| Chemiluminescence（CL）method is a trace analysis method that determinessubstance content based on the light radiation generated by chemical reactions. It hasmany advantages such as high sensitivity, wide linear range, rapid analysis, simpleequipment, easy operation, and so on. Currently, it has been widely used in the traceanalysis of inorganic substance, organic substance, medical drug residues, biologicalmacromolecules and biological active natural substances.Many studies have showed that oxidative DNA damage is closely related withthe development of tumor.8-hydroxy-2’-deoxyguanosine (8-OHdG), the majorproduct of oxidative DNA damage, can be used as a biomarker to investigate themechanism of the development of tumor induced by oxidative DNA damage.A new method of chemiluminescence analysis was established for thedetermination of8-OHdG in this study. Combining with the sample pretreatmentmethods that used commercial column solid phase extraction and8-OHdG imprintedpolymers solid phase extraction, the8-OHdG content in the urine of common humanand breast cancer human were determined and compared. Further, based on thecapture ability of antioxidant activity of natural products in vitro, achemiluminescence method was developed for study the DNA protection byprocyanidins extracted from camellia shell.The thesis includes the following five parts:Chapter1: Introduction. The first part of preface introduces the history, the basictheory and the characteristics of chemiluminescence. Then, it gave a generalintroduction of the detection methods for8-OHdG and the study method for theantioxidant activity of proanthocyanidin. Based on the above, some existed problemsin the current study were briefly analyzed, and the significance of the present studywas presented. Finally, the purposes and contents of this research were raised.Chapter2: A chemiluminescence method was developed for determination of8-OHdG for its characteristics of enhancement the light signal of Acid phenanthroline(Phen)-Cu chemiluminescence system. Under the optimum conditions, the linearityrange of this method is from1.67μg/L to333.33μg/L, the work line isΔI=23.96c(μg/L)+860.85(ΔI is the relative chemiluminescence intensity, c is theconcentration of8-OHdG) with the correlation coefficient of0.9995, the detection limit (S/N=3) is0.83μg/L.Chapter3: A SPE-Chemiluminescence method was developed for thedetermination of8-OHdG in human urine samples. The urine samples were pretreatedby C18/OH solid extraction column and then was analysed by Chemiluminescencemethod. The urine samples of a large number of breast cancer patients and normalwere both prepared by SPE and following analysied by Chemiluminescence. Theconcentrations of8-OHdG determined in normal urine and breast cancer patientsurine were5.77±1.56and67.60±27.99μg/L, respectively. The results showed that theconcentrations of8-OHdG in breast cancer patients urine was10times higher thanthat in normal one. The results of the determination are consistent with the reported.Chapter4: Molecularly imprinted polymers(MIPs) were synthesized inacetonitrile and water systems by bulk polymerization. The method used the4-vinylpyridine and acrylamide as the functional monomer,N,N’-Methylenebisacrylamide as the cross-linker, guanosine and deoxyguanosine(dG), which has the similar structures with8-OHdG, as the template molecule and2,2’-azobis-isobutyronitrile as the initiator. The MIP was then applied to separate andenrich8-OHdG in urine samples in solid extraction. The recoveries of the extractionmethod was86.8%-105.9%and the relative standard derivation (R.S.D.) of themethod was3.0%-3.5%. Compared with the commercial C18and C18/OH solid phaseextraction column, molecularly imprinted polymer as the stationary phase was betterfor pretreatment of the human urine which analyzed by chemiluminescence method orHPLC..Chapter5: A chemiluminescence method was established for study of theantioxidant activity and DNA protection by procyanidins extracted from teaoil shell(TSPC) based on the scavenging abilities of TSPC to reactive oxygen(ROS)in vitro.Hydroxide radical scavenging ability of TSPC was determined by Thechemiluminescence system of CuSO4-Phen-Vc-H2O2-DNA was used in this study,and the addition of the TSPC has great inhibitory effect on the chemiluminescenceintensity. |
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