EU ( Iii ) Sensitized Chemiluminescence Method And Its Application In Pharmaceutical Analysis

Chemiluminescence (CL) was defined as electromagnetic (ultraviolet, visible or near-infrared) radiation of the reactant, the production or the intermediate as a result of a chemical reaction. The CL analytical method was established based on the detection of the radiation of the moment intensity or the total radiation quantities. The advantages of CL analysis include high sensitivity, wide linear range, rapidity, simple instrumentation, easy automation and continuous analysis. CL analysis has been successfully explored for the analysis of a great number of important inorganic and organic substances in various fields including biotechnology, pharmacology, molecular biology, clinical medicine and environmental detection.Depending on the origin of the CL, CL reactions have been classified as direct and indirect or sensitized or energy transfer CL. The indirect CL reaction is important for improving the sensitivity of the detection and broadening the analytical range of the CL analysis. Trivalent lanthanide species display high photoluminescence efficiencies (>5% in H₂O), large Stokes' shifts (230nm～300 ran), long excited-state lifetimes (on the order of several hundred microseconds), and narrow emission spectra. Recently, these characteristic of lanthanide species have been introduced into CL analysis to develop sensitive method for the determination of several fluoroquinolone antibiotics.This thesis consists of review and research section. In the review section, the fluorescent characteristic of lanthanide species and their complex was briefly introduced. The application of lanthanide ions, Eu(III) and Tb(III), in fluorescence and CL analysis was summarized. In the research section, it was found that in the presence of Eu(III), the weak CL signal from KMnO₄-Na₂SO₃ reaction was greatly enhanced by atenolol, metoprolol, and ibuprofen and KIO₄-H₂O₂ reaction was greatly enhanced by naproxen. The experenment conditions were optimized. Based on these facts, new flow injection CL methods were developed for the determination of atenolol, metoprolol, ibuprofen and naproxen. The proposed methods were applied to the analysis of real samples. A possible mechanism on CL reaction was also proposed. The research section includes four parts.Part one, it was found that the weak CL signal from KMnO₄-Na₂SO₃ reaction was greatly enhanced when atenolol-Eu(III) complex was present into the system. Based on this finding, a new flow injection CL method for the determination of atenolol was proposed. The effects of the experimental conditions were examined. The linear range is 8.0×10^-9～1.0×10^-5 g/mL atenolol and the detection limit is 3×10^-9g/mL. The relative standard deviation is 2.4% for 1.0×10^-7g/mL atenolol solution(n=l 1). This method was applied to the determination of atenolol in urine and plasma samples satisfactory results. The mechanism of CL reaction was discussed briefly.Part two, the weak signal from KMnO₄-Na₂SO₃ reaction was greatly enhanced when Eu(III)-metoprolol tartrate complex was present into the system. Basted on this fact, a new flow injection CL method for the determination of metoprolol tartrate was proposed. Under the optical experiment conditions, the linear range is 5.0×10^-7～1.0×10^-5g/mL metoprolol tartrate and the detection limit is 1×10^-7g/mL. The relative standard deviation for 1.0×10^-6g/mL metoprolol tartrate solution is 1.4%(n=11). This method was applied to the determination of metoprolol tartrate in human plasma and urine samples.Part three, it was found that the weak CL signal from KMnO₄-Na₂SO₃ reaction could be enhanced by ibuprofen. The enhancement of ibuprofen was greatly enhanced when Eu(III) was also present in the system. Basted on this investigation, a new flow injection CL method for the determination of ibuprofen was proposed. The linear range for ibuprofen is 5.0×10^-8～5.0×10^-6g/mL and the detection limit is 1×10^-8g/mL ibuprofen. The relative standard deviation is 1.7%(1.0×10^-7g/mL ibuprofen, n=11). This method was applied to the determination of ibuprofen in table and plasma samples.Part four, it was found that naproxen displayed a weak sensitizing effect on the CL signal of KIO₄-H₂O₂ system. This sensitizing effect was greatly enhanced in the presence of Eu(III). Based on this investigation, a new CL method for the determination of naproxen was proposed combined with flow injection. The linear range of the method is 5.0×10^-8～5.0×10^-6g/mL naproxen. The detection limit is 1×10^-8g/mL naproxen. The relative standard deviation is 2.1% the 11 repleated determinations of 1.0×10^-7g/mL naproxen solution. This method was applied to the determination of naproxen in table and urine samples. The result is satisfiable. Meanwhile, possible chemiluminescence mechanism was discussed.