Study On The Separation Of Amino Acids And Prediction Of Bioactivity And Toxicity Parameters Of Drugs By Micellar Liquid Chromatography

Micellar liquid chromatography mode is formed by adding surfactant above its critical micelle concentrations to the traditional organic solvent-water reverse phase chromatography. There are many advantages in micellar liquid chromatography including that little consumption of organic solvent, high selectivity, the ability to effectively simulate biological membrane, biological samples direct injection without treatment and no need for column re-balance in gradient elution. Therefore, it is widely used in pharmaceutical analysis. Micellar liquid chromatography has the potential to solve the problems of high organic modifier consumption, unreasonable analysis time, and the case where satisfactory separation could not been obtained by other methods, which is also the situation that amino acids analysis faces. Therefore, this thesis applied micellar liquid chromatography to the separation of amino acids. In addition, we established a mixed micellar system for quantitative retention-activity relationships studies based on the ability of micellar liquid chromatography to simulating biological membrane. The main research works are as follows:1) A method for rapid separation of free amino acids using gradient elution from micellar liquid chromatography to high submicellar liquid chromatography conditions was established and applied successfully to the determination of amino acids in Compound Amino Acid Injection. 15 amino acids could be separated by baseline within 30 min. The developed method was rapid in both ways of shorter column re-equilibration time and shorter elution time compared with conventional HPLC and isocratic MLC, respectively. We investigated the influence effects including the types and concentrations of surfactants, the types and content of organic solvents and the p H value on the retention behavior of the derived amino acids. Finally, the method is simple, rapid and only short time for column re-balance, providing an efficient method for the separation of amino acids.2) A method for the separation of amino acids in medicines by mixed anionic and nonionic micellar chromatography was established, and was also applied to the analysis of Amino Acid Injection. The effect of different factors on the chromatographic behavior of the compounds was studied systematically. Besides, possible separation mechanism was discussed. In addition, chromatographic peak performance was evaluated by the parameters of the half-peak width, which indicated that the hybrid system has the potential to make weak acidic compounds peaks show good symmetry. The method has solved the problems of high consumption of organic solvents and complex gradient elution program as an alternative way for amino acids analysis.3) A mixed anionic and nonionic biopartitioning micellar chromatography model was established for the prediction of the physiological activity parameters of fourteen drugs. Micellar liquid chromatography can simulate biological membranes, and meanwhile, anionic surfactant added to the single nonionic system will better simulate the resting membrane potential. Based on these two points, we established a quantitative retention-activity relationships model for prediction physiological parameters of drugs by fitting the relationship between the retention factors and the corresponding activity and toxicity parameters. The predictive ability of the model was evaluated. It also indicated that this hybrid system is better to simulate biofilm.