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AbstractThis research validates RP-HPLC fluorimeter and OPA-IBLC derivatization parameters to lower limits of detection (LOD) and quantitation (LOQ). A 3-dimensional fluorometric scan in pH 10.4 sodium borate buffer showed peak OPA-IBLC derivatized histidine and glutamic acid fluorescence occurs at 368- to 445-nm and 336- to 452-nm λ¬ex-λ¬em, respectively. HPLC-FLD response was refined with systematic FLD analysis in pH 4 sodium acetate buffer after column separation. Derivatization reaction completion was ensured by increasing injector mixing time until peak integration area stabilized. Results inform future FLD parameters and determine racemic OPA-IBLC derivatized amino acid ratios in ostrich eggshells.
