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

研制了三种用于同时测定单个样品中有机化合物的化学发光流通式传感器。首先，将固定化尿酸酶(EC1.7.3.3)、固定化过氧化物酶(EC1.11.1.7)、载体材料、共固定化L-谷氨酸酶(EC1.4.3.11)/过氧化物酶、载体材料、共固定化L-赖氨酸氧化酶(EC1.4.3.14)/过氧化物酶依次装入1 mm内径、75 cm长的透明FEP管中，作为流动细胞放置在光电倍增管前，用于同时测定血清中尿酸、L-谷氨酸和L-赖氨酸。样品溶液一次进样即可获得三峰记录。进样波长为50~1000 nm，采样速率为11次/h，无残留。方法检出限分别为5 nM尿酸、20 nM L谷氨酸和40 nM L赖氨酸。接下来，为了同时测定脑组织匀浆中的氯化和乙酰胆碱，固定化氯化酶(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)和共固定化氯化/过氧化物酶依次填充在FEP管中作为流动细胞。测定波长范围为3~1000 nm，进样量为25/h，检出限分别为0.5 nM氯气和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，将固定化的3-羟基丁酸脱氢酶/NADH氧化酶/过氧化物酶和葡萄糖脱氢酶/NADH氧化酶/过氧化物酶依次填充在FEP管中作为流动细胞。进样量为30μ/h，检出限为10 nm 3-羟基丁酸酯和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.