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Determination of Ammonium Ion Using a Reagentless Amperometric Biosensor Based on Immobilized Alanine Dehydrogenase

The use of the enzyme alanine dehydrogenase (AlaDH) for the determination of ammonium ion (NH(4)(+))usually requires the addition of pyruvate substrate and reduced nicotinamide adenine dinucleotide (NADH) simultaneously to effect the reaction. This addition of reagents is inconvenient when an enzyme...

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Main Authors: Ling Ling, Tan,, Yook Heng, Lee,, Ahmad, Musa,
Format: Article
Language:English
Published: Mdpi Ag 2015
Subjects:
Biosensor
Ammonium Ion
Reduced Nicotinamide Adenine Dinucleotide
Poly(2-Hydroxyethyl Methacrylate
Alanine Dehydrogenase
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spelling my.usim-81012017-02-23T04:46:40Z Determination of Ammonium Ion Using a Reagentless Amperometric Biosensor Based on Immobilized Alanine Dehydrogenase Ling Ling, Tan, Yook Heng, Lee, Ahmad, Musa, Biosensor Ammonium Ion Reduced Nicotinamide Adenine Dinucleotide Poly(2-Hydroxyethyl Methacrylate Alanine Dehydrogenase The use of the enzyme alanine dehydrogenase (AlaDH) for the determination of ammonium ion (NH(4)(+))usually requires the addition of pyruvate substrate and reduced nicotinamide adenine dinucleotide (NADH) simultaneously to effect the reaction. This addition of reagents is inconvenient when an enzyme biosensor based on AlaDH is used. To resolve the problem, a novel reagentless amperometric biosensor using a stacked methacrylic membrane system coated onto a screen-printed carbon paste electrode (SPE) for NH(4)(+) ion determination is described. A mixture of pyruvate and NADH was immobilized in low molecular weight poly(2-hydroxyethyl methacrylate) (pHEMA) membrane, which was then deposited over a photocured pHEMA membrane (photoHEMA) containing alanine dehydrogenase (AlaDH) enzyme. Due to the enzymatic reaction of AlaDH and the pyruvate substrate, NH(4)(+) was consumed in the process and thus the signal from the electrocatalytic oxidation of NADH at an applied potential of +0.55 V was proportional to the NH(4)(+) ion concentration under optimal conditions. The stacked methacrylate membranes responded rapidly and linearly to changes in NH(4)(+) ion concentrations between 10-100 mM, with a detection limit of 0.18 mM NH(4)(+) ion. The reproducibility of the amperometrical NH(4)(+) biosensor yielded low relative standard deviations between 1.4-4.9%. The stacked membrane biosensor has been successfully applied to the determination of NH(4)(+) ion in spiked river water samples without pretreatment. A good correlation was found between the analytical results for NH(4)(+) obtained from the biosensor and the Nessler spectrophotometric method. 2015-05-18T03:26:25Z 2015-05-18T03:26:25Z 2011 Article 1424-8220 en Mdpi Ag
institution Universiti Sains Islam Malaysia
building USIM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universit Sains Islam i Malaysia
content_source USIM Institutional Repository
url_provider http://ddms.usim.edu.my/
language English
topic Biosensor
Ammonium Ion
Reduced Nicotinamide Adenine Dinucleotide
Poly(2-Hydroxyethyl Methacrylate
Alanine Dehydrogenase
spellingShingle Biosensor
Ammonium Ion
Reduced Nicotinamide Adenine Dinucleotide
Poly(2-Hydroxyethyl Methacrylate
Alanine Dehydrogenase
Ling Ling, Tan,
Yook Heng, Lee,
Ahmad, Musa,
Determination of Ammonium Ion Using a Reagentless Amperometric Biosensor Based on Immobilized Alanine Dehydrogenase
description The use of the enzyme alanine dehydrogenase (AlaDH) for the determination of ammonium ion (NH(4)(+))usually requires the addition of pyruvate substrate and reduced nicotinamide adenine dinucleotide (NADH) simultaneously to effect the reaction. This addition of reagents is inconvenient when an enzyme biosensor based on AlaDH is used. To resolve the problem, a novel reagentless amperometric biosensor using a stacked methacrylic membrane system coated onto a screen-printed carbon paste electrode (SPE) for NH(4)(+) ion determination is described. A mixture of pyruvate and NADH was immobilized in low molecular weight poly(2-hydroxyethyl methacrylate) (pHEMA) membrane, which was then deposited over a photocured pHEMA membrane (photoHEMA) containing alanine dehydrogenase (AlaDH) enzyme. Due to the enzymatic reaction of AlaDH and the pyruvate substrate, NH(4)(+) was consumed in the process and thus the signal from the electrocatalytic oxidation of NADH at an applied potential of +0.55 V was proportional to the NH(4)(+) ion concentration under optimal conditions. The stacked methacrylate membranes responded rapidly and linearly to changes in NH(4)(+) ion concentrations between 10-100 mM, with a detection limit of 0.18 mM NH(4)(+) ion. The reproducibility of the amperometrical NH(4)(+) biosensor yielded low relative standard deviations between 1.4-4.9%. The stacked membrane biosensor has been successfully applied to the determination of NH(4)(+) ion in spiked river water samples without pretreatment. A good correlation was found between the analytical results for NH(4)(+) obtained from the biosensor and the Nessler spectrophotometric method.
format Article
author Ling Ling, Tan,
Yook Heng, Lee,
Ahmad, Musa,
author_facet Ling Ling, Tan,
Yook Heng, Lee,
Ahmad, Musa,
author_sort Ling Ling, Tan,
title Determination of Ammonium Ion Using a Reagentless Amperometric Biosensor Based on Immobilized Alanine Dehydrogenase
title_short Determination of Ammonium Ion Using a Reagentless Amperometric Biosensor Based on Immobilized Alanine Dehydrogenase
title_full Determination of Ammonium Ion Using a Reagentless Amperometric Biosensor Based on Immobilized Alanine Dehydrogenase
title_fullStr Determination of Ammonium Ion Using a Reagentless Amperometric Biosensor Based on Immobilized Alanine Dehydrogenase
title_full_unstemmed Determination of Ammonium Ion Using a Reagentless Amperometric Biosensor Based on Immobilized Alanine Dehydrogenase
title_sort determination of ammonium ion using a reagentless amperometric biosensor based on immobilized alanine dehydrogenase
publisher Mdpi Ag
publishDate 2015
_version_ 1645152340799389696
score 13.222552

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