A new silica-gel nanospheres (SiO(2)NPs) composition was formulated, followed by biochemical surface functionalization to examine its potential in urea biosensor development. The SiO(2)NPs were basically synthesized based on sol-gel chemistry using a modified Stober method. The SiO(2)NPs surfaces were modified with amine (-NH2) functional groups for urease immobilization in the presence of glutaric acid (GA) cross-linker. The chromoionophore pH-sensitive dye ETH 5294 was physically adsorbed on the functionalized SiO(2)NPs as pH transducer. The immobilized urease determined urea concentration reflectometrically based on the colour change of the immobilized chromoionophore as a result of the enzymatic hydrolysis of urea. The pH changes on the biosensor due to the catalytic enzyme reaction of immobilized urease were found to correlate with the urea concentrations over a linear response range of 50-500 mM (R-2 = 0.96) with a detection limit of 10 mM urea. The biosensor response time was 9 min with reproducibility of less than 10% relative standard deviation (RSD). This optical urea biosensor did not show interferences by Na+, K+, Mg2+ and NH4+ ions. The biosensor performance has been validated using urine samples in comparison with a non-enzymatic method based on the use of p-dimethylaminobenzaldehyde (DMAB) reagent and demonstrated a good correlation between the two different methods (R-2 = 0.996 and regression slope of 1.0307). The SiO(2)NPs-based reflectometric urea biosensor showed improved dynamic linear response range when compared to other nanoparticle-based optical urea biosensors.

One of the major problems in the analysis of ammonia (NH3) using spectrophotometric method is highly coloured water sample. A reflectometric NH3 sensor based on a facile single-step immobilisation of cobalt(II) (Co2+) ion onto high capacity Dowex HCR-W2 microspheres has been developed to enable the direct NH3 determination in highly coloured water samples. The combination of Co2+ ion-immobilised microspheres via cation exchange chemical reaction and reflectometric technique has improved the optode sensing performance with respect to linear response range and response time. Immobilised Co2+ ion, when complexed with NH3 gave a blue colouration of hexamminecobaltate(II) ([Co(NH3)(6)](2+)) complex at pH 13. The NH3 sensor reached maximum colour intensity in similar to 6 min at ambient conditions. The linear calibration curve was 60-290 mM NH3 with a detection limit of 16 mM NH3, which is suitable for direct determination of NH3-enriched leachate water samples. A good reproducibility between 5.4 and 5.9% relative standard deviation (RSD) was obtained with the probe and the sensing performance can be regenerated using buffer pH 1. The effect of potential interference ions has been investigated during the quantification of NH3 in heavily polluted sewage water. The results obtained were in good agreement with those produced by acidimetric reference method. Crown Copyright (C) 2012 Published by Elsevier B.V. All rights reserved.

A regenerable electrochemical DNA biosensor based on a new type of acrylic microspheres and gold nanoparticles (AuNPs) composite coated onto a screen printed electrode (SPE) has been successfully developed for specific determination of the 35 S promoter from cauliflower mosaic virus (CaMV 35S) gene in soybean. DNA probe was immobilised onto acrylic microspheres via covalent bonding. The presence of modified gene in soybean can be detected via hybridisation of CaMV 35S gene-modified DNA with immobilised DNA probe, which was monitored by differential pulse voltammetry of anthraquinone-2sulfonic acid monohydrate sodium salt (AQMS) as redox indicator during hybridisation event. The peak current signal of AQMS was linearly related to the target CaMV 35S gene concentration over the range of 2 x 10(-15) to 2 x 10(-9) M (R-2 = 0.982) with a very low concentration detect limit (7.79 x 10(-16) M). The recovery test showed satisfactory results of 94.6 +/- 5.1-105.4 +/- 4.9% (n = 5) when the biosensor was used for the determination of genetically modified (GM) DNA sequences extracted from GM soybean samples. The DNA biosensor showed good reproducibility (relative standard deviation (RSD) below 5.0%, n = 5) and regenerability (RSD below 5.0%, n = 7). The biosensor response was stable up to 45 days of storage period at 4 degrees C. The main advantages of this biosensor design are very low detection limit and capability of reusing the biosensor for at least seven times after regeneration with mild sodium hydroxide. (c) 2013 The Authors. Published by Elsevier B. V. All rights reserved.

An amperometric electrochemical biosensor has been developed for ammonium (NH4+) ion detection by immobilising alanine dehydrogenase (AlaDH) enzyme in a photocurable methacrylic membrane made up of poly(2-hydroxyethyl methacrylate) (pHEMA) on a screen-printed carbon paste electrode (SPE). The current detected was based on the electrocatalytic oxidation of nicotinamide adenine dinucleotide reduced (NADH) that is proportional to the consumption of NH4+ ion whilst enzymatic amination of AlaDH and pyruvate is taking place. The biosensor was operated amperometrically at a potential of +0.6V and optimum pH 7. The NH4+ biosensor demonstrated linear response to NH4+ ion concentration in the range of 0.03-1.02 mg/L with a limit of detection (LOD) of 8.52 mu g/L. The proposed method has been successfully applied to the determination of NH4+ ion in river water samples without any pretreatment. The levels of possible interferents in the waters were negligible to cause any interference on the proposed method. The analytical performance of the biosensor was comparable to the colorimetric method using Nesslerisation but with much lower detection limit and linear response range at ppb level.

The use of multilayer of gold nanoparticles (AuNPs) attached on gold electrode surface via thiol chemistry to fabricate an ammonium (NH4+) ion biosensor based on alanine dehydrogenase (AlaDH) was investigated. The approach of the study was based on construction of biosensor by direct deposition of AuNPs and 1,8-octanedithiol (C8-DT) onto the gold electrode surface. For the immobilisation of enzyme, 2-mercaptoethanol (2BME) was first covalently attached to AlaDH via esther bonding and then followed by chemically attached the 2BME-modified AlaDH (2BME-AlaDH) moiety onto the AuNPs electrode via the exposed thiol group of 2BME. The resulting biosensor response was examined by means of amperometry for the quantification of NH4+ ion. In the absence of enzyme attachment, the use of three layers of AuNPs was found to improve the electrochemistry of the gold electrode when compared with no AuNPs was coated. However, when more than three layers of AuNPs were coated, the electrode response deteriorated due to excessive deposition of C8-DT. When AlaDH was incoporated into the AuNPs modified electrode, a linear response to NH4+ ion over the concentration range of 0.1-0.5 mM with a detection limit of 0.01 mM was obtained. In the absence of AuNPs, the NH4+ ion biosensor did not exhibit any good linear response range although the current response was observed to be higher. This work demonstrated that the incorporation of AuNPs could lead to the detection of higher NH4+ ion concentration without the need of dilution for high NH4+ ion concentration samples with a rapid response time of < 1 min.

A new bioanalytical method for urea determination by using the immobilized urease on the highly luminescent ZnS quantum dots (QDs), which functioned as the pH fluorescent label has been developed. Bioconjugation of ZnS QDs to urease via amide bond was performed to obtain the ZnS QDs-urease bioconjugate. Upon enzymatic hydrolysis of urea by the immobilized urease, a pH change to a more alkaline condition has led to the deprotonation of ZnS QDs, and an increase in the fluorescence intensity can be observed. The fluorescence intensity of the urease conjugated ZnS QDs changed linearly with the urea concentrations between 4 x 10(-9) M and 4 x 10(-3) M (R-2 = 0.992) at pH 6 with a calibration sensitivity of 179.46 intensity/decade. No noticeable influence by the Ca2+, Mg2+, K+ and Na+ ions on the response of the fluorescent pH bioprobe. Urea determination in soil sample by using the urea bioprobe was in good agreement with the standard DMAB (p-dimethylaminobenzaldehyde) UV-vis spectrophotometric method. By employing the strategy of ZnS QDs-urease bioconjugation, the stable covalent link between the two substances has appeared to widen the dynamic range and lower the detection limit for urea compared to free enzyme and QDs in solution for bioassay of urea concentration. (C) 2016 Elsevier B.V. All rights reserved.

A new optical sensor for fast screening of ammonium (NH4+) ion was developed based on immobilisation of p-nitrobenzene-diazonium chloride (Riegler's reagent) onto XAD-7 microbeads. In aqueous solution, the diazonium salt is unstable and will spontaneously decompose at room temperature but we have successfully stabilised this compound via immobilisation using physical adsorption to create a solid-state Riegler's reagent for NH4+ ion determination with a storage period of one month. The quantification of NH4+ ion concentration was possible by using reflectance spectrophotometry method with an optical fibre probe. The solid-state Riegler's reagent based optical sensor yielded a wide linear response range for NH4+ ion of 10-60 ppm and a fast response time of 2 min when compared with many commonly used coloured reagents. The limit of detection (LOD) of the optical NH4+ ion sensor was 7.9 ppm NH4+ ion with optimum response at pH 7. The response of the sensor was reproducible (4.2-5.7% relative standard deviation, n = 3) and can be regenerated using buffer pH 1. Except for Fe3+ ions, most common ions showed no serious interference. The sensor was also used for the determination of NH4+ ion in river water and the results obtained were comparable to those obtained by a standard titrimetric method. (c) 2012 Elsevier B.V. All rights reserved.

This paper reports the results for the quantitative determination of ammonia (NH3) in aqueous solution by a UV-vis spectrophotometric method and artificial neural network (ANN) intelligence tool. Quantitation of NH3 was based on the chemical reaction of NH3 with cobalt(II) (Co2+) ions in basic medium to form a blue hexamminecobaltate(II) ([Co(NH3)(6)](2+)) complex. Characterizations of Co2+ ion in solution included photostability, pH effect, response time, Co2+ ion concentration effect, dynamic linear range and reproducibility, which were performed using a UV-vis spectrophotometer. The pink cobalt species gradually changed to blue with increasing NH3 concentration. The absorption calibration curve was linear over the NH3 concentration range of 0.6-3.5 mM at optimum pH 8 with a reproducibility relative standard deviation (RSD) of <4.0%. The interference effect was found to be negligible for a number of foreign ions present in the reaction medium during NH3 determination in an aqueous environment. A set of absorbance data for the [Co(NH3)(6)](2+) complex at selected wavelengths was input for ANN training using a back-propagation algorithm. The trained network with 22 hidden neurons, a 28 500 epoch number and 0.001% learning rate has extended the dynamic NH3 concentration range to 0.6-5.9 mM with a calibration error as low as 0.0649 x 10(-3). The proposed ANN electronic sensor shows promise for NH3 estimation in unknown water samples based on pattern recognition.