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Evaluation of conditions for the HPLC determination of plasma amino acids using precolumn derivatization

✍ Scribed by Halfpenny, A. P. ;Brown, Ph. R.

Publisher: John Wiley and Sons
Year: 1985
Tongue: English
Weight: 440 KB
Volume: 8
Category: Article
ISSN: 0935-6304
DOI: 10.1002/jhrc.1240080505

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✦ Synopsis

Abstract

For the determination of free amino acids in plasma, the conditions for precolumn derivatization of the amino acids and the chromatographic separation were examined. The isoindole products, formed by the reaction of the primary amino acids with orthophthalaldehyde (OPA), were readily separated by RPLC and detected spectrofluorometrically using an excitation wave‐length of 300 nm and an emission cut‐off filter of 440 nm. Since the sensitivity of this method permits determination of amino acids in the femtomole range, the analysis can be performed on samples as small as 10 μl of filtered plasma or serum. The separation is achieved in approximately 35 minutes with good precision for the majority of the amino acids.

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