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�:	Synthesis of Chitosan/CdS nanocomposite, characterization and Detection Heavy Metal Ion Application 
Ernest Ravindran R. S1( and Subha V2
1Department of Electronics and Communication Engineering, K L University, Vaddeswaram, Guntur, Andhra Pradesh, India-522502.
2Department of Biotechnology, A.C. Tech. Anna University, Chennai, Tamilnadu, India-600025.
Abstract
Cadmium Sulfide (CdS) Quantum Dots prepared using chemical synthesis method with surface modified with the help of Chitosan. Chitosan/CdS nanocomposites is also used as a simple, rapid, sensitive and specific material for the detection of silver metal ion. In this present investigation chitosan was used as matrix to fabricate CdS QDs with narrow size distribution under mild conditions, resulting in a novel Chitosan/CdS nanocomposites. CdS Quantum Dots of 8 nm size was confirmed using Transmission Electron Microscope (TEM) and mono distribution of quantum dots was also observed. Size of CdS QDs controlled by varying reaction time and precursor concentration and the resultant products were also characterized by UV-vis spectrophotometer and X-ray Diffraction (XRD). Chitosan/CdS exhibit high aqueous solubility with stability. Silver metal ion was detected and reduced to nano size due to the reducing property of chitosan. The mechanism involved in fluorescence quenching of Chitosan/CdS by metal ion (Ag+) was identified using Photo Luminous (PL) analysis.
Keywords: Chitosan, CdS, nanocomposite, luminescence, metal detection. 



1. Introduction
Nanomaterial plays a major role in our day-to-day life. In recent years, metal nanoparticles such as silver, copper, iron, zinc and titanium were being used in various fields like medical, food package, electrical, fuel and advance transport. Various number of methods were also adapted to synthesize these nanoparticles according to the area where it performs likewise either in green or by chemical methods [1-8]. Semiconductor materials synthesized in nano size were also employed in storage of electrical energy, superconductors and transistors, etc. 
Quantum dots made up of semiconducting materials which were in nano range tends to have larger band gap energy and this peculiar property improved the necessity of the materials overall [9]. QDs is a semiconductor material that have all three dimensions confined to nano scale. Quantum dots with zero dimension is applicable in Light Emitting Diodes (LEDs), sensors and as an indicator in medical field used along with drugs for targeted drug delivery. The interest in these materials has been primarily motivated by their fundamental quantum properties, but the technological aspects of these materials are being given full consideration [10]. 
The unique optical and electronic properties owing to the presence of quantum confinement which is in built in the material. QDs exhibit remarkable advantages, including large fluorescence quantum yields, high photo bleaching threshold and excellent photo stability [11-13]. Among variety of QDs, CdS has been intensively studied due to the band-gap energy existing in the visible region, which is suitable for practical applications [11]. Modified core-shell QDs as fluorescence labels to stain biological samples were also used in medical application [12, 14]. It is reasonable, to assume that the changes of surface charges of QDs caused by the chemical or physical interaction between metal ions which affects the efficiency of core electron-hole recombination [15]. 
CdS embedded with different matrixes such as polystyrene, polyvinyl alcohol, and polyethylene glycol have been employed for various applications [16-22].  Chen and Rosenzweig et al., (2002) demonstrated copper and zinc ion analysis by utilizing a CdS luminescent quantum dot capped by different ligands in aqueous media [23]. Gattas-Asfura and Leblanc et al., (2003) synthesized the peptide coated CdS for optical detection of the copper (Cu2+) and silver (Ag+) metal ions [24].  CdS showed wider emission peaks indicating that these particles contain larger number of surface defects [25]. The analysis and measurement of heavy ion, especially Cu2+, in environmental and biological samples are extremely important for designing robust CdS fluorescence sensor [27]. 
Chitosan has good chelating ability with transition metal ions, that makes it possible for its metal ion complexes to be used as precursors to synthesize QDs. Chitosan also recognized as excellent metal ligands, forming stable complexes with many metal ions [28]. Chitin, poly (b-(1-4)-N-acetyl-D-glucosamine), is a natural polysaccharide, first identified in 1884. This biopolymer is synthesized by an enormous number of living organisms and it is the most abundant polymer after cellulose. Chitosan is the only pseudo-natural cationic polymer and thus, it finds many applications that follow from its unique character. Being soluble in aqueous solutions, it is largely used in different applications as solutions, gels, or films and fibers [29]. Chitosan polymers are natural amino polysaccharides having unique structures, multidimensional properties, highly sophisticated functions and wide ranging applications in biomedical and other industrial aspects [30-32]. As a functional material chitosan offers a unique set of characteristics namely hydrophilicity, biocompatibility and biodegradability [33]. 
In this present investigation, CdS/Chitosan quantum dots is synthesized under mild condition using chitosan as chelating metal ions in less reaction time. Chitosan act as the stabilizing agent and allow the growth of quantum dots. By varying the concentration of the reducing agent different size of CdS/Chitosan and variation in luminescence were observed. The detection of silver ions by CdS/Chitosan and formation of silver nanoparticles on CdS/chitosan can be applied for waste water treatment and detection of metal ions in drinking water.   
2. Material and Method
2.1 Materials 
Cadmium acetate, Sodium Sulfide and silver nitrate were purchased from Merck, chitosan with 85% of deacetylation, with molecular weight and viscosity of 200 cps was purchased from Sigma Aldrich. Acetic acid glacial extra pure was purchased from SDFC. Throughout the reaction millipore water was used. 
2.2 Formation of Chitosan Matrix 
Chitosan was dissolved in Acetic acid glacial of 1 wt% 1 ml was added to 49 ml of millipore water and stir for 10 min. Once the solution was thoroughly mixed, 1 g of chitosan was added to the solution under stirring condition. After 30 min of stirring a clear solution was obtained indicating the complete dissolved chitosan in diluted acidic acid. The obtained chitosan solution was well sonicated for 20 min at room temperature using ultra sonication bath. Then the solution was filtered using whattman filter paper. Cadmium acetate of 10 mM in 20 ml of millipore water was stirred separately under room temperature. Then Cadmium acetate solution was added drop by drop into the chitosan solution and stirred for 10 h. Long period of stirring reaction is to make well chelating of Cd2+ ions in the chitosan matrix. 
After chelated Cd2+ ions in chitosan matrix, sodium sulfide was prepared for 10 mM added in drop wise to the mixture. Cadmium ions has been absorbed in chitosan matrix, S2- ions from sodium sulfide moves towards cadmium ions in the matrix and combine with Cd2+ and initiate the growth of quantum dots. The formation of CdS QDs was observed by color change from colorless to light greenish yellow. The overall reaction mechanism is shown in Fig.1.
2.3 Detection of Silver ions
Silver nitrate of 10 mM was prepared and few drops is added to chitosan/CdS nanocomposite solution. An immediate color change was observed indicating that fluorescent quenches had taken place. The light greenish yellow color was completely quenches to golden brown color when silver nitrate solution added directly. Chitosan consist of OH bond and NH2 which forms bond with cadmium ions completely under stirring. Due to formation of bonds with cadmium ions it is not able to release quickly from the chitosan matrix as shown in Fig.2.
2.4. Characterisation
UV-Visible spectral analysis was done by using Jasco V-650 series Spectrophotometer with a wave length of 200 to 800 nm possessing with a scanning speed of 400 nm/min. X-ray diffraction was carried out by X-Pert Pro Diffractometer using step scan technique and with Cu-K� radiation (1.500�, 40KV, 30mA) in �-2� configuration. Pl measurement was performed using Perkin Elmer lambda 55 with UV light. TEM analysis of the metal nanoparticles was performed on a JEOL model PHILIPS instrument operating at an accelerating voltage of 120 KV. SEM was analyzed using JSM-6390 instrument with accelerating voltage of 20 v and magnification of 23 K. 
3. Result and Discussion
3.1. Synthesis of CdS/chitosan nanocomposites	
Synthesis of chitosan/CdS composites comprised of two steps: firstly, chelating cadmium ions to chitosan and secondly, the rapid addition of Na2S. Fig.3 shows the change in the absorption spectra of the chitosan/Cd2+ complex and CdS/chitosan, respectively. After addition of Na2S, the peak observed at 277 nm in the chitosan/Cd2+ complex disappears. This peak is associated with the electronic transition of nitrogen on the secondary amide and primary amine groups present in chitosan (Figure. 1) and its disappearance implies an interaction is occurring between the newly formed CdS which as peak at 430 nm and chitosan. 
3.2 Effect of precursor concentration with CdS QDs size
A study on the effects of concentration of both cadmium acetate (Cd(Ac)2) and Na2S was performed to establish which had the greatest effect on size of the QD. It was found that, by changing the Cd(Ac)2 concentration to 1, 5, and 10 mM, with Na2S constant at 5 mM, that there was little change in the CdS QD size synthesized (Fig.4) [33]. However, when the Na2S concentration was varied to 1, 5, and 10 mM, with the Cd(Ac)2 concentration at 10 mM, a significant change in CdS QD size was observed (Fig.5). These results suggest that the sizes of the QDs in the chitosan matrix are controlled primarily by the Na2S. Huang et al (1996) have recorded similar results using CdS QDs synthesized by chelating cadmium to sulfonated bisvinyl-A (BSS) followed by the injection of hydrogen sulfide (H2S) [34].
It is believed that, this effect occurs because the nucleation is limited at lower concentrations of Na2S resulting in a more rapid depletion of precursors. In combination with Ostwald ripening, subsequent growth through diffusion of the precursors from the surrounding environment to the surface of growth will only continue until the concentration reduces to the equilibrium concentration. The decreased concentrations result in a smaller equilibrium concentration, and so smaller QD nanocrystals. Furthermore, the presence of the chitosan polymer hinders further growth and aggregation of the nanocrystals. Here only variation was in concentration but the amount of addition of Na2S was same (of 1 ml), since increase in the volume of the Na2S increase the particle size and lead to aggregation.
3.3 Detection of silver ions 
Fluorescence achieved at a Na2S precursor of 5 mM and silver was well absorbed when compared to other concentrations. Different concentration of silver nitrate was added to the reaction and color changes were observed in the reaction mixture (light greenish yellow to golden brown color) indicating the presence of silver ions on the chitosan matrix. UV visible spectra shown in Fig.6 clearly shows �max at 419 nm indicating the presence of silver on CdS/chitosan. 
3.4 Quenching Mechanism 
The luminescence of CdS/chitosan nanocomposites was observed at the range of very high intensity of less reducing agent Na2S of 10 mM. The excitation given at 360 nm so that the first emission at 440 nm and thus intensity reduces with increases in silver nitrate concentration (Fig.7). The PL spectra of CdS were recorded at room temperature with excitation wavelength of 360 nm. PL spectra are observed to be broad and emission occurred at a lower energy value than that corresponding to the excitonic emission band. Change in the color after adding silver nitrate into CdS/chitosan quantum dots shows clearly quenching in fluorescence of quantum dots. 
3.5 X- ray Diffraction analysis
The broad peaks that observed in the XRD pattern of CdS is attributed to hexagonal structure of CdS nanocrystals (Fig. 8). CdS quantum dots could possess the hexagonal phase with plane (002). Hailin Hu et al., had reported that the hexagonal wurtzite structure of CdS for the plane (002) [35]. Braggs reflection observed in XRD pattern at 2� = 28.56 and 32.456 corresponds to (0 0 2) and (1 1 0) facets of crystalline of cadmium sulfide quantum dots (JCPDS No: 41-1049). 
X-Ray diffraction pattern of silver ions after being absorbed by CdS/chitosan was shown in Fig.9. XRD confirms the existence of both silver and CdS. Braggs reflection observed in XRD pattern at 2� = 38.245 correspond to the (1 1 1) facets of crystalline silver (JCPDS No: 87-0720). The sample contains mixed phase both cubic and hexagonal structures of silver. 
3.5 Scanning Electron Microscopy (SEM) and EDAX 
SEM micrograph of CdS/chitosan is depicted in Fig.10(A) and it resembles a dot like structure of average size 55.39 nm. The average size of Ag-chitosan/CdS QDs was 83.22 nm with rod shape which is observed in Fig.10(B). EDAX spectra was used to find the elemental composition of Ag capped CdS/Chitosan QDs. From the spectra shown in Fig.11(A) it was found that elements Cadmium and Sulphide are present in the composite of chitosan/CdS. After silver nitrate was added (Fig.11(B)) to the composite, it was found that the presence of silver in the chitosan/CdS nanocomposites which confirms that silver ions were been detected by chitosan/CdS.
3.6 Transmission Electron Microscopy (TEM)
TEM images of functional chitosan/CdS nanocomposites before and after adding silver ion are shown in Fig.12(A). The average size of CdS quantum dots was observed to be 7.51 nm with narrow size variations. The matrix formed by chitosan was clearly visible in TEM which shows quantum dots were also closely packed. From the TEM micrograph, the size of the QDs was observed to be slightly increased after addition of silver ions, which attribute to the complex formation of silver ion absorbed on the surface of CdS QDs. The appearance of nano spheres is due to the formation of nanovesicles as the chitosan has dried on the TEM grid. Ag-chitosan/CdS (Fig.12 (B)) appear as dark spots of agglomerated and over layed QDs. Not every chitosan nanovesicles appears to have QDs associated with them which again may be a result of the diffusion limited process of QD formation. From figure 14 it was clearly observed that chitosan matrix is light before the addition of silver ions and it turns dense after the addition of silver ions which indirectly responsible for quenching of fluorescence. 
4. Conclusions
Chitosan/CdS nanocomposites had been successfully synthesized and characterized using UV- visible spectroscopy, PL, XRD, AFM, SEM with EDAX and TEM. The crystalline structure of CdS quantum dots was hexagonal in shape (002) as deduced from XRD analysis. In this investigation, silver metal is successfully detected using CdS/chitosan nanocomposites. Detection was confirmed from the fluorescence quenching by PL, XRD and EDAX. Detection of Ag2+ ions by using chitosan/CdS and formation of silver nanoparticles on the surface of CdS/chitosan nanocomposites was confirmed using XRD. Silver ions was not only detected but also reduced to nano size which also confirmed from TEM and from SEM analysis the particle shape was found to be spherical shape of CdS in the flakes of chitosan and rod structure for Ag capped chitosan/CdS nanocomposites (3-phase nanocomposites). By increasing the concentration of the chitosan further strip can be formed where the detection can be made visible for easy confirmation of presence of metal ions. This also can be extended for sensor application in the food packing industry where chitosan/CdS nanocomposite strip is biocompatible and will not affect the food materials. 
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(Corresponding author: Tel. +918903800143
 E-mail: ravindran.ernest@kluniversity.in (R. S. Ernest Ravindran)














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