A new enzyme electrode for quantification of salicylic acid in a FIA system

This work presents an amperometric biosensor incorporated into a flow configuration comprising salicylate hydroxylase that catalyses the irreversible hydroxylation of salicylate to catechol in the presence of NADH and molecular oxygen, and tyrosinase that further oxidises catechol giving o-quinone which is electrochemically reduced at -100 mV vs. Ag/AgCl yielding catechol and entering the catalytic oxidation and electrochemical reduction cycling which results in signal amplification and, consequently, low limits of detection. Additional incorporation of glucose dehydrogenase in the enzymatic sequence results in regeneration of NADH provided that glucose is present in the carrier stream and incorporation of a dialysis membrane provides operational stability to the biosensor. The analytical characteristics of this catalytic and electrochemical transduction sequence in a FIA system are: a limit of detection of 3.5 10(-6) M (S/N = 3), a sensitivity of 22.6 nA microM(-1) cm(-2), no loss of response at least after 5 h of continuous operation, and a sample frequency of 15 h(-1). Monitoring of salicylate after ingestion of 500 mg of acetylsalicylic acid has been followed in non-pretreated urine samples and the amount of salicylate in several drugs has been also successfully quantified.