Available online at www.sciencedirect.

com

Colloids and Surfaces B: Biointerfaces 62 (2008) 136142

Facile synthesis of Ag and Au nanoparticles utilizing chitosan as a mediator agent

Dongwei Wei, Weiping Qian
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, PR China Received 28 May 2007; received in revised form 22 September 2007; accepted 23 September 2007 Available online 29 September 2007

Abstract We present here a facile route for the synthesis of crystal Ag and Au nanoparticles on the basis of chitosan as a mediator agent under quiescent conditions. The resulting Ag nanoparticles displayed controllable structural and optical properties depending on the experimental parameters. Fourier transform infrared (FTIR) study combined with zeta potential measurement disclosed that the amino groups were bound to the particles, which was accountable for the stability of nanoparticles. The Ag and Au nanoparticles prepared in our present method is a promising candidate for applications in catalysis, antimicrobial and surface-enhanced Raman scattering. 2007 Elsevier B.V. All rights reserved.
Keywords: Nanoparticles; Chitosan; A mediator agent; Structural and optical properties

1. Introduction Noble metal nanoparticles (Ag, Au, Pt and Pd) are of continuing interest because of their unusual properties compared to bulk metals and their potential applications in novel electrical, optical, magnetic, catalytic, and sensing technologies [16]. Thereinto, more attention has been focused upon preparing Ag and Au nanoparticles due to their promising applications in modern science and technology, for example, electron microscopy [7], analysis [8,9], electronics [10], materials [11], and catalysis [12]. In general, the synthesis of Ag and gold nanoparticles covers three important research themes: (1) The chemical reduction of metal precursors by a reducing agent such as NaBH4 , citrate, and ascorbate [1315]. (2) The irradiation of the solution containing metal ions with ultraviolet or visible light, and microwave and ultrasound irradiation [1621]. (3) The utilization of biomolecules and bioorganisms, such as bacteria, proteins, and polysaccharide, for the synthesis of Ag and gold nanoparticles [2226]. Recently, more and more researchers have turned to theme 3, now that the other two themes may be associated with environmental toxicity or biological hazards, while the biosynthesis

Corresponding author. Tel.: +86 25 83795719; fax: +86 25 83795719. E-mail address: wqian@seu.edu.cn (W. Qian).

could be easily controlled under mild and eco-friendly conditions. So far, one of the most studied biological macromolecules for such approach is chitosan due to some interesting features: (i) it is derivative of the polysaccharide chitin widely distributed in nature, especially in marine invertebrates, insects, fungi, and yeasts. (ii) The presence of largely free amino and hydroxyl groups in chitosan chains offers it unique physicochemical properties including the polycationic, chelating, and lm-forming characteristics [27,28]. It has been demonstrated that chitosan can be adopted as a stabilizer [29] and, more recently, as a reducing agent to synthesize gold nanoparticles [30,31]. The size and stability of the prepared nanoparticles depend on the concentration of chitosan. However, the possible interactions between Au species and chitosan in the preparation of gold nanoparticles are not clearly known. In our current paper, we present a facile route for the synthesis of crystal Ag and Au nanoparticles based on chitosan under quiescent conditions. This is also the rst preparation of Ag nanoparticles by reduction of cationic silver using polycationic polysaccharide chitosan. Our route combined the rst preparation of reaction mixture composed of metal precursor (AgNO3 or HAuCl4 ) and chitosan, and then left them to stand for reduction at designated temperature. The resulting Ag nanoparticles were stable and displayed controllable structural and optical properties depending on the experimental parameters. The possible interactions between the

0927-7765/$ see front matter 2007 Elsevier B.V. All rights reserved. doi:10.1016/j.colsurfb.2007.09.030

D. Wei, W. Qian / Colloids and Surfaces B: Biointerfaces 62 (2008) 136142

137

precursors and chitosan molecules are also examined, which could be answerable for the reduction of metal ions and capping nanoparticles. Presented below are the details of the investigation. 2. Experimental Chitosan akes, from crab shells, (Practical grade >85% deacetylated; Brookeld viscosity >200,000 cps) were purchased from Aldrich. Silver nitrate, hydrochloroauric acid, and acetic acid were analytical grade. Solutions were prepared with tri-distilled water. All compounds were used as received. Chitosan was dissolved in 1% acetic acid solution. Due to the poor solubility of chitosan, the mixture was kept overnight until a clear solution was obtained. For the synthesis of Ag nanoparticles, 4 mL of 8.1624.0 mM AgNO3 were added to 10 mL of 3.466.92 mg/mL chitosan solution under stirring at room temperature. After 15 min, the mixture was transferred to a 12.5 cm 2 cm cuvette and then left them to stand for reduction in a water bath at 4595 C for 612 h. For the synthesis of gold nanoparticles, 4 mL of 18.1 mM HAuCl4 were added to 10 mL of 3.466.92 mg/mL chitosan solution, the subsequent disposal was similar to that presented for the preparation of Ag nanoparticles except that 3-h is enough for the synthesis of Au nanoparticles. Transmission electron microscopy (TEM) was carried out on a JEM-2000EX microscope at an accelerating voltage of 120.0 kV. The optical properties of the products were measured by a SHIMADZU UVvis-NIR spectrophotometer (UV-3150). Fourier transform infrared spectroscopy was recorded on a Nicolet Nexus 870 FTIR instrument. Surface charge measurements were performed with a Zeta Potential Analyzer (BECKMAN, Delsa 440SX). 3. Results and discussion Silver nanoparticles were synthesized using chitosan from silver nitrate. The formation of Ag nanoparticles is accompanied by color changes with time from colorless to pale yellow and nally to light pink, reddish brown, yellow green or gray blue depending on the experimental parameters, as will be discussed later in this paper. The UVvis absorption spectra recorded from the resulting solutions show the characteristic surface plasmon (SP) resonance band of Ag nanoparticles centered at about 420 nm (Fig. 1) [32]. Fig. 1 shows the UVvis absorption spectra from the samples synthesized by reduction of 4 mL of different concentrations of AgNO3 with chitosan. It is clear from each gure that as the concentration of silver salt increases, there is a progressive enhancement in the intensity of the SP band of Ag nanopartilces, which indicates the increased content of Ag nanoparticles in the solution that yields the high absorbance features [33]. In Fig. 1c, one signicant observation is that the 78.0 and 156.0-mM absorption peaks at 420 nm gradually decrease in intensity relative to that of 52.0 mM, but this is not observed in other cases (Fig. 1a and b). Probably, at high reaction temperature, the as-formed higher concentration of silver nanoparticles produces clusters decreasing the nanoparticle concentration causing a decrease in the SP band intensity.

Fig. 1. UVvis spectra of the samples prepared by reduction of 4 mL of different concentrations of AgNO3 with (a) 10 mL of 3.46 mg/mL chitosan at 80 C for 6 h, (b) 10 mL of 6.92 mg/mL chitosan at 80 C for 12 h, and (c) 10 mL of 6.92 mg/mL chitosan at 95 C for 12 h.

During this period, the plasmon peak position shifts from 419 to 424 nm, supporting the increasing polydispersity of nanoparticle [34]. Fig. 2 shows the typical TEM images of the as-prepared silver nanoparticles. It is evident from Fig. 2 that approximately spherical silver nanoparticles are formed inside the nanoscopic polysaccharide templates, which provide strong evidence of UVvis results (Fig. 1a). Nanoparticles prepared with relatively high concentration of silver salts have a stronger tendency to aggregate. It may be that in the case of a high concentration

138

D. Wei, W. Qian / Colloids and Surfaces B: Biointerfaces 62 (2008) 136142

Fig. 2. TEM images of the samples synthesized by reduction of 4 mL of (a) 52.0 mM, (b) 156.0 mM, and (c) 624.0 mM of AgNO3 with 10 mL of 3.46 mg/mL chitosan at 80 C for 6 h. The inset in (c) is the electron diffraction pattern produced by crystalline Ag nanoparticles.

of metal salts, few chitosan molecules work as stabilizer by adsorbing on the surface of nanoparticles, which resulted in the formation of lower stability of nanoparticles, and vice versa. Electron diffraction pattern (the inset in Fig. 2c) exhibits rather sharp spots with diffuse rings, indicating that the as-prepared Ag nanoparticles are crystalline. We speculate that the coordination behaviors between amino groups in chitosan and Ag+ ions is responsible for the stabilization of the silver nanoparticles, which is proved by FTIR and zeta potential measurements discussed later. To explore the reduction process of silver nitrate with chitosan, FTIR measurements were carried out to identify possible interactions between silver salts and chitosan molecules, which could be accountable for the reduction of Ag ions and stabilizing silver nanoparticles. Fig. 3 presents FTIR spectra of the chitosan and Ag/chitosan samples from the reduction of 4 mL of different concentrations of AgNO3 with 10 mL of 3.46 mg/mL chitosan at 80 C for 6 h. Fig. 3b shows a ngerprint region from Fig. 3a. Although there is the possibility of overlapping between the N H and the O H stretching vibrations, the strong broad band at 33003500 cm1 (peak 1, Fig. 3a) is characteristic of the N H stretching vibration. The signicant decrease of transmittance in this band region indicates that the N H vibration was affected duo to the silver attachment [35]. On the other hand, the N H bending vibration band (peak 3, Fig. 3b) at about 1578.1 cm1 were shifted to 1567.3, 1570.8, and 1560.5, cm1 respectively, accompanied by a gradual decrease in intensity with the increased silver concentrations. This also suggests the attachment of silver to nitrogen atoms, which reduced the vibration intensity of the N H bond due to the molecule weight becoming heavier after silver binding. All in all, changes in the transmittances related to the bonds with N atoms reveal that nitrogen atoms are the binding sites for gold on chitosan. In other words, nitrogen atoms are bound to Ag nanoparticles during reduction of AgNO3 with chitosan. It is notable that Ag/chitosan samples shows a new band at about 1760 cm1 (peak 2, Fig. 3b) corresponding to carbonyl stretch vibrations in ketones, aldehydes and carboxylic acids [24]. The presence of the 1760 cm1 signal in the Ag/chitosan samples indicate that the reduction of the silver ions is coupled to the oxidation of the hydroxyl groups in chitosan molecular and/or its hydrolyzates. To determine further the species bound to Ag nanoparticles and investigate the reduction process of silver nitrate with chitosan, zeta potential of Ag nanoparticles was examined. The nanoparticles was separated from the reactive medium by centrifugation and rinsed with deionized water before measurement. Result indicated that the surfaces of Ag nanoparticles have a positive charge about 38.2 mV. Combined with FTIR analysis, it can be concluded that the stability of Ag nanoparticles prepared by the present method should be attributed to the binding of amino groups to the particles.As briey mentioned earlier, the resulting solution from the reduction of AgNO3 with chitosan shows a wide range of colors from light pink to gray blue depending on the experimental parameters, such as reactive temperatures, and concentrations of silver salt and chitosan, as demonstrated further in Fig. 4. It is known that SP band of nanoparticles is sensitive to the size, shape, and spatial

D. Wei, W. Qian / Colloids and Surfaces B: Biointerfaces 62 (2008) 136142

139

Fig. 3. (a) FTIR spectra of chitosan and Ag/chitosan samples and (b) ngerprint region (between 1800 and 700 cm1 ).

distribution of particles [34]. The different fascinating colors of Ag colloids suggest the controllable optical and structural properties of the produced particles, since the color of metal colloids is related to the SP excitation of nanoparticles. The series of UVvis absorbance spectra shown in Fig. 5 clearly exhibit unique optical absorbance of the Ag nanoparticles produced under different reaction conditions. Each of the seven spectra (Fig. 5ag) has an absorption band within 390500 nm attributing to the SP resonance of Ag nanoparticles, which can be obviously observed. Except for that, in Fig. 5e and g, an absorption band located at longer wavelengths (500800 nm) is also evidently seen, which could be found in the strong interparticle interaction among aggregates and the deviation from

spherical geometry of Ag nanoparticles [36]. Fig. 6 presents typical TEM images of three samples in Fig. 5 (ac corresponding to Fig. 5a, e, and g, respectively). It is apparent from Fig. 6a that spherical Ag nanoparticles with wide size distribution are obtained, accordance with its broad non-symmetrical spectra band (Fig. 5a), which may attribute to the slow reduction at low concentration of silver nitrate with chitosan. The resulting structures in Fig. 6b resemble a type of pine tree and display exquisite fractal features, which are generally formed by hierarchical self-assembly under nonequilibrium conditions [37]. Similar Ag nanostructure was also observed by other groups in different reaction systems [38,39]. In Fig. 6c, small linear aggregates of Ag nanoparticles are presented, accountable for

140

D. Wei, W. Qian / Colloids and Surfaces B: Biointerfaces 62 (2008) 136142

Fig. 4. Digital photograph of the samples produced from seven different syntheses, demonstrating control over optical properties. (a) (8.1 mM AgNO3 , 6.92 mg/mL chitosan, 95 C for 12 h); (b) (156 mM AgNO3, 6.92 mg/mL chitosan, 60 C for 12 h); (c) (26.0 mM AgNO3 ; 6.92 mg/mL chitosan, 95 C for 12 h); (d) (52.0 mM AgNO3 , 6.92 mg/mL chitosan, 95 C for 12 h); (e) (78 mM AgNO3 , 3.46 mg/mL chitosan, 95 C for 9 h); (f) (8.1 mM AgNO3 , 6.92 mg/mL chitosan, 60 C for 12 h); (g) the sample of (f) aged for 14 days.

the extended plasmon band (500800 nm) in UVvis spectra (Fig. 5g). In this case, the formation of silver aggregates may be due to their decreased stability originating from the degradation of chitosan during aging, which led to fewer chitosan molecules as stabilizer absorbing on the surface of preformed particles. Thus, the optical and structural variability of Ag nanoparticles produced by the reduction of silver nitrate with chitosan can be realized through variation of the experimental parameters, which is promising for applications such as catalysis [40], antimicrobial [41] and surface-enhanced Raman scattering [42]. Gold nanoparticles were synthesized by reduction of AuCl4 with chitosan. The observation of red color after reaction for 30 min suggested the formation of gold nanoparticles. The reaction mixture exhibited an absorbance band at about 520 nm (Fig. 7a) characteristic of gold nanoparticles due to its SP absorbance, which is responsible for the vivid rose-like color as shown in the inset of Fig. 7a [43]. TEM images in Fig. 7b shows that the as-formed gold nanoparticles are predominantly spherical. Ring-like diffraction pattern (the inset in Fig. 7b) reveals that the particles are crystalline. Zeta potential measurement presents that the resulting gold nanoparticles take positive charge similar to the case of prepared Ag particles. As demonstrated above, Ag and Au nanoparticles were synthesized using chitosan as nanoscopic templates. A 3-h reaction time was enough for the preparation of gold nanoparticles, however, the reduc-

Fig. 6. Representative TEM images of Ag nanoparticles synthesized using chitosan. (a) (8.1 mM AgNO3 , 6.92 mg/mL chitosan, 95 C for 12 h), (b) (78 mM AgNO3 , 3.46 mg/mL chitosan, 95 C for 9 h), and (c) (8.1 mM AgNO3 , 6.92 mg/mL chitosan, 60 C for 12 h and aged for 14 days). Fig. 5. The corresponding UVvis spectra of the samples with the same label in Fig. 4.

D. Wei, W. Qian / Colloids and Surfaces B: Biointerfaces 62 (2008) 136142

141

Fig. 7. (a) Reprehensive UVvis spectra of gold nanoparticles prepared using chitosan (8.1 mM HAuCl4 , 6.92 mg/mL chitosan, 95 C for 3 h). The inset in (a) shows the vividly rose-like color of gold nanoparticles. (b) The corresponding TEM images of gold nanoparticles in (a) after centrifugation. The inset in (b) shows the electron diffraction pattern recorded from the particles shown in (b). (For interpretation of the references to color in this gure legend, the reader is referred to the web version of the article.)

tion of cationic silver with chitosan proceeded much longer time (12 h). Although a more detailed study is required to fully understand the difference in the interactions between chitosan and two kinds of metal precursors, we suggest that two possible factors could explain this result. Firstly, the standard redox potential of Ag+ /Ag (+0.80 V) was lower than that of AuCl4 /Au (+1.002 V), which may be a key factor for the slow reduction of silver ions [44]. Secondly, the electrostatic repulsion between amino groups in chitosan and Ag+ in solution might make the reductive sites in chitosan molecules are less accessible to cationic silver than anionic tetrachloroauric ions, which also induces the slow formation of Ag nanoparticles. 4. Conclusion Synthesis of crystal Ag and Au nanoparticles was carried out utilizing nontoxic and biodegradable chitosan as a mediator agent under quiescent crystallization conditions. The resulting Ag nanoparticles displayed controllable structural and optical properties as demonstrated by UVvis spectroscopic studies and transmission electron microscopy (TEM) observations. Fourier transform infrared (FTIR) study combined with zeta potential measurement revealed the binding of amino groups to the particles, which was responsible for the stability of nanoparticles. Our demonstration that chitosan provides a redox-active template to prepare nanostructured Ag and Au crystallites is signicant for the direction of organicinorganic advanced materials building. This facile route may be extended to the preparation of some other metal nanoparticles such as platinum or palladium. Acknowledgment This research is supported by the National Nature Science Foundation of China (No. 20475009, 60121101, and 904060244), the Foundation for the Author of National Excellent Doctoral Dissertation of PR China (No. 200252), and the Foundation for Excellent Doctoral Dissertation of Southeast University.

References
[1] J.H. He, T. Kunitake, A. Nakao, Chem. Mater. 15 (2003) 4401. [2] D.D. Evanoff, G. Chumanov, Chem. Phys. Chem. 6 (2005) 1221. [3] B. Wiley, Y.G. Sun, B. Mayers, Y.N. Xia, Chem. Eur. J. 11 (2005) 454. [4] J.M. Sun, D. Ma, H. Zhang, X.M. Liu, X.W. Han, X.H. Bao, G. Weinberg, N. Pf nder, D.S. Su, J. Am. Chem. Soc. 128 (2006) 15756. a [5] X. Wang, J. Zhuang, Q. Peng, Y.D. Li, Nature 437 (2005) 121. [6] Y. Hamanaka, K. Fukuta, A. Nakamura, Appl. Phys. Lett. 84 (2004) 4938. [7] D.H. Dahanayaka, J.X. Wang, S. Hossain, L.A. Bumm, J. Am. Chem. Soc. 128 (2006) 6052. [8] L.R. Hirsch, J.B. Jackson, A. Lee, N.J. Halas, J.L. West, Anal. Chem. 75 (2003) 2377. [9] S. Jin, K. Ye, Biotechnol. Prog. 23 (2007) 32. [10] D.L. Klein, P.L. McEuen, Appl. Phys. Lett. 68 (1996) 2574. [11] Z. Li, D. Lee, X.X. Sheng, R.E. Cohen, M.F. Rubner, Langmuir 22 (2006) 9820. [12] C.C. You, S.S. Agasti, M. De, M.J. Knapp, V.M. Rotello, J. Am. Chem. Soc. 128 (2006) 14612. [13] Y. Tan, W.P. Qian, S.H. Ding, Y. Wang, Chem. Mater. 18 (2006) 3385. [14] S.H. Ding, W.P. Qian, Y. Tan, Y. Wang, Langmuir 22 (2006) 7105. [15] P. Setua, A. Chakraborty, D. Seth, M.U. Bhatta, P.V. Satyam, N. Sarkar, J. Phys. Chem. C 111 (2007) 3901. [16] T.C.R. Rocha, H. Winnischofer, E. Westphal, D. Zanchet, J. Phys. Chem. C 111 (2007) 2885. [17] D.W. Wei, W.P. Qian, J. Nanosci. Nanotechnol. 6 (2006) 2508. [18] A. Callegari, D. Tonti, M. Chergui, Nano Lett. 3 (2003) 1565. [19] K. Kurihara, J. Kizling, P. Stenius, J.H. Fendler, J. Am. Chem. Soc. 105 (1983) 2574. [20] N.N. Mallikarjuna, R.S. Varma, Cryst. Growth Des. 7 (2007) 686. [21] T.C. Chiu, S.H. Chiou, M.M. Hsieh, Y.T. Chen, H.T. Chang, J. Nanosci. Nanotechnol. 5 (2005) 2128. [22] S.S. Shankar, A. Rai, B. Ankamwar, A. Singh, A. Ahmad, M. Sastry, Nat. Mater. 3 (2004) 482. [23] S.F. DSouza, K.C. Bhainsa, Colloids Surf. B 47 (2006) 160. [24] S.P. Chandran, M. Chaudhary, R. Pasricha, A. Ahmad, M. Sastry, Biotechnol. Prog. 22 (2006) 577. [25] I. Willner, R. Baron, B. Willner, Adv. Mater. 18 (2006) 1109. [26] M.F. Lengke, M.E. Fleet, C. Southam, Langmuir 23 (2007) 2694. [27] H. Yi, L.Q. Wu, W.E. Bentley, R. Ghodssi, G.W. Rubloff, J.N. Culver, G.F. Payne, Biomacromolecules 6 (2005) 2881. [28] J. Bergera, M. Reista, J.M. Mayera, O.F.R.R. Gurnyb, N.A.P. Gurnyb, Eur. J. Pharm. Biopharms. 57 (2004) 19. [29] K. Esumi, N. Takei, T. Yoshimura, Colloids Surf. B 32 (2003) 117.

142

D. Wei, W. Qian / Colloids and Surfaces B: Biointerfaces 62 (2008) 136142 [39] L.H. Lu, A. Kobayashi, Y. Kikkawa, K. Tawa, Y. Ozaki, J. Phys. Chem. B 110 (2006) 23234. [40] Z.P. Qu, M.J. Cheng, W.X. Huang, X.H. Bao, J. Catal. 229 (2005) 446. [41] A. Pan cek, L. Kvtek, R. Prucek, M. Kol , R. Ve eov , N. Piz rov , a ar c r a u a V.K. Sharma, T. Nev cn , R. Zboil, J. Phys. Chem. B 110 (2006) e a r 16248. [42] D.S. dos Santos, P.J.G. Goulet, N.P.W. Pieczonka, O.N. Oliverira, R.F. Aroca, Langmuir 20 (2004) 10273. [43] T. Yonezawa, T. Nomura, T. Kinoshita, K. Koumoto, J. Nanosci. Nanotechnol. 6 (2006) 1649. [44] J.A. Dean, Lang Handbook of Chemistry, fteenth ed., McGraw-Hill, New York, 1998.

[30] H.Z. Huang, X.R. Yang, Biomacromolecules 5 (2004) 2340. [31] B. Wang, K. Chen, S. Jiang, F. Reincke, W. Tong, D. Wang, C.Y. Gao, Biomacromolecules 7 (2006) 1203. [32] M. Chen, L.Y. Wang, J.T. Han, J.Y. Zhang, Z.Y. Li, D.J. Qian, J. Phys. Chem. B 110 (2006) 11224. [33] P.L. Kuo, W.F. Chen, J. Phys. Chem. B 107 (2003) 11267. [34] S. Manna, S.K. Batabyal, A.K. Nandi, J. Phys. Chem. B 110 (2006) 12318. [35] L. Jin, R. Bai, Langmuir 18 (2002) 9765. [36] X.W. Lou, C.L. Yuan, L.A. Archer, Chem. Mater. 18 (2006) 3921. [37] M. Cao, T.F. Liu, S. Gao, G.B. Sun, X.L. Wu, C.W. Hu, Z.L. Wang, Angew. Chem. Int. Ed. 44 (2005) 4197. [38] J.X. Fang, H.J. You, P. Kong, Y. Yi, X.P. Song, B.J. Ding, Cryst. Growth Des. 7 (2007) 864.