A critical understanding of the formation and growth process of gold nanoparticles (GNPs) is crucial, in order to synthesize monodisperse gold nanocrystals of controllable size and shape in a predictable way. GNPs of different shapes and sizes can be produced by altering different physicochemical reaction conditions, such as pH, temperature, and the ratio of reactants. The significant variations in the morphological features of GNPs can be monitored in situ in a time-dependent manner by using UV–visible spectroscopy. In this study, we have synthesized seedless GNPs by reduction of HAuCl4 by using various molar ratios of HEPES and Na2HPO4. The shape and the geometry of the gold nanostructures were optimized by varying the pH (5, 7, and 9) of the HEPES, molar concentrations of disodium phosphate to HAuCl4, and reaction temperatures (20 °C, 40 °C, and 60 °C). The changes in the color of the reaction mixtures over time were recorded in situ in terms of the absorbance of the UV–visible light to tune and investigate the growth process of gold nanostructures. Transmission electron microscopic (TEM) images indicated that the gold nanostructures are anisotropic, stable, and exist in the size range of < 1–100 nm. The present study also confirms that the change in the color of nanostructure reaction mixtures is a function of the surface plasmon resonance bands under the influence of physicochemical reaction conditions and corroborates with the size and shape of nano-gold. Physicochemical parameters such as temperature, pH, and the molar concentration of the reactants act synergistically to influence the growth, molecular mechanics, and reaction thermodynamics that aid to affect the particle size, shape, and surface corona of the GNPs. By tracking the growth process, we can confirm that the nucleation, growth process, size, and shape of the gold nanostructures depend on the HEPES free radicals and phosphate ions which cluster to form polymeric chains on the gold nanocore. The present study's findings explain the growth process of the GNPs of various colors that can be observed by the naked eye and have promising applications, for example in the development of the biosensing rapid lateral flow immunoassays (LFIA) for the detection of mycotoxins in food samples.
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