Development Of Chemiluminometric Flow-Through Sensor For Simultaneous Determination of Neurotransmitters

研制了三种可同时测定单一样品中有机化合物的流动荧光传感器。首先，为了同时测定血清中的尿酸盐、L-谷氨酸盐和L-赖氨酸，（EC1.7.3.3），固定化过氧化物酶（EC1.11.1.7），支持材料，共固定化L-谷氨酸氧化酶（EC 1.4.3.11）/过氧化物酶、支持材料和共固定化L-赖氨酸氧化酶（EC 1.4.3.14）/过氧化物酶依次装入直径为1 mm，长75 cm的FEP透明管中，置于光电倍增管前作为流动池。通过进样一次样品溶液获得三峰记录。分析物的测定范围分别为50 - 1000 nM，采样速率为11/h，无残留。检测限为5 nM尿酸盐、20 nM L-谷氨酸盐和40 nM L-赖氨酸。接下来，为了同时测定脑组织匀浆中的氯和乙酰胆碱，将固定化氯氧化酶（EC1.1.3.7）、固定化过氧化物酶、固定化乙酰胆碱酯酶（EC3.1.1.7）和共固定化氯氧化酶/过氧化物酶依次装入FEP管中并用作流动池。分析物的测定范围分别为3 - 1000 nM，样品通量为25/h。检测限为0.5 nM氯和2 nM乙酰胆碱。最后，为了同时测定血清中的D-葡萄糖和3-羟基丁酸，共固定化3-羟基丁酸脱氢酶/NADH氧化酶/过氧化物酶，一种支持材料，和共固定化葡萄糖脱氢酶/NADH氧化酶/过氧化物酶，依次填充在FEP管中，并用作流动池。分析物的测定范围分别为0.1 - 30 μM，样品通量为30/h。检测限为10 nM 3-羟基丁酸和50 nM D-葡萄糖。

Three kinds of chemiluminometric flow-through sensors for simultaneous determination of organic compounds in a single sample have been developed. First, for simultaneous determination of urate, L-glutamate, and L-lysine in serum, immobilized uricase (EC1.7.3.3), immobilized peroxidase (EC1.11.1.7), support material, coimmobilized L-glutamate oxidase (EC1.4.3.11)/peroxidase, support material, and coimmobilozed L-lysine oxidase (EC1.4.3.14)/peroxidase were packed sequentially in a transparent FEP tube (1mm i.d, 75 cm length), and the tube was placedin front of a photomultiplier tube as a flow cell. A three-peak recording was obtained by one injection of the sample solution. The analytes were determined from 50 to 1000 nM, respectively, and sampling rate was 11/h without carryover. The detection limits were 5 nM urate, 20 nM L-glutamte, and 40 nM L-lysine. Next, for simultaneous determination of chorine and acetylcholine in brain tissue homogenates, immobilized chorine oxidase(EC1.1.3.7), immobilized peroxidase, immobilized acetylcholine esterase(EC3.1.1.7), and coimmobilized chorine oxidase/peroxidase were packed, in turn, in the FEP tube and used as a flow cell. The analytes were determined from 3 to 1000 nM, respectively, and the sample throughput was 25/h. The detection limit were 0.5 nM chorine and 2 nM acetylcholine. Finally, for simultaneous determination of D-glucose and 3-hydroxybutyrate in serum, coimmobilized 3-hydroxybutyrate dehydrogenase/NADH oxidase/peroxidase, a support material, and coimmobilized glucose dehydrogenase/NADH oxidase/peroxidase were packed, in turn, in the FEP tube and used as a flow cell. The analytes were determined from 0.1 to 30 μM, respectively, and sample throughput was 30/h. The detection limit were 10nM 3-hydorxybutyrate and 50 nM D-glucose.