An Electrochemical Enzyme Biosensor for 3-Hydroxybutyrate Detection Using Screen-Printed Electrodes Modified by Reduced Graphene Oxide and Thionine.

Gonzalo Martínez-García, Elena Pérez-Julián, Lourdes Agüí, Naomí Cabré, Jorge Joven, Paloma Yáñez-Sedeño, José Manuel Pingarrón

Journal: Biosensors 2018;7(4):50

PMID: 29137135

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Abstract

A biosensor for 3-hydroxybutyrate (3-HB) involving immobilization of the enzyme 3-hydroxybutyrate dehydrogenase onto a screen-printed carbon electrode modified with reduced graphene oxide (GO) and thionine (THI) is reported here. After addition of 3-hydroxybutyrate or the sample in the presence of NAD⁺ cofactor, the generated NADH could be detected amperometrically at 0.0 V vs. Ag pseudo reference electrode. Under the optimized experimental conditions, a calibration plot for 3-HB was constructed showing a wide linear range between 0.010 and 0.400 mM 3-HB which covers the clinically relevant levels for diluted serum samples. In addition, a limit of detection of 1.0 µM, much lower than that reported using other biosensors, was achieved. The analytical usefulness of the developed biosensor was demonstrated via application to spiked serum samples.

Address: Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain. [email protected].; Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain. [email protected].; Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain. [email protected].; Unitat de Recerca Biomèdica (URB-CRB), Antic Hospital Universitari de Sant Joan c/Sant Joan s/n, 43201 Reus, Spain. [email protected].; Unitat de Recerca Biomèdica (URB-CRB), Antic Hospital Universitari de Sant Joan c/Sant Joan s/n, 43201 Reus, Spain. [email protected].; Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain. [email protected].; Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain. [email protected].

DOI: 10.3390/bios7040050

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MeSH Terms: 3-Hydroxybutyric Acid, Biosensing Techniques, Electrodes, Graphite, Phenothiazines