New enzyme sensors for morphine and codeine based on morphine dehydrogenase and laccase

Two new enzymatic methods have been developed to quantify morphine and code ine simultaneously in a flow injection system (FIA). The first enzyme senso r for morphine or codeine is based on immobilizing morphine dehydrogenase ( MDH) and salicylate hydroxylase (SHL) on top of a Clark-type oxygen electro de. Morphine or codeine oxidation by MDH leads to a consumption of oxygen b y SHL via the production of NADPH. This decreases the oxygen current of the Clark electrode. Concentrations of codeine and morphine are detected betwe en 2 and 1000 mu M and between 5 and 1000 mu M, respectively. The second en zyme sensor for morphine is based on laccase (LACC) and PQQ-dependent gluco se dehydrogenase (GDH) immobilized at a Clark oxygen electrode. Morphine is oxidized by laccase under consumption of oxygen and regenerated by glucose dehydrogenase. Since laccase cannot oxidize codeine, this sensor is select ive for morphine. Morphine is detected between 32 nM and 100 mu M. Both sen sors can be operated simultaneously in one flow system (FIA) giving two sig nals without the requirement for a separation step. This rapid and technica lly simple method allows discrimination between morphine and codeine in les s than 1 min after injection. The sampling rate for quantitative measuremen ts is 20 h(-1). The method has been applied to the quantitative analysis of codeine or morphine in drugs.