Highly sensitive detection of glucose via glucose oxidase immobilization onto conducting polymer-coated composite polyacrylonitrile nanofibers.

Current study introduces composite polyacrylonitrile - multiwall carbon nanotubes nanofibers (PAN-MWCNTs NFs) coated with conducting polymers (polypyrrole (PPy) or poly(3,4-ethylenedioxythiophene) (PEDOT)) by chemical vapor deposition for efficient glucose detection. The potential of nanofibrous assemblies and nano-conducting elements in biosensing was explored as pre-processing of NFs with MWCNTs and post-processing with CPs were both employed. These 'core-shell' conducting NFs were further employed as platforms for glucose oxidase immobilization for enzymatic detection of glucose. The performance of the biosensors was closely correlated with the concentration of immobilized enzyme and with the type of conducting polymer. The biosensors showed high sensitivities of 92.94 and 81.72 µA/mM.cm-2 for (PAN-MWCNTs)/ PEDOT and (PAN-MWCNTs)/ PPy accompanied by low limit of detection values of 2.30 and 2.38 µM, respectively. Good operational stability was observed throughout twenty-five consecutive measurements, over 90% activity was maintained for both sensors. This study represents proof of concept for the methodology, showcasing the advantages of nanomaterial synthesis for bio-applications. The work was compared thoroughly with previously reported biosensors showing some of the best results reported to date in terms of analytical characteristics.

Trace-Level Phenolics Detection Based on Composite PAN-MWCNTs Nanofibers.

In view of major concerns regarding toxicity (genotoxic, mutagenic, hepatotoxic) of phenolics, there is an on-going necessity for sensitive and accurate analytical procedures for detection and measurements in environmental field, water, and food quality control. The current study proposes composite polyacrylonitrile nanofibrous assemblies enriched with multi-wall carbon nanotubes (PAN-MWCNTs NFs) as suitable immobilization platforms for cross-linking of tyrosinase in detection of both diphenols and monophenols, which are of much interest in water contamination. The prepared biosensor (Pt/PAN-MWCNTs/Tyr) showed high sensitivity in catechol detection (362.5 mA/M.cm2 ) and moderate sensitivity for the two monophenols (phenol, p-chlorophenol) with 14.79 and 25.46 mA/M.cm2 , respectively. The LOD values of the sensor were in the nanomolar range: 4.42×10-9 , 4.06×10-7 and 1.05×10-7  M for catechol, phenol and p-chlorophenol, respectively, allowing detection of trace amounts of phenolics in spiked water samples.

Next step in 2nd generation glucose biosensors: Ferrocene-loaded electrospun nanofibers.

Glucose determination is one of the most common analyses in clinical chemistry. Employing biosensors for this purpose has become the method of choice for home use for diabetic patients. To limit the impact of dissolved O2 concentration or possible interferences (known hindrances in the classical glucose detection approach), a variety of mediated pathways have been explored. Herein, an ingenious, facile and low-cost approach for immobilization of redox mediator within nanofibrous mats is presented. This '2nd generation' biosensor is able to avoid common issues such as leaching or diffusion barriers whilst providing the necessary close contact between the enzyme and the redox shuttle, for enhancing the detection accuracy and accelerate the response. Polyacrylonitrile nanofibers loaded with carbon nanotubes and ferrocene (PAN/Fc/MWCNT-COOH NFs) have been successfully prepared and applied as biosensing matrices upon cross-linking of glucose oxidase (GOD). The morphology of the NFs was investigated by means of scanning electron microscopy (SEM-EDX) and correlated to the kinetics of mediated electron transfer and to the efficiency in glucose detection, which were evaluated through cyclic voltammetry (CV) and amperometric measurements. The content of Fc was varied from 0.5 to 5.0 wt%, with optimum biosensor performance at 1.0 wt% exhibiting a linear range up to 8.0 × 10-3 M with sensitivity of ~27.1 mAM-1 cm-2 and 4.0 µM LOD. Excellent stability (RSD 2.68%) during 40 consecutive measurements along with insignificant interference and accurate recovery in real sample analysis (~100%) make for a very reliable sensor that can easily render itself to miniaturization and has the potential for a wide range of practical applications.

Facile copper-based nanofibrous matrix for glucose sensing: Eenzymatic vs. non-enzymatic.

The current study aims to provide a valid comparison between glucose detection efficiency with an enzymatic and a non-enzymatic sensing platform. A low-cost nano-matrix for glucose sensing was developed by drop-coating copper nanoparticles (Cu NPs) onto a polyacrylonitrile (PAN) electrospun nanofibrous assembly. The PAN NFs/Cu NPs matrix was optimized regarding electrospinning time and Cu NPs content and employed as a non-enzymatic sensor or further modified by cross-linking of glucose oxidase (GOD) for the development of an enzymatic sensor. The non-enzymatic glucose sensor was three times more sensitive (300 mAM-1cm-2) than the enzymatic one (81 mAM-1cm-2) with similar limit of detection values (5.9 and 5.6 µM, respectively). Incorporation of MWCNTs improved both the LOD (3.3 µM) and the operational stability of the non-enzymatic configuration (RSD 7.3%). The interference effect proved insignificant for the enzymatic sensor due to the innate catalytic selectivity whilst the non-enzymatic sensor acquired selectivity due to the nanofibrous PAN matrix and Nafion coating. The non-enzymatic PAN NFs/Cu NPs sensor was chosen for the detection of glucose in real blood serum samples whilst the PAN NFs/Cu NPs/GOD sensor was applied for glucose detection in fruit juices, both proving recovery results close to 100%.

The effect of copolymerization and carbon nanoelements on the performance of poly(2,5-di(thienyl)pyrrole) biosensors.

The development of biosensing interfaces based on copolymerization of benzenamine-2,5-di(thienyl)pyrrole (SNS-An) with 3,4-ethylenedioxythiophene (EDOT) is reported. Both homopolymer P(SNS-An) and copolymer P(SNS-An-co-EDOT) films were prepared and evaluated, in terms of biosensing efficiency, upon incorporation of carbon nanoelements (carbon nanotubes and fullerene) and cross-linking of glucose oxidase. The copolymer revealed superior performance as a biosensing interface as compared to the homopolymer structure or previously reported P(SNS) biosensors. The analytical characteristics and stability studies were performed both at cathodic potential, monitoring O2 consumption, as a result of catalytic reaction of glucose oxidase towards glucose and at anodic potential, following the oxidation of the H2O2 produced during the catalytic reaction. Whilst the measurements on the positive side offered an extended linear range (0.01-5.0 mM), the negative side provided sensitivity up to 104.96 µA/mMcm-1 within a shorter range. Detection limits were as low as 1.9 µM with Km value of 0.49 mM. Lastly, the most performant biosensing platforms, including copolymeric structure and CNTs were employed for analysis in real samples.

Utilization of enzyme extract self-encapsulated within polypyrrole in sensitive detection of catechol.

Utilization of polyphenol oxidases (laccase, tyrosinase) in biosensor technology is an efficient approach towards phenol detection, which is significant in numerous fields such as environmental monitoring, food industry etc. The use of crude extract instead of pure enzyme eliminates the need for costly and laborious processes of enzyme separation and purification. This study employs polyphenol oxidase extract, biosynthesized by white-rot fungi Trametes pubescens (TP) for the development of amperometric biosensors for catechol detection. The catalytic activity of the crude extract was firstly used to induce the bio-synthesis of conducting polymer - polypyrrole (Ppy), resulting in the self-encapsulation of the enzyme extract within the conducting material. The viability and biological integrity of the enzyme extract was preserved after the synthesis and was able to efficiently detect phenolic compounds such as catechol. Comparative evaluations between the biosynthesized Ppy based biosensor (bio-Ppy) and the biosensor based on bio-PPy with additional enzyme extract (bio-Ppy-TP) were performed. Lastly, the performance of these two biosensors was compared with that of a third one, based on chemically synthesized Ppy with enzyme extract (chem-Ppy-TP). All three types of biosensors proved high efficiency for catechol detection at low concentration (1-60 µM) and were employed for real sample detection in fruit wines showing linear correlation with the spectrophotometric results obtained with the Folin-Ciocalteau standard test.

An amperometric glucose biosensor based on PEDOT nanofibers.

Here we present a simple, low cost approach for the production of PEDOT nanofiber biosensors using simple techniques. Firstly, nanofibers of PEDOT were produced by the chemical vapor polymerization of EDOT on FeCl3 containing electrospun PAN nanofiber mats. The nanofibers were characterized by SEM, FTIR, CV and conductivity studies, which indicated the formation of homogeneous, porous, electroactive PEDOT nanofibers. The fabrication of biosensors was achieved through the loading of various amounts of GOx on the nanofibers. To uncover their capability, the biosensors were operated under both hydrogen peroxide production and oxygen consumption conditions. For each biosensor current response versus glucose concentration calibration curves were plotted. The sensitivity, linear range, LOD, K m and I max values of the biosensors were determined and the stabilities of all the sensors were investigated. The biosensor operating at 0.6 V revealed a lower LOD with a wider linear range, higher stability, good sensitivity and selectivity. For example, the PEDOT-NFs/GOx-3 nanofiber biosensor showed good sensitivity (74.22 µA mM-1 cm-2) and LOD (2.9 µM) with a response time of 2-3 s without any interference effects. The PEDOT-NFs/GOx-2 biosensor operating at -0.6 V exhibited extreme sensitivity of 272.58 µA mM-1 cm-2. Our studies have shown that having good sensitivity, LOD and stability makes these interference-free, easy to construct sensors viable candidates for commercialization.

Construction of ferrocene modified conducting polymer based amperometric urea biosensor.

Herein, an electrochemical urea sensing bio-electrode is reported that has been constructed by firstly electropolymerizing 4-(2,5-Di(thiophen-2-yl)-1H-pyrrol-1-yl)aniline monomer (SNS-Aniline) on Pencil Graphite Electrode (PGE), then modifying the polymer coated electrode surface with di-amino-Ferrocene (DAFc) as the mediator, and lastly Urease enzyme through glutaraldehyde crosslinking. The effect of pH, temperature, polymer thickness, and applied potential on the electrode current response was investigated besides performing storage and operational stability experiments with the interference studies. The resulting urea biosensor's amperometric response was linear in the range of 0.1-8.5mM with the sensitivity of 0.54µA/mM, detection limit of 12µM, and short response time of 2s. The designed bio-electrode was tested with real human blood and urine samples where it showed excellent analytical performance with insignificant interference.

Calixarene assembly with enhanced photocurrents using P(SNS-NH2)/CdS nanoparticle structure modified Au electrode systems.

Two novel calix[n]arene-adorned gold electrodes producing high photocurrent intensities were successfully constructed by embedding gold electrode surfaces with both P(4-(2,5-di(thiophen-2-yl)-1H-pyrrol-1-yl)benzenamine) conducting polymer and 4-mercaptoboronic acid-functionalized semiconductor CdS nanoparticles to facilitate the binding of calix[n]arene sulfonic acids with nanoparticles. This structure enabled an electron transfer cascade that both induced effective charge separation and efficiently generated photocurrent. The prepared electrodes were used to generate photocurrent by relying on the host-guest interactions of guests Br3(-) and I3(-), which if positioned well in the system was able to fill electron-hole pairs of CdS nanoparticles. As a result, host calixarene derivatives crucially held Br3(-) and I3(-) ions at a substantial distance from CdS nanoparticles. Furthermore, the effects of various calixarenes on the photocurrent obtained indicate that the generation of photocurrent intensities by the system depends on the cavity sizes of calixarene derivatives, which provide an essential center for Br3(-) and I3(-) ions.

A neutral state green polymer with a superior transmissive light blue oxidized state.

This report highlights the synthesis of only the second green polymer in the literature, which possesses superior properties over the first: a highly transmissive light blue color in the oxidized state with high optical contrast and excellent switching properties.