New enzyme sensors for morphine and codeine based on morphine dehydrogenase and laccase
New enzyme sensors for morphine and codeine based on morphine dehydrogenase and laccase
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DOI：
10.1007/s002160051320
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发表时间：
1999-05-01
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影响因子：
--
通讯作者：
Scheller, FW
中科院分区：
文献类型：
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作者：
Bauer, CG;Kühn, A;Scheller, FW
两种新的酶法已被开发用于在流动注射系统（FIA）中同时定量吗啡和可待因。第一个吗啡或可待因的酶传感器是基于将吗啡脱氢酶（MDH）和水杨酸羟化酶（SHL）固定在Clark型氧电极上。吗啡或可待因被MDH氧化导致SHL通过产生NADPH消耗氧气。这降低了克拉克电极的氧电流。可待因和吗啡的浓度分别在2和1000 μ M之间和5和1000 μ M之间检测。第二种吗啡酶传感器是基于固定在Clark氧电极上的漆酶（LACC）和PQQ依赖的葡萄糖脱氢酶（GDH）。吗啡被漆酶在耗氧的情况下氧化，并被葡萄糖脱氢酶再生。由于漆酶不能氧化可待因，该传感器对吗啡具有选择性。在32 nM和100 μ M之间检测到吗啡。两个传感器可以在一个流动系统（FIA）中同时运行，提供两个信号，而不需要分离步骤。这种快速且技术简单的方法可以在注射后不到1分钟内区分吗啡和可待因。定量测量的采样率为20 h（-1）。该方法已用于药物中可待因或吗啡的定量分析。
Two new enzymatic methods have been developed to quantify morphine and codeine simultaneously in a flow injection system (FIA). The first enzyme sensor for morphine or codeine is based on immobilizing morphine dehydrogenase (MDH) and salicylate hydroxylase (SHL) on top of a Clark-type oxygen electrode. Morphine or codeine oxidation by MDH leads to a consumption of oxygen by SHL via the production of NADPH. This decreases the oxygen current of the Clark electrode. Concentrations of codeine and morphine are detected between 2 and 1000 mu M and between 5 and 1000 mu M, respectively. The second enzyme sensor for morphine is based on laccase (LACC) and PQQ-dependent glucose 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 selective for morphine. Morphine is detected between 32 nM and 100 mu M. Both sensors can be operated simultaneously in one flow system (FIA) giving two signals without the requirement for a separation step. This rapid and technically simple method allows discrimination between morphine and codeine in less than 1 min after injection. The sampling rate for quantitative measurements is 20 h(-1). The method has been applied to the quantitative analysis of codeine or morphine in drugs.