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Development of a disposable screen-printed amperometric biosensor based on glutamate dehydrogenase, for the determination of glutamate in clinical and food applications

Hughes, Gareth; Pemberton, Roy; Fielden, P.; Hart, John P.

Authors

Gareth Hughes gareth6.hughes@uwe.ac.uk

P. Fielden



Abstract

A screen printed carbon electrode (SPCE) containing the electrocatalyst Meldola’s Blue (MB) has been investigated as the base transducer for a glutamate biosensor.
The sandwich biosensor was fabricated by firstly depositing a chitosan (CHIT) layer onto the surface of the transducer (MB-SPCE), followed by glutamate dehydrogenase (GLDH): this device is designated GLDH-CHIT-MB-SPCE. NAD+ was added to buffer solutions prior to the measurement of glutamate.
This biosensor was used in conjunction with amperometry in stirred solution at an applied potential of +0.1 V (vs. Ag/AgCl). Optimum conditions for the analysis of glutamate were found to be as follows: temperature, 35°C; buffer, pH 7; ionic strength, 75 mM; NAD+, 4 mM; CHIT 0.05% in 0.05 M HCl; GLDH, 30 U.
The linear range of the biosensor was found to be 12.5 µM to 150 µM, the calculated limit of detection (based on three times signal to noise ratio) was 1.5 µM and the sensitivity was 0.44 nA/µM.
The proposed biosensor was used to measure glutamate in serum before and after fortification with glutamate. The endogenous concentration of glutamate was found to be 1.68 mM and the coefficient of variation (CV) was 4.1%. The serum was then fortified with 2 mM of glutamate, and the resulting mean recovery was 96% with a CV of 3.3% (n = 6).
An unfiltered beef OXO cube was analysed for monosodium glutamate (MSG) content. The endogenous content of MSG was 124.80 mg/g with a CV of 8.06%.
The OXO cube solution was fortified with 0.935g (100 mM) of glutamate, the resulting mean recovery was 91% with a CV of 6.40 %.

Journal Article Type Article
Publication Date Aug 31, 2014
Journal Analytical and Bioanalytical Electrochemistry
Print ISSN 2008-4226
Peer Reviewed Peer Reviewed
Volume 6
Issue 4
Pages 435-449
APA6 Citation Hughes, G., Pemberton, R., Fielden, P., & Hart, J. P. (2014). Development of a disposable screen-printed amperometric biosensor based on glutamate dehydrogenase, for the determination of glutamate in clinical and food applications. Analytical & Bioanalytical Electrochemistry, 6(4), 435-449
Keywords glutamate, biosensor, screen-printed biosensor, amperometric, clinical, food
Publisher URL http://abechem.com/No.%204-2014/2014,6_4_,435-449.pdf
Related Public URLs http://abechem.com/

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