Optical enzymatic formaldehyde biosensor based on alcohol oxidase and pH-sensitive methacrylic-acrylic optode membrane

Optical biosensor for the detection of formaldehyde has been developed based on the transparent enzymatic stacked membranes system on the glass substrate, and employing optical absorption transducer with H+ ion-selective Nile Blue chromoionophore (NBCM) dye-doped methacrylic acrylic (MB28) copolymer membrane as the optode membrane. Alcohol oxidase (AOx) enzymes were entrapped within the biocompatible sol-gel matrix and deposited on top of the pH optode membrane. As the uppermost catalytic membrane catalyzes the oxidative conversion of formaldehyde to formic acid and hydrogen peroxide, the immobilized NBCM undergoes protonation reaction and forms HNBCM+, the dark blue ion-chromoionophore complex via H+ ion transfer reaction within the soft and flexible MB28 polymeric membrane. This rendered the enzymatic optode membrane absorbed a high yellow light intensity from the light source and exhibited maximum absorption peaks at 610 and 660 nm. Optical evaluation of formaldehyde by means on UV-vis absorption transduction of the enzymatic stacked membranes demonstrated rapid response time of 10 min with high sensitivity, good linearity and high reproducibility across a wide formaldehyde concentration range of 1 × 10-3-1 × 103 mM (R2 = 0.9913), and limit of detection (LOD) at 1 × 10-3 mM, which could be useful for formaldehyde assay in industrial, agricultural, environmental, food and beverages as well as medical samples. The formaldehyde concentration in snapper fish, pomfret fish and threadfin fish samples determined by the proposed optical enzymatic biosensor were very much close to the formaldehyde concentration values determined by the UV-vis spectrophotometric NASH standard method based on the statistical t-test. This suggests that the optical biosensor can be used as a reliable method for quantitative determination of formaldehyde levels in food samples.