Electroanalysis overview: additive manufactured biosensors using fused filament fabrication.

Additive manufacturing (3D-printing), in particular fused filament fabrication, presents a potential paradigm shift in the way electrochemical based biosensing platforms are produced, giving rise to a new generation of personalized and on-demand biosensors. The use of additive manufactured biosensors is unparalleled giving rise to unique customization, facile miniaturization, ease of use, economical but yet, still providing sensitive and selective approaches towards the target analyte. In this mini review, we focus on the use of fused filament fabrication additive manufacturing technology alongside different biosensing approaches that exclusively use antibodies, enzymes and associated biosensing materials (mediators) providing an up-to-date overview with future considerations to expand the additive manufacturing biosensors field.

Recycled PETg embedded with graphene, multi-walled carbon nanotubes and carbon black for high-performance conductive additive manufacturing feedstock.

The first report of conductive recycled polyethylene terephthalate glycol (rPETg) for additive manufacturing and electrochemical applications is reported herein. Graphene nanoplatelets (GNP), multi-walled carbon nanotubes (MWCNT) and carbon black (CB) were embedded within a recycled feedstock to produce a filament with lower resistance than commercially available conductive polylactic acid (PLA). In addition to electrical conductivity, the rPETg was able to hold >10 wt% more conductive filler without the use of a plasticiser, showed enhanced temperature stability, had a higher modulus, improved chemical resistance, lowered levels of solution ingress, and could be sterilised in ethanol. Using a mix of carbon materials CB/MWCNT/GNP (25/2.5/2.5 wt%) the electrochemical performance of the rPETg filament was significantly enhanced, providing a heterogenous electrochemical rate constant, k0, equating to 0.88 (±0.01) × 10-3 cm s-1 compared to 0.46 (±0.02) × 10-3 cm s-1 for commercial conductive PLA. This work presents a paradigm shift within the use of additive manufacturing and electrochemistry, allowing the production of electrodes with enhanced electrical, chemical and mechanical properties, whilst improving the sustainability of the production through the use of recycled feedstock.

Electroanalytical Overview: The Sensing of Mesalamine (5-Aminosalicylic Acid).

Mesalamine, known as 5-aminosalicylic acid, is a medication used primarily in the treatment of inflammatory bowel disease, including ulcerative colitis and Crohn's disease. 5-Aminosalicylic acid can be measured using various benchtop laboratory techniques which involve liquid chromatography-mass spectroscopy, but these are sophisticated and large, meaning that they cannot be used on-site because transportation of the samples, chemicals, and physical and biological reactions can potentially occur, which can affect the sample's composition and potentially result in inaccurate results. An alternative approach is the use of electrochemical based sensing platforms which has the advantages of portability, cost-efficiency, facile miniaturization, and rapid analysis while nonetheless providing sensitivity and selectivity. We provide an overview of the use of the electroanalytical techniques for the sensing of 5-aminosalicylic acid and compare them to other laboratory-based measurements. The applications, challenges faced, and future opportunities for electroanalytical based sensing platforms are presented in this review.

All-in-one continuous electrochemical monitoring of 2-phenylphenol removal from water by electro-Fenton treatment.

The biggest allure of heterogeneous electro-Fenton (HEF) processes largely fails on its high efficiency for the degradation of a plethora of hazardous compounds present in water, but still challenging to search for good and cost-effective electrocatalyst. In this work, carbon black (CB) and oxidised carbon black (CBox) materials were investigated as cathodes in the electrochemical production of hydrogen peroxide involved in HEF reaction for the degradation of 2-phenylphenol (2PP) as a target pollutant. The electrodes were fabricated by employing carbon cloth as support, and the highest H2O2 production yields were obtained for the CBox, pointing out the beneficial effect of the hydrophilic character of the electrode and oxygen-type functionalization of the carbonaceous surface. HEF degradation of 2PP was explored at -0.7 V vs. Ag/AgCl exhibiting the best conversion rates and degradation grade (total organic carbon) for the CBox-based cathode. In addition, the incorporation of an electrochemical sensor of 2PP in line with the HEF reactor was accomplished by the use of screen-printed electrodes (SPE) in order to monitor the pollutant degradation. The electrochemical sensor performance was evaluated from the oxidation of 2PP in the presence of Fe2+ ions by using square wave voltammetry (SWV) technique. The best electrochemical sensor performance was based on SPE modified with Meldola Blue showing a high sensitivity, low detection limit (0.12 ppm) and wide linear range (0.5-21 ppm) with good reproducibility (RSD 2.3 %). The all-in-one electrochemical station has been successfully tested for the degradation and quantification of 2PP, obtaining good recoveries analysing spiked waters from different water matrices origins.

Multi-walled carbon nanotubes/carbon black/rPLA for high-performance conductive additive manufacturing filament and the simultaneous detection of acetaminophen and phenylephrine.

The combination of multi-walled carbon nanotubes (MWCNT) and carbon black (CB) is presented to produce a high-performance electrically conductive recycled additive manufacturing filament. The filament and subsequent additively manufactured electrodes were characterised by TGA, XPS, Raman, and SEM and showed excellent low-temperature flexibility. The MWCNT/CB filament exhibited an improved electrochemical performance compared to an identical in-house produced bespoke filament using only CB. A heterogeneous electrochemical rate constant, [Formula: see text] of 1.71 (± 0.19) × 10-3 cm s-1 was obtained, showing an almost six times improvement over the commonly used commercial conductive CB/PLA. The filament was successfully tested for the simultaneous determination of acetaminophen and phenylephrine, producing linear ranges of 5-60 and 5-200 µM, sensitivities of 0.05 µA µM-1 and 0.14 µA µM-1, and limits of detection of 0.04 µM and 0.38 µM, respectively. A print-at-home device is presented where a removable lid comprised of rPLA can be placed onto a drinking vessel and the working, counter, and reference components made from our bespoke MWCNT/CB filament. The print-at-home device was successfully used to determine both compounds within real pharmaceutical products, with recoveries between 87 and 120% over a range of three real samples. This work paves the way for fabricating new highly conductive filaments using a combination of carbon materials with different morphologies and physicochemical properties and their application to produce additively manufactured electrodes with greatly improved electrochemical performance.

Electroanalytical overview: the sensing of carbendazim.

Carbendazim is a broad-spectrum systemic fungicide that is used to control various fungal diseases in agriculture, horticulture, and forestry. Carbendazim is also used in post-harvest applications to prevent fungal growth on fruits and vegetables during storage and transportation. Carbendazim is regulated in many countries and banned in others, thus, there is a need for the sensing of carbendazim to ensure that high levels are avoided which can result in potential health risks. One approach is the use of electroanalytical sensors which present a rapid, but highly selective and sensitive output, whilst being economical and providing portable sensing platforms to support on-site analysis. In this minireview, we report on the electroanalytical sensing of carbendazim overviewing recent advances, helping to elucidate the electrochemical mechanism and provide conclusions and future perspectives of this field.

Mixed Graphite/Carbon Black Recycled PLA Conductive Additive Manufacturing Filament for the Electrochemical Detection of Oxalate.

Mixing of graphite and carbon black (CB) alongside recycled poly(lactic acid) and castor oil to create an electrically conductive additive manufacturing filament without the use of solvents is reported herein. The additively manufactured electrodes (AMEs) were electrochemically benchmarked against a commercial conductive filament and a bespoke filament utilizing only CB. The graphite/CB produced a heterogeneous rate constant, k0, of 1.26 (±0.23) × 10-3 cm s-1 and resistance of only 155 ± 15 Ω, compared to 0.30 (±0.03) × 10-3 cm s-1 and 768 ± 96 Ω for the commercial AME. Including graphite within the filament reduced the cost of printing each AME from £0.09, with the CB-only filament, to £0.05. The additive manufacturing filament was successfully used to create an electroanalytical sensing platform for the detection of oxalate within a linear range of 10-500 µM, achieving a sensitivity of 0.0196 µA/µM, LOD of 5.7 µM and LOQ of 18.8 µM was obtained. Additionally, the cell was tested toward the detection of oxalate within a spiked synthetic urine sample, obtaining recoveries of 104%. This work highlights how, using mixed material composites, excellent electrochemical performance can be obtained at a reduced material cost, while also greatly improving the sustainability of the system.

Toward the rapid diagnosis of sepsis: dendritic copper nanostructure functionalized diazonium salt modified screen-printed graphene electrode for IL-6 detection.

Sepsis, an infectious disease affecting millions of people's health worldwide each year, calls for urgent attention to an improvement of analytical devices. Chemiluminescence immunoassay is a typical diagnostic method utilized to assess the risk development of sepsis. However, due to its high-cost, delayed, and complicated procedure, the practical utilization is therefore undoubtedly limited, especially for point-of-care test. Herein, we fabricated for the first time an immunosensor based on dendritic copper nanostructures (CuNSs) combined with 4-aminobenzoic acid (4-AB, the diazonium salt) as antibody linker modified on a screen-printed graphene electrode for the early detection of the sepsis biomarker interleukin-6 (IL-6). The electrode fabrication is made by electrodeposition, thus eliminating the multistep of nanomaterial synthesis and time wasting. The resulting dendritic CuNSs significantly increase the effective surface area (1.2 times) and the sensor's performance. The morphology of this combination was characterized using CV, EIS, SEM, EDX, and FTIR techniques. In the detection process, the appearance of IL-6 suppresses the current response of the redox probe indicator measured by differential pulse voltammetry due to the antibody-antigen complex. The subtraction of signal (ΔI) was interpreted as IL-6 concentration. This sensor exhibited a linear range from 0.05 to 500 pg mL-1 with low detection limit of 0.02 pg mL-1, proving a possibility for early sepsis screening. In addition, the established immunosensor can successfully quantify IL-6 in human serum sample, in which the results agreed well with those achieved using the standard approach, further showing high practical applicability of this developed immunosensor.

Novel Additive Manufactured Multielectrode Electrochemical Cell with Honeycomb Inspired Design for the Detection of Methyl Parathion in Honey Samples.

The development and increase in the number of crops recently have led to the requirement for greater efficiency in world food production and greater consumption of pesticides. In this context, the widespread use of pesticides has affected the decrease in the population of pollinating insects and has caused food contamination. Therefore, simple, low-cost, and quick analytical methods can be interesting alternatives for checking the quality of foods such as honey. In this work, we propose a new additively manufactured (3D-printed) device inspired by a honeycomb cell, with 6 working electrodes for the direct electrochemical analysis of methyl parathion by reduction process monitoring in food and environmental samples. Under optimized parameters, the proposed sensor presented a linear range between 0.85 and 19.6 µmol L-1, with a limit of detection of 0.20 µmol L-1. The sensors were successfully applied in honey and tap water samples by using the standard addition method. The proposed honeycomb cell made of polylactic acid and commercial conductive filament is easy to construct, and there is no need for chemical treatments to be used. These devices based on 6 working electrodes array are versatile platforms for rapid, highly repeatable analysis in food and environment, capable of performing detection in low concentrations.

Circular Economy Electrochemistry: Recycling Old Mixed Material Additively Manufactured Sensors into New Electroanalytical Sensing Platforms.

Recycling used mixed material additively manufactured electroanalytical sensors into new 3D-printing filaments (both conductive and non-conductive) for the production of new sensors is reported herein. Additively manufactured (3D-printed) sensing platforms were transformed into a non-conductive filament for fused filament fabrication through four different methodologies (granulation, ball-milling, solvent mixing, and thermal mixing) with thermal mixing producing the best quality filament, as evidenced by the improved dispersion of fillers throughout the composite. Utilizing this thermal mixing methodology, and without supplementation with the virgin polymer, the filament was able to be cycled twice before failure. This was then used to process old sensors into an electrically conductive filament through the addition of carbon black into the thermal mixing process. Both recycled filaments (conductive and non-conductive) were utilized to produce a new electroanalytical sensing platform, which was tested for the cell's original application of acetaminophen determination. The fully recycled cell matched the electrochemical and electroanalytical performance of the original sensing platform, achieving a sensitivity of 22.4 ± 0.2 µA µM-1, a limit of detection of 3.2 ± 0.8 µM, and a recovery value of 95 ± 5% when tested using a real pharmaceutical sample. This study represents a paradigm shift in how sustainability and recycling can be utilized within additively manufactured electrochemistry toward promoting circular economy electrochemistry.

Electroanalytical overview: the sensing of hydroxylamine.

One of the principal raw ingredients used in the manufacturing of pharmaceuticals, nuclear fuel, and semiconductors is hydroxylamine, a mutagenic and carcinogenic substance, ranking high on the list of environmental contaminants. Electrochemical methods for monitoring hydroxylamine have the advantage of being portable, quick, affordable, simple, sensitive, and selective enough to maintain adequate constraints in contrast with conventional yet laboratory based quantification methods. This review outlines the most recent advancements in electroanalysis directed toward the sensing of hydroxylamine. Potential future advancements in this field are also offered, along with a discussion of method validation and the use of such devices in real samples for the determination of hydroxylamine.

Analysis of Cellular Damage Resulting from Exposure of Bacteria to Graphene Oxide and Hybrids Using Fourier Transform Infrared Spectroscopy.

With the increase in antimicrobial resistance, there is an urgent need to find new antimicrobials. Four particulate antimicrobial compounds, graphite (G), graphene oxide (GO), silver-graphene oxide (Ag-GO) and zinc oxide-graphene oxide (ZnO-GO) were tested against Enterococcus faecium, Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus. The antimicrobial effects on the cellular ultrastructure were determined using Fourier transform infrared spectroscopy (FTIR), and selected FTIR spectral metrics correlated with cell damage and death arising from exposure to the GO hybrids. Ag-GO caused the most severe damage to the cellular ultrastructure, whilst GO caused intermediate damage. Graphite exposure caused unexpectedly high levels of damage to E. coli, whereas ZnO-GO exposure led to relatively low levels of damage. The Gram-negative bacteria demonstrated a stronger correlation between FTIR metrics, indicated by the perturbation index and the minimal bactericidal concentration (MBC). The blue shift of the combined ester carbonyl and amide I band was stronger for the Gram-negative varieties. FTIR metrics tended to provide a better assessment of cell damage based on correlation with cellular imaging and indicated that damage to the lipopolysaccharide, peptidoglycan and phospholipid bilayers had occurred. Further investigations into the cell damage caused by the GO-based materials will allow the development of this type of carbon-based multimode antimicrobials.

Electroanalytical Overview: The Determination of Levodopa (L-DOPA).

L-DOPA (levodopa) is a therapeutic agent which is the most effective medication for treating Parkinson's disease, but it needs dose optimization, and therefore its analytical determination is required. Laboratory analytical instruments can be routinely used to measure L-DOPA but are not always available in clinical settings and traditional research laboratories, and they also have slow result delivery times and high costs. The use of electroanalytical sensing overcomes these problems providing a highly sensitivity, low-cost, and readily portable solution. Consequently, we overview the electroanalytical determination of L-DOPA reported throughout the literature summarizing the endeavors toward sensing L-DOPA, and we offer insights into future research opportunities.

Electrochemical and thermal detection of allergenic substance lysozyme with molecularly imprinted nanoparticles.

Lysozyme (LYZ) is a small cationic protein which is widely used for medical treatment and in the food industry to act as an anti-bacterial agent; however, it can trigger allergic reactions. In this study, high-affinity molecularly imprinted nanoparticles (nanoMIPs) were synthesized for LYZ using a solid-phase approach. The produced nanoMIPs were electrografted to screen-printed electrodes (SPEs), disposable electrodes with high commercial potential, to enable electrochemical and thermal sensing. Electrochemical impedance spectroscopy (EIS) facilitated fast measurement (5-10 min) and is able to determine trace levels of LYZ (pM) and can discriminate between LYZ and structurally similar proteins (bovine serum albumin, troponin-I). In tandem, thermal analysis was conducted with the heat transfer method (HTM), which is based on monitoring the heat transfer resistance at the solid-liquid interface of the functionalized SPE. HTM as detection technique guaranteed trace-level (fM) detection of LYZ but needed longer analysis time compared to EIS measurement (30 min vs 5-10 min). Considering the versatility of the nanoMIPs which can be adapted to virtually any target of interest, these low-cost point-of-care sensors hold great potential to improve food safety.

Low-cost, facile droplet modification of screen-printed arrays for internally validated electrochemical detection of serum procalcitonin.

This manuscript presents the design and facile production of screen-printed arrays (SPAs) for the internally validated determination of raised levels of serum procalcitonin (PCT). The screen-printing methodology produced SPAs with six individual working electrodes that exhibit an inter-array reproducibility of 3.64% and 5.51% for the electrochemically active surface area and heterogenous electrochemical rate constant respectively. The SPAs were modified with antibodies specific for the detection of PCT through a facile methodology, where each stage simply uses droplets incubated on the surface, allowing for their mass-production. This platform was used for the detection of PCT, achieving a linear dynamic range between 1 and 10 ng mL-1 with a sensor sensitivity of 1.35 × 10-10 NIC%/ng mL-1. The SPA produced an intra- and inter-day %RSD of 4.00 and 5.05%, with a material cost of £1.14. Internally validated human serum results (3 sample measurements, 3 control) for raised levels of PCT (>2 ng mL-1) were obtained, with no interference effects seen from CRP and IL-6. This SPA platform has the potential to offer clinicians vital information to rapidly begin treatment for "query sepsis" patients while awaiting results from more lengthy remote laboratory testing methods. Analytical ranges tested make this an ideal approach for rapid testing in specific patient populations (such as neonates or critically ill patients) in which PCT ranges are inherently wider. Due to the facile modification methods, we predict this could be used for various analytes on a single array, or the array increased further to maintain the internal validation of the system.

Flexible Label-Free Platinum and Bio-PET-Based Immunosensor for the Detection of SARS-CoV-2.

The demand for new devices that enable the detection of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) at a relatively low cost and that are fast and feasible to be used as point-of-care is required overtime on a large scale. In this sense, the use of sustainable materials, for example, the bio-based poly (ethylene terephthalate) (Bio-PET) can be an alternative to current standard diagnostics. In this work, we present a flexible disposable printed electrode based on a platinum thin film on Bio-PET as a substrate for the development of a sensor and immunosensor for the monitoring of COVID-19 biomarkers, by the detection of L-cysteine and the SARS-CoV-2 spike protein, respectively. The electrode was applied in conjunction with 3D printing technology to generate a portable and easy-to-analyze device with a low sample volume. For the L-cysteine determination, chronoamperometry was used, which achieved two linear dynamic ranges (LDR) of 3.98-39.0 µmol L-1 and 39.0-145 µmol L-1, and a limit of detection (LOD) of 0.70 µmol L-1. The detection of the SARS-CoV-2 spike protein was achieved by both square wave voltammetry (SWV) and electrochemical impedance spectroscopy (EIS) by a label-free immunosensor, using potassium ferro-ferricyanide solution as the electrochemical probe. An LDR of 0.70-7.0 and 1.0-30 pmol L-1, with an LOD of 0.70 and 1.0 pmol L-1 were obtained by SWV and EIS, respectively. As a proof of concept, the immunosensor was successfully applied for the detection of the SARS-CoV-2 spike protein in enriched synthetic saliva samples, which demonstrates the potential of using the proposed sensor as an alternative platform for the diagnosis of COVID-19 in the future.

Exploring the Role of the Connection Length of Screen-Printed Electrodes towards the Hydrogen and Oxygen Evolution Reactions.

Zero-emission hydrogen and oxygen production are critical for the UK to reach net-zero greenhouse gasses by 2050. Electrochemical techniques such as water splitting (electrolysis) coupled with renewables energy can provide a unique approach to achieving zero emissions. Many studies exploring electrocatalysts need to "electrically wire" to their material to measure their performance, which usually involves immobilization upon a solid electrode. We demonstrate that significant differences in the calculated onset potential for both the hydrogen evolution reaction (HER) and oxygen evolution reaction (OER) can be observed when using screen-printed electrodes (SPEs) of differing connection lengths which are immobilized with a range of electrocatalysts. This can lead to false improvements in the reported performance of different electrocatalysts and poor comparisons between the literature. Through the use of electrochemical impedance spectroscopy, uncompensated ohmic resistance can be overcome providing more accurate Tafel analysis.

Circular Economy Electrochemistry: Creating Additive Manufacturing Feedstocks for Caffeine Detection from Post-Industrial Coffee Pod Waste.

The recycling of post-industrial waste poly(lactic acid) (PI-PLA) from coffee machine pods into electroanalytical sensors for the detection of caffeine in real tea and coffee samples is reported herein. The PI-PLA is transformed into both nonconductive and conductive filaments to produce full electroanalytical cells, including additively manufactured electrodes (AMEs). The electroanalytical cell was designed utilizing separate prints for the cell body and electrodes to increase the recyclability of the system. The cell body made from nonconductive filament was able to be recycled three times before the feedstock-induced print failure. Three bespoke formulations of conductive filament were produced, with the PI-PLA (61.62 wt %), carbon black (CB, 29.60 wt %), and poly(ethylene succinate) (PES, 8.78 wt %) chosen as the most suitable for use due to its equivalent electrochemical performance, lower material cost, and improved thermal stability compared to the filaments with higher PES loading and ability to be printable. It was shown that this system could detect caffeine with a sensitivity of 0.055 ± 0.001 µA µM-1, a limit of detection of 0.23 µM, a limit of quantification of 0.76 µM, and a relative standard deviation of 3.14% after activation. Interestingly, the nonactivated 8.78% PES electrodes produced significantly better results in this regard than the activated commercial filament toward the detection of caffeine. The activated 8.78% PES electrode was shown to be able to detect the caffeine content in real and spiked Earl Grey tea and Arabica coffee samples with excellent recoveries (96.7-102%). This work reports a paradigm shift in the way AM, electrochemical research, and sustainability can synergize and feed into part of a circular economy, akin to a circular economy electrochemistry.

Novel and highly stable strategy for the development of microfluidic enzymatic assays based on the immobilization of horseradish peroxidase (HRP) into cotton threads.

The use of biological components in the development of new methods of analysis and point-of-care (POC) devices is an ever-expanding theme in analytical chemistry research, due to the immense potential for early diagnosis of diseases and monitoring of biomarkers. In the present work, the evaluation of an electrochemical microfluidic device based on the immobilization of horseradish peroxidase (HRP) enzyme into chemically treated cotton threads is described. This bioreactor was used as a channel for the build of the microfluidic device, which has allowed to use of a non-modified screen-printed electrode (SPE) as an amperometric detector. Cotton threads were treated using citric acid, and the immobilization of HRP has been performed by EDC/NHS crosslinking, connecting amine groups of the enzymes to carboxylic acids in the cellulosic structure. For the analytical evaluation, an amperometric assay for hydrogen peroxide detection was performed after the injection of H2O2 and hydroquinone (HQN) concomitantly. The enzymatic reaction consumes H2O2 leading to the formation of O-quinone, which is readily reducible at non-modified SPE. Several experimental parameters related to enzyme immobilization have been investigated and under the best set of conditions, a good analytical performance was obtained. In addition, the threads were freezer-stored and, after 12 weeks, 84% of hydrogen peroxide sensitivity was maintained, which is very reasonable for enzyme-based systems and still offers good analytical precision. Therefore, a simple and inexpensive microfluidic system was reported by crosslinking carboxylic groups to amine-containing macromolecules, suggesting a new platform for many other protein-based assays.

Functionalized screen-printed electrodes for the thermal detection of Escherichia coli in dairy products.

Accurate and fast on-site detection of harmful microorganisms in food products is a key preventive step to avoid food-borne illness and product recall. In this study, screen-printed electrodes (SPEs) were functionalized via a facile strategy with surface imprinted polymers (SIPs). The SIP-coated SPEs were used in combination with the heat transfer method (HTM) for the real-time detection of Escherichia coli. The sensor was tested in buffer, with a reproducible and sensitive response that attained a limit of detection of 180 CFU/mL. Furthermore, selectivity was assessed by analyzing the sensor's response to C. sakazakii, K. pneumoniae and S. aureus as analogue strains. Finally, the device was successfully used for the detection of E. coli in spiked milk as proof-of-application, requiring no additional sample preparation. These results suggest the proposed thermal biosensor possesses the potential of becoming a tool for routine, on-site monitoring of E. coli in food safety applications.