Abstract

A comparative study of two post-column derivatization techniques for amino acid determination has been completed. Using conventional high-performance liquid chromatography, amino acids were separated on either sodium- or lithium-form, polystyrene-based, strong cation exchangers, then derivatized with either o-phthalaldehyde or ninhydrin. Amino acid detection limits with o-phthalaldehyde are near 5 pmoles for primary amines and 100 pmoles for secondary amines such as proline. Detection limits with ninhydrin are near 100 pmoles. The o-phthalaldehyde system should be the system chosen whenever detection of secondary amino acids is unimportant. Retention time reproducibility averaged 1–4% relative standard deviation for both systems with a median peak area relative standard deviation of 3%. A substantial benefit of amino acid determination by high-performance liquid chromatography is the ease with which one can apply this type of chromatography to solve other separation problems.

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