Studies On The Determination Methods Of Anticoagulation Warfarin And The Interaction Between Warfarin And Bovine Serum Albumin By Fluorescence Spectrometry

Warfarin is the prevalent oral anticoagulation that taken by patients following mechanical heart valve replacement avoiding thromboembolism. Once the anticoagulation monitoring is improper, serious clinical complications will occur. So it is crucial for clinical anticoagulation monitoring during the course of anticoagulation following mechanical heart valve replacement. And it is the significance tache to predominate anticoagulation therapy accurately for decrease clinical complications.The dissertation includes six parts.Chapter 1: ReviewResearches of determining warfarin in late years both here and abroad were reviewed in this part. And the contents of the dissertation were proposed.Chapter 2: Fluorescence Analysis of Anticoagulant Warfarin in the Medium of Aqueous-EthanolA new analytical method was proposed for determining warfarin in the medium of aqueous-ethanol, the conditions of determination were discussed. The fluorescence intensity was linearly related to warfarin sodium concentration in the range of 0.3～7.1μg·mL^-1, and the detection limit was 0.1μg·mL^-1. The method was applied to the determination of warfarin sodium in warfarin sodium tablets with satisfactory. Recoveries were 96.7 104.9%.Chapter 3: Determination of Warfarin with Fluorescence Spectrometry Usingβ-Cyclodextrin as the Reinforcing AgentBased onβ-cyclodextrin can enhance the fluorescence intensity of warfarin, a new fluorescence analysis method was developed in this paper. Under the optimized experiment conditions, the fluorescence intensity of warfarin sodium was linearly related to the concentration in the range of 0.1～10.2μg·mL^-1, and the detection limit was 0.04μg·mL^-1. The method had been applied to the determination of warfarin sodium in medicine tablets. Recoveries were 96.7～102.4%.Chapter 4: The Inclusion Interaction between Anticoagulant Warfarin andβ-CyclodextrinThe inclusion interaction between warfarin andβ-cyclodextrin was investigated by fluorescence spectrometry. The results shown that the ratio of inclusion complex between warfarin andβ-cyclodextrin was 1:1, which was measured by Benesi - Hildebrand equation improved upon direct fluorescence analytical method. And the corresponding inclusion constant was 610 L·mol^-1.Chapter 5: Determining Warfarin by High Performance Capillary ElectrophoresisThe different conditions of determining warfarin by high performance capillary electrophoresis were studied. The results shown that the detecting wavelength of warfarin was 210 run; buffer solution was phosphate of 15 mmol·L^-1 (pH 9.18); and the load voltage was 9 kV. Under the optimized experiment conditions, the linear range of warfarin was 0.3～15.4μg·mL^-1, and the detection limit was 0.1μg·mL^-1 .Chapter 6: Study on the Interaction between Warfarin and Bovine Serum Albumin by Ultraviolet and Fluorescence SpectrometryThe interaction between warfarin and bovine serum albumin under the physiology conditions was investigated by using ultraviolet and fluorescence spectrometry. The quenching mechanism of bovine serum albumin by warfarin was discussed via the effect of temperature on quenching constant and the change on ultraviolet spectrum of bovine serum albumin before and after adding warfarin. The results shown that the quenching of bovine serum albumin by warfarin was a dynamic quenching, and the quenching constants were 6.05×10⁴ L·mol^-1 (16℃) and 6.14×10⁴ L·mol^-1 ( 37℃). Based on the theory of Forster energy transfer, the energy transfer efficiency and the distance between warfarin and bovine serum albumin were 0.37 and 3.15 nm, respectively.