Analytical Sciences


Abstract − Analytical Sciences, 32(8), 815 (2016).

3,3′,5,5′-Tetramethylbenzidine Oxidation on Paper Devices for Horseradish Peroxidase-based Assays
Lori Shayne Alamo BUSA,*1,*2 Takeshi KOMATSU,*1 Saeed MOHAMMADI,*1 Masatoshi MAEKI,*3 Akihiko ISHIDA,*3 Hirofumi TANI,*3 and Manabu TOKESHI*3,*4,*5,*6
*1 Graduate School of Chemical Sciences and Engineering, Hokkaido University, Kita 13 Nishi 8, Kita, Sapporo 060-8628, Japan
*2 Physical Sciences Department, College of Arts and Sciences, Nueva Vizcaya State University, Bayombong, Nueva Vizcaya 3700, Philippines
*3 Division of Applied Chemistry, Faculty of Engineering, Hokkaido University, Kita 13 Nishi 8, Kita, Sapporo 060-8628, Japan
*4 ImPACT Research Center for Advanced Nanobiodevices, Nagoya University, Furo-cho, Chikusa, Nagoya 464-8603, Japan
*5 Innovative Research Center for Preventive Medical Engineering, Nagoya University, Furo-cho, Chikusa, Nagoya 464-8601, Japan
*6 Institute of Innovative for Future Society, Nagoya University, Furo-cho, Chikusa, Nagoya 464-8601, Japan
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.