Computer-assisted optimization of HPLC with post-column reaction for the determination of amino-acids

Abstract Computer-assisted optimization of a high-performance liquid chromatograph and associated post-column reactor is reported for the determination of six amino-acids. First six, then seven, experimental variables were considered. Non-standard experimental conditions were found which gave significantly improved colour development in the ninhydrin reaction. The composite modified simplex method for experimental optimization and a novel system-response function facilitated rapid and simultaneous improvement of separation, sensitivity and analysis time. The approach described is directly applicable to many similar systems.