Abstract
The o-phthalaldehyde method for pre-column derivatization of amino acids for compositional analysis has been optimized for quantitation at the low picomole level. As little as 20 pmol of peptide are hydrolyzed and 6 pmol can be derivatized for analysis. The method results in quantitative recovery of the amino acids and gives reliable quantitation even at the 6 pmol level. The study details the procedures involved in hydrolyzing small amounts of peptides. The effects of purity and stability of the o-phthalaldehyde reagent, pH, and composition of the mobile phase are discussed with a view to adaptation for use with various high-performance liquid chromatographic systems. Optimization of the derivatization conditions and strategies for lengthening column life are described. The relative performances of different reversed-phase columns from two different manufacturers are compared.
Published Version
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