Title: Rapid analysis of amino acids by HPLC using pre-column derivatization with PITC.
Abstract: A rapid quantitative analysis was devised for phenylthiocarbamyl (PTC) derivatives of common amino acids with phenylisothiocyanate (PITC)using reversed-phase HPLC. This study was performed with an HPLC system of Waters Associates. The HPLC conditions were: column, NOVA-PAK cartridge C/118, 5×100mm using a Z-module; column temp., 45±1°C; mobile phase, (A) 0.1M ammonium acetate buffer (pH 6.5) containing 5% acetonitrile and (B) 60% acetonitrile in water; gradient conditions, 0% B to 50% B for 5min on a linear gradient and 50% B to 100% B for the next 1min on a linear gradient; flow-rate, 2.0ml/min; detection, UV at 254 nm; analysis time, 6minutes. The retention time had a relative average percent age standard deviation of ±0.39. Thecorrelation coefficients between the peak areas and injected amounts (25 to 500 pmol) were more than 0.999. The coefficients of variation in peak areas were 0.68-1.80.