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<p>Microfluidic paper-based analytical devices (μPADs) offer affordable and accessible diagnostic solutions, particularly for resource-limited settings. Recent advancements, such as multichannel μPAD arrays for multi-analyte testing and paper microzone plates mimicking traditional well plates, enhance their compatibility with laboratory workflows. This study focuses on the enzymatic responses of chitosan and GSH on paper microzone plates, using absorbance-based colorimetric measurements performed via a microplate reader. The paper microzone plates were fabricated by screen-printing PMMA solution to create hydrophobic barriers, defining hydrophilic microzones. The enzymatic behaviour of chitosan in the TMB oxidation reaction was monitored for TMB concentrations ranging from 2 to 10&#xa0;mM. Kinetic analysis on the paper microzone plates yielded a Michaelis–Menten constant (K<sub>m</sub>) of 2.26&#xa0;mM and a maximum reaction rate (V<sub>max</sub>) of 0.67 µMmin<sup>−1</sup>. The obtained K<sub>m</sub> value for chitosan on the paper-based platform was lower than that of the 96-well plate. A strong linear correlation (R<sup>2</sup> = 0.97) was observed between the absorbance values from the paper microzone plate and the 96-well plate at 652&#xa0;nm for TMB oxidation. The detection limit for GSH on the paper microzone plates was determined to be 1.23&#xa0;mM based on absorbance analysis and 1.45&#xa0;mM based on RGB analysis. These findings highlight the effectiveness of paper microzone plates for absorbance-based kinetic monitoring, offering a versatile and efficient platform for assay development and analyte detection.</p>

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Paper microzone plates as a platform for absorbance-based kinetic monitoring using enzymatic responses of chitosan and glutathione

  • 期刊论文

  • 发表日期：

    2025年10月2日

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  • Diksha Maheswari,
  • Hemant Sankar Dutta

摘要

Microfluidic paper-based analytical devices (μPADs) offer affordable and accessible diagnostic solutions, particularly for resource-limited settings. Recent advancements, such as multichannel μPAD arrays for multi-analyte testing and paper microzone plates mimicking traditional well plates, enhance their compatibility with laboratory workflows. This study focuses on the enzymatic responses of chitosan and GSH on paper microzone plates, using absorbance-based colorimetric measurements performed via a microplate reader. The paper microzone plates were fabricated by screen-printing PMMA solution to create hydrophobic barriers, defining hydrophilic microzones. The enzymatic behaviour of chitosan in the TMB oxidation reaction was monitored for TMB concentrations ranging from 2 to 10 mM. Kinetic analysis on the paper microzone plates yielded a Michaelis–Menten constant (Km) of 2.26 mM and a maximum reaction rate (Vmax) of 0.67 µMmin−1. The obtained Km value for chitosan on the paper-based platform was lower than that of the 96-well plate. A strong linear correlation (R2 = 0.97) was observed between the absorbance values from the paper microzone plate and the 96-well plate at 652 nm for TMB oxidation. The detection limit for GSH on the paper microzone plates was determined to be 1.23 mM based on absorbance analysis and 1.45 mM based on RGB analysis. These findings highlight the effectiveness of paper microzone plates for absorbance-based kinetic monitoring, offering a versatile and efficient platform for assay development and analyte detection.