Paper-Based Enzymatic Colorimetric Assay for Rapid Malathion Detection

Due to their unique properties, paper-based biosensors have attracted attention as inexpensive devices for on-site analysis. To achieve fast and sensitive detection of analytes, immobilization of enzymes with high apparent activities on paper is highly desirable; however, this is challenging. Herein, we report an improved approach to attach a malathion degrading enzyme, PoOPHM9, on paper via an interlocking network of Pluronic F127 (PF127)-poly(acrylic acid)-enzyme conjugates. The addition of PF127 improved retention of enzymatic activity as the apparent kinetic constant V-max of the immobilized enzyme increased two-fold compared with the paper prepared without PF127. The PF127-poly(acrylic acid)-PoOPHM9 papers provided rapid colorimetric detection of malathion at 0.1-50 mM. The detection was completed within 5 min using a smartphone and image analysis software. As a proof-of-concept, malathion-contaminated water, plant, and apple samples were analyzed with the papers successfully. This material is promising for on-site rapid analysis of malathion-contaminated samples.