3,3′,5,5′-Tetramethylbenzidine Oxidation on Paper Devices for Horseradish Peroxidase-based Assays

Lori Shayne Alamo BUSA; Takeshi KOMATSU; Saeed MOHAMMADI; Masatoshi MAEKI; Akihiko ISHIDA; Hirofumi TANI; Manabu TOKESHI

doi:10.2116/analsci.32.815

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3,3′,5,5′-Tetramethylbenzidine Oxidation on Paper Devices for Horseradish Peroxidase-based Assays

Lori Shayne Alamo BUSA, Takeshi KOMATSU, Saeed MOHAMMADI, Masatoshi MAEKI, Akihiko ISHIDA, Hirofumi TANI, Manabu TOKESHI

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Keywords: TMB oxidation, horseradish peroxidase, paper-based analytical devices, photolithography, wax-printing

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Supplementary material

2016 Volume 32 Issue 8 Pages 815-818

DOI https://doi.org/10.2116/analsci.32.815

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Abstract

We report on the colorimetric oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) by hydrogen peroxide using horseradish peroxidase on photolithography-fabricated (P-PAD) and wax-printed (W-PAD) paper-based analytical devices. Fabricating PADs via photolithography exposes the hydrophilic areas to polymers (photoresists) and solvents, not only reducing the hydrophilicity, but also affecting the TMB–H₂O₂ assay system with an unavoidable incomplete elimination of photoresist during fabrication. Detection signals are then observed in the presence of photoresist residues on the P-PAD, even at a blank HRP concentration.

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