Search Results (456)

Detecting small molecules in biological fluids is essential for diagnosing diseases, monitoring therapy, and studying how the body works. Traditional biosensing methods—such as amperometric, optical, or piezoelectric systems—offer excellent sensitivity but often rely on complex instruments, additional reagents, or time-consuming sample preparation. Potentiometric biosensors, by contrast, provide a simpler, low-power, and label-free alternative that can operate directly in biological environments. This review explores the latest progress in potentiometric biosensing for small-molecule detection, focusing on new solid-contact materials and advanced sensing membranes and compact device designs. We also discuss key challenges, including biofouling, matrix effects, and signal drift, together with promising strategies such as antifouling coatings, nanostructured interfaces, and calibration-free operation. Finally, we highlight how combining potentiometric sensors with artificial intelligence, digital data processing, and flexible electronics is shaping the future of personalized and point-of-care diagnostics. By summarizing recent advances and identifying remaining barriers, this review aims to show why potentiometric biosensors are becoming a powerful and versatile platform for next-generation biomedical analysis. Full article

In situ electrochemical imaging of corrosion reactions is performed directly by scanning electrochemical microscopy (SECM) in generation-collection mode. This method involves redox conversion of soluble metal ions at the amperometric tip for quantification. Unfortunately, many metals, such as copper, do not undergo redox conversion to a soluble state and are deposited on the SECM tip. They therefore modify the electrochemical behavior of the tip and require consideration of metal stripping processes. In addition, the miniaturization of the electrode to operate as a microelectrode tip can be accompanied by variations in the potential range and distribution of the redox processes related to copper deposition and redissolution, thus complicating the adequate choice of electrochemical conditions applied to the tip for the unambiguous operation of SECM in the generation-collection mode to study the corrosion of copper-based materials. Therefore, in this work, a study of different parameters for the amperometric determination of Cu²⁺ ions was conducted using gold disk electrodes of 500 and 10 μm diameter to represent the typical sizes employed in conventional and scanning microelectrochemical measurements. The investigation was performed to analyze the effect of underpotential deposition (UPD) and overpotential deposition (ODP) on the voltammetric characteristics of copper deposition and redissolution resulting from variations in the solution composition, i.e., the nature of anions and pH. The dependence and limits of the reduction and reoxidation waves were analyzed as functions of the Cu²⁺ ion concentration, the ionic strength of the electrolyte, and the pH of the solution. The results were interpreted as UPD and OPD. Under conditions close to the marine environment, the release of Cu²⁺ ions can be unambiguously detected and quantified from potentials above −0.1 V vs. Ag/AgCl. Full article

The bioanalysis of body fluids plays a crucial role in clinical diagnostics, pharmaceutical research, forensic science, and biomarker discovery. Traditional chromatographic techniques are widely used in clinical laboratories but are often costly and time-consuming. Capillary electrophoresis (CE) and its modifications, such as capillary zone electrophoresis, isotachophoresis, and micellar electrokinetic chromatography, have emerged as efficient, cost-effective, and miniaturized alternatives for analyzing small organic and inorganic molecules in biological fluids. This paper deals with the applications of CE-based electromigration techniques in the determination of various analytes in urine, blood, saliva, and cerebrospinal fluid. The study further discusses the advantages and limitations of different detection methods, including ultraviolet-visible spectroscopy, laser-induced fluorescence, mass spectrometry, conductivity, and amperometric detection. A focus is given to the identification and quantification of amino acids and their metabolites as potential biomarkers for metabolic and degenerative disorders. The work highlights recent advancements in CE methodologies and their potential to enhance sensitivity and selectivity in bioanalytical applications. Full article

Colorectal cancer (CRC) remains a significant global health burden, mainly due to late diagnosis and chemotherapy resistance. Macrophage migration inhibitory factor (MIF), a proinflammatory cytokine associated with tumor progression, has emerged as a promising biomarker in CRC. However, its clinical utility is limited by the lack of rapid and accessible detection methods. In this study, we report an electrochemical immunotechnology for the sensitive and selective quantification of MIF protein in CRC tissue samples. By combining magnetic microparticles (MMPs), antibody-based recognition, horseradish peroxidase (HRP) labeling, and amperometric transduction at disposable screen-printed carbon electrodes (SPCEs), the developed methodology displayed a linear dynamic range from 0.24 to 20 ng mL⁻¹, enabling quantification across clinically relevant MIF levels, and achieving a low limit of detection (0.07 ng mL⁻¹). In addition, the developed method is the only one reported for MIF assembled on MMPs and addresses its determination in a relevant oncological scenario (paired non-tumoral (NT) and tumoral (T) tissues from individuals diagnosed with CRC at different stages of the disease). The analysis, requiring only 100 ng of tissue extract, allowed efficient discrimination between NT and T paired tissues, and successfully differentiated between healthy, early (I–II) and advanced (III–IV) CRC stages, achieving these results in just 105 min. Full article

Alanine aminotransferase (ALT) is a key biomarker of liver function. Compared with conventional assays for ALT detection—which are expensive, time-consuming, labor-intensive, and require experienced personnel—biosensors represent a promising alternative, but it remains unclear which biorecognitive enzymatic configuration offers the best analytical performance for ALT detection. This study presents the development and comparative evaluation of two amperometric biosensors based on oxidase biorecognition elements: pyruvate oxidase (POx) and glutamate oxidase (GlOx). Enzymes were immobilized onto platinum electrodes under optimized conditions using entrapment for POx (pH 7.4, enzyme loading 1.62 U/µL, PVA-SbQ concentration 13.2%) and covalent crosslinking for GlOx (pH 6.5, enzyme loading 2.67%, glutaraldehyde concentration 0.3%). Analytical parameters were systematically assessed, including linear range (1–500 U/L for POx vs. 5–500 U/L for GlOx), limit of detection (1 U/L for both), and sensitivity (0.75 vs. 0.49 nA/min at 100 U/L). The POx-based biosensor demonstrated higher sensitivity and lower detection limits, whereas the GlOx-based biosensor exhibited greater stability in complex solutions and reduced assay costs due to a simpler working solution. Moreover, while the POx-based system is uniquely suited for ALT determination, the GlOx-based sensor can be affected by AST activity in samples but may also be adapted for targeted AST detection. Overall, the study highlights a trade-off between sensitivity, robustness, and versatility in ALT biosensor design, providing guidance for the rational development of clinically relevant devices. Full article

Background/Objectives: This study comprehensively characterized the O- and N-glycosylation profiles of bispecific antibodies (BsAbs) via advanced analytical techniques to evaluate their structural and functional implications. Methods: High-resolution MS revealed O-xylosylation at Ser468 within the (G4S)4 linker peptide, which was identified as xylose with a molecular weight of 132.042 Da. HILIC-HPLC analysis of N-glycosylation revealed glycan species engineered to eliminate Fc effector functions. O-glycosylation analysis via β-elimination followed by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) identified xylose as the predominant glycan. Results: O-xylosylation does not affect the binding of BsAbs to either antigen Programmed Death-1 (PD-1) or Vascular Endothelial Growth Factor (VEGF). Notably, O-xylosylation interactions with mannose receptor represent the first discovery highlighting potential immunomodulatory roles. Conclusions: This study highlights the critical importance of monitoring comprehensive glycosylation characterization during the development of BsAb with (G4S)n linkers to ensure optimal therapeutic efficacy, safety, and reduced immunogenic potential. Full article

A novel voltammetric sensor was constructed by modifying a glassy carbon electrode with a composite material consisting of platinum–nickel-doped tin oxide and carbon black (PtNiSnO₂-CB/GCE), enabling highly sensitive differential pulse voltammetry (DPV) determination of trazodone HCl (TRZ). The DPV experimental parameters, including the composition of the supporting electrolyte and instrumental settings, were carefully optimized to achieve maximum analytical efficiency. Within the linear range of 1–10 µM, quantification of TRZ molecules could be performed without the preconcentration step. When applying a 60 s accumulation time (in the range 0.02–0.2 µM of TRZ), the detection limit reached 4.1 nM (1.67 mg L⁻¹), indicating superior sensitivity compared to previously reported voltammetric techniques. The method demonstrated good reproducibility, with a relative standard deviation of 4.3% for 10 repeated measurements at 0.06 µM TRZ. The developed sensor exhibits excellent stability, simplicity of fabrication, and operational convenience. Its practical applicability was confirmed by the successful analysis of molecules of TRZ in diverse sample types, including pharmaceutical products, urine, plasma, river water, and artificial gastric and intestinal fluids, with recovery rates between 97.7% and 104.2%. Flow injection analysis (FIA) with amperometric detection was also performed for TRZ molecule determination. Full article

Amperometric biosensors, due to their high sensitivity, fast response time, low cost, simple control, miniaturization capabilities, and other advantages, are receiving significant attention in the field of medical diagnostics, especially in monitoring blood glucose levels in diabetic patients. In this study, an amperometric glucose biosensor based on immobilized enzyme glucose oxidase (GOx) and bimetallic platinum cobalt (PtCo) nanoparticles was developed. The PtCo nanoparticles, deposited on a graphite rod electrode, exhibited peroxidase-like catalytic properties and were able to electrocatalyze the reduction of H₂O₂. After immobilization of the GOx, an amperometric signal generated by the biosensor was directly proportional to the glucose concentration in the range of 0.04–2.18 mM. The biosensor demonstrated a sensitivity of 19.38 μA mM⁻¹ cm⁻², with a detection limit of 0.021 mM and a quantification limit of 0.064 mM. In addition to this analytical performance, the biosensor exhibited excellent repeatability (relative standard deviation (RSD) was 4.90%); operational and storage stability, retaining 98.93% and 95.33% of its initial response after 26 cycles of glucose detection and over a 14-day period, respectively; and anti-interference ability against electroactive species, as well as exceptional selectivity for glucose and satisfactory reproducibility (RSD 8.90%). Additionally, the biosensor was able to detect glucose levels in blood serum with a high accuracy (RSD 5.89%), indicating potential suitability for glucose determination in real samples. Full article

We develop a modular, wireless, solar- and battery-powered system for detecting chlorpyrifos (Lorsban^TM 2.5% DP) in water using electrochemical impedance spectroscopy (EIS). The system integrates a Raspberry Pi Zero 2W for data processing, Python-based software (version 3.12.2), and a solar charge manager to power all components via a lithium-ion battery and solar panel. A commercial EmStat Pico Module and an amperometric biosensor with acetylcholinesterase (AChE) detect chlorpyrifos. Nine water samples with varying concentrations were tested using a 20 Hz–200 kHz frequency sweep and 15 mV excitation. Bode plots and statistical analyses confirmed statistically significant impedance variation as a function of chlorpyrifos concentration, validating the system as a portable, sensitive, and effective tool for environmental monitoring. Full article

An important class of analytes are volatile organic carbons (VOCs), particularly aliphatic primary alcohols. Here, we report the straightforward modification of a commercially available carbon monoxide sensor to detect a range of aliphatic primary alcohols at room temperature. The mass transport mechanisms governing the performance of the sensor were investigated using diffusion in multiple layers of the sensor to model the response to an abrupt change in analyte concentration. The sensor was shown to have a large capacitance because of the nanoparticulate nature of the platinum working electrode. It was also shown that the modified sensor had performance characteristics that were mainly determined by the condensation of the analyte during diffusion through the membrane pores. The sensor was capable of a quantitative amperometric response (sensitivity of approximately 2.2 µA/ppm), with a limit of detection (LoD) of 17 ppm methanol, 2 ppm ethanol, 3 ppm heptan-1-ol, and displayed selectivity towards different VOC functional groups (the sensor gives an amperometric response to primary alcohols within 10 s, but not to esters or carboxylic acids). Full article

A facile fabrication method was developed for the growth of Cu_1.8Se nanosheets (NSs) on a Cu foil substrate, enabling dual-functionality as an electrochemical sensor for H₂O₂ and an active surface-enhanced Raman scattering (SERS) substrate. The process involved the preparation of Cu(OH)₂ nanowires (NWs) via electrochemical oxidation, followed by chemical conversion to Cu_1.8Se through a selenization process. The morphology, composition, and microstructure of the resulting Cu_1.8Se NSs were systematically characterized using scanning electron microscopy (SEM), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). The Cu_1.8Se NSs exhibited excellent electrocatalytic activity for H₂O₂ reduction, achieving a notably low detection limit of 1.25 μM and demonstrating rapid response and high sensitivity with a linear relationship in amperometric detection. Additionally, SERS experiments using Rhodamine B as a probe molecule and the Cu_1.8Se NS/Cu foil as a substrate displayed outstanding performance, with a detection limit as low as 1 μM. The flower-like structure of the Cu_1.8Se NSs exhibited linear dependence between analyte concentration and detection signals, along with satisfactory reproducibility in dual-sensing applications. These findings underscore the scalability and potential of this fabrication approach for advanced sensor development. Full article

Nitrite (NO₂⁻) has long been recognized as a contaminant of concern due to its detrimental effects on both human health and the environment. As a result, there is a continuing need to develop sensitive, real-time, low-cost, and portable systems for the accurate detection of trace levels of NO₂⁻ in drinking water. We present a novel, low-cost, and easy-to-fabricate amperometric sensor designed for detecting low concentrations of NO₂⁻ in drinking water. The fabrication technique involves the electrodeposition of manganese and copper oxides onto a carbon working electrode. CuO and MnO₂ act synergistically as efficient catalysts for the electrooxidation of nitrite to nitrate (NO₃⁻) thanks to their complementary redox properties. The resulting sensor exhibits high catalytic activity toward the electrooxidation of NO₂⁻, with a sensitivity of 10.83 μA/µM, a limit of detection (LOD) of 0.071 µM, and a good linear dynamic concentration range (0.2–60 µM). The sensor’s performance was evaluated against potential interfering analytes (NO₃⁻, Cl⁻, NH₄⁺, and NH₂Cl), all of which showed negligible interference. Reproducibility (maximum standard deviation 2.91%) and repeatability (usable up to three times) were also evaluated. Full article

A polyoxometalate-based metal–organic complex with the ability to treat pollutants in water was obtained under hydrothermal conditions, namely [Ni(H₂L)(HL)₂](PMo₁₂O₄₀)·3H₃O·4H₂O (1) (H₂L = 4,4′-(1H,1′H-[2,2′-biimidazole]-1,1′-diyl)dibenzoicacid). Structural analysis reveals that the [Ni(H₂L)(HL)₂] units are interconnected into a 2D layer via hydrogen bonds between adjacent carboxyl groups and water molecules of crystallization. [PMo₁₂O₄₀]³⁻ anions are embedded within the larger pores of the layer and are connected to the adjacent layers through hydrogen bonds, ultimately expanding the structure into a 3D supramolecular architecture. The intermolecular interactions were studied via Hirshfeld surface (HS) analysis. Electrochemical performance tests reveal that 1 exhibits electrocatalytic activity toward the oxidation and reduction of diverse pollutants in water, including NO₂⁻, Cr(VI), BrO₃⁻, Fe(III), and ascorbic acid (AA). Additionally, it can also serve as an amperometric sensor for the detection of BrO₃⁻ and Cr(VI). Photocatalytic studies reveal that compound 1 functions as a bifunctional photocatalyst, which not only achieves efficient degradation of organic dyes but also demonstrates remarkable reduction efficiency for toxic Cr(VI). Compound 1 demonstrates significant potential for practical water remediation applications. Full article

Flow injection analysis (FIA) is widely used in drug screening, neurotransmitter detection, and water analysis. In this study, we investigated the electrochemical sensing performance of glassy graphene electrodes derived from pyrolyzed positive photoresist films (PPFs) via rapid thermal annealing (RTA) on SiO₂/Si and polycrystalline diamond (PCD). Glassy graphene films fabricated at 800, 900, and 950 °C were characterized using Raman spectroscopy, scanning electron microscopy (SEM), and atomic force microscopy (AFM) to assess their structural and morphological properties. Electrochemical characterization in phosphate-buffered saline (PBS, pH 7.4) revealed that annealing temperature and substrate type influence the potential window and double-layer capacitance. The voltammetric response of glassy graphene electrodes was further evaluated using the surface-insensitive [Ru(NH₃)₆]^3+/2+ redox marker, the surface-sensitive [Fe(CN)₆]^3−/4− redox couple, and adrenaline, demonstrating that electron transfer efficiency is governed by annealing temperature and substrate-induced microstructural changes. FIA with amperometric detection showed a linear electrochemical response to adrenaline in the 3–300 µM range, achieving a low detection limit of 1.05 µM and a high sensitivity of 1.02 µA cm⁻²/µM. These findings highlight the potential of glassy graphene as a cost-effective alternative for advanced electrochemical sensors, particularly in biomolecule detection and analytical applications. Full article

Artificial enzymes or nanozymes (NZs) are gaining significant attention in biotechnology due to their stability and cost-effectiveness. NZs can offer several advantages over natural enzymes, such as enhanced stability under harsh conditions, longer shelf life, and reduced production costs. The booming interest in NZs is likely to continue as their potential applications expand. In our previous studies, we reported the “green” synthesis of copper hexacyanoferrate (gCuHCF) using the oxidoreductase flavocytochrome b₂ (Fcb₂). Organic–inorganic micro-nanoparticles were characterized in detail, including their structure, composition, catalytic activity, and electron-mediator properties. An SEM analysis revealed that gCuHCF possesses a flower-like structure well-suited for concentrating and stabilizing Fcb₂. As an effective peroxidase (PO) mimic, gCuHCF has been successfully employed for H₂O₂ detection in amperometric sensors and in several oxidase-based biosensors. In the current study, we demonstrated the uniqueness of gCuHCF that lies in its multifunctionality, serving as a PO mimic, a chemosensor for ammonium ions, a biosensor for L-lactate, and exhibiting perovskite-like properties. This exceptional ability of gCuHCF to enhance fluorescence under blue light irradiation is being reported for the first time. Using gCuHCF as a PO-like NZ, novel oxidase-based sensors were developed, including an optical biosensor for L-arginine analysis and electrochemical biosensors for methanol and glycerol determination. Thus, gCuHCF, synthesized via Fcb₂, presents a promising platform for the development of amperometric and optical biosensors, bioreactors, biofuel cells, solar cells, and other advanced devices. The innovative approach of utilizing biocatalysts for nanoparticle synthesis highlights a groundbreaking direction in materials science and biotechnology. Full article