Study On Amine-containing Drugs Using Capillary Electrophoresis-Electrochemiluminescence Coupled With Other Techniques

Capillary electrophoresis-elecreochemiluminescence(CE-ECL)combines CE advantages of high separation efficiency,fast speed and small sample volume and ECL performances of high sensitivity,wide linear range and simple equipment.CE-ECL has been widely used in various fields including pharmaceutical analysis,environmental analysis and immunnoassays.CE-ECL application in pharmaceutical analysis is a complement to the traditional analysis methods,which provides a low cost and high sensitivity analysis method to pharmaceutical analysis.Chemical modified electrode(CME)is made by coating the molecular,ion and polymer with excellent chemical properties onto the surface of the electrode,which will give the electrode a special function.At present,CME has become a method to improve the sensitivity of the elecreochemiluminescence detection.There are two purposes in the paper.On the one hand,it aims to expand the application of the Ru(bpy)32+-based CE-ECL technique.On the other hand,another purpose is to improve the sensitivity of CE-ECL by chemical modified electrode.The main contents and results were introduced as follows:1.A simple,fast and sensitive method was developed to study the interaction between human serum albumin(HSA)and amlodipine besylate(AML)using capillary electrophoresis with electrochemiluminescence(CE-ECL)coupled with ultrasonic technique for the first time.The CE-ECL parameters that affect separation and detection were optimized.Under the optimized conditions,the linear concentration range of AML was from 0.18 to 27 μmol/L(r2=0.9963)and the detection limit of AML was 0.050 μmol/L(S/N=3).The time required for combination equilibrium by ultrasonic was 50 min,which was far less than that by normal incubation(220 min).It took 340 min to achieve the equilibrium by traditional dialysis and only 60 min by ultrasonic microdialysis.Compared with traditional dialysis,the use of ultrasonic microdialysis greatly shortened experimental time and simplified experimental procedures.The number of binding site and binding constant were 1.28 and 6.21×1O4 L/mol,respectively.2.A new method for determination of clomipramine hydrochloride and norclomipramine hydrochloride was developed by coupling liquid-liquid microextraction and capillary electrophoresis with electrochemiluminescence technique.Clomipramine hydrochloride and norclomipramine hydrochloride were separated using acetonitrile as an additive.The CE-ECL parameters that affect separation and detection were optimized.Under the optimum conditions,the linear concentration ranges were between 0.0007 and 2.0 μg/mL for clomipramine hydrochloride with a correlation coefficient of 0.9995,and between 0.001 and 2.0μg/mL for norclomipramine hydrochloride with a correlation coefficient of 0.9983.Detection limits of clomipramine hydrochloride and norclomipramine hydrochloride were 0.23 and 0.33 ng/mL(S/N=3),respectively.The recoveries of clomipramine hydrochloride and norclomipramine hydrochloride were 98.60%and 98.80%,respectively.The RSDs of intra-and inter-assay of clomipramine hydrochloride were 3.2%and 3.8%,respectively.The RSDs of intra-and inter-assay of norclomipramine hydrochloride were 2.7%and 4.0%,respectively.3.In this experiment,a new composite rGO-CuS was synthesized by one-pot hydrothermal method.Based on the enhancement of Ru(bpy)32+ by rGO-CuS modified electrode,the developed new method was used to determine amlodipine besylate(AML)using capillary electrophoresis-chemical modified electrode-electrochemiluminescence.The CE-ECL parameters that affect separation and detection were optimized.Under the optimum conditions,the linear ranges were between 0.008 and 5.0 μg/mL for amlodipine besylate with a correlation coefficient of 0.9988,and the detection limit was 2.8 ng/mL(S/N=3).Compared with the use of bare platinum as working electrode,the sensitivity of amlodipine besylate was significantly improved by using chemical modified electrode and the method has been successfully applied in the detection of amlodipine besylate in mice plasma.The recoveries was from 95.42%to 98.50%and RSD was 3.2%.The thesis consists of four chapters.Chapter 1 is the introduction,Chapter 2 for study on the interaction between amlodipine besylate and human serum albumin by ultrasonic microdialysis coupled with capillary electrophoresis electrochemiluminescence,Chapter 3 for simultaneous determination of clomipramine hydrochloride and its metabolite norclomipramine hydrochloride in urine samples using dispersive liquid-liquid microextraction followed by capillary electrophoresis electrochemiluminescence,Chapter 4 for an ultrasensitive electrochemiluminescence sensor based on rGO-CuS composite coupled with capillary electrophoresis for determination of amlodipine besylate in mice plasma.The chapters from 2 to 4 are my research work.