A screen-printed disposable biosensor for selective determination of putrescine

We have developed screen-printed carbon electrodes for the determination of putrescine (Put) via the enzyme monoamine oxidase that was immobilized on the surface of the electrode by cross-linking it with bovine serum albumin using glutaraldehyde. A mixture of 5 % of tetrathiofulvalene (TTF) and carbon ink was used for the fabrication of the screen-printed working electrode. Put was amperometrically detected by measurement of the current due to the oxidation of the mediator TTF. The use of TTF lowers the working potential to +250 mV (vs. a screen-printed Ag/AgCl reference electrode). Response is linear in the range from 16 to 101 μM, and the detection limit is 17.2?±?4.6 μM, with a reproducibility of 9.6 % (n?=?4) in terms of relative standard deviation. The effects of potentially interfering biogenic amines such as cadaverine, histamine, spermine, spermidine and tryptamine were also evaluated. The biosensor was successfully applied to the determination of Put in zucchini and anchovies.

Mediatorless amperometric bienzyme glucose biosensor based on horseradish peroxidase and glucose oxidase cross-linked to multiwall carbon nanotubes

We report on a bienzyme-channeling sensor for sensing glucose without the aid of mediator. It was fabricated by cross-linking horseradish peroxidase (HRP) and glucose oxidase (GOx) on a glassy carbon electrode modified with multiwalled carbon nanotubes (MWNTs). The bienzyme was cross-linked with the MWNTs by glutaraldehyde and bovine serum albumin. The MWNTs were employed to accelerate the electron transfer between immobilized HRP and electrode. Glucose was sensed by amperometric reduction of enzymatically generated H₂O₂ at an applied voltage of ?50 mV (vs. Ag/AgCl). Factors influencing the preparation and performance of the bienzyme electrode were investigated in detail. The biosensor exhibited a fast and linear response to glucose in the concentration range from 0.4 to 15 mM, with a detection limit of 0.4 mM. The sensor exhibited good selectivity and durability, with a long-term relative standard deviation of <5 %. Analysis of glucose-spiked human serum samples yielded recoveries between 96 and 101 %.

Amperometric biosensor for bisphenol A based on a glassy carbon electrode modified with a nanocomposite made from polylysine, single walled carbon nanotubes and tyrosinase

We have prepared a nanocomposite consisting of single-walled carbon nanotubes and polylysine. It was characterized by transmission electron microscopy, X-ray photoelectron spectroscopy, and by UV/vis and FTIR spectroscopy. Tyrosinase was covalently immobilized on the nanocomposite, and the resulting bioconjugate deposited on a glassy carbon electrode to form a biosensor for bisphenol A. The biosensor was characterized by scanning electron microscopy and electrochemical impedance spectroscopy. Under optimized experimental conditions, the biosensor gives a linear response to bisphenol A in the 4.00 nM to 11.5 μM concentration range. Its sensitivity is 788 mA M^?1 cm^?2, and the lower detection limit is 0.97 nM (at an S/N of 3). The biosensor shows good repeatability, reproducibility and long-term stability. In a preliminary practical application, it was successfully applied to the determination of bisphenol A in leachates of plastic spoons.

Electrochemical behavior and voltammetric determination of L-tryptophan and L-tyrosine using a glassy carbon electrode modified with single-walled carbon nanohorns

We report a simple method for the direct and quantitative determination of L-tryptophan (Trp) and L-tyrosine (Tyr) using a glassy carbon electrode (GCE) modified with single-walled carbon nanohorns (SWCNHs). The SWCNH modified GCE exhibits high electrocatalytic activity towards the oxidation of both Trp and Tyr. It shows a linear response to Trp between 0.5 and 50 μM and to Tyr between 2 and 30 μM. The detection limits for Trp and Tyr are 50 nM and 400 nM, respectively. In addition, the modified GCE displays good selectivity and good sensitivity, thus making it suitable for the determination of Trp and Tyr in spiked serum samples.

Glucose biosensor based on a platinum electrode modified with rhodium nanoparticles and with glucose oxidase immobilized on gold nanoparticles

We have developed an enzymatic glucose biosensor that is based on a flat platinum electrode which was covered with electrophoretically deposited rhodium (Rh) nanoparticles and then sintered to form a large surface area. The biosensor was obtained by depositing glucose oxidase (GOx), Nafion, and gold nanoparticles (AuNPs) on the Rh electrode. The electrical potential and the fractions of Nafion and GOx were optimized. The resulting biosensor has a very high sensitivity (68.1 μA mM^?1 cm^?2) and good linearity in the range from 0.05 to 15 mM (r?=?0.989). The limit of detection is as low as 0.03 mM (at an SNR of 3). The glucose biosensor also is quite selective and is not interfered by electroactive substances including ascorbic acid, uric acid and acetaminophen. The lifespan is up to 90 days. It was applied to the determination of glucose in blood serum, and the results compare very well with those obtained with a clinical analyzer.

Novel glucose biosensor based on a glassy carbon electrode modified with hollow gold nanoparticles and glucose oxidase

A novel glucose biosensor is presented as that based on a glassy carbon electrode modified with hollow gold nanoparticles (HGNs) and glucose oxidase. The sensor exhibits a better differential pulse voltammetric response towards glucose than the one based on conventional gold nanoparticles of the same size. This is attributed to the good biological conductivity and biocompatibility of HGNs. Under the optimal conditions, the sensor displays a linear range from 2.0?×?10^?6 to 4.6?×?10^?5?M of glucose, with a detection limit of 1.6?×?10^?6?M (S/N?=?3). Good reproducibility, stability and no interference make this biosensor applicable to the determination of glucose in samples such as sports drinks.

Lactate biosensor based on a bionanocomposite composed of titanium oxide nanoparticles, photocatalytically reduced graphene, and lactate oxidase

We have developed a lactate biosensor based on a bionanocomposite (BNC) composed of titanium dioxide nanoparticles (TiO₂-NPs), photocatalytically reduced graphene, and lactate oxidase. Graphene oxide was photochemically reduced (without using any chemical reagents) in the presence of TiO₂-NPs to give graphene nanosheets that were characterized by atomic force microscopy, Raman and X-ray photoelectron spectroscopy. The results show the nanosheets to possess few oxygen functionalities only and to be decorated with TiO₂-NPs. These nanosheets typically are at least 1 μm long and have a thickness of 4.2 nm. A BNC was obtained by mixing lactate oxidase with the nanosheets and immobilized on the surface of a glassy carbon electrode. The resulting biosensor was applied to the determination of lactate. Compared to a sensor without TiO₂-NPs, the sensor exhibits higher sensitivity (6.0 μA mM^?1), a better detection limit (0.6 μM), a wider linear response (2.0 μM to 0.40 mM), and better reproducibility (3.2 %).

An enzymatic glucose biosensor based on a glassy carbon electrode modified with manganese dioxide nanowires

A glassy carbon electrode was modified with β-manganese dioxide (β-MnO₂), and glucose oxidase (GOx) was immobilized on its surface. The β-MnO₂ nanowires were prepared by a hydrothermal method and characterized by scanning electron microscopy and powder X-ray diffraction. They were then dispersed in Nafion solution and cast on the glassy carbon electrode (GCE) to form an electrode modified with β-MnO₂ nanowires that exhibits improved sensitivity toward hydrogen peroxide. If GOx is immobilized in the surface, the β-MnO₂ acts as a mediator, and Nafion as a polymer backbone. The fabrication process was characterized by electrochemical impedance spectroscopy, and the sensor and its materials were characterized by cyclic voltammetry and amperometry. The biosensor enables amperometric detection of glucose with a sensitivity of 38.2 μA?·?mM^?1?·?cm^?2, and a response time of?<?5 s. This study also demonstrates the feasibility of realizing inexpensive, reliable, and high-performance biosensors using MnO₂ nanowires.

Miniaturised enzymatic conductometric biosensor with Nafion membrane for the direct determination of formaldehyde in water samples

A new conductometric enzyme-based biosensor was developed for the determination of formaldehyde (FA) in aqueous solutions. The biosensor was prepared by cross-linking formaldehyde dehydrogenase from Pseudomonas putida with bovine serum albumin in saturated glutaraldehyde vapours (GA) at the surface of interdigitated gold microelectrodes. Nicotinamide adenine dinucleotide cofactor (NAD⁺) was added in solution at each measurement to maintain enzyme activity. Addition of a Nafion layer over the enzyme modified electrode resulted in a significant increase of biosensor signal due to enhanced accumulation of protons generated by enzymatic reaction at the electrode surface. Different parameters affecting enzyme activity or playing a role in ionic transfer through the Nafion membrane were optimised. In optimal conditions (0.045 mg enzyme, 30 min exposure to GA, 0.3 μL of a 1 % (v/v) Nafion solution deposit, measurement in 5 mM phosphate buffer pH 7 containing 20 μM NAD⁺), the biosensor signal was linear up to 10 mM FA, and the detection limit was 18 μM. Relative standard deviations calculated from five consecutive replicates of FA solutions were lower than 5 % in the 1–10 mM range. The biosensor was successfully applied to the determination of FA in spiked water samples (tap water and Rhone river water), with recoveries in the 95–110 % range.

Glucose biosensor based on glucose oxidase immobilized on unhybridized titanium dioxide nanotube arrays

A glucose biosensor has been fabricated by immobilizing glucose oxidase (GOx) on unhybridized titanium dioxide nanotube arrays using an optimized cross-linking technique. The TiO₂ nanotube arrays were synthesized directly on a titanium substrate by anodic oxidation. The structure and morphology of electrode material were characterized by X-ray diffraction and scanning electron microscopy. The electrochemical performances of the glucose biosensor were conducted by cyclic voltammetry and chronoamperometry measurements. It gives a linear response to glucose in the 0.05 to 0.65 mM concentration range, with a correlation coefficient of 0.9981, a sensitivity of 199.6 μA mM^?1 cm^?2, and a detection limit as low as 3.8 µM. This glucose biosensor exhibited high selectivity for glucose determination in the presence of ascorbic acid, sucrose and other common interfering substances. This glucose biosensor also performed good reproducibility and long-time storage stability. This optimized cross-linking technique could open a new avenue for other enzyme biosensors fabrication.

Reagentless d-sorbitol biosensor based on d-sorbitol dehydrogenase immobilized in a sol–gel carbon nanotubes–poly(methylene green) composite

A reagentless d-sorbitol biosensor based on NAD-dependent d-sorbitol dehydrogenase (DSDH) immobilized in a sol–gel carbon nanotubes–poly(methylene green) composite has been developed. It was prepared by durably immobilizing the NAD⁺ cofactor with DSDH in a sol–gel thin film on the surface of carbon nanotubes functionalized with poly(methylene green). This device enables selective determination of d-sorbitol at 0.2 V with a sensitivity of 8.7?μA?mmol^?1?L?cm^?2 and a detection limit of 0.11 mmol?L^?1. Moreover, this biosensor has excellent operational stability upon continuous use in hydrodynamic conditions.

Biosensor for bisphenol A leaching from baby bottles using a glassy carbon electrode modified with DNA and single walled carbon nanotubes

We have developed a biosensor for highly sensitive and selective determination of the endocrinic disruptor bisphenol A (BPA). It is based on glassy carbon electrode modified with calf thymus DNA and a composited prepared from single walled carbon nanotubes (SWNT) and Nafion. The interaction between BPA and DNA was studied by voltammetry. The binding constant was determined to be 3.55?×?10³ M^?1, and the binding site has a length of 4.3 base pairs. These electrochemical studies provide further information for a better understanding of the toxicity and carcinogenicity of BPA. Under optimal conditions, the biosensor displays a linear electrochemical response to BPA in the 10 nM to 20 μM concentration range, with a detection limit as low as 5.0 nM (at an S/N of 3). The method was successfully applied to the quantification of BPA in leachates from plastic baby bottles. Recoveries range from 94.0 % to 106.0 % which underpins the excellent performance of this SWNT-based DNA sensor.

Amperometric determination of sulfite using screen-printed electrodes modified with metallic nanoparticles

We report on the amperometric determination of sulfite using screen-printed carbon electrodes (SPCEs) modified with gold and silver nanoparticles that were deposited on the electrode to improve the capabilities of detection. The electrode is fairly selective and responds to sulfite with an oxidation current (at 300 mV and pH 6) in the 9.80 to 83.33 μM concentration range. The precision in terms of repeatability and reproducibility is 14.4 % and 10.7 % in the case of SPCEs modified by gold nanoparticles. The method was applied to the determination of sulfite in drinking water, pickle juice and vinegar. Recoveries ranged from 96 % to 104 %.

Direct electrochemistry of hemoglobin on an ionic liquid carbon electrode modified with zinc tungstate nanorods

We have constructed a new electrochemical biosensor by immobilization of hemoglobin (Hb) and ZnWO₄ nanorods in a thin film of chitosan (CTS) on the surface of carbon ionic liquid electrode. UV–vis and FT-IR spectra reveal that Hb remains in its native conformation in the film. The modified electrode was characterized by scanning electron microscopy, electrochemical impedance spectroscopy and cyclic voltammetry. A pair of well-defined redox peaks appears which indicates direct electron transfer from the electrode. The presence of CTS also warrants biocompatibility. The electron transfer coefficient and the apparent heterogeneous electron transfer rate constant were calculated to be 0.35 and 0.757 s^?1, respectively. The modified electrode displays good electrocatalytic activity for the reduction of trichloroacetic acid with the detection limit of 0.613 mmol L^?1 (3σ). The results extend the protein electrochemistry based on the use of ZnWO₄ nanorods.

Chronocoulometric DNA biosensor based on a glassy carbon electrode modified with gold nanoparticles, poly(dopamine) and carbon nanotubes

We describe a sensitive chronocoulometric biosensor for the sequence-specific detection of DNA. It is based on a glassy carbon electrode modified with multi-walled carbon nanotubes, polydopamine, and gold nanoparticles. The ruthenium(III)hexammine complex acts as the electrochemical indicator. Electrochemical impedance spectra and scanning electron microscopy are employed to investigate the assembly of the electrode surface. The signals of the ruthenium complex electrostatically bound to the anionic phospho groups of the DNA strands are measured by chronocoulometry before and after hybridization. The difference in signal intensity is linearly related to the logarithm of the concentration of the target DNA in the range of 1.0 nM to 10 fM with a detection limit of 3.5fM (S/N?=?3) under optimal conditions. This biosensor exhibits excellent sensitivity and selectivity and has been used for an assay of complementary target DNA in human serum sample with satisfactory results.

Application of tubular tetrapod magnesium oxide in a biosensor for hydrogen peroxide

Tubular tetrapod magnesium oxide (tt-MgO) can be synthesized by thermal evaporation of Mg metal powder with a pre-grown tetrapod ZnO template. The morphology and structure of the tt-MgO were characterized by X-ray diffraction, scanning electron microscopy and transmission electron microscopy. A composite prepared from tt-MgO, nafion and horseradish peroxidase was employed to modify a gold electrode to result in an electrochemical biosensor for hydrogen peroxide that displays excellent sensitivity and rapid response in the presence of hydroquinone as a mediator. Its sensitivity is 335.4 μA mM^-1 cm^-2, its response is linear in the range from 1.0 to 450 μM, and the detection limit is 0.3 μM. These results demonstrate that tt-MgO provides a promising material for the designs of biosensors.

A silica-dextran nanocomposite as a novel matrix for immobilization of horseradish peroxidase, and its application to sensing hydrogen peroxide

We report on a novel matrix of solgel organic–inorganic nanocomposite that was fabricated from silica sol gel and dextran. It was used for the immobilization of horseradish peroxidase (HRP) to give a biosensor for hydrogen peroxide (H₂O₂). The sensor film was characterized by Fourier transform infrared and UV–vis spectroscopy with respect to structural features and the conformation of the enzyme. The topographies of the surface of the electrode were investigated by field emission scanning electron microscopy. The biosensor was used to determine H₂O₂ quantitatively in the presence of Methylene blue as a mediator with high electron transfer efficiency. A pair of stable and well defined quasi-reversible redox peaks of the HRP [Fe (III)]/HRP [Fe (II)] redox couple was observed at pH 7.0. The biosensor responds to H₂O₂ in the 0.5 mM to 16.5 mM concentration range, and the limit of detection is 0.5 mM.

Electrochemical sensor for Baicalein using a carbon paste electrode doped with carbon nanotubes

We report on the voltammetric determination of the flavonoid Baicalein by using a carbon paste electrode that was doped with multi-walled carbon nanotubes. The resulting sensor exhibits excellent redox activity towards Baicalein due to the large surface area and good conductivity of the electrode. Cyclic voltammetry at various scan rates was used to investigate the redox properties of Baicalein. At the optimum conditions, the sensor displays a linear current response to Baicalein in the 0.02–10 μM concentration range, with a limit of detection of 4.2 n M. The method was successfully applied to the determination of Baicalein in spiked human blood serum samples and in a Chinese oral liquid.

Synthesis of multi-branched gold nanoparticles by reduction of tetrachloroauric acid with Tris base, and their application to SERS and cellular imaging

A biosensor for hydrogen peroxide was constructed by immobilizing horseradish peroxidase on chitosan-wrapped NiFe₂O₄ nanoparticles on a glassy carbon electrode (GCE). The electron mediator carboxyferrocene was also immobilized on the surface of the GCE. UV?Cvis spectra, Fourier transform IR spectra, scanning electron microscopy, and electrochemical impedance spectra were acquired to characterize the biosensor. The experimental conditions were studied and optimized. The biosensor responds linearly to H₂O₂ in the range from 1.0?×?10^?5 to 2.0?×?10^?3?M and with a detection limit of 2.0?×?10^?6?M (at S/N?=?3).

A novel, disposable, screen-printed amperometric biosensor for ketone 3-β-hydroxybutyrate fabricated using a 3-β-hydroxybutyrate dehydrogenase–mesoporous silica conjugate

A disposable amperometric biosensor for ketone 3-β-hydroxybutyrate (3HB) has been developed successfully. The sensor is based on a screen-printed carbon electrode containing Meldola’s Blue (MB) and sensing components containing nicotinamide adenine dinucleotide (NAD⁺) and 3-β-hydroxybutyrate dehydrogenase (3HBDH) immobilized in mesoporous silica (FSM8.0) using an aqueous photo-cross-linkable polymer matrix of polyvinyl alcohol (O-391), and it requires only a small sample volume of 10 μL for the measurement. The behavior of a resulting biosensor, i.e., 3HBDH–FSM8.0/NAD⁺/MB-SPCE, was examined in terms of NAD⁺ concentration for construction, pH, applied potential, operational range, selectivity, and storage stability. The sensor showed an optimum response at a pH of 7.6 and at an applied potential of ?50 mV. The determination range and the response time for 3HB were from 30 μM to 8 mM and approximately 30 s, respectively. In addition, the sensor was quite stable and maintained >90 % of its initial response after being stored for over 6 months. This result implies that our method provides a novel biosensor for ketone 3-β-hydroxybutyrate which is easy-to-use, cost-effective, and has good reproducibility, which are vital for commercial purposes.