Amperometric enzyme electrode for determination of theophylline in serum

Abstract

This paper describes an amperometric enzyme electrode for the rapid determination of theophylline in serum. The method is based on the catalysed oxidation of theophylline by the haem-containing enzyme theophylline oxidase. Results are presented for two approaches. First, ferrocene monocarboxylic acid was used as a mediator. The second-order rate constant was 1·1 × 10 3 1 mol −1 s −1. Secondly, the organic conducting salts NMP.TCNQ was used to construct enzyme electrodes. These electrodes were employed for the rapid (60 s) measurement of theophylline in serum at a working potential of +100 mV versus Ag/AgCl. Linear calibration curves were obtained over the clinically relevant range ( y = 0.13 x + 0.22, n = 8). Caffeine, theobromine and 3-methyl-xanthine at levels up to 100 mg l −1 do not interfere and 1-methylxanthine shows cross-reactivity at concentrations greater than 50 mg l −1.