| This thesis includes a review and a research section. In the review, the history of immunoassay is introduced, the basic principles, characteristics and system of electrogenerated chemiluminescence immunoassay (ECLIA) is presented. Moreover, the applications of ECLIA in recent years are reviewed. ECLIA which uses an ECL species as a label to study the reaction of antigen and antibody and determinate antigen, hapten or antibody has become an increasingly promising immunoassay method owing to its non-radiation, simple instrumentation, high sensitivity, wide dynamic range, reproducibility, simplicity and rapidity analysis, its automation from electrochemical control.The research section contains four subunits. In the research section, we have developed ECL method for determination of sulfite, norfloxacin, haloperidol in neutral solution and multilabeled homogeneous ECLIA method for digoxin.A novel and simple energy transfer ECL method for the determination sulfite has been developed. It was found that a weak ECL signal of sulfite is electrochemically generated on a platinum electrode in neutral aqueous solution and strongly enhanced by Rhodamine B as an energy receptor and further enhanced by a neutral surfactant of Tween 80. Under 0.10 M phosphate buffer solution (pH=7.5) containing 2.0 × 10-6 g mL-1 Rhodamine B and 0.4% (v/v) Tween 80, the ECL response at the potential of 0.82V(vs Ag/AgCl) to the concentration of sulfite is linear range from 1.0 × 10-7 g/mL to 8.0 × 10-6 g/mL with the detection limit of 5.0 × 10-8 g/mL. Relative standard deviation(n=11, 1.0×10-6 g/mL) was 3.8%. The proposed method has been successfully applied to the determination of sulfite in pharmaceutical injections of VK3 and white sugar.A novel ECL method for the determination of norfloxacin is developed based on the enhancement effect of norfloxacin on the ECL of sodium hydrogen sulfite at a platinum electrode in a neutral solution. The linear range for norfloxacin is from 8.0 × 10-8 g/mL to 2.0 × 10-6 g/mL and the detection limit is 4.5 × 10-8 g/mL with the relativestandard deviation of 2.9%(n=11, 5.0× 10-7 g/mL). The proposed method has been successfully applied to the determination of norfloxacin in capsule and eye drops.A simple ECL method which based on the coupled ECL reaction of tris(2,2'-bipyridy)ruthenium(II) with amine-containing analyte has been developed for the rapid and sensitive determination of haloperidol. Under optimum situation, calibration graphs were obtained over the concentration range from 1.0× 10-9 g/mL to 9.0 ×10-6 g/mL for haloperidol with the detection limit of 8.0 × 10-10 g/mL. The proposed method has been successfully applied to the determination of haloperidol in pharmaceutical formulations.A novel homogeneous ECLIA for the determination of small hapten digoxin is developed using BSA and Ru(bpy)2(dcbpy)NHS as a carrier protein and ECL electrochemical luminophor, respectively. Digoxin(Dig) conjugated BSA was modified with Ru(bpy)2(dcbpy)NHS(Ru) to form Dig-BSA-Ru conjugate. After immunoreaction between anti-digoxin antibody and Dig-BSA-Ru conjugate, the complex of Ab-Dx-BSA-Ru undergoes a less ECL quantity. Digoxin hapten was determined throughout its therapeutic range with the detection limit of 4.0 × 10-10g/mL and in the range from 1.0× 10-9g/mL to 1.0× 10-7 g/mL by competitive immunoreaction between Dig-BSA-Ru conjugate and digoxin with fixed quantity of anti-digoxin antibody. The proposed method has been applied to determine digoxin in human control serum with satisfactory results.
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