Determination of ammonium ion using a reagentless amperometric biosensor based on immobilized alanine dehydrogenase

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The use of the enzyme alanine dehydrogenase (AlaDH) for the determination of ammonium ion (NH4+) 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 NH4+ 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, NH4+ 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 NH4+ ion concentration under optimal conditions. The stacked methacrylate membranes responded rapidly and linearly to changes in NH4+ ion concentrations between 10-100 mM, with a detection limit of 0.18 mM NH4+ ion. The reproducibility of the amperometrical NH4+ biosensor yielded low relative standard deviations between 1.4-4.9%. The stacked membrane biosensor has been successfullyapplied to the determination of NH4+ ion in spiked river water samples without pretreatment. A good correlation was found between the analytical results for NH4+ obtained from the biosensor and the Nessler spectrophotometric method.

Original languageEnglish
Pages (from-to)9344-9360
Number of pages17
JournalSensors
Volume11
Issue number10
DOIs
Publication statusPublished - Oct 2011

Fingerprint

Alanine Dehydrogenase
dehydrogenases
Biosensing Techniques
alanine
Ammonium Compounds
bioinstrumentation
Biosensors
pyruvates
Ions
membranes
NAD
Membranes
enzymes
Pyruvic Acid
ion concentration
ions
Enzymes
nicotinamide
adenines
low molecular weights

Keywords

  • Alanine dehydrogenase
  • Ammonium ion
  • Biosensor
  • Poly(2-hydroxyethyl methacrylate)
  • Reduced nicotinamide adenine dinucleotide

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Atomic and Molecular Physics, and Optics
  • Analytical Chemistry
  • Biochemistry

Cite this

Determination of ammonium ion using a reagentless amperometric biosensor based on immobilized alanine dehydrogenase. / Tan @ Chong, Ling Ling; Ahmad, Musa; Lee, Yook Heng.

In: Sensors, Vol. 11, No. 10, 10.2011, p. 9344-9360.

Research output: Contribution to journalArticle

@article{a1920c770e714d218f6491cd4d019d8d,
title = "Determination of ammonium ion using a reagentless amperometric biosensor based on immobilized alanine dehydrogenase",
abstract = "The use of the enzyme alanine dehydrogenase (AlaDH) for the determination of ammonium ion (NH4+) 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 NH4+ 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, NH4+ 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 NH4+ ion concentration under optimal conditions. The stacked methacrylate membranes responded rapidly and linearly to changes in NH4+ ion concentrations between 10-100 mM, with a detection limit of 0.18 mM NH4+ ion. The reproducibility of the amperometrical NH4+ biosensor yielded low relative standard deviations between 1.4-4.9{\%}. The stacked membrane biosensor has been successfullyapplied to the determination of NH4+ ion in spiked river water samples without pretreatment. A good correlation was found between the analytical results for NH4+ obtained from the biosensor and the Nessler spectrophotometric method.",
keywords = "Alanine dehydrogenase, Ammonium ion, Biosensor, Poly(2-hydroxyethyl methacrylate), Reduced nicotinamide adenine dinucleotide",
author = "{Tan @ Chong}, {Ling Ling} and Musa Ahmad and Lee, {Yook Heng}",
year = "2011",
month = "10",
doi = "10.3390/s111009344",
language = "English",
volume = "11",
pages = "9344--9360",
journal = "Sensors (Switzerland)",
issn = "1424-8220",
publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",
number = "10",

}

TY - JOUR

T1 - Determination of ammonium ion using a reagentless amperometric biosensor based on immobilized alanine dehydrogenase

AU - Tan @ Chong, Ling Ling

AU - Ahmad, Musa

AU - Lee, Yook Heng

PY - 2011/10

Y1 - 2011/10

N2 - The use of the enzyme alanine dehydrogenase (AlaDH) for the determination of ammonium ion (NH4+) 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 NH4+ 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, NH4+ 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 NH4+ ion concentration under optimal conditions. The stacked methacrylate membranes responded rapidly and linearly to changes in NH4+ ion concentrations between 10-100 mM, with a detection limit of 0.18 mM NH4+ ion. The reproducibility of the amperometrical NH4+ biosensor yielded low relative standard deviations between 1.4-4.9%. The stacked membrane biosensor has been successfullyapplied to the determination of NH4+ ion in spiked river water samples without pretreatment. A good correlation was found between the analytical results for NH4+ obtained from the biosensor and the Nessler spectrophotometric method.

AB - The use of the enzyme alanine dehydrogenase (AlaDH) for the determination of ammonium ion (NH4+) 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 NH4+ 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, NH4+ 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 NH4+ ion concentration under optimal conditions. The stacked methacrylate membranes responded rapidly and linearly to changes in NH4+ ion concentrations between 10-100 mM, with a detection limit of 0.18 mM NH4+ ion. The reproducibility of the amperometrical NH4+ biosensor yielded low relative standard deviations between 1.4-4.9%. The stacked membrane biosensor has been successfullyapplied to the determination of NH4+ ion in spiked river water samples without pretreatment. A good correlation was found between the analytical results for NH4+ obtained from the biosensor and the Nessler spectrophotometric method.

KW - Alanine dehydrogenase

KW - Ammonium ion

KW - Biosensor

KW - Poly(2-hydroxyethyl methacrylate)

KW - Reduced nicotinamide adenine dinucleotide

UR - http://www.scopus.com/inward/record.url?scp=80054984927&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80054984927&partnerID=8YFLogxK

U2 - 10.3390/s111009344

DO - 10.3390/s111009344

M3 - Article

C2 - 22163699

AN - SCOPUS:80054984927

VL - 11

SP - 9344

EP - 9360

JO - Sensors (Switzerland)

JF - Sensors (Switzerland)

SN - 1424-8220

IS - 10

ER -