Enzymatic-spectrophotometric determination of sucrose in coffee beans

A spectrophotometric method for determining sucrose is proposed. Sucrose is hydrolyzed by invertase into glucose and fructose. Then, glucose is oxidized in presence of glucose oxidase and the produced hydrogen peroxide reacts with phenol-4-sulfonic acid sodium salt and 4-aminoantipyrine in presence of peroxidase, yielding a pink dye with an absorption maximum at 505 nm. This method was validated following the EURACHEM and VAM project guidelines for method validation. Trueness, precision, robustness, sensitivity and linearity were considered. The method was applied to the determination of sucrose in green and roasted coffee beans. A comparison with the HPLC method with pulsed amperometric detection was carried out.