Date of Award
Chemistry & Biochemistry
Three-dimensional fluorometric scans of OPA-IBLC derivatized glutamic acid and histidine solutions revealed that decreasing concentration maintained maximum λem but produced different maximum λex. In pH 0.4 M sodium borate buffer (pH 10.4), 50 μM derivatized amino acid solutions had maxima around 350- to 450-nm λex-λem where 12.5 μM solutions had maxima around 230- to 450-nm λex- λem. In the lower concentration, the λex peak at 230 nm was about 80% higher than 350 nm. Thus, collecting spectra with 230 nm λex would provide better LOD and LOQ. Results inform HPLC standard concentrations since differential quenching effects may produce non-linear calibration curves.
Jacobs, Aaron, "Finding Optimal OPA-IBLC Derivatized Amino Acid Analysis Parameters with Three-Dimensional Fluorometric Scans for HPLC Method Validation" (2021). Honors Theses. 249.
High performance liquid chromatography; Amino acids
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