Studies On Immobilized Enzyme Reactor For Sequential Injection Chemiluminescence Assay Of Glucose

In this paper, a novel immobilized enzyme reactor was developed, and it was applied to glucose detection by sequential injection chemiluminescence. The reactor featured long service life, stable performance and ease of fabrication. The glucose in plasma and urine was successfully detected with automatic operation.In chapter one, the principles, characteristics and applications of sequential injection analysis and chemiluminescence analysis were introduced, followed by an introduction of the enzymatic immobilization methods and combination of sequential injection technique with chemiluminescence analysis. Finally, the design of this work was presented.In the second chapter, the preparation of immobilized enzyme reactor, featuring large reactive surface and low flow impedance, was described. The reactor was made of a thin plastic film and two PDMS slab embedded by the reactive particles. The combining setup of the immobilized enzyme reactor with sequential injection chemiluminescence was designed. Various key parameters governing the analytical performance of the system were investigated, including the sequence of sample and reagent zones, reagent volume, reagent concentration, sample volume, flow rate and staying time. A linear range of 2.5×10-5～7.0×10-4 mol L-1, a detection limit of 3.7×10-6 mol L-1(3σ) and a relative standard deviation of 1.2%(n=11,5.0×10-4 mol L-1) were obtained. Carryover was less than 5% at a throughput of 63 samples/h. The recoveries of glucose spikes added to the plasma and urine samples ranged from 96.7 %～104.4%.In the third chapter, conclusion and prospect for the immobilized enzyme reactor and sequential injection chemiluminescence system were presented.