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

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2007-04, Vol.848 (2), p.323-328
• Main Authors: Devall, A.J., Blake, R., Langman, N., Smith, C.G.S., Richards, D.A., Whitehead, K.J.
• Format: Article
• Language: English
• Subjects: 3-Mercaptopropionic Acid - chemistry; Amino acid; Amino Acids - analysis; Amino Acids - cerebrospinal fluid; Amino Acids - chemistry; Analysis; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Dialysis Solutions - analysis; Dialysis Solutions - chemistry; Fundamental and applied biological sciences. Psychology; General pharmacology; Medical sciences; Microdialysis; Molecular Structure; Monolithic; o-Phthalaldehyde - chemistry; Ortho-phthaldialdehyde; Pharmacology. Drug treatments; Rats; Reproducibility of Results; Reversed-phase liquid chromatography
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2006.10.049

The development of a HPLC method using a monolithic C 18 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 ( r 2) for each amino acid was found to be greater than 0.99 up to a 10 μM 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.