P-38 Towards Amino Acid Analysis of Flame-treated Avian Eggshell Using RP-HPLC

We developed a method for the Agilent 1260 Infinity HPLC system to quantitatively study the extent of amino acid racemization in flame-treated ostrich (S. camelus) and emu (D. novaehollandiae) eggshell fragments. We ground the samples with a mortar and pestle, dissolved them in 7 M HCl, and heated them inside an oven at 110°C for 6 hours to isolate the eggshell proteins from the surrounding calcite and hydrolyze the peptide bonds. We then evaporated the samples and rehydrated them with an internal standard. For the analysis, we employed reversed-phase HPLC with pre-column derivatization using an OPA/IBLC reagent. We were able to analyze several amino acids while reducing the elution time to 30 minutes