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Amperometric PMMA-microchip with integrated gold working electrode for enzyme assays

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Abstract

The use of a gold film in-channel detector combined with a poly(methyl methacrylate) (PMMA) CE microchip has been tested for alkaline phosphatase (AP) enzymatic assays. Tris-borate or Tris-Gly (pH 9.0, 50 mmol L−1) buffer solutions were appropriate as running buffer. Signals for three common AP products: α-naphthol, p-nitrophenol, and ascorbic acid, were obtained. They were reproducible (RSD 4.4% for six successive electropherograms corresponding to 5 mmol L−1 α-naphthol solution) and the response was dependent on concentration (linear relationship for ascorbic acid solutions between 5 and 20 mmol L−1 concentration). Use of an end-channel gold film electrode was also investigated. If one of the reagents (substrate or enzyme) is included in the running buffer, two different types of enzymatic assay are feasible in less than 3 min.

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Acknowledgements

This work has been supported by the FICYT under project FC-03-PB02-101. Mario Castaño Álvarez thanks FICYT for the award of a Ph.D. grant.

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Authors and Affiliations

  1. Departamento de Química Física y Analítica, Universidad de Oviedo, 33006, Oviedo, Asturias, Spain

    Mario Castaño-Álvarez, M. Teresa Fernández-Abedul & Agustín Costa-García

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  1. Mario Castaño-Álvarez
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  2. M. Teresa Fernández-Abedul
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  3. Agustín Costa-García
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Correspondence to Agustín Costa-García.

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Castaño-Álvarez, M., Fernández-Abedul, M.T. & Costa-García, A. Amperometric PMMA-microchip with integrated gold working electrode for enzyme assays. Anal Bioanal Chem 382, 303–310 (2005). https://doi.org/10.1007/s00216-005-3151-2

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  • DOI: https://doi.org/10.1007/s00216-005-3151-2

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