| Flow injection (FI) Chemiluminescence(CL) analysis has made great progresses in analytical chemistry for its high sensitivity, wide linear dynamic range, rapidity, simplicity, low cost, and being appropriate for on-line analysis. It has been applied in different fields such as biotechnology, pharmacology, and environmental detection. In recent years, various biosensors based on chemiluminescence have also been developed rapidly. This thesis focused on the applications of FI-CL method in determination of biologic matters, and a novel plant tissue-based chemiluminescence biosensor was also proposed in this paper. The work mainly consists of the following four parts: Part I A novel plant-tissue sensor based on flow injection chemiluminescence for the determination of diamine The biological material pea tissue acted as the molecular recognition element that was packed in a mini-glass column, and diamine was oxidized under the catalysis of DAO to produce hydrogen peroxide, which can react with luminol in the presence of Co2+ or K3Fe(CN)6 to generate a CL signal. The CL emission intensity was linear with Putrescine concentration in the range of 0.5-80 μM; the detection limit was 0.03 μM. A complete analysis could be performed in 1.2 min with a relative standard deviation of 2.01%(n=11). It has also been found that the DAO in pea tissue possessed better selectivity for diamine. The present biosensor was applied successfully to the analysis of diamine in fish samples. Part II Determination of spermine using an inhibitory chemiluminescent flow injection method A highly sensityive flow injection chemiluminescence method has been developed for determination of spermine (Spm). It based on the inhibition of Spm to the chemiluminescence reaction of Luminol-Co2+ -H2O2 in sodium hydroxide medium. This method possesses wider linear range (2×10-9 -8×10-5 mol/L), higher sensitivity (detection limit is 8×10-10mol/L) and better producibility (RSD is 0.61%). The proposed method was used to determine Spm in lung cancer patient's urine with satisfactory results. Part III The study of chemiluminescent characteristics between hydroxyl radical and several alkaloids and applications According to Fenton reaction, the chemiluminesce behavior of several alkaloids was studied. The results showed that, among the five common bases in DNA and RNA, purine was more easily oxidized by hydroxyl radical. A possible chemiluminescent mechanism of the reaction system was discussed. Based on this, the FL-CL methods for the determination of guanine and theophylline were developed. The proposed methods have been used to determine guanine in urine and theophylline in pharmaceutical preparation with satisfactory results. Part IV The study of anti-oxidation properties of ascorbic acid and glutathione using Guanine-Co2+-H2O2 chemiluminescence system The Guanine-Co2+-H2O2 chemiluminescence system was used for the study of anti-oxidation properties of ascorbic acid and glutathione. The results showed that the two substances could scavenge hydroxyl radical and prevent guanine from injuring. This method also supplied an effectively means for the study of DNA anti-injuring mechanisms. |
PDF Full Text Request