Pablo Fanjul-Bolado

Two-dimensional materials (2MD) have emerged as a transformative platform for biomedical sensing owing to their unique properties. Over the past decade, 2DM, such as graphene and MXenes have enabled sensing concepts with the potential to surpass the sensitivity, speed, and miniaturization limits of conventional technologies. When combined with other innovative materials such as aptamers or magnetic nanoparticles (MNPs), a range of components can be developed. From electrochemical electrodes and field effect transistor channels to conductive inks and triboelectric nanogenerators, these components offer a variety of biomedical applications that have the potential to revolutionize healthcare. This Roadmap reviews the current state of 2DM-based biomedical sensing within four EU-funded Horizon Europe projects under the Graphene Flagship, identifies key scientific and technological barriers, and outlines priority research directions for the short (2027) and mid-term (2030) future. The analysis focuses on the materials, various components comprising these materials, and several biomedical applications that are currently being researched within the overall biomedical focus of the Graphene Flagship Initiative. By consolidating expert perspectives from the four consortia, this roadmap aims to guide coordinated efforts to accelerate the transition of 2DM from laboratory demonstrations to robust, scalable, and impactful biomedical sensing technologies.

Electrogenerated chemiluminescence (ECL) is a powerful analytical technique that combines the best features of both electrochemical and photoluminescence methods. In this work, we present a direct ECL-based method for the detection of fentanyl using unmodified screen-printed electrodes. The analysed system consists of tris(2,2′-bipyridyl)ruthenium(II) (Ru(bpy)₃²⁺) as the luminophore and fentanyl as the co-reactant. A comprehensive optimization of the experimental parameters, such as buffer pH, luminophore concentration and working electrode material, was performed in order to maximize the ECL response. The optimal conditions are identified as PBS buffer pH 6, 2.5 × 10⁻³ M Ru(bpy)₃²⁺ and bare gold screen-printed electrodes. Under these conditions, the system exhibited a strong and reproducible ECL signal, with a linear response to fentanyl concentration from 1 × 10⁻⁷ to 1 × 10⁻⁵ M and a limit of detection of 6.7 × 10⁻⁸ M. Notably, the proposed method does not require electrode surface modification, sample pretreatment or complex instrumentation, offering a rapid, sensitive, and cost-effective alternative for fentanyl detection. Furthermore, the storage of bare SPEs at room temperature in a dry place ensures their stability over months or even years, overcoming the limitations offered by ECL systems based on modifications of the working electrode with different nanomaterials. These findings highlight the potential of the proposed ECL approach as a robust and sensitive tool for the detection of synthetic opioids. Its simplicity, portability, and analytical performance make it particularly attractive for forensic and clinical applications where rapid and accurate opioid screening is essential.

Electrogenerated chemiluminescence (ECL) is a powerful analytical technique that combines the best features of both electrochemical and photoluminescence methods. In this work, we present a direct ECL-based method for the detection of fentanyl using unmodified screen-printed electrodes. The analysed system consists of tris(2,2'-bipyridyl)ruthenium(II) (Ru(bpy)32+) as the luminophore and fentanyl as the co-reactant. A comprehensive optimization of the experimental parameters, such as buffer pH, luminophore concentration and working electrode material, was performed in order to maximize the ECL response. The optimal conditions are identified as PBS buffer pH 6, 2.5×10⁻³ M Ru(bpy)32+ and bare gold screen-printed electrodes. Under these conditions, the system exhibited a strong and reproducible ECL signal, with a linear response to fentanyl concentration from 1×10⁻⁷ to 1×10⁻⁵ M and a limit of detection of 6.7×10⁻⁸ M. Notably, the proposed method does not requires the electrode surface modification, sample pretreatment or complex instrumentation, offering a rapid, sensitive, and cost-effective alternative for fentanyl detection. Furthermore, the storage of bare SPEs at room temperature in a dry place ensures their stability over months or even years, overcoming the limitations offered by ECL systems based on modifications of the working electrode with different nanomaterials. These findings highlight the potential of the proposed ECL approach as a robust and sensitive tool for the detection of synthetic opioids. Its simplicity, portability, and analytical performance make it particularly attractive for forensic and clinical applications where rapid and accurate opioid screening is essential.

A new spectroelectrochemical two-enzyme sensor system has been developed for the detection of acetaldehyde in wine. A combination of spectroscopy and electrochemistry improves the analytical features of the electrochemical sensor because the optical information collected with this system is only associated with acetaldehyde and avoids the interferents also present in wines as polyphenols. Spectroelectrochemical detection is achieved by the analysis of the optical properties of the K₃[Fe(CN)₆]/K₄[Fe(CN)₆] redox couple involved in the enzymatic process: aldehyde dehydrogenase catalyzes the aldehyde oxidation using β-nicotinamide adenine dinucleotide hydrate (NAD⁺) as a cofactor and, simultaneously, diaphorase reoxidizes the NADH formed in the first enzymatic process due to the presence of K₃[Fe(CN)₆]. An analysis of the characteristic UV-vis bands of K₃[Fe(CN)₆] at 310 and 420 nm allows the detection of acetaldehyde, since absorption bands are only related to the oxidation of this substrate, and avoids the contribution of other interferents.

Health monitoring is experiencing a radical shift from clinic‐based to point‐of‐care and wearable technologies, and a variety of nanomaterials and transducers have been employed for this purpose. 2D materials (2DMs) hold enormous potential for novel electronics, yet they struggle to meet the requirements of wearable technologies. Here, aiming to foster the development of 2DM‐based wearable technologies, reduced graphene oxide (rGO)‐based liquid‐gated transistors (LGTs) for cation sensing in artificial sweat endowed with distinguished performance and great potential for scalable manufacturing is reported. Laser micromachining is employed to produce flexible transistor test patterns employing rGO as the electronic transducer. Analyte selectivity is achieved by functionalizing the transistor channel with ion‐selective membranes (ISMs) via a simple casting method. Real‐time monitoring of K⁺ and Na⁺ in artificial sweat is carried out employing a gate voltage pulsed stimulus to take advantage of the fast responsivity of rGO. The sensors show excellent selectivity toward the target analyte, low working voltages (<0.5 V), fast (5–15 s), linear response at a wide range of concentrations (10 µm to 100 mm), and sensitivities of 1 µA/decade. The reported strategy is an important step forward toward the development of wearable sensors based on 2DMs for future health monitoring technologies.

In the present paper, the design, fabrication, and analytical applications of three novel cells for flow injection, thin-layer, and batch electrochemical measurements using screen-printed electrode chips (SPECs) are described. Each cell consisted of an acrylic base and a transparent acrylic cover. The essential construction feature of each cell base was a cavity to accommodate the SPEC, whereas the construction features of the clear acrylic cover determined the cell shape and its function. The presented cells offered several common advantages, which include (i) convenient electrical connection of the SPEC to any potentiostat without the need for special cables, (ii) the SPEC was completely contained within the cell body, which eliminated the risk of its breakage, (iii) suitable for use with a large number of commercially available SPECs, and (iv) excellent SPEC sealing. The flow cell offered additional advantages of convenient customization of the cell dead volume and convenient visual inspection of the surface and the vicinity of SPEs. The presented thin-layer cell is the first report on a dedicated cell which realized a near-ideal thin-layer steady-state voltammetry using SPECs. The universal batch cell (UBC) offered extreme versatility and proved suitable for all batch applications in sample volumes ranging from 25 μL to 40 mL with an optional controlled temperature and atmosphere. Moreover, a novel way to achieve stirred-solution chronoamperometry and hydrodynamic voltammetry using SPECs (with superior signal-to-noise ratios) using the UBC is described. Electrochemical measurements to demonstrate the merits and the applicability of all cells are also presented.

A new transducer based on a screen-printed carbon electrode has been developed for the quantification of thiram. Detection of this fungicide is based on the performance of two enzymes: (1) aldehyde dehydrogenase catalyzes the aldehyde oxidation using NAD⁺ as a cofactor and simultaneously, (2) diaphorase reoxidizes the NADH formed in the first enzymatic process due to the presence of hexacyanoferrate(III) which is reduced to hexacyanoferrate(II). Taking into account that aldehyde dehydrogenase is inhibited by thiram, the current decreases with pesticide concentration and thiram can be electrochemically quantified below legal limits. The transducer proposed in this work involves the modification of the carbon WE with the co-factors (NAD⁺ and hexacyanoferrate(III)) required in the enzymatic system. The new device employed in this work allows the detection of 0.09 ppm thiram, a concentration below legal limits (Maximum Residue Limits 0.1–10 ppm).

Dithiocarbamate fungicides (DTFs) are widely used to control various fungal diseases in crops and ornamental plants. Maximum residual limits in the order of ppb-ppm are currently imposed by legislation to prevent toxicity problems associated with excessive use of DTFs. The specific analytical determination of DTFs is complicated by their low solubility in water and organic solvents. This review summarizes the current analytical procedures used for the analysis of DTF, including chromatography, spectroscopy, and sensor-based methods and discusses the challenges related to selectivity, sensitivity, and sample preparation. Biosensors based on enzymatic inhibition demonstrated potential as analytical tools for DTFs and warrant further research, considering novel enzymes from extremophilic sources. Meanwhile, Raman spectroscopy and various sensors appear very promising, provided the selectivity issues are solved.

Over the last few decades, a large number of pH sensitive materials with new compositions and structures have been proposed. Solid state sensors based on organic, inorganic and composite materials are actively investigated, with an increasing interest in the performance offered by nano-scale materials. Our review provides a thorough, up-to-date knowledge of a wide range of pH measurement methods and related sensing materials, firstly by introducing well established materials and methods for pH sensing and then, by covering recent developments in inorganic, organic and nano-engineered devices. The main sensor parameters, including sensitivity, stability, response time and testing conditions are reported. Given the importance of pH sensing in environmental applications, in particular seawater monitoring, sensors tested in seawater are highlighted and discussed. Graphical Abstract