HIGH-SPEED RP-HPLC/FL ANALYSIS OF AMINO-ACIDS AFTER AUTOMATED 2-STEP DERIVATIZATION WITH O-PHTHALDIALDEHYDE/3-MERCAPTOPROPIONIC ACID AND 9-FLUORENYLMETHYL CHLOROFORMATE

This paper describes an automated high-speed RP-HPLC/FL method, including two-step precolumn derivatization with o-phthaldialdehyde/3-mercaptopropionic acid (OPA/3-MPA) for primary amino acids and with 9-fluorenylmethyl chloroformate (FMOC-Cl) for secondary amino acids. The OPA/3-MPA derivatives eluted within 6.5 min, while the retention time of the last eluted amino acid derivative (FMOC-Pro) was 8.0 min. The total analysis time, including precolumn derivatization, separation, column washing and reequilibration cycles, was only 18 min. Therefore, 75-80 samples per day can be analysed during unattended operation. The efficiency of the method was demonstrated by the separation of amino acids extracted from potato tubers.