Gold nanoparticles (AuNPs) are currently under intense investigation for biomedical and biotechnology applications, thanks to their ease in preparation, stability, biocompatibility, multiple surface functionalities, and size-dependent optical properties. The most commonly used method for AuNP synthesis in aqueous solution is the reduction of tetrachloroauric acid (HAuCl₄) with trisodium citrate. We have observed variations in the pH and in the concentration of the gold colloidal suspension synthesized under standard conditions, verifying a reduction in the reaction yield by around 46% from pH 5.3 (2.4 nM) to pH 4.7 (1.29 nM). Citrate-capped AuNPs were characterized by UV-visible spectroscopy, TEM, EDS, and zeta-potential measurements, revealing a linear correlation between pH and the concentration of the generated AuNPs. This result can be attributed to the adverse effect of protons both on citrate oxidation and on citrate adsorption onto the gold surface, which is required to form the stabilization layer. Overall, this study provides insight into the effect of the pH over the synthesis performance of the method, which would be of particular interest from the point of view of large-scale manufacturing processes.

In this article we present the development of a biosensor system that integrates nanotechnology, optomechanics and a spectral detection algorithm for sensitive quantification of antibiotic residues in raw milk of cow. Firstly, nanobiosensors were designed and synthesized by chemically bonding gold nanoparticles (AuNPs) with aptamer bioreceptors highly selective for four widely used antibiotics in the field of veterinary medicine, namely, Kanamycin, Ampicillin, Oxytetracycline and Sulfadimethoxine. When molecules of the antibiotics are present in the milk sample, the interaction with the aptamers induces random AuNP aggregation. This phenomenon modifies the initial absorption spectrum of the milk sample without antibiotics, producing spectral features that indicate both the presence of antibiotics and, to some extent, its concentration. Secondly, we designed and constructed an electro-opto-mechanic device that performs automatic high-resolution spectral data acquisition in a wavelength range of 400 to 800 nm. Thirdly, the acquired spectra were processed by a machine-learning algorithm that is embedded into the acquisition hardware to determine the presence and concentration ranges of the antibiotics. Our approach outperformed state-of-the-art standardized techniques (based on the 520/620 nm ratio) for antibiotic detection, both in speed and in sensitivity.

The misuse of antibiotics in the cattle sector can lead to milk contamination, with concomitant effects on the dairy industry and human health. Biosensors can be applied in this field; however, the influence of the milk matrix on their activity has been poorly studied in light of the preanalytical process. Herein, aptamer-conjugated gold nanoparticles (nanoaptasensors) were investigated for the colorimetric detection in raw milk of four antibiotics used in cattle. The effect of milk components on the colorimetric response of the nanoaptasensors was analyzed by following the selective aggregation of the nanoparticles, using the absorption ratio A₅₂₀/A₇₂₀. A preanalytical strategy was developed to apply the nanoaptasensors to antibiotic-contaminated raw milk samples, which involves a clarification step with Carrez reagents followed by the removal of cations through dilution, chelation (EDTA) or precipitation (NaHCO₃). The colorimetric signals were detected in spiked samples at concentrations of antibiotics as low as 0.25-fold the maximum residue limits (MRLs) for kanamycin (37.5 μg/L), oxytetracycline (25 μg/L), sulfadimethoxine (6.25 μg/L) and ampicillin (1 μg/L), according to European and Chilean legislation. Overall, we conclude that this methodology holds potential for the semiquantitative analysis of antibiotic residues in raw milk obtained directly from dairy farms.

Background: Numerous studies have proposed the use of fluorescent semiconductor nanoparticles or quantum dots (QDs) as novel tools to label cells and tumors. However, QD applications are limited by their toxicity in biological systems and little is known about whether QDs affect the capacity of cancer cells to metastasize. Previously, we described the “biomimetic” synthesis of CdTe-QDs (QDs-glutathione [GSH]) with increased biocompatibility and the potential utility in labeling cells. Purpose: In order to determine the feasibility of using QDs-GSH as a tool for tracking tumor cells during early metastasis, we characterized here for the first time, the in vitro and in vivo effects of the incorporation of green or red biomimetic QDs-GSH into B16F10 cells, a syngeneic mouse melanoma line for metastasis assays in C57BL/6 mice. Methods: B16F10 cells were labeled with green or red biomimetic QDs-GSH in the presence or absence of n-acetylcysteine. Then, migration, invasion and proliferation of labeled B16F10 were evaluated in vitro. Finally, the B16F10 cells labeled with red QDs-GSH were used to monitor in vivo lung metastasis at early time points (5 minutes to 24 hours) or after 21 days in C57BL/6 mice. Results: We developed a methodology that allows obtaining QDs-GSH-labeled B16F10 cells (nearly 100% viable labeled cells), which remained viable for at least 5 days and migrated similarly to control cells. However, proliferation, invasion, and the capacity to form metastatic nodules in the lungs were severely attenuated. Fluorescence imaging revealed that distribution/accumulation of QDs-GSH-labeled B16F10 cells could be tracked following injection into C57BL/6 mice (syngeneic preclinical metastasis model) and that these cells preferentially accumulated in the perialveolar area in lungs as early as 5 minutes post-injection. Conclusion: The methodology described here represents a useful alternative for monitoring initial events during tumor cell metastasis.

Gold nanoparticles (AuNPs) are currently under intense investigation for biomedical and biotechnology applications, thanks to their ease in preparation, stability, biocompatibility, multiple surface functionalities and size-dependent optical properties. The most commonly used method for AuNPs synthesis in aqueous solution is the reduction of tetrachloroauric acid (HAuCl4) with trisodium citrate. We observed variations in the pH and concentration of the gold colloidal suspension synthesized under standard conditions, verifying a reduction in the reaction yield by around 46% from pH 5.3 (2.4 nM) to pH 4.7 (1.29 nM). Citrate-capped AuNPs were characterized by UV-visible spectroscopy, TEM, EDS and zeta-potential measurements, revealing a linear correlation between pH and the concentration of the generated AuNPs. This result can be attributed to the adverse effect of protons both on citrate oxidation and on citrate adsorption onto the gold surface, which is required to form the stabilization layer. Overall, this study provides insight into the effect of the pH over the synthesis performance of the method, which would be of particular interest from the point of view of large-scale manufacturing processes.

Gold nanoparticles (AuNPs) exhibit unique properties that make them appealing for applications in biosensing and other emerging fields. Despite the availability of numerous synthesis methods, important questions remain to be addressed regarding the volume effect on the synthesis yield and quality of AuNPs in the light of biosensing research. The present study addresses these issues by developing a novel microvolumetric citrate-reduction method to improve the synthesis of AuNPs, which were characterized by electronic microscopy, energy dispersive spectroscopy, zeta potential and colorimetric analysis. A comparison of the novel microsynthesis method with the standard Turkevich method demonstrated its superior performance in terms of yield, monodispersity, rapidity (in one step), reproducibility, and stability. The analytical behavior of AuNPs-based aptasensors prepared by microsynthesis was investigated using kanamycin detection and showed higher reproducibility and improved detection limits (3.4 times) compared to those of Turkevich AuNPs. Finally, the effect of pH was studied to demonstrate the suitability of the method for the screening of AuNP synthesis parameters that are of direct interest in biosensing research; the results showed an optimal pH range between 5.0 and 5.5. In summary, the approach described herein has the potential to improve research capabilities in biosensing, with the added benefits of lowering costs and minimizing waste generation in line with current trends in green nanotechnology.