Date of Award
Bachelor of Science
In partition chromatography a solid is used to hold a film of liquid and the distribution of a substance is between the liquid phase and another immiscible liquid phase. Because most substances have slightly different distribution coefficient between two immiscible solvents, a series of successive extractions will cause a separation. This is the predominant factor in the paper chromatography. The paper itself is an inert support holding a stationary aqueous phase. As the immiscible solvent flows past an area containing the solute, by capillary flow, partition occurs. When the immiscible solvent reaches an area not containing any solute, partition again occurs resulting in a transfer of the solute, partition again occurs resulting in a transfer of the solute from its point of application to a point some distance down the paper. This method of separation, which has been developed to cover the separations of many organic and biological compounds, has some important limitations concerned with the fast separation and quantitative analysis of the separated products. The flow rate of the organic developing solvent is slow (2-3 sm./ hr.) and the organic developing solvent is commonly phenol, which presents problems in its use because of its corrosive nature. Also the amount of solute which can be separated per unit area of the paper is extremely small. This amount is usually in the order of 1 to 10 microliters of a 0.01 molar solution of the solute. Amounts of this size require special apparatus for the measuring and application of the solution. Because the amount of solute is so small, experimental error in its analysis is increased. The purpose of this project was to develop a method of analyzing amino acids quantitatively that could be performed by an undergraduate in introductory courses in quantitative analysis. The formost thought in developing the procedure was to keep it as simple and straight forward as possible. In connection with this was to use only apparatus which was easily obtained and simple to use. The use of ion exchange papers seemed to offer the best solution. They were proven effective in the separation of amino acids of the basic type (1) (4) and allowed a separation to be performed in a length of a lab period. The work done outlines a method of analysis and separation which could be performed in two lab periods with reproducible results.
Coutant, John Ennis, "Rapid quantitative analysis of basic amino acids by ion exchange resin paper chromatography" (1966). Honors Theses. 1783.