Separation of alpha- from beta-arylalanines by nickel nitrilotriacetate chromatography.

ABSTRACT

A method is described to separate alpha- from beta-arylalanines by ligand exchange chromatography on a nickel nitrilotriacetate agarose column with UV monitoring of the effluent. Separate mixtures containing an alpha- and beta-arylalanine pair (1 mg of each) were individually loaded onto the nickel resin pre-equilibrated with the mobile phase at room temperature, and the amino acids were eluted from the column with a gradient from pH 12.0-8.0. The beta-arylalanines eluted first, followed by the alpha-isomers. The four alpha/beta-amino acid pairs tested were well separated with baseline resolution. An aliquot of each fraction was chemically treated to derivatize the amino acids to their N-acyl methyl ester analogs, and their identities were confirmed by GC/MS analysis. The sample recovery was quantitative (>98%), and the column matrix was very resilient, as demonstrated by consistent separation of the solutes after approximately 100 preparative cycles.