| This thesis contains two parts. The first part is a review, the second part are research reports.A review is given on the new development of Chemiluminescence ( CL ) detection schemes for High Performance Liquid Chromatography ( HPLC ). It involved to the information about basic principles, instrument structure. Among them, the technique of HPLC combined with acidic CL system detection is mainly reviewed, At the same time, It covers the applications and the developing tendency of acidic HPLC - CL in pharmaceutical analysis.The thesis main introducing the two of KMnO4 and Ce(IV) acidic CL system. Research reports composed of four parts as follows.1. Determination of Cefalexin and Trimethoprim in Human Serum Using High Performance Liquid Chromatography with Chemiluminescence Detection SystemA novel high performance liquid chromatography ( HPLC ) detector based on chemiluminescence ( CL ) reaction was developed for the simultaneous determination of cefalexin ( CFX ) and trimethoprim ( TMP ). Cefalexin and Trimethoprim have obvious sensitizing effect to the potassium permanganate - formaldehyde chemiluminescence reaction in acidic medium. The separation was carried out with ZORBAX SB-C18 column ( 250 mm×4.6 mm, i.d., 5μm ) at 30℃. The mobile phase consisted of methanol - water solution ( 25 : 75, v/v ). The chemiluminescence intensity were linear with cefalexin and trimethoprim concentrations in the range of 0.50～100.0μg·mL-1 and 0.25～40.0μg·mL-1, respectively. The detection limit for cefalexin and trimethoprim were 0.16μg·mL-1 ( 3σ) and 0.08μg·mL-1 ( 3σ). The relative standard deviation ( n = 11 ) were 1.9 % for 2.0μg·mL-1 of cefalexin and 2.5 % for 1.0μg·mL-1 of trimethoprim. The recovery were in the range of 98.5 %～104.3 % and 97.8 %～103.7 %. The method has been satisfactorily applied to the determination of cefalexin and trimethoprim in human serum samples.2. Flow injection chemiluminescence determination of furosemideA novel chemiluminescence method for the determination of furosemide was developed. It was based on the fact that furosemide could enhance the chemiluminescence intensity from the reaction of Cerium ( IV ) with sodium sulfite in an acidic medium. The relationship between the enhanced intensity of chemiluminescence and the concentration of furosemide was linear with the range of 0.01～1.0 mg·L-1 and the detection limit was 4×10-3 mg·L-1 ( 3σ). The relative standard deviation ( RSD ) was 2.2 % ( n = 11, c= 0.2 mg·L-1). The method has been successfully applied to the determination of furosemide with the recoveries of 95 %～105 %.3. Cerium (IV) chemiluminescence system for determination of pefloxacin mesylateA novel chemiluminescence ( CL ) system for Pefloxacin Mesylate combined with flow injection analysis was proposed in this paper. It was based on the reaction between Cerium ( IV ) and sodium sulfite in acidic medium to give CL signal, which was enhance by Pefloxacin Mesylate. The experimental conditions for the CL reaction were optimized. Under the optimum conditions, the concentration of Pefloxacin Mesylate was proportional with the CL intensity in the range of 0.05～5.0μg·mL-1 with a RSD of 1.8 % ( n = 11, c= 0.6μg·mL-1 ). The detection limit was 0.005μg·mL-1 ( 3σ). The recovery was in the range of 97.4 %～103.7 %. The method has been successfully applied to the determination of Pefloxacin Mesylate in pharmaceutical preparation.4. Investigation of five FQNS veterinary Drug Residues in eggs by High Performance Liquid Chromatography with Chemiluminescence Detection System A novel high performance liquid chromatography ( HPLC ) detector based on Ce ( IV ) - Na2SO3 chemiluminescence ( CL ) system reaction was developed for the simultaneous determination of five FQNS veterinary drug residues could all enhance the chemiluminescence intensity in acidic medium. The separation was carried out with ZORBAX SB - C18 column ( 250 mm×4.6 mm i.d., 5μm ) at 25℃. The mobile phase consisted of acetonitrile - 0.1 mol·L-1 phosphate buffer solution ( pH 7.0 ) ( 20 : 80, v/v ). The chemiluminescence intensity were linear with PM, EN, NF, OF, CP concentration in the range of 0.2～20.0,3.0～50.0,3.0～50.0,0.3～20.0,3.0～50.0μmol·L-1, respectively. The detection limit for PM, EN, NF, OF, CP were 0.02,0.31,0.31,0.03,0.31μmol·L-1 ( 3σ). The relative standard deviation ( n = 11 ) were 2.5 %,3.4 %,2.9 %,3.2 %,2.8 % for 2.0μmol·L-1 PM,4.0μmol·L-1 EN,4.0μmol·L-1 NF,1.0μmol·L-1 OF,10.0μmol·L-1 CP. The recovery were in the range of 98.4 %～103.5 %. The method has been satisfactorily applied to the determination of five FQNS veterinary drug residues in egg samples.
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