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An electrochemical magnetic immunosensing strategy was developed for the determination of HER2-ECD, a breast cancer biomarker, and breast cancer cells in human serum. A sandwich assay was performed on carboxylic acid-functionalized magnetic beads (MBs) using a screen-printed carbon electrode (SPCE) as transducer surface. The affinity process was detected using electroactive labels; core/shell streptavidin-modified CdSe@ZnS Quantum Dots (QDs). Cd 2+ ions, released from the QDs, were determined by differential pulse anodic stripping voltammetry (DPASV). An assay time of 90 min, with an actual hands-on time of about 20 min, a linear range between 0.50–50 ng·mL −1 of HER2-ECD and a limit of detection of 0.29 ng·mL −1 were achieved. Analysis of live breast cancer cells was also performed using the optimized assay. Breast cancer cell lines SK-BR-3 (a HER2-positive cell line), MDA-MB-231 (a HER2-negative cell line) and MCF-7 (a cell line with low HER2 expression) were tested. The selectivity of the assay towards SK-BR-3 cells was confirmed. A concentration-dependent signal that was 12.5× higher than the signal obtained for the HER2-negative cells (MDA-MB-231) and a limit of detection of 2 cells·mL −1 was obtained. Graphical abstract Schematic representation of the electrochemical immunomagnetic assay for the determination of the breast cancer biomarker HER2-ECD and cancer cells using magnetic beads (MBs), a screen-printed carbon electrode (SPCE) as transducer surface and quantum dots (QD) as electroactive labels.

In this paper, it has been demonstrated that Au(III) is able to act instead of O 2 in the oxidase enzymatic reaction, so that it becomes reduced to purple gold nanoparticles (AuNPs). The plasmon band (at 540 nm) can be used as the analytical signal. Tyramine has been determined using its enzymatic reaction with tyramine oxidase (TAO). The kinetic of the AuNP formation has been studied in the light of both the Avrami equation for crystallization and the Finke-Watsy mechanism for AuNP nucleation and growth. The effects of the Au(III), TAO and tyramine concentrations on the corresponding kinetic constants have been investigated. Working at room temperature, under optimal conditions (phosphate buffer pH 7.0, TAO 0.5 U.mL −1 Au(III) 1 mM), the linear response ranges from 2.5 × 10 −5 M to 3.3 × 10 −4 M Tyramine (5.6% RSD) and the LOD is 2.9 × 10 −6 M. Under these conditions, the signal is measured after 30 min reaction (to obtain the highest sensitivity), but this time can be significantly reduced by increasing the temperature (the reaction is finished after 4 min when working at 50 °C). The method has been applied to tyramine determination in a cheese sample with good results. The new scheme proposed in this paper can be extended, in principle, to other enzymatic methods based on oxidase enzymes. Graphical abstract Tyramine is determined by measuring the plasmon band of the gold nanoparticles formed during its enzymatic reaction with Tyramine oxidase. Moreover, a mathematical model has been developed to explain the formation of the gold nanoparticles during the reaction.

Gold nanoparticles of different sizes have been synthesized and surface-functionalized with selected RNA probes in order to develop a rapid, low-cost and sensitive method for detection of microRNA146a (miR146a). The strategy is based on the change of colour that can be observed visually after aggregation of the RNA modified-gold nanoparticles (AuNPs) in presence of miR146a. Experimental conditions have been carefully selected in order to obtain a good sensitivity that allows to perform visual detection of microRNA at the nM level, achieving a detection limit of 5 nM. Good repeatability and selectivity versus other sequences that only differ from miR146a in 3 bases was achieved. miR146a has been described as one of the main microRNA involved in the immune response of bovine mastitis, being expressed in tissue, blood and milk samples. The method was successfully applied to the detection of miR146a in raw cow milk samples. The present scheme constitutes a rapid and low-cost alternative to perform highly sensitive detection of microRNA without the need of instrumentation and amplification steps for the early detection of bovine mastitis in the agrofood industry. Graphical abstract Schematic representation of the assay based on aggregation of RNA-modified gold nanoparticles (blue) in presence of microRNA146a generating a dark blue spot onto a solid support, versus a pink spot observed in absence of miR146a due to dispersed gold nanoparticles (red).

A rapid fluorometric method is described for the determination of lactate and cholesterol by using ZnO nanowires (ZnO NWs). The assay is based on the detection of the hydrogen peroxide generated during the enzymatic reactions of the oxidation of lactate or cholesterol. Taking advantage of the electrostatic interactions between the enzymes and the ZnO NWs, two bioconjugates were prepared by mixing the nanomaterial and the enzymes, viz. lactate oxidase (LOx) or cholesterol oxidase (ChOx). The enzymatically generated hydrogen peroxide quenches the fluorescence of the ZnO NWs, which have emission peaks at 384 nm and at 520 nm under 330 nm photoexcitation. H 2 O 2 quenches the 520 nm band more strongly. Response is linear up to 1.9 μM lactate concentration, and up to 1.1 μM cholesterol concentration. Relative standard deviation was found to be 5%. The detection limits for lactate and cholesterol are 0.54 and 0.24 μM, respectively. Graphical abstract Schematic representation of fluorescence assay based on ZnO nanowires photoluminiscence for lactate and colesterol detection

An electrochemical method is described for the determination of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (8-MeIQx) which is a heterocyclic aromatic amine formed in cooked food samples. The method uses a screen-printed carbon nanofiber electrode that is modified with silver nanoparticles (AgNPs) in a Nafion matrix. The surface of the modified electrode was characterized by UV-vis spectrometry, dynamic light scattering, scanning electron microscopy and Raman spectroscopy. The average size of the AgNPs is 14 nm. The modified electrode exhibits good properties in terms of reversibility, fast kinetics of electron transfer, and large electroactive area toward the reduction of 8-MeIQx. Differential pulse voltammetry is the most suitable electrochemical technique for quantification of 8-MeIQx, best at a voltage of −0.21 V (versus Ag reference electrode). The first derivative serves as the analytical signal that increases linearly in the 0.015–40 mg L −1 8-MeIQx concentration range, with a 5 μg L −1 detection limit. A dispersive liquid-liquid microextraction procedure assisted via ionic liquid was developed to isolate the analyte from real samples. The whole extraction-preconcentration and voltammetric method allows to determine 30 and 70 μg L −1 in (spiked) bouillon cube, meat broth, beer and wine, with recoveries in the 93.6–110.4% range. Graphical abstract Schematic presentation for the analysis of aromatic amine 8-MeIQx, resultant compound from cooking meat. Extracted sample solution was placed onto modified electrode surface thus obtaining voltammetric analytical signal. So, quantification atrelevant levels can be performed.

Surface-enhanced Raman spectroscopy (SERS) is gaining importance as an ultrasensitive analytical tool for routine high-throughput analysis of a variety of molecular compounds. One of the main challenges is the development of robust, reproducible and cost-effective SERS substrates. In this work, we study the SERS activity of 3D silver mirror–like micro-pyramid structures extended in the z-direction up to 3.7 μm (G0 type substrate) or 7.7 μm (G1 type substrate), prepared by Si-based microfabrication technologies, for trace detection of organophosphorous pesticides, using paraoxon-methyl as probe molecule. The average relative standard deviation (RSD) for the SERS intensity of the peak displayed at 1338 cm −1 recorded over a centimetre scale area of the substrate is below 13% for pesticide concentrations in the range 10 −6 to 10 −15  mol L −1 . This data underlies the spatial uniformity of the SERS response provided by the microfabrication approach. According to finite-difference time-domain (FDTD) simulations, such remarkable feature is mainly due to the contribution on electromagnetic field enhancement of edge plasmon polaritons (EPPs), propagating along the pyramid edges where the pesticide molecules are preferentially adsorbed. Graphical abstract

In this work, novel silver sulphide quantum dots (Ag 2 S QD) are electrochemically quantified for the first time. The method is based on the electrochemical reduction of Ag + to Ag 0 at −0.3 V on screen-printed carbon electrodes (SPCEs), followed by anodic stripping voltammetric oxidation that gives a peak of currents at +0.06 V which represents the analytical signal. The optimized methodology allows the quantification of water-stabilized Ag 2 S QD in the range of approximately 2 × 10 9 –2 × 10 12 QD·mL −1 with a good reproducibility (RSD: 5%). Moreover, as proof-of-concept of relevant biosensing application, Ag 2 S QD are evaluated as tags for Escherichia coli ( E. coli ) bacteria determination. Bacteria tagged with QD are separated by centrifugation from the sample solution and placed on the SPCE surface for quantitative analysis. The effect of two different Ag 2 S QD surface coating/stabilizing agents on both the voltammetric response and the bacteria sensing is also evaluated. 3-mercaptopropionic acid (3-MPA) is studied as model of short length coating ligand with no affinity for the bacteria, while boronic acid (BA) is evaluated as longer length ligand with chemical affinity for the polysaccharides present in the peptidoglycan layer on the bacteria cells surface. The biosensing system allows to detect bacteria in the range 10 −1 -10 3 bacteria·mL −1 with a limit of detection as low as 1 bacteria·mL −1 . This methodology is a promising proof-of-concept alternative to traditional laboratory-based tests, with good sensitivity and short time and low cost of analysis. Graphical abstract Novel silver sulphide quantum dots (Ag 2 S QD) are electrochemically quantified for the first time. Moreover, Ag 2 S QD are evaluated as tags for Escherichia coli bacteria determination. The effect of two different QD surface coating ligands is also evaluated.

New electrochemical sensors are described for the rapid quantification of silver nanoparticles (AgNPs). They are based on the immobilization of L-cysteine on a glassy carbon electrode (GCE) and on the formation of electropolymerized oligomers on the GCE. Ligands with amino, sulfur and carboxy functional groups are used that are capable of selectively retaining AgNPs. The experimental conditions for electropolymerizations were optimized for each of four monomers studied: L-lysine, thiophene-3-carboxamide, thionin, and o-phenylene-diamine. The best retention capabilities and conditions for quantification of AgNPs were found for immobilized nanoparticle voltammetry. This method is more sensitive than others based on the sorption of AgNPs from dispersions. These ligands also retain ionic silver species. Hence, Ag(I) and Ag 0 can be discriminated, and the total silver content can be quantified by stripping voltammetry. The best analytical properties (for dispersions of AgNPs of 40 nm diameter) were found with GCEs carrying electropolymerized L-lysine. Figures of merit include (a) sensitivity of 4.329 ± 0.031 μA μg −1  mL cm −2 , (b) a detection limit of 0.010 μg mL −1 , and (c) a relative standars deviation of about 7.2% (for n  = 4). The poly-L-lysine sensors can also evaluate the size of the AgNPs in the range 20 to 80 nm diameter, owing to displacements of the maximum potential of the voltammetry peaks. Graphical abstract Schematic representation of the quantification of silver nanoparticles (AgNPs) with a glassy-carbon electrode (GCE) modified with electropolymerized poly(L-lysine). AgNPs (and Ag + ions) are selectively preconcentrated in the polymer and determined by differential-pulse stripping voltammetry.

Screen-printed carbon electrodes (SPCE) were modified with nanocomposite membranes based on polystyrene sulfonate (PSS) or poly(diallyldimethylammonium) (PDDA) matrices and different nanomaterials. Carbon nano-powders (CnP), carbon nano-fibers (CnF) and multi-walled carbon nano-tubes (MWCNTs) were incorporated on PSS matrix. Nickel was incorporated by ion exchange in PSS-CnP composite membranes. Gold nanoparticles (AuNp) were photochemically and electrochemically synthesised and introduced into PDDA membranes. The electrochemical behaviour of methylisothiazolinone (MIT) using these modified electrodes was studied by cyclic voltammetry in 0.1 mol L −1 NaOH. No electrochemical response is obtained on PSS-nanocarbon transducers at the assayed conditions. The nickel-based transducers allow the MIT identification but not quantification. Using AuNp-based electrochemical transducers, it is observed that in presence of MIT, the electron transfer for AuNp reduction is inhibited, and an oxidation peak appears at + 0.45 V, indicating an interaction between MIT and AuNp on the electrode surface. These facts support the usefulness of the AuNp-based electrodes for the determination of MIT. The intensity of the anodic peak observed at + 0.45 V vs. Ag/AgCl was used as analytical signal for MIT determination. A linear relationship between anodic peak current and MIT concentration is observed in the range 8.7 to 36 mg L −1 using the transducer prepared by incorporating gold into the PDDA membrane by ion exchange and synthesising AuNp electrochemically. For this electrode, the limit of detection is 2.6 mg L −1 and the reproducibility, expressed as relative standard deviation (RSD), is lower than 7%. Graphical abstract Schematic representation of the preparation of gold nanoparticles (AuNp) and poly(diallyldimethylammonium) (PDDA)-based platforms and methylisothiazolinone (MIT) electrochemical response on these nanostructured platforms.