Optimization of separation and detection of 6-aminoquinolyl derivatives of amino acids by using reversed-phase liquid chromatography with on line UV, fluorescence and electrochemical detection

The combined use of UV-absorbance, fluorescence and electrochemical detection was proposed for the analysis of a set of thirteen amino acids by reversed-phase liquid chromatography (RP-HPLC) using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate as a precolumn derivatization reagent. The utility of using three detectors in series was demonstrated. The separation of all derivatized amino acids was optimized with the aid of a computer optimization program from only four simple linear gradient measurements. The effectiveness of a reliable retention prediction of solutes under any gradient profile using other gradient or isocratic data was also examined.