Metallic nanoparticles for surface enhancement

2. Methodology

2.2 Metallic nanoparticles for surface enhancement

Surface modification using nanomaterials was used in this study to support the adsorption of the analytes to the chemical sensor and to provide chemical signal enhancement [11]. Furthermore, such an approach allows the designing of substrates capable of adsorption towards the analytes via intermolecular forces. As such, two benchtop techniques;chemical reduction, and the self- assembly method (SAM), were selected to fabricate metallic nanoparticles and use them to chemically functionalize the coated samples from section 2.1. The chemical reduction method used to synthesize the metallic nanoparticles (MNPs) follows three key steps: reduction of metal salts using a reducing agent, stabilizing the ionic mixtures and lastly control of the size distribution using a capping agent [12-15]. The newly formed MNPs are then used to coat the thin metallic films using the self-assembly method. In SAM, molecules from the surrounding media that is the

48 MNPs are chemisorbed or physiosorbed onto the surface embedded in the solution [16]. In this manner, the nanomaterials arrange themselves on the surface via chemical interactions that foster the stereochemistry of the adsorption process, forming well-ordered structures of various shapes and sizes on the platform. Figure 2.2 below is a schematic of the chemical reduction and self-assembly methods used in this study.

Figure 2.2:Schematic of the functionalization process using chemical reduction and the self-assembly method

Two crosslinkers were investigated for this study based on the presence of their functional groups which serve as active sites for chemical adsorption. Sodium citrate tribasic and cysteamine were selected because the former contains hydroxyl (-OH) and carboxylate groups (-COO^-) while the latter possesses amine (-NH2) and possibly thiol groups (-SH) depending on preferred orientation.

The sections that follow explain the processes undertaken to synthesize MNPs of gold (Au) and silver (Ag) using the above crosslinkers as well as the coating steps followed during self-assembly on the gold slides. Calculations of the theoretical concentrations are given in the supplementary section C1.

2.2.1 Citrate (Cit) crosslinking metallic nanoparticles of gold and silver on coated slides.

Citrate stabilized MNPs were prepared using an adjusted version of the Turkevich method [17]

as follows. For gold/citrate nanoparticles (Au@Citrate), 60 mg of sodium tetrachloroaurate (III) dihydrate (NaAuCl4, Sigma: 298174) was dissolved in 125 mL deionized water and brought to a boil. Meantime, a 1 % solution sodium citrate tribasic dihydrate (Sigma, 71409) was prepared in

49 deionized water and 12.5 mL of the solution was pipetted into the AuCl4 solution. The reaction mixture was stirred vigorously on a heated magnetic stove for 1 hour. The solution turned wine red which indicates the formation of gold nanoparticles [18-19]. The chemical reaction for the Au@Cit synthesis is explained by the balanced equation below.

3𝐶₆𝐻₅𝑂₇⁻³+ 6𝑂𝐻⁻+ 2𝐴𝑢⁺³ = 2𝐴𝑢⁰+ 3𝑂_2↑+ 3𝐻₂𝑂

Silver citrate nanoparticles were synthesized using a revised version of the Lee and Meisel method [20]. Briefly, 22.5 mg of silver nitrate (AgNO3, Sigma: 101510) was dissolved in 125 mL deionized water and brought to a boil. 12.5 mL of 1 % citrate solution was pipetted into the boiling solution and stirred for 1 hour. The solution turned yellow-green which indicated the presence of silver nanoparticles. The chemical reaction of this process can be explained by the equation.

4𝐴𝑔⁺+ 𝐶₆𝐻₅𝑂₇𝑁𝑎₃+ 2𝐻₂𝑂 = 4𝐴𝑔⁰+ 𝐶₆𝐻₅𝑂₇𝐻₃+ 3𝑁𝑎⁺+ 𝐻⁺+ 𝑂_2↑

The Au@Citrate and Ag@Citrate solutions were subsequently analyzed using UV-Vis spectroscopy. After spectroscopic confirmation of MNPs, 60 mL of each solution was diluted 1:1 with water and filtered using a 0.45 µm filter and a 10 mL syringe. 20 mL of the filtered solutions were pipetted on petri dishes containing metal-coated slides: Au@Citrate on Au slide and Ag@Citrate on Ag. The MNPs were allowed to self-assemble on the substrates overnight and then subsequently rinsed with water to remove unbound MNPs. The slides were dried under vacuum for 3 hours prior to analysis with Raman microscopy.

2.2.2 Cysteamine (Cys) crosslinking metallic nanoparticles of gold and silver on coated slides.

For Au@cysteamine MNPs, 400 μL of 0.213 M cysteamine hydrochloride and 40 mL of 1.40 mM HAuCl4 were mixed in a 100 mL glass bottle. The mixture was stirred for 20 min at room temperature in the dark. 10 mL of freshly prepared NaBH4 solution (10 mM) was then quickly added into the above aqueous solution under vigorous stirring, and the mixture was further stirred for 30 min at room temperature in the dark. The resulting wine-red solution was filtered by 0.45 μm Millipore membrane filter and stored in the refrigerator (4 °C) before use [21]. The reaction equation for this synthesis is as follows:

𝐻𝐴𝑢𝐶𝑙₄+ 3𝐻𝑆(𝐶𝐻₂)₂𝑁𝐻₂ = 𝐴𝑢(𝑆(𝐶𝐻₂)₂𝑁𝐻₂)₃+ 4𝐻𝐶𝑙

In the case of Ag@cysteamine MNPs, a 30 mL portion of 0.2 mM sodium borohydride (NaBH4) was taken in an Erlenmeyer flask fitted with a magnetic stir bar and placed in an ice bath with constant stirring for 20 min. In another flask, 20 mL of 0.1 mM AgNO3 solution was placed in an ice bath and 3 mL of 0.3% of cysteamine hydrochloride was mixed with it. Then cold NaBH4

solution was added to it dropwise at a very slow rate until the solution became vivid yellow. The flask containing yellow Ag-Nps was removed from the ice bath and was allowed to come at room temperature with constant stirring. The solution was centrifuged and washed several times with dionized water [22]. The reaction for the above process is described by the following equation:

𝐴𝑔𝑁𝑂₃+ 𝐻𝑆(𝐶𝐻₂)₂𝑁𝐻₂ = 𝐴𝑔(𝐶𝐻₂)₂𝑁𝐻₂+ 𝐻𝑁𝑂₃

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