| Recent years, traditional enzymatic analysis based on isolated and purified enzyme has increasingly underwent the challenge from the novel bioactive catalysis materials including microorganism, animal and plant organ or tissue, cell and receptor, especially from animal and plant tissue. The advantages of more stable catalytic activity, longer lifetime, easy for materials getting and low cost make the kind of tissue enzyme catalysis-based biosensor or analytical system more and more attractive. Research on it thereby has become a hotspot in the study field of enzymatic analysis and biosensor. Bioluminescent microorganisms have been playing an important role in the fields of food safe inspection, environmental monitoring and the others. The finding and study on novel bioluminescent microorganisms and construction of new reagentless bioluminescence biosensors is one of the main tasks for the researchers in the research field. Attributing to the excellent characteristics of high sensitivity, wide linear range, rapidity, simple instrument and suitable for automatization and continuous analysis, chemiluminescence have attracted many people's attention and have been successfully applied for the determination of various important inorganic and organic compounds in the fields of biotechnology, pharmacology, molecule biology, clinic medicine and environmental monitoring.The present study focused on three aspects. The first is to on-line couple tissue enzymatic catalysis with spectrophotometric, fluorophotometric and chemiluminescent analysis systems and established serials of tissue enzymatic catalysis-based optical analytical systems. Thesecond involved in the finding and study on a novel and native bioluminescent fungi-Armillariaella mellea (AM). Based on the investigations on its bioluminescent behavior in gas and aqueous phase, a very simple and reagentless biosensor for gas oxygen and dissolved oxygen determination was developed. In the last aspect, some new chemiluminescence analytical systems and methods were proposed and the feasibility of coupling it to on-line microdialysis or microultrafiltration sampling for the study of drug-protein interaction and applying it for real sample analysis was examined.Chapter 1 Tissue enzymatic catalysis-based optical analytical systemsIn this chapter, correlative study advancement on tissue enzyme catalytic analysis was summarized briefly in Section 1.1 at first.In Section 1.2, a novel, simple, inexpensive and reagentless mushroom tissue-based spectrophotometric analytical system for the determination of catechol and catecholamines (isoprenaline and dopamine) combined with flow injection analysis is proposed. The mushroom tissue acts as the molecular recognition element. Catecholamines are oxidized by oxygen under the catalysis of polyphenol oxidase (PPO) in the tissue column to produce quinone. The concentrations of catecholamines are determined by monitoring the obtained products at their maximum absorption. The linear range of catechol concentration is 2xlO"6to 1 10-3 g ml-1 with the detection limit of 4 10-7 g ml-1 (3). The response to the concentration of isoprenaline is linear in the range of 4xlO"6~8*10"4 g ml"1 with a detection limit of 1.3x10"6 g mr'(3a). It is linear with the concentration of dopamine in the range 4xlO"6-8*10"4 g ml"1 with a detection limit of l.OxlO"6 g ml"1 (3a). The system has been successfully applied to the determination of isoprenaline and dopamine in pharmaceutical preparation. The obtained results are well in agreement with the values determined by the standard methods in pharmacopoeia.In Section 1.3, a novel simple and reagentless mushroom tissue-based fluorescence system for the determination of isoprenaline over the range of 3x10"8 -IxlO"5 g ml'1 combined with flow injection analysis is described. Mushroom tissue exhibits the rich activity of polyphenol oxidase (PPO), which was employed as molecular recognition elements. Isoprenaline was oxidized under the catalysis of PPO in tissue column to produce isoproteroquinone...
|
PDF Full Text Request