Development And Application Of Capillary Electrophoresis Based On Rapid Separation And High Sensitivity

Capillary electrophoresis(CE) has been widely used as a powerful separation and analytical technique in recent years because of its advantages including high performance,high speed,low analytes and sample consumptions.The aim of the thesis is to establish the rapid,sensitive capillary electrophoresis methods and apply to the drug quality control and pharmacokinetic study.The thesis is composed of four chapters.In the first chapter,the general situation of CE was introduced,and some opinions on the development hotspots were proposed.In the second chapter,a fast CZE method was established to simultaneous determine D-amphetamine and diphenhydramine in Quick-Acting Anti-Motion Capsules(QAAMC).The influence of different parameters(internal standard, injection modes,pH,concentration of the running buffer and applied voltage) was systematically studied.Running buffer:50 mmol/L phosphate-50 mmol/L borate(pH 5.5);Capillary:40.2 cm×75μm i.d.,effective length 30 cm,uncoated;Applied Voltage:20 kV(+)→(-);Column temperature:25℃;Electrokinetic injection:10 kV×4 s;Detection:200 nm.The migration times of D-amphetamine, diphenhydramine and anlodipine were 1.45,1.56 and 1.67 min,respectively.The determination was completed within 2.0 min.With the comparison of HPLC method, the analysis time was shortened at least 3 min.In the third chapter,the FASS technique was introduced to the CZE to get a high sensitivity to support the pharmacokinetic study of QAAMC.The influence of different parameters(composition of sample solvent,pH,concentration and composition of the running buffer and injection time) was systematically studied.The developed method was compared with LC/MS/MS method in literature.The results demonstrated that this simple and rapid CZE method was alternative to the LC/MS/MS method and sufficiently sensitive to follow plasma levels of D-amphetamine and diphenhydramine after oral administration of QAAMC to beagle dogs.In the fourth chapter,a 96-well SPE-CZE method was established and developed for the clinical pharmacokinetic study of prulifloxacin tablets.This paper firstly described a method for quantification of ulifloxacin,the active metabolite of prulifloxacin in human plasma by capillary zone electrophoresis using lomefloxacin as internal standard.Under the condition of 200 mmol/L borate buffer,pH 10.5, applied voltage 20 kV,hydrostatic injection 2psi×5 s,NM394 and lomefloxacin can be baseline separated.The peak area ratio of NM394 and lomefloxacin vs. concentration of NM394 was linear over the range from 0.02 to 2μg/mL.A comparison of pharmacokinetic parameters obtained by CZE and HPLC method showed no significant differences.It was confirmed that the CZE method was an alternative method for routine analysis of ulifloxacin in plasma comparing with the reference method.