| This thesis includes two sections.In first section,the informations such as history,principles and equipment about capillary electrophoresis with laser-induced fluorescence and Capillary clectrophoresis with chemiluminescence detection are reviewed.In the research report section,four applied reports about CE-CL for determination of carcinoembryonic antigen,interferon alpha, prolactin,human choriogonadotropin were reported respectively.Capillary elcctrophoresis(CE) emerged in the early 80s as a highly efficient means of separation for the analysis of complex mixtures in extremely small samples with outstanding resolving power.The relatively short separation times,simple instrumentation and low operational costs make it an intensive exploitation.Currently,UV-absorbance detectors are employed in most commercially available CE instruments owing to their broad applicability and relatively low cost. However,the optical path lengths are limited by the capillary column internal diameter to usually less than 100pm,which in turn seriously limits the sensitivity.Laser-induced fluorescence(LIF) detectors for CE have enjoyed increasing popularity owing to its high sensitivity and low detection limit.However,apart from the considerable extra investment of costly equipment,additional efforts are also required for introducing fluorophores to nonfluorescent analytes through pre or post column derivation.The sensitivity of electrochemical detectors for CE is less dependent on the sample volume,and highly sensitive detection could be achieved,but the contamination of electrode surfaces might also pose problems in the analysis of real samples.Chemiluminescence analysis is a trace analysis method which determines the content of substance according to the light emission of the chemical reaction.In 1877,Radziszeuski found that lophine emitted green light when it reacted with oxygen in the presence of a base.This is the first example of chemiluminescence using a synthetic organic compound.Since then,a number of chemiluminescent compounds have synthesized and studied for their chemiluminescent properties. Chemiluminescence analysis has high sensitivity(Detection limit can be 10-12—10-18mol/L),wide linear range(three to six magnitude order),low background and simpler,lower instruments which is suited to post-column detection for very low-volume sample in CE.1 A homogeneous immunoassay for determination of carcinoembryonic antigen in human serum using capillary electrophoresis with chemiluminescence detectionA sensitive and rapid homogeneous immunoassay based on capillary electrophoresis(CE) with enhanced chemiluminescence(CL) detection has been developed for the determination of carcinoembryonic antigen(CEA) in human serum.The chemiluminescence reaction of luminal and hydrogen catalyzed by HRP can be enhanced by Sodium tetraphenylboron,Under the optimal conditions,the linear range was form 2.0 to 80.0μg/L(R=0.9921),and the detection limit was 0.1μg/L.In this study,the free HRP-labeled CEA(HRP-CEA) and the bound HRP-labeled CEA(HRP-CEA-Ab) were separated in the separation capillary within 12 min using a borate run buffer.This method has been used for assaying CEA in human serum using a noncompetitive format.2 A homogeneous immunoassay for determination of interferon alpha in human serum using capillary electrophoresis with chemiluminescence detectionInterferon alpha(α-IFN) in human serum studied in rabbit by capillary electrophoresis(CE) with enhanced chemiluminescence(CL) technique in this paper.This method has been used for assayingα-IFN in human serum using a noncompetitive format.After optimize separation conditions. the free HRP-labeledα-IFN(HRP-α-IFN) and the bound HRP-labeledα-IFN(HRP-α-IFN-Ab) were separated in the separation capillary within 8 min using a borate run buffer.the linear range was form14.5 to 312 pg/ml(R=0.9939),and the detection limit was 7.25 pg/ml.The results confirm that the method simple,fast and accurate for the measurements ofα-IFN in human serum.3 A homogeneous immunoassay for determination of prolactin in human serum using capillary electrophoresis with chemiluminescence detectionA rapid homogeneous immunoassay based on capillary electrophoresis(CE) with enhanced chemiluminescence(CL) detection has been developed for the determination of prolactin(PRL) in human serum.The chemiluminescence reaction of luminal and hydrogen catalyzed by HRP can be enhanced by Sodium tetraphenylboron.The separation was carried out on a 50cm×75μm i.d. fused-silica capillary.,the applied voltage was 15 KV,injection time was 10 s and all separations were performed in 50mmol/L borate(pH 9.5) buffer.Under the optimal conditions,the free HRP-labeled PRL(HRP-PRL) and the bound HRP-labeled PRL(HRP-PRL-Ab) were separated in the separation capillary within 6 min.the linear range was form 16 to 1600μIU/ml(R=0.9918),and the detection limit was 2.3μIU/ml.The assay also provided good reproducibility and accuracy.This method can be applied successfully to determine PRL in human serum analysis.4 A homogeneous immunoassay for determination of human choriogonadotropin in human serum using capillary electrophoresis with chemiluminescence detectionThe aim of this study to develop a rapid and sensitive homogeneous immunoassay method for human choriogonadotropin(β-HCG) in human serum using capillary electrophoresis(CE) coupled with enhanced chemiluminescence(CL) detection.The chemiluminescence reaction of luminal and hydrogen catalyzed by HRP can be enhanced by Sodium tetraphenylboron,Under the optimal conditions,the free HRP-labeledβ-HCG(HRP-β-HCG) and the bound HRP-labeledβ-HCG(HRP-β-HCG-Ab) were separated in the separation capillary and be detected,the linear range was form 0.5 to 20 mIU/ml(R=0.9937),and the detection limit was 0.1mIU/ml.All separation could be achieved within 11 min.This method has been successfully detected theβ-HCG in human serum.
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