Electrochemical (Bio)Sensors Enabled by Fused Deposition Modeling-Based 3D Printing: A Guide to Selecting Designs, Printing Parameters, and Post-Treatment Protocols.

The 3D printing (or additive manufacturing, AM) technology is capable to provide a quick and easy production of objects with freedom of design, reducing waste generation. Among the AM techniques, fused deposition modeling (FDM) has been highlighted due to its affordability, scalability, and possibility of processing an extensive range of materials (thermoplastics, composites, biobased materials, etc.). The possibility of obtaining electrochemical cells, arrays, pieces, and more recently, electrodes, exactly according to the demand, in varied shapes and sizes, and employing the desired materials has made from 3D printing technology an indispensable tool in electroanalysis. In this regard, the obtention of an FDM 3D printer has great advantages for electroanalytical laboratories, and its use is relatively simple. Some care has to be taken to aid the user to take advantage of the great potential of this technology, avoiding problems such as solution leakages, very common in 3D printed cells, providing well-sealed objects, with high quality. In this sense, herein, we present a complete protocol regarding the use of FDM 3D printers for the fabrication of complete electrochemical systems, including (bio)sensors, and how to improve the quality of the obtained systems. A guide from the initial printing stages, regarding the design and structure obtention, to the final application, including the improvement of obtained 3D printed electrodes for different purposes, is provided here. Thus, this protocol can provide great perspectives and alternatives for 3D printing in electroanalysis and aid the user to understand and solve several problems with the use of this technology in this field.

Batch injection analysis with amperometric detection for fluoroquinolone determination in urine, pharmaceutical formulations, and milk samples using a reduced graphene oxide-modified glassy carbon electrode.

In this work, the batch injection analysis system with amperometric detection using reduced graphene oxide as a modifier of glassy carbon electrode (GCE) was investigated for the simple, fast, and sensitive monitoring of levofloxacin (LEVO) and ciprofloxacin (CIPRO) in samples of pharmaceutical formulations, synthetic urine, and milk (low- and high-fat content). LEVO and CIPRO were quantified in seven samples using amperometric measurements at +1.10 V vs Ag/AgCl, KCl(sat). The developed methods showed excellent analytical performance with limits of detection of 0.30 and 0.16 µmol L-1, linear range from 3.0 to 50 µmol L-1 and 1.0 to 50 µmol L-1, relative standard deviation below 9.7 and 3.1%, and recovery ranges ranging from 80 to 107% and from 78 to 109% for LEVO and CIPRO, respectively. In addition, the minimum sample preparation (simple dilution) combined with a high analytical frequency (130 to 180 analyses per hour) can be highlighted. Thus, the methods are promising for implementation in routine analysis and quality control to different samples.

Electrochemical methods for the determination of antibiotic residues in milk: A critical review.

Several antibiotics have been applied to veterinary medicine due to their broad-spectrum of antibacterial activity and prophylactic power. Residues of these antibiotics can be accumulated in dairy cattle, in addition to promoting contamination of the environment and, in more serious cases, in milk, causing a public health problem. Different regulatory agencies establish maximum residue limits for these antibiotics in milk, so it becomes important to develop sensitive analytical methods for monitoring these compounds. Electrochemical techniques are important analytical tools in analytical chemistry because they present low cost, simplicity, high sensitivity, and adequate analytical frequency (sample throughput) for routine analyses. In this sense, this review summarizes the state of the art of the main electrochemical sensors and biosensors, instrumental techniques, and sample preparation used for the development of analytical methods, published in the last five years, for the monitoring of different classes of antibiotics: aminoglycosides, amphenicols, beta-lactams, fluoroquinolones, sulfonamides, and tetracyclines, in milk samples. The different strategies to develop electrochemical sensors and biosensors are critically compared considering their analytical features. The mechanisms of electrochemical oxidation/reduction of the antibiotics are revised and discussed considering strategies to improve the selectivity of the method. In addition, current challenges and future prospects are discussed.

An indirect electrochemical method for aqueous sulfide determination in freshwaters using a palladium chelate as a selective sensor.

Sulfide anion is a highly toxic and corrosive compound and its presence above the threshold concentrations (i.e. µmol L-1) in freshwaters may indicate environmental pollution. Besides, the increase in sulfide concentration results in modifications of the organoleptic proprieties of water and air. Many analytical methodologies have been designed for aqueous sulfide quantification, however, due to the high reactivity and instability of sulfide, the pursue of a simple, sensitive, selective, and portable analytical method is still a current demand. In this study, an indirect electrochemical method for the determination of sulfide based on its interaction with a palladium complex - bis(2-aminobenzoate) palladium(II) - acting as a selective chemosensor is described. The reaction leads to the demasking of the electroactive ligand 2-aminobenzoic acid (i.e. anthranilic acid) and square wave voltammetry is employed to monitor its concentration using a glassy carbon electrode (GCE). Experimental conditions were optimized and the reaction was performed in Britton-Robinson (BR) buffer at pH 5 for 4 min, providing the higher magnitude of the analytical signal. A linear relation (r2 > 0.99) from 3 to 30 µmol L-1 of sulfide was obtained with a limit of detection of 0.10 µmol L-1. Recovery experiments using freshwater samples spiked with sulfide revealed overall satisfactory results for the limit concentration levels permitted by regulatory agencies. Therefore, the proposed methodology shows advantages in terms of portability, selectivity, sensitivity, low-cost, and easiness-to-use enabling monitoring of sulfide in a variety of waters.

Direct analysis of ascorbic acid in food beverage samples by flow injection analysis using reduced graphene oxide sensor.

In this paper, a simple, sensitive and precise electroanalytical method was developed using flow injection analysis (FIA) with amperometric detection and reduced graphene oxide sensor for ascorbic acid determination in samples of multivitamin beverages, milk, fermented milk, and milk chocolate. The advantages of this sensor include a potential displacement of 450 mV and a 2-fold peak current increase for electrochemical oxidation of ascorbic acid, which resulted in a highly sensitive method. No interference of sample matrix was observed, avoiding solvent extraction procedures (samples were only diluted). The FIA allowed a high analytical frequency, approximately 96 injections per hour, together with adequate detection limit of 4.7 µmol L-1. Good precision (RSD < 7%) and accuracy (recoveries between 91 and 108%) evidenced the robustness of the method. The method was compared with ultra-fast liquid chromatography (UFLC) obtaining statistically similar results (95% confidence level). The ascorbic acid content in samples varied from 0.065 to 2.53 mmol L-1.

Boron Doped Diamond Electrodes in Flow-Based Systems.

Boron-doped diamond (BDD) electrodes present several notable properties, such as the largest potential window of all electrode materials (especially in anodic potentials), low background and capacitive currents, reduced fouling compared to other electrodes, mechanical robustness, and good stability over time. On the other hand, flow-based systems are known as well-established approaches to minimize reagent consumption and waste generation and with good compromise between sample throughput and analytical performance (mechanization of chemical assays). This review focuses on the use of BDD electrodes for electrochemical detection in flow systems, such as flow injection analysis (FIA), batch injection analysis (BIA), high performance liquid chromatography (HPLC), and capillary electrophoresis (CE). The discussion deals with the historical evolution of BDD, types of electrochemical pre-treatments (cathodically/H-terminated or anodically/O-terminated), cell configurations, and analytical performance. Articles are discussed in chronological order and subdivided according to the type of flow system: FIA, BIA, HPLC, and CE.

Ultrafast capillary electrophoresis method for the simultaneous determination of ammonium and diphenhydramine in pharmaceutical samples.

Ammonium and diphenhydramine are active ingredients commonly found in the same pharmaceutical preparations. We report, for the first time, a sub-minute method for the simultaneous determination of ammonium and diphenhydramine. The method is based on capillary electrophoresis with capacitively coupled contactless conductivity detection. Both analytes can be quantified in a single run (∼80 injections/h) using 30 mmol/L 2-(N-morpholino)ethanesulfonic acid and 15 mmol/L lithium hydroxide (pH 6.0) as background electrolyte. The separation by capillary electrophoresis was achieved on a fused-silica capillary (50 cm total length, 10 cm effective length, and 50 µm inside diameter). The limits of detection were 0.04 and 0.02 mmol/L for ammonium and diphenhydramine, respectively. The proposed method also provided adequate recovery values for spiked samples (100-106 and 97-104% for ammonium and diphenhydramine, respectively). The results obtained with the new capillary electrophoresis method were compared with those of the high-performance liquid chromatography method for diphenhydramine and the Kjeldahl method for ammonium and no statistically significant differences were found (95% confidence level).

Use of pyrolyzed paper as disposable substrates for voltammetric determination of trace metals.

The possibility of using pyrolyzed paper as disposable working electrodes for trace metals determination is reported for the first time. A small piece of pyrolyzed paper (0.7×0.7cm) was positioned at the bottom side of the electrochemical cell using a rubber O-ring, which defined the electrode area (0.48cm; 0.18cm2). A large number of electrodes can be obtained from a single piece of standard dimensions (2.5cm×7.5cm) of paper, therefore minimizing the cost per unit. The electrochemical performance of the pyrolyzed paper was demonstrated by cyclic voltammetry, electrochemical impedance spectroscopy and by the determination of Zn, Cd, and Pb by square-wave anodic stripping voltammetry. The unmodified pyrolyzed paper showed excellent performance for Pb and Cd detection (LOD =0.19 and 0.16 ppb, respectively). In the presence of Bi3+(in-situ film formation), the simultaneous determination of Zn, Cd and Pb was also possible (LOD=0.26, 0.25, and 0.39 ppb, respectively).

A batch injection analysis system with square-wave voltammetric detection for fast and simultaneous determination of naphazoline and zinc.

In this work, a batch-injection analysis system with square-wave voltammetric (BIA-SWV) detection was applied for the first time to the simultaneous determination of inorganic (zinc) and organic (naphazoline) species. Both compounds were detected in a single run (70 injections h(-1)) with a small injection volume (∼100 µL). The calibration curves exhibited linear response range between 3.0 and 21.0 µmol L(-1) (r=0.999) for naphazoline and between 10.0 and 60.0 µmol L(-1) (r=0.992) for zinc. The detection limits were 0.13 and 0.04 µmol L(-1) for zinc and naphazoline, respectively. Good reproducibility was achieved for multiple measurements of a solution containing both species (RSD<1.0%; n=20). The results obtained with the BIA-SWV method for the simultaneous determination of naphazoline and zinc were compared to those obtained by HPLC (naphazoline) and by FAAS (zinc); no statistically significant differences were observed (95% confidence level).

Simultaneous determination of three species with a single-injection step using batch injection analysis with multiple pulse amperometric detection.

In this work, the possibility of simultaneous determination of three compounds with a single-injection step using batch injection analysis with multiple pulse amperometric detection (BIA-MPA) is demonstrated for the first time. A sequence of three potential pulses (+1.25 V, +1.60 V, and +1.80 V) was applied with the acquisition of three separate amperograms. 8-Chlorotheophylline was detected selectively at +1.25 V, both 8-chlorotheophylline and pyridoxine at +1.60V and 8-chlorotheophylline, pyridoxine, and diphenhydramine at +1.80 V. Subtraction between the currents detected at the three amperograms (with the help of correction factors) was used for the selective determination of pyridoxine and diphenhydramine. The proposed method is simple, inexpensive, fast (60 injections h(-1)), and present selectivity for the determination of the three compounds in pharmaceutical samples, with results similar to those obtained by HPLC (95% confidence level).

Ultra-fast determination of caffeine, dipyrone, and acetylsalicylic acid by capillary electrophoresis with capacitively coupled contactless conductivity detection and identification of degradation products.

Capillary electrophoresis with capacitively coupled contactless conductivity detection (CE-C(4)D) was used for fast, simultaneous determination of dipyrone (DIP), caffeine (CAF), and acetylsalicylic acid (ASA). In the same run and in less than 1min, the degradation products from DIP and ASA were also detected. In addition, the usage of the CE-C(4)D system allowed, for the first time, the detection of methylamine as a degradation product of DIP. Capillary electrophoresis with electrospray mass spectrometry experiments were carried out in order to confirm the formation of methylamine. The limits of detection by CE-C(4)D were 5, 5, and 6µmolL(-1) for CAF, DIP, and ASA, respectively. The proposed method was applied to the analysis of these compounds in pharmaceutical formulations with similar results to those achieved by HPLC (p<0.05).

Two new electrochemical methods for fast and simultaneous determination of codeine and diclofenac.

In this paper, we present two new electrochemical methods for fast and simultaneous determination of codeine (CO) and diclofenac (DCF). The first one is based on batch injection analysis with amperometric detection (BIA-MPA) and the second one is based on capillary electrophoresis with capacitively coupled contactless conductivity detection (CE-C(4)D). The proposed BIA-MPA method is highly-precise (RSD of 1.1% and 0.9% for DCF and CO, respectively; n=10), fast (300 injections h(-1)) and has low detection limits (1.1 and 1.0 µmol L(-1) for DCF and CO, respectively). The proposed CE-C(4)D method allows the determination of CO and DFC in less than 1 min with high precision (RSD of 0.3% and 0.7% for DCF and CO, respectively; n=10) and low detection limits (11 and 21 µmol L(-1) for DCF and CO, respectively). Both proposed methods were applied to the determination of CO and DCF in pharmaceutical samples with similar results to those achieved by high-performance liquid chromatography (HPLC) at a 95% confidence level.

Fast and direct determination of butylated hydroxyanisole in biodiesel by batch injection analysis with amperometric detection.

We report here, for the first time, application of batch injection analysis (BIA) with amperometric detection for determination of the phenolic antioxidant butylated hydroxyanisole (BHA) in biodiesel. A sample plug was directly injected onto a boron-doped diamond electrode immersed in 50% v/v hydroethanolic solution with 0.1 mol L(-1) HClO(4) using an electronic micropipette. Importantly, the only preparation step required for biodiesel analysis is dilution in the same hydroethanolic electrolyte solution. Our proposed method has several advantages for routine biodiesel analysis, including: a low relative standard deviation between injections (0.29%, n=20), high analytical frequency (120 h(-1)), adequate recovery values (93-101%) for spiked samples, satisfactory accuracy (based on comparative determinations by high-performance liquid-chromatography), and a low detection limit (100 ng of BHA per g of biodiesel). Finally, our method can be adapted for the determination of other antioxidants in biodiesel samples.