| Chemiluminescence (CL) is defined as the emission of light from an electronically excited species which produced during the course of a chemical reaction, and is observed when the electronically excited product or intermediate formed decays to the ground state with a photon emitted. Due to its high sensitivity, wide linear range, rapidity, simplicity and so on, the chemiluminescence method can he combined with other separate method especially electrophoresis and high performance liquid chromatogram etc, which can avoid the disadvantage of CL (poor selectivity). This has been successfully applied in environmental science, pharmaceutical science,and life science, and has become one of the most powerful techniques in analytical chemistry. The chemiluminescence (CL) detection system combined with HPLC separation method can offer excellent analytical selectivity and sensitivity. No doubtable, chemiluminescence linked to HPLC will be widely applied in biological science, clinical science environmental science and pharmaceutical science.The thesis consists of two parts. One is a review related to the application of HPLC with post-column chemiluminescence detection to flavones and antibiotics. Emphasis is taken on the development in the recent ten years.The other is a research report which is composed of 4 components as follows. 1) The determination of Fisten in human serum by high performance liquid chromatography with chemiluminescence detection; 2) RP-HPLC determination of a new intermediate S-AMOSA of antibiotic; 3) Determination of S-AMOSA by high performance liquid chromatography after derivatization with 9-flurenylmethyl chloroformate; 4) Determination of the water soluble vitamins in animal livers by revise-phase high performance liquid chromatography.1. The Determination of Fisten in Human Serum by High Performance Liquid Chromatography with Chemiluminescence DetectionA novel chemiluminescence determination of fisetin in human serum by high performance liquid chromatographic (HPLC) separation with chemiluminescence detector was described. The signal produced by the chemiluminescent reaction (CL) between luminol and ferricyanide was greatly increased in presence of fisetin. The linear range of fisetin concentration is 7.7×10-6～9.6×10-4g/L (r=0.9998) and the detection limit is 2.7×10-7g/L. The method is reliable, accurate and suitable to the determination of fisetin.2. RP-HPLC Determination of a New Intermediate S-AMOSA of AntibioticA rapid and sensitive RP-HPLC method was established for the determination of intermediate S-AMOSA of antibiotic. A Hypersil C18(4.6 mm×150 mm, 5.0μm) column and SPD-6AV detector were used. The mobile phase was composed of phosphate buffer. Detection wave length was 211 nm and the temperature of the column was room one. The linearity of the calibration curve was well correlated(r=0.9999) in the range of 4.8～120μg/mL. The recovery was 98.0%～103.0%. The method has been successfully applied to the determination of S-AMOSA, which was proved to be accurate, simple, effective and reproductive.3. Determination of S-AMOSA by High Performance Liquid Chromatography after Derivatization with 9-flurenylmethyl ChloroformateObjective to establish a high performance liquid chromatographic (HPLC) method for the determination of S-AMOSA. After derivatization with FMOC, S-AMOSA was separated and quantified by HPLC. A mobile phase of methanol-H2O, an analytical column of C18 and UV-VIS detection wavelength of 263 nm were used in the study. The linearity of the calibration curve was well correlated(r=0.9996) in the range of 0.01125～0.36 g/L. The recovery was 97.78%～102.33%. The method is reliable, accurate and suitable to the determination of S-AMOSA.4. Determination of the Water Soluble Vitamins in Animal Livers by Revise-phase High Performance Liquid ChromatographyTo establish a method for assaying four water-soluble vitamins in animal livers by high performance liquid chromatographic method. Gradient elution was used for the separation of the four water-soluble vitamins in 12.5 min. The chromatographic conditions were: analytical column, Shimadzu VP-ODS(150 mm×4.6 mm i.d., 5.0μm); column temperature, ambient; mobile phase with gradient elution, 0.01 mol/L potassium dihydrogen phosphate buffer(pH6.2)/methanol from 13: 87 to 28: 72 in 12.5 min; flow rate, 1.2 mL/min. The standard curve of VitC, VitB1, VitB6, VitB2 were linear in the concentration range of 22.3～116.0 , 2.4～152.5 , 2.2～340.0 , 1.7～272.0μg/mL respectively. The average of recovery of VitC, VitB1, VitB6, VitB2 was in the range of 98.6～102.3% and the RSD was 1.14%, 1.56%, 1.34%, 1.38%respectively. This method is accurate, simple, rapid and reproducible and can assay simultaneously for VitC, VitB1, VitB6, VitB2 in animal livers.
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