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Applied Surface Science 317 (2014) 914–922

Contents lists available at ScienceDirect

Applied Surface Science

journa l h om epa ge: www.elsev ier .com/ locate /apsusc

ynthesis and characterization of amoxicillin derived silveranoparticles: Its catalytic effect on degradation of someharmaceutical antibiotics

. Junejoa,b, A. Günerc,∗, A. Baykalb

National Center of Excellence in Analytical Chemistry, University of Sindh Jamshoro, Jamshoro 76080, PakistanDepartment of Chemistry, Fatih University, Buyukcekmece, 34500 Istanbul, TurkeyDepartment of Biology, Fatih University, Buyukcekmece, 34500 Istanbul, Turkey

r t i c l e i n f o

rticle history:eceived 22 April 2014eceived in revised form 21 August 2014ccepted 23 August 2014vailable online 1 September 2014

eywords:ntibioticsilver nanoparticleseducing agentntimicrobial activityastewater

a b s t r a c t

We synthesized novel amoxicillin derived silver nanoparticles (Amp-Ag (0) NPs) in aqueous solutionby one-pot simple synthetic method by reducing silver nitrate by the help of amoxicillin antibiotic asa reducing/capping agent and NaOH as the catalyst for reaction enhancement. The formation of theAmp-Ag (0) NPs was monitored using UV–Vis absorption spectroscopy which confirmed the formationof Amp-Ag (0) NPs by exciting the typical surface plasmon absorption maxima at 404 nm. Transmissionelectron microscopy (TEM) confirmed the spherical morphology and monodispersed Amp-Ag (0) NPswith particle size 6.87 ± 2.2 nm. The antibacterial activities of the antibiotics were evaluated againstGram-negative bacteria Escherichia coli, Salmonella enteritidis, Pseudomonas aeruginosa and Gram-positivebacteria Streptococcus pneumonia, Streptococcus pyogenes, Staphylococcus aureus by the disk diffusionmethod. Whereas standard antibiotics showed normal zone of inhibition, the reduced ones with Amp-Ag(0) NPs showed no inhibition zone. The antimicrobial results therefore reveal that newly synthesizedAmp-Ag (0) NPs had an excellent catalytic activity as catalyst for the 100% reduction of antibiotics i.e.

cefdinir, cefditoren, cefiximee, ceftriaxone sodium and doxycycline, which was carried out in just 2–5 min.They were recovered easily from reaction medium and reused with enhanced catalytic potential fivetimes. Based upon these results it has been concluded that Amp-Ag (0) NPs are novel, rapid, and highlycost-effective for environmental safety against pollution by antibiotics in wastewater and extendable forcontrol of other reducible contaminants as well.

© 2014 Elsevier B.V. All rights reserved.

. Introduction

In the recent years, there has been an increasing interest aboutiffusion of pharmaceuticals into the environment and related risks

n highly developed countries [1]. A large diversity of these pharma-eutical compounds has been repeatedly found in river streams andewage treatment plants effluents [2]. These compounds enter intoquatic environment after their ingestion and subsequent excre-ion without any modifications or in the form of non-metabolizedarent compounds [3]. Along with diverse pharmaceutical com-ounds present in the environment, special emphasis has been

iven to antibiotics, which are the most often discussed pharma-euticals because of their potential role in the development ofntibiotic resistant bacteria [4]. Antibiotics are widely used for

∗ Corresponding author. Tel.: +90 212 8863300x2099; fax: +90 212 8663402.E-mail addresses: aguner@fatih.edu.tr, aguner@outlook.com (A. Güner).

ttp://dx.doi.org/10.1016/j.apsusc.2014.08.133169-4332/© 2014 Elsevier B.V. All rights reserved.

human medicine and agriculture which can easily enters in theaquatic environment via wastewater and other sources, where theyhave been found at measurable concentrations (according to datasupplied by the European Federation of Animal Health in 1999 therewere a total of 13,216 ton of antibiotics used in the European Union)[5]. An antibiotic does not metabolize 100% by humans and ani-mals body [6]. Some of the active quantity is excreted after bodymetabolism and may find their way to municipal sewage treatmentplants from the excretions [7].

Alaton and Dogruel [8] reported that the concentration ofpenicillin formulation in real antibiotics wastewater may be upto 400 mg/L. Antibiotics wastewater has high chemical oxygendemand (COD) and low biochemical oxygen demand (BOD), andhence biological processes are unsuitable for the wastewater treat-

ment. Advanced oxidation processes (AOPs) have proved to behighly effective in the degradation of most of the pollutantsin wastewaters [9]. Large amount of antibiotics in the environ-ment could affect terrestrial and aquatic organisms [10–12], alter

face Sc

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Y. Junejo et al. / Applied Sur

icrobial activity and community composition [13], and lead torevalence of bacterial resistance to antibiotics [14–16]. In ordero reduce the negative impacts on the environment and humanealth, it is necessary to understand the input sources for variouslasses of antibiotics.

Amoxicillin derived silver nanoparticles were synthesized byur research group and used as catalyst for the degradation somezo dyes and degradation of some pharmaceutical Antibiotics forhe first time. The synthesis of Ag (0) NPs does not required sophisti-ated apparatus and expensive chemicals. The duration of synthesiss very short [17–19].

Up to now, most studies on antibiotics in wastewater treatmentlant focused on the concentration levels of certain antibiotics suchs sulfamethoxazole, tetracycline and ciprofloxacin in influent,ffluent and sludge [20–23]. Several recent studies reported theegradation of some antibiotics in activated sludge systems in lab-ratory conditions [24–26]. However, little is known about theccurrence and removal of different classes of antibiotics together.urthermore, aqueous phase removal percentage is often useds the only parameter for elimination efficiencies of antibioticsn wastewater treatment plant [27]. In fact an aqueous removalercentage cannot comprehensively evaluate the elimination ofntibiotics in wastewater treatment plant, since some antibioticsave a tendency to partition into the sludge [28]. Mass balancepproach would be an effective way to understand the fate ofntibiotics in wastewater treatment plant and their mass loading tohe receiving environment [29]. Elimination of organic compoundsn the environment is the result of different processes. These pro-esses can be biotic ones, i.e. biodegradation by bacteria and fungi.on-biotic elimination processes are sorption, hydrolysis, photo-

ysis, oxidation and reduction.A number of methods are existing for the synthesis of Ag (0)

Ps; hydrothermal [30], sonochemical [31], electron beam irradia-ion [32], extraction of leave [33], seed extract methods [34] and son. As compared these synthesis routes, one of the green, cheapernd simplest methods for the synthesis of Ag (0) NPs is the use ofntibiotic as reducing and capping agent.

Here, we report a very simple one-pot method for the syn-hesis of silver nanoparticles by green and biological methody amoxicillin antibiotic and their application as heterogeneous,ecoverable and reusable catalyst for remarkably faster and com-lete reduction of antibiotic from wastewater in the presencef NaBH4. The entire research work is based on excellent econ-my of the process in terms of using cheaper chemicals, facilend simpler synthesis of catalyst, with shorter time for productormation (quicker procedure) and easy recovery and recycle ofatalyst.

. Materials and methods

.1. Chemicals

All chemicals and reagents used in this study were of high puritynd Analytical grade. Erythromycin was purchased from Fluka,aOH, NaBH4, and AgNO3 from Merck. Standards of all antibiotics

uch as amoxicillin, ceftriaxone sodium, doxycycline, cefiximee,efdinir, cefditoren were gifted from Turkish medicine manufac-ure (DEVA).

.2. Synthesis of amoxicillin-derived Ag (0) NPs

Amp-Ag (0) NPs were synthesized by taking 0.4 ml solution ofgNO3 in a 20 ml test tube, followed by deionized water then 0.3 mlmpicillin standard and then 0.2 ml NaOH, final volume was 10 ml.

ience 317 (2014) 914–922 915

2.3. Instrumentations

X-ray powder diffraction (XRD) analysis was conducted on aRigaku Smart Lab Diffractometer operated at 40 kV and 35 mA usingCu K� radiation.

High resolution transmission electron microscopy (HRTEM)analysis was performed using a JEOL JEM 2100 microscope. A dropof diluted sample in alcohol was dropped on a TEM grid.

Fourier transform infrared (FT-IR) spectra were recorded intransmission mode with a Bruker ATR-FT-IR spectrometer. FT-IRspectra in the range 4000–400 cm−1 were recorded in order toinvestigate the nature of the chemical bonds formed.

The UV–Vis measurement was done using a Shimadzu UV–Vis2600.

2.4. Catalysis study of silver nanoparticles

The catalytic effect of silver nanoparticles was monitored for theceftriaxone sodium, doxycycline, cefiximee, cefdinir, cefditoren.The catalytic reduction of these antibiotics was carried out in astandard quartz cell with 1 cm path length and about 3 ml volumecontaining Ag (0) NPs encumbered pieces of glass. The reactionwas performed in the presence of small quantity of NaBH4 onlyand Amp-Ag (0) NPs. Under optimized conditions, the catalyticreaction procedures were as follows: ceftriaxone sodium, doxycy-cline, cefiximee, cefdinir, and cefditoren. The amount of individualreagent was taken as 0.2 ml from 100 �M antibiotic was taken inquartz cell followed by the addition of 2.80 ml of milli Q water thenby 0.1 ml of 0.1 M NaBH4. Carefully and immediately after additionof 0.10 mg, Amp-Ag (0) NPs were immobilized on pieces of glassand put in quartz cell for reduction process. The absorption spec-tra were monitored by a UV/Vis spectrophotometer with a timeinterval of 50 s in a scanning range of 200–600 nm.

2.5. Antimicrobial activity

All the compounds have been screened for antimicrobial activityusing the qualitative disk diffusion method by measuring the inhi-bition zone in mm against Escherichia coli ATCC 25922, Salmonellaenteritidis ATCC 4931, Pseudomonas aeruginosa ATCC 27853 (asGram-negative bacteria) and Streptococcus pneumonia ATCC 49619,Streptococcus pyogenes ATCC 21060, and Staphylococcus aureusATCC 25923 (as Gram-positive bacteria) in Tryptic soy agar (BDDifcoTM) medium. The sterilized agar media were poured into Petriplates and allowed to solidify. On the surface of the media freshmicroorganism cultures (108 CFU ml−1, CFU, colony-forming units)were spread with the help of sterilized L-shaped glass loop. A cylin-der glass pipette of 5 mm diameter (presterilized) was used to borecavities. Both the standard antibiotics such as cefdinir, cefditoren,cefixime, ceftriaxone sodium and doxycycline and their reducedwith Amp-Ag (0) NPs (10 �g/ml each) were placed serially in thecavities with the help of micropipette and allowed to diffuse for 1 h.The standard antibiotics were used as controls. These plates wereincubated at 37 ◦C for 18–24 h for antibacterial activity. The inhi-bition zone of the compounds were measured and evaluated. Eachprepared sample was measured in three replicates.

2.6. Recovery and reuse of Amp-Ag (0) NPs as catalyst

After completion of catalytic reaction, the broken pieces con-taining immobilized Amp-Ag (0) NPs were taken out of the cell,

washed three times by deionized water and dried under N2 asbefore. These glass pieces having Amp-Ag (0) NPs were reused ina fresh antibiotic solution for checking its second performance ascatalyst in a solution prepared by procedure given in 2.3. Similar

916 Y. Junejo et al. / Applied Surface Science 317 (2014) 914–922

300 35 0 40 0 45 0 50 0 55 0 60 0 65 0 70 00.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1.0

Abs

orba

nce

Waveleng th (nm)

(a)

(b)

(c)

(d)

Fig. 1. Time study of ampicillin derived silver nanoparticles until 1 month observa-tion ((a) 10 min, (b) 24 h, (c) 1 week and (d) 1 month).

pm

3

3

cUttoowtwaayN

5001000150020002500300035004000

% T

rans

mitt

ance

wavenumber (cm-1 )

a

b

Amp-Ag (0) NPs was calculated as 6.87 ± 2.2 nm and spherical in

rocess was repeated five times to see any change in the perfor-ance of catalytic activity of reused Amp-Ag (0) NPs.

. Results and discussions

.1. UV–visible study

UV–Vis spectrometry was performed to know the effect of con-entration of silver nitrate, amoxicillin, and NaOH in solution. TheV–Vis spectra were recorded after 10 min of mixing the solution,

he color of solution was yellow, which is the indication of syn-hesized Ag (0) NPs [18,19]. UV–Vis spectra show absorbance peakf Amp-Ag (0) NPs at 404 nm as shown in Fig. 1. The time studyf Amp-Ag (0) NPs was observed until one month, the wavelengthas shifted to words blue shift from 408 to 404 nm. The �max after

he formation of 10 min of Amp-Ag (0) NPs was at 408 nm whichas slowly increase in absorbance and shifted to words blue shift

fter 24 h until one weak it reached at 405 nm. Finally, the �max wast 404 nm after one month and the color of the solution was dark

ellow, it shows the stability with blue shifted formation of Ag (0)Ps.

Fig. 3. HRTEM images and particle size dist

Fig. 2. FT-IR spectra of (a) amoxicillin antibiotic and (b) Amp-Ag (0) NPs.

3.2. FT-IR study

The FT-IR spectra of amicilin standard and amicilin derivedAmp-Ag (0) NPs were presented in Fig. 2a and b respectively. FT-IR spectra amoxicillin standard showed the two broad and strongsignals at 1666 cm−1 and 1390 cm−1 which can be assigned tohydroxyl groups [35,36] (Fig. 2a). Related peaks have been seen at1651 cm−1 and 1382 cm−1 by other workers [37]. That one broadband above 3000 cm−1 is present with no changes correspondingto the presence of O H (3400 cm−1) and NH (3166 cm−1) stretch-ing frequencies experienced by COOH and NH2 groups [38].The broad band at 3166 cm−1, 3036 cm−1 and 2966 cm−1 repre-sents the N H and O H stretching frequencies [39]. On the otherhand, the bands of the C H and N H in-plane bend vibrations areat 1034 cm−1 [40] but in our case this band appears at 1027 cm−1

which shows slight shift of the band in sample (Fig. 2b).

3.3. HRTEM study

The morphology of Amp-Ag (0) NPs was investigated by HRTEM.It was observed that small size and monodispersed Amp-Ag (0)NPs were synthesized by the help of antibiotic. The particle size of

shape as shown in Fig. 3. A number of methods have been reportedfor synthesis of Amp-Ag (0) NPs and characterized by TEM [18,19].

ribution diagram of Amp-Ag (0) NPs.

Y. Junejo et al. / Applied Surface Science 317 (2014) 914–922 917

30 40 50 60 70

Inte

nsity

(111 )

(200)

(220 )

TN

3

abccf(t(

3

af

3

aesntA2a

3

woticNwtT

200 25 0 30 0 35 0 40 0

0.5

1.0

1.5

2.0

abso

rban

ce

Waveleng th (nm)

a(i)

(ii)

(iii )

200 25 0 30 0 35 0 40 00.0

0.5

1.0

1.5

2.0

Abs

orba

nce

wavelength (nm)

b(i)

(ii )

(iii )

Fig. 5. (a) Reduction of cefdinir in the presence of strong reducing agent NaBH4, (i)

2 The ta (deg .)

Fig. 4. XRD powder pattern of amoxicillin derived Amp-Ag (0) NPs.

ypical TEM images and a corresponding histogram for Amp-Ag (0)Ps are given in Fig. 3.

.4. XRD study

XRD pattern of Amp-Ag (0) NPs obtained by using solid products the result of drying of Amp-Ag (0) NPs (Fig. 4). Nearly 2 mg of darklack solid sample was analyzed Amp-Ag (0) NPs with face centeredubic (FCC) structure. The average crystallite size of the product wasalculated by using Line profile fitting [41] and found as 7.2 ± 1.1 nmor observed three peaks with the following miller indices: (1 1 0),2 0 0), and (2 2 0) [42]. These observed miller indices matched withhe ICDD card no. 99-200-4306 and proved the presence of Amp-Ag0) NPs [18,19]. Other peaks belong to amoxicillin.

.5. Catalysis study of Amp-Ag (0) NPs

The reduction of each antibiotic was analyzed in the absencend presence of Amp-Ag (0) NPs respectively. Sample preparationor the UV–Vis study was outlined in Section 2.4.

.5.1. Reduction of cefdinir antibioticMonodispersed Amp-Ag (0) NPs were checked for the catalytic

ctivity against cefdinir, it was observed that Amp-Ag (0) NPsxhibit excellent catalytic activity. Here, we used rich hydrogenource NaBH4 as reducing agent in the absence of catalyst, it wasoticed that in the absence of Amp-Ag (0) NPs NaBH4 was unableo reduce cefdinir (Fig. 5a). On the other hand, in the presence ofmp-Ag (0) NPs catalyst, the reduction was very fast and just within

min reduction was completed. Cefdinir shows absorbance bandt 284 and 224 nm (Fig. 5b) both peak was continuously decreased.

.5.2. Reduction of cefditoren antibioticThe catalytic activity of newly synthesized Amp-Ag (0) NPs

as investigated for reduction/degradation of cefditoren antibi-tic as illustrated in Fig. 6a and b the reaction was monitored byhe use of NaBH4 as strong reducing agent. It was observed thatn the absence of Amp-Ag (0) NPs, NaBH4 was unable to reduceefditoren (Fig. 6a). Conversely, in the presence of Amp-Ag (0)

Ps, the reduction reaction was fast and the complete reductionas within 5 min after the addition of Amp-Ag (0) NPs. Cefdi-

oren showed the absorption bands at 267 and 235 nm (Fig. 6b).he peak at 267 was showed the shifting of the wavelength to

0 min, (ii) 10 min and (iii) 24 h; (b) reduction of cefdinir in the presence of NaBH4

and catalyst Amp-Ag (0) NPs, (i) 0 min, (ii) 1 min and (iii) 2 min.

297 with decreasing of the peak intensity, and another peak at235 nm was continuously after the addition of decreased Amp-Ag(0) NPs.

3.5.3. Reduction of ceftriaxone sodium antibioticThe catalytic activity of small sized Amp-Ag (0) NPs was

checked against the reduction of ceftriaxone sodium, by NaBH4as reducing agent. The mixture of ceftriaxone sodium antibioticand NaBH4 was examined until 24 h very small reduction wasoccurred as shown in Fig. 7a. The small sized Amp-Ag (0) NPsexhibits excellent catalytic response within 4 min complete reduc-tion takes place. Ceftriaxone sodium antibiotic shows characteristicpeak at 240 and 271 nm, this band decreases very fast by theaddition of Amp-Ag (0) NPs in 5 min. The consecutive absorp-tion spectra of the reduction of ceftriaxone sodium antibioticcatalyzed by the immobilized silver nanoparticles are shown inFig. 7b.

3.5.4. Reduction of doxycycline antibioticUV–Vis spectral studies were carried out to check the per-

formance of freshly prepared Amp-Ag (0) NPs for the catalyticreduction/degradation of doxycycline antibiotic as illustrated in

Fig. 8a and b. We observed high catalytic efficiency of theamoxicillin capped Amp-Ag (0) NPs fabricated by our newlyintroduced rapid and inexpensive synthetic route. Fig. 8a revealsthe absorption profile of doxycycline antibiotic in the presence

918 Y. Junejo et al. / Applied Surface Science 317 (2014) 914–922

200 25 0 300 350 40 0 4500.0

0.5

1.0

1.5

2.0

2.5A

bsor

banc

e

Waveleng th (nm)

a

(i)(ii)

(iii )

200 250 300 350 400

0.5

1.0

1.5

2.0

2.5

Abs

orba

nce

Wavelengt h (n m)

b(i)

(ii)

(iii)

(iv)

(v)

Fig. 6. Reduction of cefditoren antibiotic (a) cefditoren in the presence of reducingagent NaBH , (i) 0 min, (ii) 10 min and (iii) 24 h; (b) cefditoren in the presence ofr3

oua

aTswAbw

3

tst(lrreeTN5

200 25 0 30 0 35 0 4000.0

0.5

1.0

1.5

2.0

Abs

orba

nce

Wavelength (n m)

a(i) (ii)(iii)

200 250 300 350 4000.0

0.5

1.0

1.5

2.0

2.5

Abs

orba

nce

Wavelength (nm)

b(i)

(ii)

(iii )

(iv)

(v)

Fig. 7. Reduction of ceftriaxone sodium antibiotic (a) ceftriaxone sodium antibioticin the presence of reducing agent NaBH4, (i) 0 min, (ii) 10 min and (iii) 24 h; (b)

4

educing agent and catalyst Amp-Ag (0) NPs kinetic study, (i) 0 min, (ii) 1 min, (iii) min, (iv) 3 min and (v) 4 min.

f NaBH4 and it was observed that peaks at 274 and 245 nmndergoes very minute change in absorption until 24 h of irradi-tion.

However, we observed the process of degradation of antibioticfter addition of Amp-Ag (0) NPs catalyst in the presence of NaBH4.he spectra are shown with respect to time (Fig. 8b). The UV–Vispectral studies suggest the complete degradation of the antibioticithin 4 min which highlights the excellent catalytic potential ofmp-Ag (0) NPs. The degradation of antibiotic was characterizedy a continuously decrease in absorption at 274 and 345 nm, whichas monitored by the UV–Vis spectra.

.5.5. Reduction of cefiximee antibioticTo explore the efficiency of Amp-Ag (0) NPs in aqueous sys-

em for the reduction of cefiximee antibiotic which showed theurface plasmon resonance at 232 and 287 nm, there is small reduc-ion of antibiotic was observed with NaBH4 in absence of Amp-Ag0) NPs for an hour, it was observed that in the absence of cata-yst, strong reducing agent NaBH4 is unable to reduce antibiotic. Iteduces cefiximee up to small extent, 5.7% as shown (Fig. 9a) theeduction of antibiotic by NaBH4 did not occur to an appreciablextent in the absence of Amp-Ag (0) NPs, however, the catalytic

fficiency of Amp-Ag (0) NPs can be confirmed from (Fig. 9b).his indicates the reduction of dye in the presence of Amp-Ag (0)Ps. Cefiximee antibiotic showed complete reduction reaction in

min.

ceftriaxone sodium antibiotic in the presence of reducing agent and catalyst Amp-Ag(0) NPs kinetic study, (i) 0 min, (ii) 1 min, (iii) 3 min, (iv) 3 min and (v) 4 min.

3.6. Recycling and reuse of Ag (0) NPs

Glass supported Amp-Ag (0) NPs (0.15 mg) were removedwashed sequentially and reused five times for catalytic reductionof these antibiotics. The reducing potential of recovered and reusedAmp-Ag (0) NPs for ceftriaxone sodium, doxycycline, cefiximee,cefdinir, cefditoren was calculated, which is given (Fig. 10a–e). Theslow deactivation of catalysts Amp-Ag (0) NPs also confirms thatpoisoning of catalysts is insignificant. This work promises a soundenvironmental safety for several water based systems against dyepollution.

3.6.1. Antimicrobial efficacyApplying the disk diffusion it was observed that standard

antibiotics such as cefdinir, cefditoren, cefixime, ceftriaxonesodium and doxycycline have a high inhibitory action on thesix microorganisms used for investigation, E. coli ATCC 25922,S. enteritidis ATCC 4931, P. aeruginosa ATCC 27853, S. pneumo-nia ATCC 49619, S. pyogenes ATCC 21060, and S. aureus ATCC25923.

Initially, different concentration of antibiotics was performed

on S. aureus in order to determine. Fig. 11 shows that almostall the antibiotics with different concentrations (1, 10, 50, and100 �g/ml) gave a very close inhibition zone each other on agarplates. Hence, 10 �g/ml was selected and used throughout the

Y. Junejo et al. / Applied Surface Science 317 (2014) 914–922 919

200 25 0 30 0 350 400 450 500

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b

(i)

(ii )

(iii)

(iv)(v)

Fig. 8. Reduction of doxycycline antibiotic (a) doxycycline antibiotic in the presenceof reducing agent NaBH4, (i) 0 min, (ii) 10 min and (iii) 24 h; (b) doxycycline antibioticin the presence of reducing agent and catalyst Amp-Ag (0) NPs kinetic study, (i)0

aoas

or

b

200 250 300 350 400

0.0

0.2

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Abs

orba

nce

Wavelength (nm)

a(i)(ii )

(iii)

200 25 0 30 0 35 0 40 00.0

0.2

0.4

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0.8

1.0

1.2

1.4

1.6

1.8

Abs

orba

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Wavelength (n m)

b(i)

(ii)

(iii )

(iv)

(v)(vi)

Fig. 9. Reduction of cefixime antibiotic (a) cefixime antibiotic in the presence ofreducing agent NaBH4, (i) 0 min, (ii) 10 min and (iii) 24 h; (b) cefixime antibiotic inthe presence of reducing agent and catalyst Amp-Ag (0) NPs kinetic study, (i) 0 min,

TB

C

min, (ii) 1 min, (iii) 3 min, (iv) 3 min and (v) 4 min.

ntimicrobial study. Meanwhile, different concentration of antibi-tics reduced with Amp-Ag (0) NPs was also performed on S.ureus and no inhibition zone process was observed (data nothown).

The experiments were done in comparison with standard antibi-tics by measuring inhibition zone diameters and the experimental

esults are shown in Fig. 12 and Table 1.

Almost all the standard antibiotics clearly showed antimicro-ial ability against tested microorganisms (Fig. 12) because of

able 1iological activity of the prepared samples on the microorganisms tested by disk diffusio

Microorganisms Diameter of zone of inhibition (mm)

CFDN/CFDN* CDTR/CDTR*

E. coli 30/0 18/0

S. enteritidis 27/0 9/0

P. aeruginosa 10/0 0/0

S. pneumonia 35/0 15/0

S. pyogenes 27/0 12/0

S. aureus 32/0 28/0

FDN, cefdinir; CDTR, cefditoren; CFIX, cefixim; CTRX, ceftriaxone; DOXY, doxycycline.* Antibiotics reduced with Ag (0) NPs.

(ii) 1 min, (iii) 3 min, (iv) 3 min, (v) 4 min and (vi) 5 min.

known bactericidal effects of these antibiotics. The zones aroundthe bore cavities on the agar plates were clearly observed as weexpected. On the other hand, it was observed that the standardantibiotics reduced with Amp-Ag (0) NPs do not involve in theinhibition zone process (Table 1). The bactericidal effects ofthese standard antibiotics were eliminated successfully. There-fore, newly synthesized Amp-Ag (0) NPs were showed excellent

catalytic activity as catalyst for the 100% reduction of theseantibiotics.

n method.

CFIX/CFIX* CTRX/CTRX* DOXY/DOXY*

28/0 35/0 23/030/0 28/0 25/00/0 27/0 20/017/0 30/0 27/025/0 30/0 20/020/0 30/0 28/0

920 Y. Junejo et al. / Applied Surface Science 317 (2014) 914–922

1 2 3 4 50

20

40

60

80

100

Effic

ienc

y %

# of cycles

100% 99% 98% 97% 95%

a

1 2 3 4 50

20

40

60

80

100

Effic

ienc

y %

# of cycles

100% 98% 96% 95% 93%

b

1 2 3 4 50

20

40

60

80

100

95%97% 96%98%

Effic

ienc

y %

# of cycles

100%c

1 2 3 4 50

20

40

60

80

100

92 %94 %96 %98 %

Effic

ienc

y %

# of cycle s

d100 %

1 2 3 4 50

20

40

60

80

100

98% 91%93%96%

Effic

ienc

y %

# of cycles

100%e

Fig. 10. The efficiency of Amp-Ag (0) NPs for the reduction of (a) cefdinir, (b) cefditoren, (c) ceftriaxone sodium, (d) doxycycline, and (e) cefixime.

Fig. 11. The antimicrobial activity of standard antibiotics with different concentrations against S. aureus.

Y. Junejo et al. / Applied Surface Science 317 (2014) 914–922 921

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ig. 12. The antimicrobial activity of standard antibiotics cefdinir, cefditoren, cefigainst S. enteritidis, S. pneumonia, P. aeruginosa, S. pyogenes, E. coli and S. aureus by

. Conclusion

Based upon our findings, nanosized spherical Amp-Ag (0)Ps have been synthesized by the help of antibiotic (amoxi-illin) as a reducing agent as well as capping and stabilizinggent. These nanoparticles are stable in solution for a very longime more than one month without any aggregation. The syn-hesized nanoparticles were confirmed by many characterizationechniques. UV–Vis spectrophotometer was used as first charac-erization. The interaction of bonding formation was confirmedy FT-IR spectrophotometer. Morphology of Amp-Ag (0) NPs wasbserved by TEM analysis and crystalline pattern was confirmedy powder XRD. The synthesized Amp-Ag (0) NPs have proved ashe remarkably efficient catalysts with enhanced rate of reductionor cefdinir, cefditoren, cefixime, ceftriaxone sodium and doxycy-line. This study provides an economical solution to protect aquaticnvironment in terms of time saving and can be equally useful foreduction of several other antibiotics. The present study is a firsteport about the use of Amp-Ag (0) NPs as reduction catalysts forifferent antibiotics and probable to open new doors for their fur-her catalytic applications in analogous and other experiments ofnvironmental and industrial importance.

cknowledgements

Authors would like to thank to TUBITAK for Research Fellow-hip program for foreign citizens (BIDEB 2216) and this work wasupported by Fatih University under BAP Grant No: P50021301-Y3146).

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