High Pressure Liquid Chromatography (HPLC) Method Development: Determine Multiple Amino Acids Detection Limits.
L-amino acids are the building blocks of proteins in eukaryotic organisms; these amino acids will undergo racemization over time to reach equilibrium with their D counterpart. We would like to apply this information and using reversed-phase High Pressure Liquid Chromatography (HPLC), analyze protein degradation within samples of avian eggshells by observing the racemization of amino acids from eggshells that have been treated under various conditions to replicate possible environmental effects. We continue efforts to develop a robust HPLC methodology to apply to different samples provided by our collaborators. To date we have determined a solvent system and information about detection limits specifically with fluorescence detection. We have a method for sample preparation from the calcium carbonate matrix of the eggshell that will need to be validated with our experimental HPLC methods. We plan to add a chiral column method and ultraviolet detection.