| The major research field of pharmaceutical analysis involves the analysis of effectivecomponents in Chinese medicine, the monitoring of active ingredient of drugs in the body,chiral drugs separation, the interaction between drug molecule and biomacromolecule and soon. Due to the superior seperation efficiency, fast speed, low cost, and wide application,capillary electrophoresis (CE) is becoming a rapid development technique for pharmaceuticalanalysis. In this paper, based on three separation modes including capillary zoneelectrophoresis (CZE), non-aqueous capillary electrophoresis (NACE), and affinity capillaryelectrophoresis (ACE), coupled with gas chromatography-mass spectrometry (GC-MS),chemiluminescence (CL), and three-dimensional fluorescence spectra, several simple andsensitive analysis methods for the determination of some drugs were developed. The proposedmethods had been successfully applied for the dermination of various drugs such as effectivecomponents in Chinese medicine, the monitoring of active ingredient of drugs in the body.Moreover, the interaction mechanism between an antitumor drug and human serum albuminwas investigated by ACE, and satisfactory results were obtained.In Chapter1, after demonstration the basic principle of CE, different seperation modesand their application in drugs analysis were reviewed.In Chapter2, the analytical methods for the determination of anthraquinones and fattyacids in Semen Cassiae were studied. Several anthraquinone compounds in Semen Cassiaewere successfully seperated by NACE, and the influence factors including the types ofbuffer, non-aqueous media and additive were investigated in detail. Under the optimizedconditions, a new method for simultaneous determination of emodin and aurantio-obtusinwas established. The linear ranges were2.04—204and3.32—332μg/mL, respectively,with the limit of detection (LOD)0.61μg/mL for emodin and1.0μg/mL foraurantio-obtusin. Furthermore, fatty acids were extracted from cassiae seed by usingultrasonic-assisted method with petroleum ether as solvent, and the extracts were analyzedby GC-MS. The results showed that ultrasonic-assisted extraction was more effective forunsaturated fatty acid than conventional refluxing method. There were seven fatty acidswere characterized and determined. Unsaturated fatty acids especially linoleic acid are richin the extracts with a content of mass fraction44.45%.Chapter3described the establishment of novel methods for the determination of cephalosporins and medicines combined with them in the body fluids by CE. Firstly, theapplicability of capillary zone electrophoresis for simultaneous determination ofcefamandole and probenecid in human serum and urine samples has been studied. Underthe optimized conditions, the analytes can be separated in6min effectively. When salicylicacid was chosen as the internal standard，the standard curves of cefamandole andprobenecid showed good linearity between10～200and5～110mg/L respectively with thecorrelation coefficients r＞0.999. Secondly, a rapid and sensitive method for simultaneousdetermination of cefoperazone, sulbactam and probenecid by CZE was also proposed. Theexperiment results showed that the standard curves for three drugs had good linearity，andthe LODs were3.2,15and1.6μg/mL(S/N=3), respectively.Chapter4presented the development of methods for the determination offluoroquinolones in pharmaceutical preparation and biological fluids withCE-electrochemiluminescence(CE-ECL) and flow injection chemiluminescence(FI-CL). First,fleroxacin and proline in urine were simultaneously determined by CE coupled withelectrochemiluminescence. The linear ranges were0.4～80and0.1～60μg/mL, respectively,with the LOD of0.06μg/mL for fleroxacin and0.02μg/mL for proline. Moreover, the uniquebehaviors of enoxacin belongs to quinolones drugs in different chemiluminescence systemswere investigated by flow injection chemiluminescence. Based on it, a new sensitive methodof CL was proposed for determination of enoxacin. The LOD was as low as10-10g/mL,which is more sensitive than CE-ECL（10-8g/mL）and CE（10-6g/mL）.In chapter5, the characterization of interaction between antitumor drug5-fluorouracil(FU) and human serum albumin (HSA) was studied by affinity capillary electrophoresis(ACE). The binding of FU drug to HSA at near-physiological conditions was evaluated. FUwas found to show low affinity toward HSA, with binding constant of9.19×104M-1. Thepositive H and S values obtained by ACE showed that the binding reaction was anendothermic process, the entropy drived the binding, and hydrophobic interaction playedmajor roles in the binding of FU to HSA. The replacement test with warfarin and ketoprofenshowed that binding site of FU at the protein HSA is believed to be site I. Three-dimensionalfluorescence studies showed that the presence of FU could change the conformation of BSAduring the binding process.In chapter6, the main work, findings and innovation of this thesis were illustrated. At theend, the development trend of CE in the application of drugs analysis was presented. |
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