Measurement of Sulfite Using Sulfite Oxidase and Luminol Chemiluminescence

Abstract

Measurement of sulfite using sulfite oxidase and luminol chemiluminescence was performed. Sulfite oxidase from chicken liver was immobilized in a reactor column filled with polysaccharide gel beads. The effects of buffer solution, hydrogen peroxide concentration and sulfite concentration on the chemiluminescence intensity were examined. For the same concentration of hydrogen peroxide, carbonate buffer solution system showed about 3 times higher chemiluminescence intensity than is the case with borate buffer solution. Hydrogen peroxide produced through the enzymatic reaction reacted with luminol to produce chemiluminescence. The intensity of the light emitted was proportional to the sulfite concentration at 3 × 10^–9– 10^–6M of sulfite. With the use of peroxidase from Arthromyces ramosus, the lower limit of sulfite detection was 3 × 10^–10M, which is the lowest concentration detectable using a biosensor. Relative standard deviation (RSD) of the response for this sulfite concentration was within 5% (n = 5).