DEVELOPMENT OF CHEMILUMINOMETRIC FLOW-THROUGH SENSOR WITH IMMOBILIZED ENZYMES

流动注射系统中固定化酶的化学发光通流传感器已被开发。过氧化物酶”。将亮氨酸脱氢酶、nadh氧化酶、甲醛脱氢酶、尿酸酶、谷氨酸氧化酶和赖氨酸氧化酶共价固定在三烷基化的亲水性乙烯基聚合物珠上，装入透明聚四氟乙烯管（1.0 mm I.D. x 1.5 mm o.d）中，并将该管置于光电倍增管前作为流动细胞。由于这些酶是喜热酶，因此固定酶的稳定性优于先前使用的酶。传感器稳定了一个月。研究发现，节霉菌的过氧化物酶很适合作为鲁米诺-过氧化氢反应的化学发光-流动注射测量催化剂。研制了一种测定血清中l -谷氨酸的选择性流式传感器。摘要采用四种免疫反应偶联的方法，建立了l -谷氨酸的选择性检测原理，利用酶/过氧化物酶/谷氨酸氧化酶/过氧化物酶，在流动细胞中固定化尿酸酶分解血清中鲁米诺-过氧化氢化学发光反应的主要干扰成分之一的尿酸。尿酸酶反应产生的过氧化氢容易与还原性组分如抗坏血酸和谷胱甘肽反应，多余的过氧化氢被固定化过氧化物酶分解。样品塞中的l -谷氨酸用固定化谷氨酸氧化酶酶促转化为过氧化氢，过氧化氢在固定化过氧化物酶上与鲁米诺反应，产生与谷氨酸浓度成正比的化学发光。研制了一种用于测定鱼肉中组胺含量的流式传感器。从土壤中分离的节杆菌结晶石kaitb -007细胞中发现了组胺氧化酶，氧化酶与过氧化物酶共价固定在聚合物珠上，并装入tptfe管中。每隔2分钟进行一次组胺测定，无残留。在1.25%的反应是可再生的相对标准偏差ll-REPLICATE注射50 MM组胺。少

CHEMILUMINOMETRIC FLOW-THROUGH SENSORS WITH IMMOBILIZED ENZYMES IN FLOW INJECTION SYSTEMS HAVE BEEN DEVELOPED. PEROXIDASE." LEUCINE DEHYDROGENASE, NADH OXIDASE, FORMALDEHYDE DEHYDROGENASE, URICASE, GLUTAMATE OXIDASE, AND LYSINE OXIDASE WERE IMMOBILIZED COVALENTLY ON TRESYLATED HYDROPHILIC VINYL POLYMER BEADS AND PACKED INTO TRANSPARENT PTFE TUBE (1.0 MM I.D. X 1.5 MM O.D.), AND THE TUBE WAS PLACED IN FRONT OF A PHOTOMULTIPLIER TUBE AS A FLOW CELL THE STABILITIES OF THE IMMOBILIZED ENZYMES WERE SUPERIOR TO THOSE PREVIOUSLY USED, BECAUSE THESE WERE THERMOPHILIC ENZYMES. THE SENSORS WERE STABLE FOR A MONTH. IT WAS FOUND THAT THE PEROXIDASE FROM ARTHROMYCES RAMOSUS IS WELL SUITED AS CATALYST FOR CHEMILUMINOMETRIC FLOW-INJECTION MEASUREMENTS OF LUMINOL-HYDROGEN PEROXIDE REACTION.A SELECTIVE FLOW-THROUGH SENSOR FOR THE DETERMINATION OF L-GLUTAMATE IN SERUM WAS DEVELOPED. THE PRINCIPLE FOR THE SELECTIVE DETECTION FOR L-GLUTAMATE WAS BASED ON COUPLED RACTIONS OF FOUR SEQUENTIALLY ALIGNED IMMOB … More ILIZED OXJDASES, UIRCASE/PEROIXIDASE/ GLUTAMATE OXIDASE/PEROXIDASE IN A FLOW CELL THE IMMOBILIZED URICASE WAS USED EMPLOYED TO DECOMPOSE URATE, WHICH IS ONE OF THE MAJOR INTERFERING COMPONENTS IN SERUM FOR LUMINOL-HYDROGEN PEROXIDE CHEMILUMINESCENCE REACTION. THE HYDROGEN PEROXIDE PRODUCED FROM THE URICASE REACTION READILY REACTED WITH REDUCING COMPONENTS, SUCH AS ASCORBATE AND GLUTATHIONE, AND THEN THE EXCESS HYDROGEN PEROXIDE WAS DECOMPOSED BY THE IMMOBILIZED PEROXIDASE. L-GLUTAMATE IN THE SAMPLE PLUG WAS ENZYMATICALLY CONVERTED TO HYDROGEN PEROXIDE WITH IMMOBILIZED GLUTAMATE OXIDASE SUBSEQUENTLY THE PEROXIDE REACTED WITH LUMINOL ON THE IMMOBILIZED PEROXIDASE TO PRODUCE CHEMILUMINESCENCE, PROPORTIONAL TO GLUTAMATE CONCENTRATION.A FLOW-THROUGH SENSOR FOR THE DETERMINATION OF HISTAMINE IN FISH MEAT WAS DEVELOPED. HISTAMINE OXIDASE WAS FOUND IN CELLS OF ARTHROBACTER CRYSTALLOPOITES KAIT-B-007 ISOLATED FROM SOIL THE OXIDASE AND PEROXIDASE WERE COIMMOBILIZED COVALENTLY ON POLYMER BEADS AND PACKED INTO THE TPTFE TUBE. ONE ASSAY FOR HISTAMINE WAS DONE AT INTERVALS OF 2 MIN WITHOUT CARRYOVER. THE RESPONSE WAS REPRODUCIBLE WITHIN 1.25 % OF THE RELATIVE STANDARD DEVIATION FRO ll-REPLICATE INJECTIONS OF 50 MM HISTAMINE. Less