Amino acid analysis by HPLC: optimized conditions for chromatography of phenylthiocarbamyl derivatives.
A method for analysis of phenylthiocarbamyl-amino acids by HPLC is described. Solvent A contains 0.05 M sodium acetate plus triethylamine (2.75 ml/liter) adjusted to pH 6.40 with phosphoric acid. Solvent B consists of 50% solvent A, 40% acetonitrile, and 10% methanol. The stationary phase is a 3-micron Spherisorb ODS-2 column (4.6 X 100 mm). A new sample can be injected every 27 min. Variation of the triethylamine concentration from 1 to 4 ml/liter is shown to affect column selectivity in a predictable manner, and this provides a rational basis for optimizing chromatographic conditions. Resolution of certain hydrophilic amino acids (Arg, Thr, Ala, and Pro), and mobile phase stability are improved over previously described procedures.
D000596:Amino Acids / Q000032:analysis*; D001322:Autoanalysis; D002851:Chromatography, High Pressure Liquid; D004529:Egg White / Q000032:analysis; D005021:Ethylamines; D005344:Fibrinopeptide A / Q000032:analysis; D017879:Isothiocyanates; D009113:Muramidase / Q000032:analysis; D013861:Thiocyanates
D000596:Amino Acids; D005021:Ethylamines; D017879:Isothiocyanates; D013861:Thiocyanates; C005441:phenylisothiocyanate; D005344:Fibrinopeptide A; D009113:Muramidase; C016162:triethylamine