (mg of metal

Available Online at http://www.recentscientific.com

International Journal

of Recent Scientific

ResearchInternational Journal of Recent Scientific ResearchVol. 4, Issue, 6, pp.1022 1026, July, 2013

Article History:

Received 14th, June, 2013

Received in revised form 26th, June, 2013

Accepted 16

th

, July, 2013Published online 30th July, 2013

Copy Right, IJRSR, 2013, Academic Journals. All rights reserved.

Key words:

Green synthesis,Murraya paniculata, ElectronMicroscopy, Silver nanoparticles

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International Journal of Recent Scientific Research, Vol. 4, Issue, 7, pp. 1022 - 1026, July, 2013

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nanoparticles synthesized by one gram of leaf tissue) is the metalspecific uptake, C0 is the initial metal concentration (mg l

-1), C is

the residual metal concentration (mg l-1) and X is the biomass

concentration of the leaf tissue (g).

RESULTS AND DISCUSSION

UV- Visible spectrum of silver nanoparticles

UV-Visible spectroscopy analysis was carried out on a Labomed(Model UV- D3200) UV-Visible spectrophotometer. The leafbroth reduced aqueous silver nitrate into silver ions and formed

into silver nanoparticles. The leaf broth had pale yellow colourbefore the addition of silver nitrate solution which was colorless(Figure 2). After the addition of aqueous silver nitrate, the leafbroth (reaction medium) gradually changed into dark brown colorwithin one hour of reaction (Figure 2). It indicates that the silver

nitrate is rapidly reduced into silver ions within one hour. UV-Visible spectra of the reaction media taken at different timeintervals explicit that the Surface Plasmon Resonance (SPR)

vibrations are found between 410nm and 460nm with the max at420 nm which is blue shifted (Figure 3). The maxin SPR bands of

silver nanoparticles varies with the substrate or an organism bywhich they are synthesized. For example the max in SPR band isat 400 nm byE.coli (Natarajan et al., 2010); 410 nm byBacillussubtilis (Saifuddin et al., 2009); 430nm by leaves ofEuphorbiahirta (Elumalai et al., 2010); 405nm and 480nm by leaves ofEnicostema hysopifolium andRauvolfia tetraphylla respectively(Veena et al., 2011). In the present study, the absorbance of the

reaction medium increases up to 1.4 a.u. at 420nm in one hour ofreaction.

Figure 1Murraya paniculata leaves

Thereafter the absorbance gradually decreases (Figure 3). It

indicates that the capping agents like secondary metabolites orenzymes present in the leaf broth reduce the aqueous silver nitraterapidly within one hour. The saturation of capping agents in the

reaction medium after one hour started to decrease in thereduction process of silver nitrate into silver ions. The time takenfor the change in colour of the reaction medium varies from plant

to plant. Interestingly, the leaf extracts ofBoswellia ovalifoliolataandShorea tumbuggaia made the colour change within 10 and 15minutes respectively (Savithramma et al., 2011). The leaf extract

ofMerrimia tridendata reduced the silver nitrate and observedthis colour change within 24hrs (Ganesan et al., 2013) while theleaf extract ofAcalypha indica andCamellia sinesis reduced the

silver nitrate and made the colour change from pale yellow tobrown within 30 minutes (Krishnaraj et al., 2010; Kalayan et al.,

2010). The Blue shift is related to a decrease in the particle sizeand the red shift due to the increased size of silver nanoparticles(Mulvaney, 1996).

Figure 2 Colour change of the reaction medium (leaf broth ofMurraya

paniculata and 1mM aqueous silver nitrate during the biological synthesis of

silver nanoparticles). A- Control (Aqueous silver nitrate), B- leaf broth of

Murraya paniculata C, D, E, F, G and H are the reaction media at different

time intervals of reaction such as 5 minutes, 30 minutes, one hour, two hours,

4 hours and 24hours respectively.

The UV-visible spectroscopic analysis explains that the aqueoussilver nitrate is reduced silver ions and in turn to form silvernanoparticles by the biomolecules present in the leaf broth of

Murraya paniculata.

Figure 3 UV-Visible spectra of silver nanoparticles synthesized by leaf

aqueous extract ofMurraya paniculata as a function of time

FT-IR Spectroscopic Analysis

FT-IR measurements (Shimadzu FTIR spectrophotometer, using

KBr pellet method) identified the biomolecules in the leaf broth ofMurraya paniculata which were responsible for reducing silvernitrate and the synthesized silver nanoparticles. The nature ofchemical bonds present in the reaction medium was characterized.

The FTIR spectrum of leaf broth before reaction, showed severalabsorption peaks at 461, 538, 601, 655, 720, 914, 1102, 1191,

1304, 1402, 1456, 1668 , 2850, 2922, 3201 and 3314 cm-1

(Figure4a). The FT-IR spectrum of purified and dried reaction mediumwith silver nanoparticles (Figure 4b) shows the absorbance peakat 461, 605, 653, 752, 806, 1112, 1195, 1386, 1400, 1452, 1670,

2108, 2975, 3193 and 3314 cm-1. The strong absorbance band at

1386 cm-1

was associated with the stretch of functional groupssuch as C-O-C-, -C=O-, -C=C, -C=O- (Elavazhagan and

Arunachalam, 2011). The absorbance bands are known to beassociated with the stretching vibrations for -C-C-O, -C-C-, -C=C-, C=O (esters, ethers) and C-O (polyols) respectively (Bar et al.,

2009).

Figure 4a FT-IR spectrum of leaf broth ofMurraya paniculata

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The total disappearance of the bands at 538, 914, 2850 and2922 cm-1after bio-reduction may be ascribed to the reduction

of silver ions into silver nanoparticles, leading to unsaturatedcarbonyl groups with broad peak at 1670 cm

-1.

Figure 4b FT-IR spectrum of silver nanoparticles using leaf

extracts ofMurraya paniculata

XRD Analysis

Crystalline metallic silver nanoparticles were examined by X-raydiffractometer Shimadzu (XRD 6000)). Figure 5 shows the X-ray

diffraction peak obtained for the synthesized silver nanoparticles

using Murraya paniculata leaf extract. The Full-Width HalfMaximum (FWHM) values measured for the plane of reflection

were measured using Debye-Scherrers equation, t=0.9/Cos(Narashima et al., 2011). The mean size of the nanoparticles wasestimated as 31nm using the observed XRD pattern at 2= 27.7marked within (111), 29.2 marked within (200), 32.1 markedwithin (220) and 38.05 marked within (311). The XRD patternrevealed that the silver nanoparticles formed are face centered

cubic (fcc) structures (Mohsen et al., 2011).

Figure 5 XRD pattern of silver nanoparticles formed after

reaction of leaf broth ofMurraya paniculata

The XRD pattern thus showed that the silver nanoparticlesformed are crystalline in nature (Dubey et al., 2009). This alsosuggested that the crystallization of bio-organic phase occurred

on the surface of silver nanoparticles (Sathyavathi et al.,2010).

Figure 6 SEM images of silver nanoparticles synthesized using

Murraya paniculata leaf broth (at 20000X magnification).

SEM and EDAX Analysis

Scanning electron microscopy (SEM) analysis of silvernanoparticles was done using Hitachi S-4500 Scanning Electron

Microscopy.The surface morphology (ie. shape and size) of thesilver nanoparticles was shown in Figure 6. The uniform sphericalshape nanoparticles were obtained with the sized ranging from 20-50nm (~). Similarly, the spherical shaped silver nanoparticles with

a diameter ranging from 30-40nm were synthesized usingBoswellia ovalifoliolata (Savithramma et al., 2011); 30-50nm

usingMerremia tridendata (Ganesan et al., 2013); Plant extractsof Carcia papaya (Jain et al., 2009) andEmblica officinalis(Ankamvaret al., 2005).

Figure 7 EDX image of silver nanoparticles synthesized

usingMurraya paniculata leaf broth

The strong silver peak in the EDX- spectrum (Figure 7) confirmsthe presence of elemental silver. Metallic silver nanoparticlesgenerally show typical optical absorption peaks approximately at3KV due to Surface Plasmon Resonance (Magudapathi et al.,2001). The EDX peak of Ag along with Cl, K and Ca as themixed components present in the reaction medium. The strongelemental signal along with weak oxygen that may be originated

from the biomolecules bound to the surface of the nanoparticles

(Song et al., 2009).

Figure 8a TEM image of silver nanoparticles synthesized

from theMurraya paniculata leaf

TEM Analysis

Transmission electron microscopy (TEM) analysis of the samplewas carried out using Philips-Techno 10 instrument operated at an

acceleration voltage of 200KV with resolution of 0.3nm. A dropof leaf broth was placed on carbon coated copper grid andexposed to infrared light for solvent evaporation. Fig. 8a shows

the TEM image at high resolution of silver nanoparticles that arewell dispersed with sizes ranging from 5-50 nm (~).The sizedistribution of silver nanoparticles which ranges between 5 and 50is shown in Fig. 8b. However, the size of the silver nanoparticles

synthesized usingElaeagnus latifoliawas found to range from 30

to 50 nm (Probin et al., 2012). Interestingly, the size of the silvernanoparticles synthesized using Vitex negundo ranged from 10 to

Position [2Theta] (Copper (Cu))

20 30 40 50 60 70

Counts

0

100

200

10

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20nm, some larger and uneven shapes in range of 25-30nm werereported (Veena et al., 2011).

Atomic Absorption Spectroscopy

The amount of silver nanoparticles synthesized was analyzed with

Atomic Absorption Spectroscopy (Schimadzu AAS 6300) aftertermination of the reduction of silver nitrate in order to find outresidual concentration of silver. Interestingly, one gram dry

weight ofMurraya paniculata leaves could synthesize 1.24 mg ofsilver nanoparticles synthesized from this eco- friendly and costeffective method. This is an alternate and simple method for the

synthesis of silver nanoparticles by plants due to its rapidreduction within less time ie., one hour.

10 20 30 4 0 5 0

0

5

10

15

20

25

30

35

No.ofParticles

Size of the nanopart ic les (nm)

Fig. 8b Histogram of silver nanoparticles synthesized from

the Murraya paniculata leaf

CONCLUSION

Thus the present study, the synthesis of silver nanoparticles usingleaf broth ofMurraya paniculatahas been achieved with the rapidreduction of silver nitrate into silver nanoparticles. Interestingly,

the reaction medium changed its colour from pale yellow to dark

brown within one hr of reaction. The UV-Visible spectrum of thereaction medium with silver nanoparticles synthesized using

Murraya paniculata has max at 420 nm and absorbance wasraised upto 1.4 a.u. The FTIR spectrum showed the totaldisappearance of the bands at 538, 914, 2850 and 2922 cm

-1after

bio-reduction may be ascribed to the reduction of silver nitrate

into silver nanoparticles, leading to unsaturated carbonyl groupswith broad peak at 1670 cm

-1. The SEM images obtained in the

present study elucidate the existence of very small and uniformly

spherical nanoparticles with size ranging from 5-50 nm. The XRDand TEM analyses determine the average size of the nanoparticlesis of 31 nm. The strong silver peak obtained from the EDX

spectrum confirms the significant presence of elemental silver.

Finally the Atomic Absorption Spectroscopy analysis of the silvernanoparticles synthesized by the leaf tissue ofMurraya paniculata

brings out the ability of one gram of dry weight ofMurrayapaniculata leaves to synthesize 1.24 mg of silver nanoparticles.Thus the silver nanoparticles are synthesized using a green

resource, Murraya paniculata which are an alternate method tophysical and chemical syntheses due to its cost effective and eco-friendly nature.

Acknowledgements

Authors acknowledge the financial support by Science andEngineering Research Board, Department of Science andTechnology, Government of India, New Delhi to carry out this

work.

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