Enzymatic spectrophotometric determination of nitrites in beer

A colorimetric assay for nitrite determination in beer based on c-type multiheme enzyme Nitrite reductase (NiR) isolated from Desulfovibrio desulfuricans ATCC 27774, was developed. Using the enzyme in solution, nitrite assay was linear in the 10^-8 - 10^-2M range with a detection limit of 10^-8 M and a recovery ranging from 90 to 107%. The imprecision ranged from 4 to 10% on the entire calibration curve. With NiR immobilised onto a nylon coil, a flow reactor was developed which showed a narrower linear range (10^-5 - 10^-2 M) and a higher detection limit (10^-5 M) than with the enzyme in solution, but made it possible to reuse the enzyme up to 100 times (50% residual activity). Sample preparation was simple and fast: only degassing and beer dilution by buffer was needed. This enzymatic assay was in good agreement with the results obtained using commercial nitrite determination kits.