Study On Analysis Of Pharmaceutical, Food And DNA Hybridization Using Flow Injection-Chemiluminescence

Chemiluminescence (CL) is defined as the emission of electromagenetic radiation produced by a chemical reaction. The reaction can take place in gas, liguid, and solid phase. CL analysis is characterized by sensitivity, simplicity and low-cost. It requires no light sources and thus voids the effects of stray light and the instability of light source. Most CL reactions can finish in several seconds and are prone to the environmental effect. Thus, the use of CL assay in quantitative analysis is limited.The Flow injection analysis is a solution treatment method in non equational state. Its combination with Chemiluminescence provides high efficiency and accuracy. Flow injection chemiluminescence becomes more and more important in many fields.The emergence of nanotechnology is opening new horizon for the application of nanoparticles in analytical chemistry. The unique physical and chemical properties of nanoparticles offer excellent prospects for chemical and biological sensor. Coupling such unique properties and the high sensitivity offers great potential for the application of CL analysis.In this dissertation, we coupled the flow injection analysis with sensitive chemiluminescence to research the application of CL in pharmaceutic analysis, food analysis and the specific sequence DNA analysis. The dissertation is composed of four chapters as followings:Chapter 1: Introduced the main CL principle, often used reaction system, the application of CL in the analysis of prote(?) and DNA, food and medicine, and some coupling techniques with CL, including flow-injection analysis, high performance liquid chromatography, capillary electrophoresis and Micro Total Analysis Systems. At last pointed out the p(?)pose and meaning of the dissertation.Chapter 2: Flow injection-chemiluminescence determination of sulfadiazine compound naristillaeA simple, sensitive and selective flow injection-chemiluminescence method for the determination of sulfadiazine in compound naristillae has been investigated. It is based upon the chemilimunescence reaction of sulfadiazine, formaldehyde and potassium permanganate in polyphosphate acid medium. The optimum conditions for the chemiluminescence emission were investigated. Under the optimum conditions, the linear range for the determination of sulfadiazine was 8.0×10^-7 to 2.0×10^-4 mol/L with a detection limit of 2.0×10^-7mol/L calculated as proposed by IUPAC and a relative standard deviation of 2.53% for 11 solutions of 5.0×10^-5 mol/L sulfadiazine on the same day. It was also found that the coexisting ephedrine hydrochloride did not interfere with this determination. This led to the successful application of the proposed method for the direct and selective determination of sulfadiazine in compound naristillae.Chapter 3: Determination of Resveratrol in Red Wine by Solid Phase Extraction-Flow Injection Chemiluminescence MethodA sensitive flow injection chemiluminescence method has been developed for the detection of resveratrol in red wine based on the fact that resveratrol can greatly enhance chemiluminescence reaction between KMnO₄ and HCHO in sulfuric acid medium. Analyte were preconcentrated on solid sorbent (C₁₈ solid-phase extraction cartridge). Under the optimum conditions, the proposed method allows the measurement of resveratrol over the range of 1.32×10^-8 to 1.32×10^-5 mol/L with a detection limit of 3.30×10^-9mol/L, and the relative standard deviation for 1.32×10^-5 mol/L resveratrol (n = 11) is 3.8%. This method has been successfully applied for the determination of the resveratrol in red wine. Furthermore, the possible reaction mechanism was also discussed.Chapter 4: DNA Hybridization at Magnetic Nanoparticles with Flow injection chemiluminescent DetectionA DNA optical sensor system for DNA hybridization assay has been developed. The method involves magnetic nanoparticles for ssDNA immobilization and copper sulfide nanoparticle as oligonucleotide label and Flow injection chemiluminescence (FI-CL) for assay detection. The magnetic nanoparticles have the advantages of easy preparation, easy surface modification and low cost. The ssDNA with the amino group at the 5' end was covalently immobilized to the carboxyl-terminated magnetic beads in the presence of l-ethyl-3-(3-dimeth-ylaminopropyl)carbodiimide (EDAC). The copper sulfide (CuS) nanoparticle-labeled oligonucleotides probe was used to identify the ssDNA immobilized on the magnetic nanoparticles based on a specific hybridization reaction. After being anchored on the hybrids, copper sulfide nanoparticles are dissolved to Cu²⁺ in HNO₃ solution and sensitively determined based on the catalyze reaction of Cu²⁺- alkaline luminol-H₂O₂ in FI-CL system. The proposed system coupled the sensitive CL method, effective magnetic separation for eliminating nonspecific adsorption effects with the abundant Cu²⁺ released from each hybrid and allows the detection of specific sequence DNA targets at levels as low as 0.02pmol. Meanwhile, it offers great promise for DNA hybridization analysis.