Monolithic column-based reversed-phase liquid chromatography separation for amino acid assay in microdialysates and cerebral spinal fluid.

Adam Devall, R Blake, N Langman, CG Smith, Douglas Richards, Kevin Whitehead

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The development of a HPLC method using a monolithic C18 column is described using fluorescence detection for the assay of 21 amino acids and related substances with derivatisation using ortho-phthaldialdehyde (OPA) in the presence of 3-mercaptopropionic acid (3-MPA). The method employs a tertiary gradient and has a run time of 24 min. Linearity (r2) for each amino acid was found to be greater than 0.99 up to a 10 microM concentration; reproducibility across all analyses (relative standard deviation (R.S.D.)) was between 0.97 and 6.7% and limit of detection (LOD) between 30 and 300 fmol on column. This method has been applied to the analysis of amino acids in both spinal microdialysis and cerebral spinal fluid samples.
Original languageEnglish
Pages (from-to)323-8
Number of pages6
JournalJournal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Volume848
Issue number2
DOIs
Publication statusPublished - 1 Apr 2007

Keywords

  • monolithic
  • amino acid
  • Ortho-phthaldialdehyde
  • reversed-phase liquid chromatography

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