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Research ArticleAntibacterial Activity of Green Synthesis ofIron Nanoparticles Using Lawsonia inermis andGardenia jasminoides Leaves Extract

Tayyaba Naseem and Muhammad Akhyar Farrukh

Nanochemistry Laboratory Department of Chemistry GC University Lahore Lahore 54000 Pakistan

Correspondence should be addressed to Muhammad Akhyar Farrukh akhyar100gmailcom

Received 22 November 2014 Accepted 29 December 2014

Academic Editor Mallikarjuna N Nadagouda

Copyright copy 2015 T Naseem and M A Farrukh This is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

Recently development of reliable experimental protocols for synthesis of metal nanoparticles with desired morphologies and sizeshas become amajor focus of researchers Green synthesis ofmetallic nanoparticles has accumulated an ultimate interest over the lastdecade due to their distinctive properties that make them applicable in various fields of science and technologyMetal nanoparticlesthat are synthesized by using plants have emerged as nontoxic and ecofriendly In this study a very cheap and simple conventionalheating method was used to obtain the iron nanoparticles (FeNPs) using the leaves extract of Lawsonia inermis and Gardeniajasminoides plant The iron nanoparticles were characterized by thermal gravimetric analysis (TGA) Fourier transform infraredspectroscopy (FT-IR) transmission electron microscopy (TEM) scanning electron microscopy (SEM) atomic force microscopy(AFM) and X-ray diffraction (XRD) The antibacterial activity was studied against Escherichia coli Salmonella enterica Proteusmirabilis and Staphylococcus aureus by using well-diffusion method

1 Introduction

Nanoparticles are considered as important structural massesof nanotechnologyThe unique andmost important propertyof the nanoparticles is that they unveil superior activityThere are remarkable applications of metal nanoparticles inthe areas of diagnostic biological probes catalysis displaydevices and optoelectronics [1] The widespread practicalapplication of metal nanoparticles (lt100 nm) is attributableto a number of their unique properties [2ndash5] Metal nanopar-ticles are widely synthesized using physical and chemical pro-cesses which allow one to acquire particles with the preferredcharacteristics [6ndash8] Several methods like hydrothermal [9]conventional heating [10] anodization [11] deposition pre-cipitation [12] wet oxidation [13] electrodeposition [14] andsonication [15] are being applied to synthesize the nanoparti-cles However these production methods are usually expen-sive and labor-intensive and are potentially hazardous to theenvironment and living organisms

Green synthesis has advances over chemical and physicalmethod as it is cost operative atmosphere friendly and easily

scrabbled up for large scale synthesis and in thismethod thereis no need to use high energy temperature and toxic chemi-cals Green synthesis offer better influence control over crys-tal growth and their steadiness Green synthesized nanopar-ticles are cheap and economical and have many applicationsin science [16ndash19]

From the dawn of civilization human beings have usedvariousmedicinal plants to fight diseases [20] Lawsonia iner-mis is a dwarf shrub commonly known as ldquoMehndi orHennardquoin Pakistan It is renowned worldwide due to its cosmeticuse for the reason of exclusive active principles in the leavesIt contains different variety of molecules which are bioactiveIt is believed to decrease body temperature in situation of highfever and give beautiful and healthy hair Lawsonia inermis isgrown in various dry tropical and subtropical areas of NorthAfrica South Asia South East Asia and theMiddle East [21]Strong antimicrobial anticancer anti-inflammatory anal-gesic antiparasitic and virucidal properties of this plant havebeen reported [22] Lawsonia inermis leaves were studied fortheir antimicrobial prospective and they exhibited notable

Hindawi Publishing CorporationJournal of ChemistryVolume 2015 Article ID 912342 7 pageshttpdxdoiorg1011552015912342

2 Journal of Chemistry

antibacterial activity against Gram-negative bacterial strains[23]

Gardenia jasminoides Ellis is a flowering plant whichhas its place in genus Gardenia and family Rubiaceae Tra-ditionally in many Asian countries it has been used as a folkmedicine [24] This plant has numerous medicinal uses fortreating hemorrhage jaundice toothaches hepatitis sprainswounds and skin conditions [25ndash27] As a hemostatic agentGardenia is very active as well in handling injuries of thejoints tendons and muscles ldquoCrocetinrdquo is an extractedchemical compound from the Gardenia berry from which ayellow-silk dye has been made for this treatment [28]

In nanometer size metallic nanoparticles iron hasreceived special attention because of its physical and chemicalproperties which are determined by its size shape compo-sition crystallinity and structure [29] Bimetallic iron andsilver containing nanoparticles (Fe-Ag NPs) have numerousapplications in optical medical and remediation fields [30]Iron nanoparticles can also be used as oxidant for the synthe-sis of multiwalled carbon nanotubes- (MWCNTs-) corethio-phene polymer-sheath composite nanocables in the presenceof cationic surfactant decyl trimethyl ammonium bromide(DTAB) [31] Against the bacterial strains causing digestiveproblems iron nanoparticles of corn flakes-like morphologygave excellent antibacterial activity [32]

Bacterial resistance to various antibiotics is a seriousclinical dilemma so different antimicrobial activities wereperformed using plants as a source The development in thefield of green chemistry has delivered different nanomaterialsas substitute antibacterial agents In this present study aneffort is made to synthesize iron nanoparticles using leavesextract of Lawsonia inermis and Gardenia jasminoides asreducing agent The characterization of green synthesizediron nanoparticles was characterized by thermal gravimetric(TGA) transmission electron microscopy (TEM) scanningelectronmicroscopy (SEM) Fourier transform infrared spec-troscopy (FT-IR) X-ray diffraction (XRD) and atomic forcemicroscopy (AFM) Also antibacterial activity was studiedagainst human pathogenic Gram-negative (Escherichia coliSalmonella enterica andProteusmirabilis) andGram-positive(Staphylococcus aureus) bacterial strains

2 Materials and Methods

Fresh leaves of Henna and Gardenia plants were collectedfrom various botanical gardens of Lahore Pakistan Sulfuricacid (conc) and iron sulphate (FeSO

4) were purchased from

Merck and all chemicals were used short of any further purifi-cation CORNING (PC-420D) hot plate was used tomaintainthe temperature in the synthesis process Various techniqueswere used to characterize the synthesized samples that isFourier transform infrared spectroscopy (FTIR) on MIDACM2000 which identified various functional groups presentin extract and which determined the presence of metalpowder X-ray diffractometer (XRD) of Xrsquopert PRO andPANalytical equippedwith a copper anode source generatingX-rays having wavelength equal to 154 A The elementalcomposition and morphology were investigated by using

scanning electron microscope-energy-dispersive X-ray spec-troscopy (SEM-EDX) on Hitachi S3400 on an acceleratingvoltage of 150 kVThe size of synthesized sample particles wasdetermined by transmission electron microscope (TEM) ofPhillip CM12 microscope

21 Preparation of Powder Fresh leaves of Henna and Gar-denia plants were softly eroded in deionized water by whichthe dust particles were removed and plants material was thenplaced to dry under sunlight for seven days All of the driedleaves of plantswere groundusing grinder pastel andmortarAfter the process of grinding the leaves powder went throughsieving to get very fine particles of uniform size Nanoparti-cles were synthesized by using sieved powder

22 Preparation of FeNPs A simple conventional heatingmethod was used in the synthesis of iron nanoparticles(FeNPs) by using plant extract Plant extract was prepared bydissolving 2 gm of the sieved powder in 50mL of deionizedwater and the resultingmixture kept on stirring for 3 hours byusing the magnetic stirrer The resulting solution was placedto stable for 1 hour and then filtered 10mL of 001M FeSO

4

solution was used in which plant extract (filtrate) was addedafter every interval of 5 minutes using 2mL in each intervaluntil 50mL resulting mixture was stirred at 70∘CThe differ-ence of temperature was noted after every interval of 5 min-utes The solution was placed to cool down and the productwas parted by centrifugation (10000 rpm) for 2 minutes Theproduct was dried at 50∘C for 3 hours The plant extract(filtrate) acts as reducing capping and stabilizing agent iniron nanoparticles synthesis [33]

23 Antibacterial Studies Three human pathogenic Gram-negative (Escherichia coli Salmonella enterica and Proteusmirabilis) and one Gram-positive (Staphylococcus aureus)bacterial strains were used for antimicrobial study of ironnanoparticles by well-diffusion method [34ndash36] The bacte-rial strains were grown in Luria-Bertani (LB) at 37∘C withcontinuous shaking at 200 rpm for 24 hours 100120583L fromeachbacterial culture was spread on LB agar plates with the help ofL-shaped glass spreader Three wells were developed in eachplate with the help of sterilized steel borer of 8mm diameterand 30 120583L sample suspension was loaded in each well Theplates were incubated for 24 hours at 37∘C Diameter of theinhibition zones was recorded in mm The experiment wasrepeated thrice and the average values were calculated forantibacterial activity

3 Results and Discussion

31 TGA of FeNPs Synthesized Using Henna and GardeniaLeave Extract The weight losses were 12 at 60ndash205∘C and31 at 280ndash510∘Cwhich were due to the removal of moisturehydrogen and three oxygen molecules present in coumaricacid (chemical constituent in Henna) respectively as shownin Figure 1

Figure 2 represents the TGA for the FeNPs synthesizedusing Gardenia plant extract Initial weight was lost at 7

Journal of Chemistry 3

0 200 400 600 800 1000

0

10

20

40

60

80

100

Wei

ght (

)

0

TGA

DSC

Henna leaves

minus30

minus20

minus10

60ndash205∘C

12

280ndash510∘C

31

Removal ofH2O + H2 and3O2 present incoumaric acid

Hea

t flow

(Wg

)

Temperature (∘C)

Figure 1 TGA of FeNPs synthesized using Henna leaves extract

0 200 400 600 800 1000

0

5

20

40

60

80

100

0

Gardenia leaves

90ndash205∘C

7

380ndash690∘C22

2H2 present incatechin

Wei

ght (

)

Hea

t flow

(Wg

)

Temperature (∘C)

minus5

minus10

minus15

minus20

minus25

Removal of H2O and

Figure 2 TGA of FeNPs synthesized using Gardenia plant extract

at 90ndash205∘C because of removal of moisture two hydro-gen molecules present in catechin (chemical constituent ofGardenia) Second weight loss was 22 at 340ndash690∘C andrepresents the removal of two oxygen molecules present inferric sulphate used

32 FTIRAnalysis of FeNPs SynthesizedUsingHenna andGar-denia Leaves Extract Themain constituent of Henna extractis Lawson (2-hydroxy-14-naphthoquinone) It contains ben-zene unit p-benzoquinone unit and phenolic group TheHenna extract was evaporated to dryness to get a solid massIts FTIR spectrum is shown in Figure 3 [37]

The phenolic OndashH stretch appeared at 330238ndash326484 cmminus1 The aromatic C=C stretching frequency whichappeared at 153981 cmminus1 is due to p-coumaric acid presentin the sample The C=O stretching frequency appeared at162264 cmminus1 Thus Lawson was characterized by IR spectro-scopy It was inferred that Lawson has coordinated with Fe0through the phenolic oxygen aromatic ring and C=O groupof the p-benzoquinone resulting in the formation of Fe0which have Lawson as capping and stabilizing agent on theanodic sites of the metal surface The band at 61263 cmminus1

Tran

smitt

ance

()

95

90

85

80

3500 3000 2500 2000 1500 1000

3302

1622

1539

612

(C=O)

(C=C)

(FendashO)

(OndashH)

75

Wavenumber (cmminus1)

33023

1539

612

(C=C)

(FendashO)

(OndashH)

Figure 3 FTIR spectra of FeNPs synthesized using Henna leavesextract

(COOH)

3500 3000 2500 2000 1500 1000

80

85

90

95

3465

1024

622

2950

1720

16201178


Tran

smitt

ance

()

(OndashH) (CndashH)

(CndashO)

(CndashN) (FendashO)

(C=O)

(COOH)

3465

622

2950

16201178

(OndashH) (CndashH)


Figure 4 FTIR spectra of FeNPs synthesized using Gardenia leavesextract

was due to Fe as shown in Figure 3 Thus FTIR spectral studyleads to the conclusion that the fingermark film consists ofFe0 Lawson complex [38]

The FTIR spectra of Fe nanoparticles synthesized withGardenia leave extract are given in Figure 4 The broadabsorption band in the region from 3400 to 3200 cmminus1 rep-resents ndashOH group stretching and another peak at 2700 cmminus1represents CndashH stretching The bands at 164728 and152171 cmminus1 may be assigned respectively to a C=O stretch-ing vibration band (C=O) and a coupled vibration involvingthe bending and theCndashN stretchingmodes of the amido bondof the biomass The peak at 103247 cmminus1 is corresponding tothe vibrations of CndashO inCndashCCOORTheband at 61263 cmminus1was due to Fe vibrations

33 XRD Analysis of FeNPs Synthesized Using Henna andGardenia Leaves Extract The nature and phase compositionof FeNPswere identified byX-ray powder diffractometerwithBraggrsquos angle ranging from 10∘ to 70∘ The presence of Fe innanopowder was confirmed by a series of reflection angles(2120579) at 4434∘ and 6443∘ having hkl values (111) (200) and(202) [39] respectively with cubic plane of Fe [40] as shownin Figure 5


30 40 50 60 70

111

202

400

600

800

200

2120579 (deg)

Cou

nts (

sminus1)

111

202200

Figure 5 XRD pattern of FeNPs synthesized using Henna leavesextract

30 40 50 60 70

111

202

400

600

800

200

2120579 (deg)

Cou

nts (

sminus1)

Figure 6 XRD pattern of FeNPs synthesized using Gardenia leavesextract

The presence of Fe in nanopowder synthesized by Garde-nia leave extract was confirmed by a series of reflection angles(2120579) at 4434∘ and 6443∘ having hkl values (111) (200) and(202) [39] respectively with cubic plane of Fe [40] as shownin Figure 6

34 TEM Image of FeNPs Using Henna and Gardenia LeavesExtract TEM analysis of the FeNPs was performed whichwere formed by using the Henna leaves extract and FeSO

4

salt solutionThe size of iron nanoparticles synthesized usingHenna leaves extract was calculated as 21 nm as shown inFigure 7 While particle size of the same was observed as32 nm when synthesized using the Gardenia leaves extract asshown in Figure 8

35 SEM-EDXAnalysis of FeNPs SynthesizedUsingHenna andGardenia Leaves Extract FeNPs synthesized using extract ofleaves of Henna plant are studied under SEM It indicatesthat nanoparticles formed are agglomerated because of theadhesive nature having morphology of distorted hexagonal-like appearance as shown in Figure 9(a)

Elemental composition of FeNPs synthesized usingHenna leaves extract was determined by using EDX analysisIt was observed that the percentage of iron is 686 carbonis 5459 oxygen is 3657 magnesium and phosphorus are068 and potassium is 063 as shown in Figure 9(b) Mgand carbon are due to plant constituents

100 nm

Figure 7 TEM analysis of FeNPs synthesized using Henna leavesextract

100 nm

Figure 8 TEM image of FeNPs synthesized using Gardenia leavesextract

FeNPs synthesized using extract of leaves of Gardeniaplant are studied under SEM and shown in Figure 10(a)It indicates that nanoparticles formed are agglomeratedbecause of the adhesive nature having morphology of shat-tered rock-like appearance

Elemental composition of FeNPs synthesized using Gar-denia leaves extract was also determined by using EDXanalysis

Elemental composition was found as percentage of ironis 468 carbon is 5079 oxygen is 4137 aluminium is076 silicon is 157 and potassium is 083 as shown inFigure 10(b)

351 Antibacterial Results Iron nanoparticles were synthe-sized using five different plants that is Lawsonia inermisGardenia jasminoides Azadirachta indica and Camellia sin-ensis leaves extract and Cinnamon zeylanicum barks extractand it was found that all are susceptible to all bacterial strainsHere we are representing the results of mainly the two ofthem that is iron nanoparticles synthesized using Lawsoniainermis and Gardenia jasminoides (Table 1) and the compar-ison is given in Figure 11 FeNPs of Gardenia jasminoideswere more potent against Staphylococcus aureus with zoneof inhibition (ZOI) 16mm whereas for Lawsonia inermis itwas 15mm as shown in Figure 12 Against Escherichia coliSalmonella enterica and Proteus mirabilis the ZOI of ironnanoparticles of Lawsonia inermis and Gardenia jasminoidesleaves extract was 14mmand 15mm 9mmand 12mm 11mmand 13mm respectively


(a)

0 05 10 15 20 25 30(keV)

O

Mg

Fe

(b)

Figure 9 (a) SEM image and (b) EDX graph of FeNPs synthesizedusing Henna leaves extract

Table 1 Antibacterial activity of iron nanoparticles of Lawsoniainermis and Gardenia jasminoides leaves extract (30 120583LmL)

MicroorganismsLeaves extract

Lawsonia inermis Gardenia jasminoidesZone of inhibition (mm)

Bacterial strainsE coli (minus) 14 15S enterica (minus) 09 12P mirabilis (minus) 11 13S aureus (+) 15 16

4 Conclusion

Due to the rich biodiversity of plants the green world haspotential for the synthesis of noble metal nanoparticles Ironnanoparticles with an average size of 21 nm and 32 nm weresynthesized using Lawsonia inermis and Gardenia jasmi-noides leaves extract respectively Green synthesized ironnanoparticles in the present study show good antibacterial

(a)

0 05 10 15 20 25 30(keV)

Fe

SiAl

O

Fe

SiAl

(b)

Figure 10 (a) SEM image and (b) EDX graph of FeNPs synthesizedusing Gardenia leaves extract

Zone

of i

nhib

ition

(ZO

I)

Esch

erich

ia co

li

Salm

onell

a en

teric

a

Prot

eus m

irabi

lis

Stap

hylo

cocc

us a

ureu

s

Lawsonia inermisGardenia jasminoides

Bacterial strains

0

2

4

6

8

10

12

14

16

Figure 11 Comparison of antibacterial activity of iron nanoparticlesof Lawsonia inermis andGardenia jasminoides leaves extract againstbacterial strains

activity against the human pathogens Escherichia coli andStaphylococcus aureus As Lawsonia inermis and Gardenia


(a) (b)

Figure 12 (a) Zone inhibition of FeNPs against P4= Escherichia coli P

5= Salmonella enterica B = Lawsonia inermis and D = Gardenia

jasminoides (b) Zone inhibition of FeNPs against P4= Staphylococcus aureus P

5= Proteus mirabilis B = Lawsonia inermis and D =Gardenia

jasminoides

jasminoides have been used in folk medicines these greeniron nanoparticles of Lawsonia inermis and Gardenia jasmi-noides have potential biomedical activities and have severalpaybacks such as suitability for medical and pharmaceuticalsubmissions

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

References

[1] J Wagner T Kirner G Mayer J Albert and J M KohlerldquoGeneration of metal nanoparticles in a microchannel reactorrdquoChemical Engineering Journal vol 101 no 1mdash3 pp 251ndash2602004

[2] M C Roco ldquoNanotechnology convergence with modern biol-ogy and medicinerdquo Current Opinion in Biotechnology vol 14no 3 pp 337ndash346 2003

[3] L Zhang F X Gu J M Chan A Z Wang R S Langer and OC Farokhzad ldquoNanoparticles in medicine therapeutic applica-tions and developmentsrdquo Clinical Pharmacology andTherapeu-tics vol 83 no 5 pp 761ndash769 2008

[4] M-C Daniel and D Astruc ldquoGold nanoparticles assemblysupramolecular chemistry quantum-size-related propertiesand applications toward biology catalysis and nanotechnologrdquoChemical Reviews vol 104 no 1 pp 293ndash346 2004

[5] T S Wong and U Schwaneberg ldquoProtein engineering inbioelectrocatalysisrdquo Current Opinion in Biotechnology vol 14no 6 pp 590ndash596 2003

[6] J H Fendler Ed Nanoparticles and Nanostructured FilmsPreparation Characterization and Applications John Wiley ampSons New York NY USA 1998

[7] A Panacek L Kvıtek R Prucek et al ldquoSilver colloid nanoparti-cles synthesis characterization and their antibacterial activityrdquoThe Journal of Physical Chemistry B vol 110 no 33 pp 16248ndash16253 2006

[8] S Kundu V Maheshwari S Niu and R F Saraf ldquoPoly-electrolyte mediated scalable synthesis of highly stable silvernanocubes in less than a minute using microwave irradiationrdquoNanotechnology vol 19 no 6 Article ID 065604 2008

[9] H Perveen M A Farrukh M Khaleeq-ur-Rahman B Munirand M A Tahir ldquoSynthesis structural properties and catalytic

activity of MgO-SnO2nanocatalystsrdquo Russian Journal of Physi-

cal Chemistry A vol 89 no 1 pp 99ndash107 2015[10] M Shahid M A Farrukh A A Umar and M Khaleeq-Ur-

Rahman ldquoSolvent controlled synthesis of CaO-MgOnanocom-posites and their application in the photodegradation of organicpollutants of industrial wasterdquo Russian Journal of PhysicalChemistry A vol 88 no 5 pp 836ndash844 2014

[11] M A Farrukh C-K Thong R Adnan and M A Kamar-ulzaman ldquoPreparation and characterization of zinc oxide nano-flakes using anodization method and their photodegradationactivity on methylene bluerdquo Russian Journal of Physical Chem-istry A vol 86 no 13 pp 2041ndash2048 2012

[12] H Yazid R Adnan and M A Farrukh ldquoGold nanoparticlessupported on titania for the reduction of p-nitrophenolrdquo IndianJournal of ChemistrymdashSection A Inorganic Physical Theoreticaland Analytical Chemistry vol 52 no 2 pp 184ndash191 2013

[13] K M A Saron M R Hashim and M A Farrukh ldquoStresscontrol in ZnO films on GaNAl

2O3via wet oxidation of Zn

under various temperaturesrdquo Applied Surface Science vol 258no 13 pp 5200ndash5205 2012

[14] E M Mkawi K Ibrahim M K M Ali M A Farrukh and AS Mohamed ldquoSynthesized and characterization of Cu

2ZnSnS

4

(CZTS) thin films deposited by electrodeposition methodrdquoApplied Mechanics and Materials vol 343 pp 85ndash89 2013

[15] IMuneerM A Farrukh S JavaidM Shahid andMKhaleeq-ur-Rahman ldquoSynthesis of Gd

2O3Sm2O3nanocomposite via

sonication and hydrothermal methods and its optical prop-ertiesrdquo Superlattices and Microstructures vol 77 pp 256ndash2662015

[16] R Brayner F Fievet and T Coradin ldquoSynthesis of organicand bioorganic nanoparticles an overview of the preparationmethodsrdquo inNanomaterials ADanger or a Promise AChemicaland Biological Perspective J Allouche Ed pp 27ndash74 SpringerLondon UK 2013

[17] V Parashar R Parashar B Sharma and A C Pandey ldquoParthe-nium leaf extract mediated synthesis of silver nanoparticlesa novel approach towards weed utilizationrdquo Digest Journal ofNanomaterials and Biostructures vol 4 no 1 pp 45ndash50 2009

[18] T Klaus-Joerger R Joerger E Olsson and C-G GranqvistldquoBacteria as workers in the living factory metal-accumulatingbacteria and their potential for materials sciencerdquo Trends inBiotechnology vol 19 no 1 pp 15ndash20 2001

[19] K S Kavitha S Baker D Rakshith et al ldquoPlants as green sourcetowards synthesis of nanoparticlesrdquo International Research Jour-nal of Biological Sciences vol 2 no 6 pp 66ndash76 2013


[20] U Bandyopadhyay K Biswas I Chattopadhyay and R KBanerjee ldquoBiological activities and medicinal properties ofneem (Azadirachta indica)rdquo Current Science vol 82 no 11 pp1336ndash1345 2002

[21] W-H Chun Y-C Chang L-J Yang et al ldquoClinicopathologicfeatures of skin reactions to temporary tattoos and analysis ofpossible causesrdquoArchives of Dermatology vol 138 no 1 pp 88ndash92 2002

[22] O A Habbal A A Al-Jabri and A G El-Hag ldquoAntimicrobialproperties of Lawsonia inermis (henna) a reviewrdquo AustralianJournal of Medical Herbalism vol 19 no 3 pp 114ndash125 2007

[23] I Abulyazid E M E Mahdy and R M Ahmed ldquoBiochemicalstudy for the effect of henna (Lawsonia inermis) on Escherichiacolirdquo Arabian Journal of Chemistry vol 6 no 3 pp 265ndash2732013

[24] T Debnath P J Park N C Deb Nath N B Samad HW Parkand B O Lim ldquoAntioxidant activity of Gardenia jasminoidesEllis fruit extractsrdquo Food Chemistry vol 128 no 3 pp 697ndash7032011

[25] S J Choi M-J Kim H J Heo et al ldquoAmeliorating effect ofGardenia jasminoides extract on amyloid beta peptide-inducedneuronal cell deficitrdquoMolecules and Cells vol 24 no 1 pp 113ndash118 2007

[26] R A A Lelono S Tachibana and K Itoh ldquoIsolation ofantifungal compounds from Gardenia jasminoidesrdquo PakistanJournal of Biological Sciences vol 12 no 13 pp 949ndash956 2009

[27] P S George G A Ravishankar and L V VenkataramanldquoClonal multiplication of Gardenia jasminoides Ellis throughaxillary bud culturerdquo Plant Cell Reports vol 13 no 1 pp 59ndash621993

[28] R FarzinebrahimiTissue culture and biological activities of Gar-denia jasminoides Ellis [MS thesis] University ofMalaya KualaLumpur Malaysia 2012

[29] S AMahdyQ J Raheed and P T Kalaichelvan ldquoAntimicrobialactivity of zero valent iron nanoparticlesrdquo International Journalof Modern Engineering Research vol 2 no 1 pp 578ndash581 2012

[30] V K Sharma K M Siskova and R Zboril ldquoMagnetic bimetal-lic FeAg nanoparticles decontamination and antimicrobialagentsrdquo in Interactions of Nanomaterials with Emerging Environ-mental Contaminants vol 1150 of ACS Symposium Series chap-ter 11 pp 193ndash209 American Chemical Society WashingtonDC USA 2013

[31] K R Reddy H M Jeong and A V Raghu ldquoSynthesis ofMWCNTs-corethiophene polymer-sheath composite nanoca-bles by a cationic surfactant-assisted chemical oxidative poly-merization and their structural propertiesrdquo Journal of PolymerScience Part A Polymer Chemistry vol 48 no 7 pp 1477ndash14842010

[32] OMahapatra S Ramaswamy S V KNune T Yadavalli andCGopalakrishnan ldquoCorn flake-likemorphology of iron nanopar-ticles and its antibacterial propertyrdquoThe Journal of General andApplied Microbiology vol 57 no 1 pp 59ndash62 2011

[33] V V Makarov A J Love O V Sinitsyna et al ldquoldquoGreenrdquo nan-otechnologies synthesis of metal nanoparticles using plantsrdquoActa Naturae vol 6 no 1 pp 35ndash44 2014

[34] S Ahmad M A Farrukh M Khan M Khaleeq-ur-Rahmanand M A Tahir ldquoSynthesis of iron oxidendashtin oxide nanoparti-cles and evaluation of their activities against different bacterialstrainsrdquo Canadian Chemical Transactions vol 2 no 2 pp 122ndash133 2014

[35] S Shah M N Chandraprabha and K Samrat ldquoSynthesisand characterization of zero valent iron nanoparticles and

assessment of its antibacterial activityrdquo International Review ofApplied Biotechnology andBiochemistry vol 2 no 1 pp 145ndash1512014

[36] M Sampath R Vijayan E Tamilarasu A Tamilselvan and BSengottuvelan ldquoGreen synthesis of novel jasmine bud-shapedcopper nanoparticlesrdquo Journal of Nanotechnology vol 2014Article ID 626523 7 pages 2014

[37] D P Suhas H M Jeong T M Aminabhavi and A V RaghuldquoPreparation and characterization of novel polyurethanes con-taining 441015840-oxy-14-diphenyl bis(nitromethylidine)diphenolschiff base diolrdquo Polymer Engineering and Science vol 54 no1 pp 24ndash32 2014

[38] S Rajendran M Agasta R B Devi B S Devi K Rajam andJ Jeyasundari ldquoCorrosion inhibition by an aqueous extract ofHenna leaves (Lawsonia Inermis L)rdquo Zastita Materijala vol 50pp 77ndash84 2009

[39] M A Farrukh S Ali and M K Rahman ldquoPhotodegradationof 246-trinitrophenol catalyzed byZnMgOnanoparticles pre-pared in aqueous-organicmediumrdquoKorean Journal of ChemicalEngineering vol 30 no 11 pp 2100ndash2107 2013

[40] Z S Basinski W Hume-Rothery and A L Sutton ldquoThe latticeexpansion of iron locality synthetic sample at 119879 = 1033KrdquoProceedings of the Royal Society of London vol 229 pp 459ndash4671955

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Analytical ChemistryInternational Journal of


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antibacterial activity against Gram-negative bacterial strains[23]

Gardenia jasminoides Ellis is a flowering plant whichhas its place in genus Gardenia and family Rubiaceae Tra-ditionally in many Asian countries it has been used as a folkmedicine [24] This plant has numerous medicinal uses fortreating hemorrhage jaundice toothaches hepatitis sprainswounds and skin conditions [25ndash27] As a hemostatic agentGardenia is very active as well in handling injuries of thejoints tendons and muscles ldquoCrocetinrdquo is an extractedchemical compound from the Gardenia berry from which ayellow-silk dye has been made for this treatment [28]

In nanometer size metallic nanoparticles iron hasreceived special attention because of its physical and chemicalproperties which are determined by its size shape compo-sition crystallinity and structure [29] Bimetallic iron andsilver containing nanoparticles (Fe-Ag NPs) have numerousapplications in optical medical and remediation fields [30]Iron nanoparticles can also be used as oxidant for the synthe-sis of multiwalled carbon nanotubes- (MWCNTs-) corethio-phene polymer-sheath composite nanocables in the presenceof cationic surfactant decyl trimethyl ammonium bromide(DTAB) [31] Against the bacterial strains causing digestiveproblems iron nanoparticles of corn flakes-like morphologygave excellent antibacterial activity [32]

Bacterial resistance to various antibiotics is a seriousclinical dilemma so different antimicrobial activities wereperformed using plants as a source The development in thefield of green chemistry has delivered different nanomaterialsas substitute antibacterial agents In this present study aneffort is made to synthesize iron nanoparticles using leavesextract of Lawsonia inermis and Gardenia jasminoides asreducing agent The characterization of green synthesizediron nanoparticles was characterized by thermal gravimetric(TGA) transmission electron microscopy (TEM) scanningelectronmicroscopy (SEM) Fourier transform infrared spec-troscopy (FT-IR) X-ray diffraction (XRD) and atomic forcemicroscopy (AFM) Also antibacterial activity was studiedagainst human pathogenic Gram-negative (Escherichia coliSalmonella enterica andProteusmirabilis) andGram-positive(Staphylococcus aureus) bacterial strains

2 Materials and Methods

Fresh leaves of Henna and Gardenia plants were collectedfrom various botanical gardens of Lahore Pakistan Sulfuricacid (conc) and iron sulphate (FeSO

4) were purchased from

Merck and all chemicals were used short of any further purifi-cation CORNING (PC-420D) hot plate was used tomaintainthe temperature in the synthesis process Various techniqueswere used to characterize the synthesized samples that isFourier transform infrared spectroscopy (FTIR) on MIDACM2000 which identified various functional groups presentin extract and which determined the presence of metalpowder X-ray diffractometer (XRD) of Xrsquopert PRO andPANalytical equippedwith a copper anode source generatingX-rays having wavelength equal to 154 A The elementalcomposition and morphology were investigated by using

scanning electron microscope-energy-dispersive X-ray spec-troscopy (SEM-EDX) on Hitachi S3400 on an acceleratingvoltage of 150 kVThe size of synthesized sample particles wasdetermined by transmission electron microscope (TEM) ofPhillip CM12 microscope

21 Preparation of Powder Fresh leaves of Henna and Gar-denia plants were softly eroded in deionized water by whichthe dust particles were removed and plants material was thenplaced to dry under sunlight for seven days All of the driedleaves of plantswere groundusing grinder pastel andmortarAfter the process of grinding the leaves powder went throughsieving to get very fine particles of uniform size Nanoparti-cles were synthesized by using sieved powder

22 Preparation of FeNPs A simple conventional heatingmethod was used in the synthesis of iron nanoparticles(FeNPs) by using plant extract Plant extract was prepared bydissolving 2 gm of the sieved powder in 50mL of deionizedwater and the resultingmixture kept on stirring for 3 hours byusing the magnetic stirrer The resulting solution was placedto stable for 1 hour and then filtered 10mL of 001M FeSO

4

solution was used in which plant extract (filtrate) was addedafter every interval of 5 minutes using 2mL in each intervaluntil 50mL resulting mixture was stirred at 70∘CThe differ-ence of temperature was noted after every interval of 5 min-utes The solution was placed to cool down and the productwas parted by centrifugation (10000 rpm) for 2 minutes Theproduct was dried at 50∘C for 3 hours The plant extract(filtrate) acts as reducing capping and stabilizing agent iniron nanoparticles synthesis [33]

23 Antibacterial Studies Three human pathogenic Gram-negative (Escherichia coli Salmonella enterica and Proteusmirabilis) and one Gram-positive (Staphylococcus aureus)bacterial strains were used for antimicrobial study of ironnanoparticles by well-diffusion method [34ndash36] The bacte-rial strains were grown in Luria-Bertani (LB) at 37∘C withcontinuous shaking at 200 rpm for 24 hours 100120583L fromeachbacterial culture was spread on LB agar plates with the help ofL-shaped glass spreader Three wells were developed in eachplate with the help of sterilized steel borer of 8mm diameterand 30 120583L sample suspension was loaded in each well Theplates were incubated for 24 hours at 37∘C Diameter of theinhibition zones was recorded in mm The experiment wasrepeated thrice and the average values were calculated forantibacterial activity

3 Results and Discussion

31 TGA of FeNPs Synthesized Using Henna and GardeniaLeave Extract The weight losses were 12 at 60ndash205∘C and31 at 280ndash510∘Cwhich were due to the removal of moisturehydrogen and three oxygen molecules present in coumaricacid (chemical constituent in Henna) respectively as shownin Figure 1

Figure 2 represents the TGA for the FeNPs synthesizedusing Gardenia plant extract Initial weight was lost at 7


0 200 400 600 800 1000

0

10

20

40

60

80

100

Wei

ght (

)

0

TGA

DSC

Henna leaves

minus30

minus20

minus10

60ndash205∘C

12

280ndash510∘C

31


Hea

t flow

(Wg

)

Temperature (∘C)


0 200 400 600 800 1000

0

5

20

40

60

80

100

0

Gardenia leaves

90ndash205∘C

7

380ndash690∘C22


Wei

ght (

)

Hea

t flow

(Wg

)

Temperature (∘C)

minus5

minus10

minus15

minus20

minus25

Removal of H2O and





Tran

smitt

ance

()

95

90

85

80

3500 3000 2500 2000 1500 1000

3302

1622

1539

612

(C=O)

(C=C)

(FendashO)

(OndashH)

75


33023

1539

612

(C=C)

(FendashO)

(OndashH)


(COOH)

3500 3000 2500 2000 1500 1000

80

85

90

95

3465

1024

622

2950

1720

16201178


Tran

smitt

ance

()

(OndashH) (CndashH)

(CndashO)


(C=O)

(COOH)

3465

622

2950

16201178

(OndashH) (CndashH)







30 40 50 60 70

111

202

400

600

800

200

2120579 (deg)

Cou

nts (

sminus1)

111

202200


30 40 50 60 70

111

202

400

600

800

200

2120579 (deg)

Cou

nts (

sminus1)




4




100 nm


100 nm







(a)

0 05 10 15 20 25 30(keV)

O

Mg

Fe

(b)






4 Conclusion


(a)

0 05 10 15 20 25 30(keV)

Fe

SiAl

O

Fe

SiAl

(b)


Zone

of i

nhib

ition

(ZO

I)

Esch

erich

ia co

li

Salm

onell

a en

teric

a

Prot

eus m

irabi

lis

Stap

hylo

cocc

us a

ureu

s


Bacterial strains

0

2

4

6

8

10

12

14

16




(a) (b)





jasminoides




References



















2ZnSnS

4









































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


0 200 400 600 800 1000

0

10

20

40

60

80

100

Wei

ght (

)

0

TGA

DSC

Henna leaves

minus30

minus20

minus10

60ndash205∘C

12

280ndash510∘C

31


Hea

t flow

(Wg

)

Temperature (∘C)


0 200 400 600 800 1000

0

5

20

40

60

80

100

0

Gardenia leaves

90ndash205∘C

7

380ndash690∘C22


Wei

ght (

)

Hea

t flow

(Wg

)

Temperature (∘C)

minus5

minus10

minus15

minus20

minus25

Removal of H2O and





Tran

smitt

ance

()

95

90

85

80

3500 3000 2500 2000 1500 1000

3302

1622

1539

612

(C=O)

(C=C)

(FendashO)

(OndashH)

75


33023

1539

612

(C=C)

(FendashO)

(OndashH)


(COOH)

3500 3000 2500 2000 1500 1000

80

85

90

95

3465

1024

622

2950

1720

16201178


Tran

smitt

ance

()

(OndashH) (CndashH)

(CndashO)


(C=O)

(COOH)

3465

622

2950

16201178

(OndashH) (CndashH)







30 40 50 60 70

111

202

400

600

800

200

2120579 (deg)

Cou

nts (

sminus1)

111

202200


30 40 50 60 70

111

202

400

600

800

200

2120579 (deg)

Cou

nts (

sminus1)




4




100 nm


100 nm







(a)

0 05 10 15 20 25 30(keV)

O

Mg

Fe

(b)






4 Conclusion


(a)

0 05 10 15 20 25 30(keV)

Fe

SiAl

O

Fe

SiAl

(b)


Zone

of i

nhib

ition

(ZO

I)

Esch

erich

ia co

li

Salm

onell

a en

teric

a

Prot

eus m

irabi

lis

Stap

hylo

cocc

us a

ureu

s


Bacterial strains

0

2

4

6

8

10

12

14

16




(a) (b)





jasminoides




References



















2ZnSnS

4









































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


30 40 50 60 70

111

202

400

600

800

200

2120579 (deg)

Cou

nts (

sminus1)

111

202200


30 40 50 60 70

111

202

400

600

800

200

2120579 (deg)

Cou

nts (

sminus1)




4




100 nm


100 nm







(a)

0 05 10 15 20 25 30(keV)

O

Mg

Fe

(b)






4 Conclusion


(a)

0 05 10 15 20 25 30(keV)

Fe

SiAl

O

Fe

SiAl

(b)


Zone

of i

nhib

ition

(ZO

I)

Esch

erich

ia co

li

Salm

onell

a en

teric

a

Prot

eus m

irabi

lis

Stap

hylo

cocc

us a

ureu

s


Bacterial strains

0

2

4

6

8

10

12

14

16




(a) (b)





jasminoides




References



















2ZnSnS

4









































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


(a)

0 05 10 15 20 25 30(keV)

O

Mg

Fe

(b)






4 Conclusion


(a)

0 05 10 15 20 25 30(keV)

Fe

SiAl

O

Fe

SiAl

(b)


Zone

of i

nhib

ition

(ZO

I)

Esch

erich

ia co

li

Salm

onell

a en

teric

a

Prot

eus m

irabi

lis

Stap

hylo

cocc

us a

ureu

s


Bacterial strains

0

2

4

6

8

10

12

14

16




(a) (b)





jasminoides




References



















2ZnSnS

4









































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


(a) (b)





jasminoides




References



















2ZnSnS

4









































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

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