The Use of a 3-Factor Statistical Mixiure Design and a 2-Factor Chromatograhic Response Function to the RP-HPLC Separation of Several Free α-Amino Acids

A 3-factor statistical mixture design and a 2-factor chromatographic response function were used in the solvent optimization of the RP-HPLC separation of a mixture containing free histidine, methionine, tyrosine, phenylalanine and tryptophan. The influences of phosphoric acid, methanol and acetonitrile on the separation were compared. A solvent condition resulting in a satisfactory separation of the amino acid mixture was predicted.