Development of alcohol biosensor with porphyrin for alcoholic beverage

Hydrometry and rudimentary testing are conventional methods for ethanol detection. These methods are limited by lacking precision and accuracy resulting inability to detect trace ethanol concentrations. The data derived from those methods provide less suiTable for quality control and regulatory in the current scenario.The researcher was trying to develop new method to measure the ethanol content. Biosensors are being considered promising approaches for the detection of ethanol due to their enhanced specificity and simplified sample treatment. Normally the ethanol was identified through the reaction of alcohol in the presence of alcohol oxidase (AOX), which is operated by oxygen (O2) consumption and hydrogen peroxide (H2O2) production. The amount of H2O2 is directly proportional to the ethanol content. Furthermore, the H2O2 react with colorless form of the reduced 3,3',5,5'-tetramethylbenzidine (TMB), turning into blue oxidized TMB under the influence of peroxidase. To further improve the detection of ethanol content, the researcher investigated porphyrin and metalloporphyrin complexes as mimetic catalyst inspired by nature and possessing peroxidase-like properties.This study investigates the catalytic properties of ethanol reduction using AOX, porphyrin and metalloporphyrin complexes. The innovative approach offers potential benefits in terms of ethanol stability, and specificity when compared with traditional approaches. In this study, a representative porphyrin and metalloporphyrin complexes were selected including TPP, CuTPP, TOBPP, CuTOBPP, TOMPP, CuTOMPP and FeTOMPP, which were utilized for reacting with oxidized TMB in order to quantifying ethanol content in local beverages. All compounds were tested for their peroxidase-like activity. The obtained results showed that TPP and FeTOMPP exhibited better performance than CuTPP, TOBPP, CuTOBPP, TOMPP, and CuTOMPP. Furthermore, the FeTOMPP demonstrated outstanding detection properties than TPP, due to the higher activity (0.0977 µM-1) lower H2O2 detection limit (2.4 µM) and low amount at 35 µg/ml by the mean of absorbances at 652 nm. By harnessing the Alcohol oxidase (AOX)/FeTOMPP/TMB system, a novel colorimetric biosensor for alcohol detection was developed. This biosensor demonstrated good response and sensitivity towards ethanol detection within the range of 0-15% (v/v), with a lower detection limit of 0.0524% (v/v) and sensitivity at 0.0535% (v/v). The comparison of ethanol content in different market alcoholic beverages sample using gas chromatography (GC) and this developed biosensor reveal that ethanol values were differed less than 5%. These findings emphasize the potential applications of the AOX/FeTOMPP/TMB system for alcohol detection especially with a local beverage with rapid response (30 minutes) and precision method, moreover, it provides the possibility usage for other ethanol analytical in many applications.