Abstract
Hexavalent chromium is a toxic heavy metal getting discharged into the environment and water bodies through various industrial processes. Conventional analysis methods call for expensive equipment and complicated sample pretreatment that made unsuitable for onsite detection. Paper is used as an enzyme immobilization platform because of its property to wick the liquid by capillary action; lightweight, cheap and can be easily patterned or cut according to the requirements for developing biosensor. In this study, enzyme immobilization of glucose oxidase (GOx) on filter paper were examined using three polysaccharides such as chitosan, sodium alginate and dextran for entrapment efficiency, activity and stability of the immobilized enzyme. Among the three, chitosan proved efficient for enzyme entrapment with about 90% efficiency at 0.3% (w/v) chitosan. The stability was checked after 1 week at 4 °C and room temperature, where the chitosan entrapped enzyme retained nearly 97% stability at 4 °C. Enzyme inhibition study of GOx and Cr(VI) was carried out using chronoamperometry shown uncompetitive type of inhibition. A paper-based electrochemical biosensor strip was developed by immobilizing GOx enzyme on filter paper using chitosan as an entrapping agent and associating it with a screen-printed carbon electrode for amperometric measurements. The linear range of detection was obtained as 0.05-1 ppm with the limit of detection as 0.05 ppm for Cr(VI), which is the standard permissible limit in potable water. The relative standard deviation (5.6%) indicates good reproducibility of the fabricated biosensor.