Analysis of optically pure .BETA.-phenylalanine produced by penicillin G acylase through HPLC

Feasible HPLC methods were developed to measure some compounds in reaction of optically pure β-phenylalanine (BPA) produced by penicillin G acylase. Four substances,i.e. phenylacetamide,BPA,N-phenylacetyl-BPA and phenylacetic acid could be detected effectively using Zorbax XDB-C18 column under the following elution conditions:mobile phase was 7mmol/L phosphate (pH 3.0),containing acetonitrile 35% (v/v) and SDS 0.68g/L,1mL/min flow rate with detection wavelength 210nm. When all their concentrations were under 0.2mg/mL,contents of four substances had good linear correlation with their own peak area. Enantiomers of two chiral compounds,i.e. N-phenylacetyl-BPA and BPA could be detected under the optimum elution conditions using Chirobiotic T and Chirobiotic R columns,respectively.