Smartphone-enabled flow injection amperometric glucose monitoring based on a screen-printed carbon electrode modified with PEDOT@PB and a GOx@PPtNPs@MWCNTs nanocomposite.

This study presents a modified screen-printed carbon electrode (SPCE) to determine glucose in a custom-built flow injection system. The biosensor was constructed by immobilizing glucose oxidase on porous platinum nanoparticles decorated on multi-walled carbon nanotubes (GOx@PPtNPs@MWCTNs). The fabrication of the biosensor was completed by coating the GOx@PPtNPs@MWCTNs nanocomposite on an SPCE modified with a nanocomposite of poly(3,4-ethylenedioxythiophene) and Prussian blue (GOx@PPtNPs@MWCTNs/PEDOT@PB/SPCE). The fabricated electrode accurately measured hydrogen peroxide (H2O2), the byproduct of the GOx-catalyzed oxidation of glucose, and was then applied as a glucose biosensor. The glucose response was amperometrically determined from the PB-mediated reduction of H2O2 at an applied potential of -0.10 V in a flow injection system. Under optimal conditions, the developed biosensor produced a linear range from 2.50 µM to 1.250 mM, a limit of detection of 2.50 µM, operational stability over 500 sample injections, and good selectivity. The proposed biosensor determined glucose in human plasma samples, achieving recoveries and results that agreed with the hexokinase-spectrophotometric method (P > 0.05). Combining the proposed biosensor with the custom-built sample feed, a portable potentiostat and a smartphone, enabled on-site glucose monitoring.

Flow injection amperometric uric acid biosensor based on AuNPs-GO-CS porous composite cryogel coated on PB-PEDOT:PSS modified screen-printed carbon electrode.

An enzymatic amperometric uric acid (UA) biosensor was successfully developed by modifying a screen-printed carbon electrode (SPCE) with Prussian blue-poly(3,4-ethylene dioxythiophene) polystyrene sulfonate composite (PB-PEDOT:PSS). The modified SPCE was coated with gold nanoparticles-graphene oxide-chitosan composite cryogel (AuNPs-GO-CS cry). Uricase (UOx) was directly immobilized via chemisorption on AuNPs. The nanocomposite was characterized by scanning electron microscopy, transmission electron microscopy, ultraviolet-visible spectroscopy, and Fourier transform infrared spectroscopy. The electrochemical characterization of the modified electrode was performed by cyclic voltammetry and electrochemical impedance spectroscopy. UA was determined using amperometric detection based on the reduction current of PB which was correlated with the amount of H2O2 produced during the enzymatic reaction. Under optimal conditions, the fabricated UA biosensor in a flow injection analysis (FIA) system produced a linear range from 5.0 to 300 µmol L-1 with a detection limit of 1.88 µmol L-1. The proposed sensor was stable for up to 221 cycles of detection and analysis was rapid (2 min), with good reproducibility (RSDs < 2.90 %, n = 6), negligible interferences, and recoveries from 94.0 ± 3.9 to 101.1 ± 2.6 %. The results of UA detection in blood plasma were in agreement with the enzymatic colorimetric method (P > 0.05).

Disposable Polyaniline/m-Phenylenediamine-Based Electrochemical Lactate Biosensor for Early Sepsis Diagnosis.

Lactate serves as a crucial biomarker that indicates sepsis assessment in critically ill patients. A rapid, accurate, and portable analytical device for lactate detection is required. This work developed a stepwise polyurethane-polyaniline-m-phenylenediamine via a layer-by-layer based electrochemical biosensor, using a screen-printed gold electrode for lactate determination in blood samples. The developed lactate biosensor was electrochemically fabricated with layers of m-phenylenediamine, polyaniline, a crosslinking of a small amount of lactate oxidase via glutaraldehyde, and polyurethane as an outer membrane. The lactate determination using amperometry revealed the biosensor's performance with a wide linear range of 0.20-5.0 mmol L-1, a sensitivity of 12.17 ± 0.02 µA·mmol-1·L·cm-2, and a detection limit of 7.9 µmol L-1. The developed biosensor exhibited a fast response time of 5 s, high selectivity, excellent long-term storage stability over 10 weeks, and good reproducibility with 3.74% RSD. Additionally, the determination of lactate in human blood plasma using the developed lactate biosensor was examined. The results were in agreement with the enzymatic colorimetric gold standard method (p > 0.05). Our developed biosensor provides efficiency, reliability, and is a great potential tool for advancing lactate point-of-care testing applications in the early diagnosis of sepsis.

A Novel Microfluidic-Based OMC-PEDOT-PSS Composite Electrochemical Sensor for Continuous Dopamine Monitoring.

Fast and precise analysis techniques using small sample volumes are required for next-generation clinical monitoring at the patient's bedside, so as to provide the clinician with relevant chemical data in real-time. The integration of an electrochemical sensor into a microfluidic chip allows for the achievement of real-time chemical monitoring due to the low consumption of analytes, short analysis time, low cost, and compact size. In this work, dopamine, used as a model, is an important neurotransmitter responsible for controlling various vital life functions. The aim is to develop a novel serpentine microfluidic-based electrochemical sensor, using a screen-printed electrode for continuous dopamine detection. The developed sensor employed the composite of ordered mesoporous carbon (OMC) and poly (3,4 ethylenedioxythiophene)-poly (styrene sulfonate) (PEDOT-PSS). The performance of a microfluidic, integrated with the sensor, was amperometrically evaluated using a computer-controlled microfluidic platform. The microfluidic-based dopamine sensor exhibited a sensitivity of 20.2 ± 0.6 µA µmol L-1, and a detection limit (LOD) of 21.6 ± 0.002 nmol L-1, with high selectivity. This microfluidic-based electrochemical sensor was successfully employed to determine dopamine continuously, which could overcome the problem of sensor fouling with more than 90% stability for over 24 h. This novel microfluidic sensor platform provides a powerful tool for the development of a continuous dopamine detection system for human clinical application.

An enzymatic histamine biosensor based on a screen-printed carbon electrode modified with a chitosan-gold nanoparticles composite cryogel on Prussian blue-coated multi-walled carbon nanotubes.

A histamine biosensor was developed based on a screen-printed carbon electrode modified with Prussian blue (PB) electrodeposited on multi-walled carbon nanotubes covered with a macroporous layer of chitosan-gold nanoparticles composite cryogel (CS-AuNPs Cry). With its high specific surface area and conductivity, CS-AuNPs Cry proved an excellent supporting material for diamine oxidase (DAO) immobilization. PB acted as a redox mediator to promote electron transfer between hydrogen peroxide and the electrode surface. The PB reduction current was measured during the hydrogen peroxide-releasing oxidation of histamine catalyzed by DAO. The proposed biosensor displayed two linear ranges: 2.50-125.0 µmol L-1 and 125.0-400.0 µmol L-1. The limit of detection was 1.81 µmol L-1. Reproducibility was good (RSD = 5.46%), operational stability high, long-term stability excellent, and selectivity good. The biosensor determined histamine levels in fish and shrimps with satisfactory recoveries, and the obtained results agreed with those obtained by ELISA.

A highly sensitive flow injection amperometric glucose biosensor using a gold nanoparticles/polytyramine/Prussian blue modified screen-printed carbon electrode.

A novel oxidase enzyme sensor was fabricated based on the chemisorption of highly active glucose oxidase (GOx) on gold nanoparticles that were adsorbed on a polytyramine layer (AuNPs/Pty). The GOx/AuNPs/Pty assembly was coated on a Prussian blue (PB)-modified screen-printed carbon electrode (SPCE) to produce the GOx/AuNPs/Pty/PB/SPCE biosensor. The amperometric glucose biosensor response was measured at -0.10 V using a Ag pseudo-reference electrode through the reduction current of the PB mediator in a flow injection analysis system. Under optimised experimental conditions, the developed biosensor displayed linearity over the 1.0 µM-1.0 mM glucose concentration range and a limit of detection of 1.0 µM (S/N ≥ 3). A low value for the Michaelis constant of 0.21 mM indicated that the immobilised GOx had high affinity for glucose. The developed biosensor exhibited excellent operational stability over 374 injections, long-term stability over 3 weeks, good reproducibility (relative standard deviations = 1.9%-4.3%) and high selectivity. The results obtained analysing glucose in blood plasma samples were satisfactory when compared with the corresponding results recorded implementing the standard hexokinase-spectrophotometric method (P > 0.05).

An ultrasensitive label-free electrochemical immunosensor based on 3D porous chitosan-graphene-ionic liquid-ferrocene nanocomposite cryogel decorated with gold nanoparticles for prostate-specific antigen.

A highly sensitive and selective label-free electrochemical immunosensor was successfully fabricated for measuring prostate-specific antigen (PSA). A composite of chitosan, graphene, ionic liquid and ferrocene (CS-GR-IL-Fc) was drop casted onto a screen-printed carbon electrode (SPCE) and frozen to create a layer of 3D porous cryogel (CS-GR-IL-Fc cry) which was decorated with gold nanoparticles (AuNPs). The biocompatibility and porosity of the cryogel increased the surface area available for AuNPs loading via amino groups and the population of anti-PSA, immobilized on the AuNPs via chemisorption, could be increased. The CS-GR-IL-Fc cry displayed excellent conductivity, enhancing electron transfer and amplifying the current signal. Differential pulse voltammetry was employed to determine PSA by measuring the reduction in the Fc oxidation peak current in response to the formation of PSA/anti-PSA immunocomplex. Under the optimized incubation time and electrolyte pH, the developed immunosensor displayed excellent analytical performances, including a wide linear range at concentrations from 1.0 × 10-7 to 1.0 × 10-1 ng mL-1, with a very low limit of detection of 4.8 × 10-8 ng mL-1 and good reproducibility (relative standard deviation of <4.6%, n = 6), stability (90% sensitivity within 20 days), repeatability (12 cycles of binding-rebinding, the sensitivity > 90%) and selectivity. The results obtained from the device for the determination of PSA in human serum were consistent with results from the enzyme-linked immunosorbent assay (P > 0.05), and indicated the promising potential of the proposed immunosensor in clinical diagnosis.

Simultaneous Detection of Ammonium and Nitrate in Environmental Samples Using on Ion-Selective Electrode and Comparison with Portable Colorimetric Assays.

Simple, robust, and low-cost nitrate- and ammonium-selective electrodes were made using substrate prepared from household materials. We explored phosphonium-based ILs and poly (methyl methacrylate)/poly(decyl methacrylate)(MMA-DMA) copolymer as matrix materials alternative to classical PVC-based membranes. IL-based membranes showed suitability only for nitrate-selective electrode exhibiting linear concentration range between 5.0 × 10-6 and 2.5 × 10-3 M with a detection limit of 5.5 × 10-7 M. On the other hand, MMA-DMA-based membranes showed suitability for both ammonium- and nitrate-selective electrodes, and were successfully applied to detect NO3- and NH4⁺ in water and soil samples. The proposed ISEs exhibited near-Nernstian potentiometric responses to NO3- and NH4⁺ with the linear range concentration between 5.0 × 10-5 and 5.0 × 10-2 M (LOD = 11.3 µM) and 5.0 × 10-6 and 1.0 × 10-3 M (LOD = 1.2 µM), respectively. The power of ISEs to detect NO3- and NH4⁺ in water and soils was tested by comparison with traditional, portable colorimetric techniques. Procedures required for analysis by each technique from the perspective of a non-trained person (e.g., farmer) and the convenience of the use on the field are compared and contrasted.

Phage-based capacitive biosensor for Salmonella detection.

This article reports the detection of Salmonella spp. based on M13 bacteriophage in a capacitive flow injection system. Salmonella-specific M13 bacteriophage was immobilized on a polytyramine/gold surface using glutaraldehyde as a crosslinker. The M13 bacteriophage modified electrode can specifically bind to Salmonella spp. via the amino acid groups on the filamentous phage. An alkaline solution was used to break the binding between the sensing surface and the analyte to allow renewable use up to 40 times. This capacitive system provided good reproducibility with a relative standard deviation (RSD) of 1.1%. A 75 µL min-1 flow rate and a 300 µL sample volume provided a wide linear range, from 2.0 × 102 to 1.0 × 107 cfu mL-1, with a detection limit of 200 cfu mL-1. Bacteria concentration can be analyzed within 40 min after the sample injection. When applied to test real samples (raw chicken meat) it provided good recoveries (100-111%). An enrichment process was also explored to increase the bacteria concentration, enabling a quantitative detection of Salmonella spp. This biosensor opens a new opportunity for the detection of pathogenic bacteria using bacteriophage.

Pyrrolidinyl PNA polypyrrole/silver nanofoam electrode as a novel label-free electrochemical miRNA-21 biosensor.

A label-free electrochemical miRNA biosensor was developed based on a pyrrolidinyl peptide nucleic acid (acpcPNA)/polypyrrole (PPy)/silver nanofoam (AgNF) modified electrode. The AgNF was electrodeposited as redox indicator on a gold electrode, which was then functionalized with an electropolymerized layer of PPy, a conducting polymer, to immobilize the PNA probes. The fabrication process was investigated by electrochemical impedance spectroscopy. The biosensor was used to detect miRNA-21, a biomarker abnormally expressed in most cancers. The signal was monitored by the change in current of the AgNF redox reaction before and after hybridization using cyclic voltammetry. Two PNA probe lengths were investigated and the longer probe exhibited a better performance. Nucleotide overhangs on the electrode side affected the signal more than overhangs on the solution side due to the greater insulation of the sensing surface. Under optimal conditions, the electrochemical signal was proportional to miRNA-21 concentrations between 0.20fM and 1.0nM, with a very low detection limit of 0.20fM. The biosensor showed a high specificity which could discriminate between complementary, single-, doubled-base mismatched, and non-complementary targets. Three out of the seven tested plasma samples provided detectable concentrations (63 ± 4, 111 ± 4 and 164 ± 7fM). The sensor also showed good recoveries (81-119%). The results indicated the possibilities of this biosensor for analysis without RNA extraction and/or amplification, making the sensor potentially useful for both the prognosis and diagnosis of cancer in clinical application.

Capacitive antibacterial susceptibility screening test with a simple renewable sensing surface.

A simple renewable surface for a rapid antibacterial susceptibility test has been demonstrated. The 3-aminophenylboronic acid (3-APBA) modified electrode bind with cis-diol groups on the cell wall of both gram positive and gram negative bacteria. The detection of antibacterial susceptibility response by a capacitive system can be done within a short time, 2.5h for the whole process, with good repeatability of the electrode's preparation. An acid solution, could break the bonding between 3-APBA and the bacteria, which were then easily removed by the fluid flow, renewing the sensing surface for the next test. This modified electrode can be reused up to 35 times. This sensor is useful for testing the susceptibility of bacteria to antibacterial agents that affect their cell wall. Results from the capacitive sensor corresponded well with the antimicrobial information in the literature and to the morphology of the treated bacteria revealed by scanning electron microscopy. Antimicrobial susceptibility to natural products could also be easily tested.

4-mercaptophenylboronic acid functionalized gold nanoparticles for colorimetric sialic acid detection.

A simple and selective colorimetric sensor for sialic acid detection, based on the aggregation of 4-mercaptophenylboronic acid functionalized gold nanoparticles (4-MPBA-AuNPs) was developed. The color of the solution changed from wine-red to blue after binding with sialic acid. The colorimetric sensor provided good analytical performances with a linear dynamic range of 80µM to 2.00mM and a 68±2µM limit of detection without any effect from possible interferences and sample matrix. In addition, the quantitative results were obtained within only 10min. This developed sensor was used to detect sialic acid in blood serum samples and the results were in good agreement with those from the current periodate-resorcinol method (P>0.05) thus indicating that this developed colorimetric sensor can be used as an alternative method for sialic acid detection with a shorter analysis time and a high accuracy.

Enhancing capacitive DNA biosensor performance by target overhang with application on screening test of HLA-B*58:01 and HLA-B*57:01 genes.

A highly sensitive label-free DNA biosensor based on PNA probes immobilized on a gold electrode was used to detect a hybridization event. The effect of a target DNA overhang on the hybridization efficiency was shown to enhance the detected signal and allowed detection at a very low concentration. The sensors performances were investigated with a complementary target that had the same length as the probe, and the signal was compared to the target DNAs with different lengths and overhangs. A longer target DNA overhang was found to provide a better response. When the overhang was on the electrode side the signal enhancement was greater than when the overhang was on the solution side due to the increased thickness of the sensing surface, hence produced a larger capacitance change. Using conformationally constrained acpcPNA probes, double stranded DNA was detected sensitively and specifically without any denaturing step. When two acpcPNA probes were applied for the screening test for the double stranded HLA-B*58:01 and HLA-B*57:01 genes that are highly similar, the method differentiated the two genes in all samples. Both purified and unpurified PCR products gave comparable results. This method would be potentially useful as a rapid screening test without the need for purification and denaturation of the PCR products.

Highly-sensitive label-free electrochemical carcinoembryonic antigen immunosensor based on a novel Au nanoparticles-graphene-chitosan nanocomposite cryogel electrode.

For the first time, a simple and highly sensitive label-free electrochemical carcinoembryonic antigen (CEA) immunosensor based on a cryogel electrode has been developed and tested. The as-prepared nanocomposite combined the advantages of the graphene, AuNPs and chitosan (AuNPs-GP-CS) together with the ease of preparing a cryogel coupled to a silver deposition, to act as a redox mediator, on a Au electrode. Under the optimal conditions, the decrease of the cyclic voltammetry (CV) silver peak current was proportional to the CEA concentration over a range of from 1.0×10(-6) to 1.0 ng mL(-1) with a detection limit of 2.0×10(-7) ng mL(-1). This AuNPs-GP-CS cryogel electrode gave a 1.7 times higher sensitivity and 25 times lower detection limit than the non-cryogel electrode. Moreover, the proposed electrochemical immunosensor exhibited good selectivity, reproducibility and stability. When applied to analyse clinical serum samples, the data determined by the developed immunosensor were in agreement with those obtained by the current hospital analysis system (enzyme linked fluorescent assay) (P>0.05), to indicate that the immunosensor would be potentially useful for clinical diagnostics.

Electrochemical detection of the disease marker human chitinase-3-like protein 1 by matching antibody-modified gold electrodes as label-free immunosensors.

Tissue inflammation, certain cardiovascular syndromes and the occurrence of some solid tumors are correlated with raised serum concentrations of human chitinase-3-like protein 1 (YKL-40), a mammalian chitinase-like glycoprotein, which has become the subject of current research. Here we report the construction and characterization of an electrochemical platform for label-free immunosensing of YKL-40. Details of the synthesis of YKL-40 and production of anti-YKL-40 immunoglobulin G (IgG) are provided and cross-reactivity tests presented. Polyclonal anti-YKL-40 IgG was immobilized on gold electrodes and the resulting immunosensors were operated in an electrochemical flow system with capacitive signal generation. The strategy offered a wide linear detection range (0.1µg/L to 1mg/L) with correlation coefficients (R(2)) above 0.99 and good sensitivity (12.28±0.27nF/cm(2) per decade of concentration change). Additionally, the detection limit of 0.07±0.01µg/L was well below that of optical enzyme-linked immunosorbent assays (ELISAs), which makes the proposed methodology a promising alternative for YKL-40 related disease studies.

A novel molecularly imprinted chitosan-acrylamide, graphene, ferrocene composite cryogel biosensor used to detect microalbumin.

A novel highly sensitive and selective molecularly imprinted polymer (MIP) cryogel biosensor for determination of microalbumin in urine samples was fabricated. The MIP gel was prepared based on the graft copolymerization of acrylamide with N,N'-methylenebisacrylamide on chitosan using human serum albumin (HSA) as the template. The sub-zero polymerization allowed the solvent to form ice crystals and left a macroporous cryogel structure when it was thawed. After removing the template, the specific imprinted surface on cryogel pore walls was used to detect HSA via a redox mediator (ferrocene), entrapped in the cryogel, using differential pulse voltammetry (DPV). The electrochemical detection was improved by the presence of graphene that has been composited within the polymer. For determination of albumin, the fabricated MIP cryogel biosensor showed a high sensitivity with a wide linear range of 1.0 × 10(-4) to 1.0 × 10(1) mg L(-1) and a low limit of detection of 5.0 × 10(-5) mg L(-1) (S/N = 3). The sensor also provided a very good reusability, i.e., the sensitivity remained >90% after 9 cycles of binding-rewashing (18 analyses per cycle), while the sensitivity only decreased to 90% after 6 weeks of storage at room temperature. The biosensor also showed a good selectivity, both against bovine serum albumin (BSA) and some common possible interfering compounds normally present in urine (ascorbic acid, uric acid, urea, sodium, chloride, potassium and creatinine). The excellent performance of the biosensor was confirmed by analyzing microalbumin in urine samples, and results were in good agreement with those obtained by the standard immunoturbidimetric method (P > 0.05).

An amperometric uric acid biosensor based on chitosan-carbon nanotubes electrospun nanofiber on silver nanoparticles.

A novel amperometric uric acid biosensor was fabricated by immobilizing uricase on an electrospun nanocomposite of chitosan-carbon nanotubes nanofiber (Chi-CNTsNF) covering an electrodeposited layer of silver nanoparticles (AgNPs) on a gold electrode (uricase/Chi-CNTsNF/AgNPs/Au). The uric acid response was determined at an optimum applied potential of -0.35 V vs Ag/AgCl in a flow-injection system based on the change of the reduction current for dissolved oxygen during oxidation of uric acid by the immobilized uricase. The response was directly proportional to the uric acid concentration. Under the optimum conditions, the fabricated uric acid biosensor had a very wide linear range, 1.0-400 µmol L(-1), with a very low limit of detection of 1.0 µmol L(-1) (s/n = 3). The operational stability of the uricase/Chi-CNTsNF/AgNPs/Au biosensor (up to 205 injections) was excellent and the storage life was more than six weeks. A low Michaelis-Menten constant of 0.21 mmol L(-1) indicated that the immobilized uricase had high affinity for uric acid. The presence of potential common interfering substances, for example ascorbic acid, glucose, and lactic acid, had negligible effects on the performance of the biosensor. When used for analysis of uric acid in serum samples, the results agreed well with those obtained by use of the standard enzymatic colorimetric method (P > 0.05).

Label-free capacitive DNA sensor using immobilized pyrrolidinyl PNA probe: effect of the length and terminating head group of the blocking thiols.

This paper reports, for the first time, the influence of the length and the terminating head group of blocking thiols on the sensitivity and specificity of a label-free capacitive DNA detection system using immobilized pyrrolidinyl peptide nucleic acid (acpcPNA) probes. A C-terminal lysine-modified acpcPNA was immobilized through four different alkanethiol self-assembled monolayers (SAMs), i.e., 3-mercaptopropionic acid (MPA), thioctic acid (TA), thiourea (TU) and mercaptosuccinic acid (MSA). The hybridization between the acpcPNA probes and the target DNA was directly measured using the capacitive system. Five blocking thiols of various lengths (C=3, 6, 8, 9 and 11), with the -OH terminating head group, i.e., 3-mercapto-1-propanol (3-MPL), 6-mercapto-1-hexanol (6-MHL), 8-mercapto-1-octanol (8-MOL), 9-mercapto-1-nonanol (9-MNL), 11-mercapto-1-undecanol (11-MUL) and another blocking thiol (C=11) with a -CH(3) terminating head group, and 1-dodecanethiol (1-DDT) were investigated. The blocking thiol with the same length as the total spacer of the immobilized acpcPNA gave the highest sensitivity and specificity with the -OH terminating head group providing a slightly better signal than the -CH(3) group. Under the optimized conditions, the immobilized acpcPNA probes provided a wide linear range for DNA detection (1.0 × 10(-11)-1.0 × 10(-8)M) with a very low detection limit in the picomolar range. The modified acpcPNA electrode could be reused through at least 58 cycles. The high sensitivity and very low detection limits are potentially useful for the analysis of ultra-trace levels of DNA in samples. Preliminary studies were also performed to see the effect of probe concentration and target length.

Interference compensation for thin layer coulometric ion-selective membrane electrodes by the double pulse technique.

Ion-selective membranes operated in a thin layer coulometric detection mode have previously been demonstrated to exhibit attractive characteristics in view of realizing sensors without the need for frequent recalibration. In this methodology, the analyte ion is exhaustively removed across an ion-selective membrane by an applied potential, and the resulting current is integrated to yield the coulomb number and hence the amount of analyte originally present in the sample. This exhaustive process, however, places greater demands on the selectivity of the membrane compared to direct potentiometry, since the level of interference will increase as the analyte depletes. We evaluate here a double pulse protocol to reduce the level of interference, in which the sample is electrolyzed once again after the initial coulometric detection pulse. Since the analyte ion is no longer present at significant concentrations during the second pulse, but an interfering ion of high concentration did not appreciably deplete, the second electrolysis step may be used to partially compensate for undesired interference. These processes are here evaluated by numerical simulation for ions of the same charge, demonstrating that the resulting coulomb number may indeed be reduced for systems of limited selectivity. The improvement in operational selectivity relative to uncompensated coulometry is found to be ca. 6-fold. The methodology is successfully demonstrated experimentally with a calcium selective membrane and tetraethylammonium as a model interfering agent, and the observed relative errors after background compensation can be favorably compared to that in direct potentiometry where no sample depletion occurs.

Ultra trace analysis of small molecule by label-free impedimetric immunosensor using multilayer modified electrode.

A multilayer electrode modified with a self-assembled thiourea monolayer (SATUM) followed by gold nanoparticles (AuNPs), mercaptosuccinic acid (MSA) and antibody was investigated for the detection of ultra trace amount of a small molecule (chloramphenicol) in an impedimetric system. The formation of the antibody-antigen complex at the electrode surface caused the impedance to increase. Under optimum conditions three modified electrodes were compared the SATUM/AuNPs/MSA electrode provided a wide linear range (0.50-10) × 10⁻¹6 M, and a very low determination limit of 1.0 × 10⁻¹6 M. This determination limit was much lower than the SATUM/AuNPs electrode, 1.0 × 10⁻¹5 M, and SATUM electrode, 4.7 × 10⁻¹4 M. The modified electrode provided good selectivity for chloramphenicol detection and can be reused up to 45 times with a relative standard deviation of lower than 4%. When applied to determine chloramphenicol in shrimp samples, the results agreed well with those obtained by the high-performance liquid chromatography coupled with a photo diode array detector (P > 0.05). The developed system can be applied to detect other small molecules using appropriate affinity binding pairs.