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

被引：23

作者：

Devall, A. J. ^{[1
]}

Blake, R. ^{[1
]}

Langman, N. ^{[1
]}

Smith, C. G. S. ^{[1
]}

Richards, D. A. ^{[1
]}

Whitehead, K. J. ^{[1
]}

机构：

[1] Univ Birmingham, Sch Med, Div Neurosci, Dept Pharmacol,Pain Signalling Grp, Birmingham B15 2TT, W Midlands, England

来源：

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2007年 / 848卷 / 02期

关键词：

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

D O I：

10.1016/j.jchromb.2006.10.049

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 mu M 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. (c) 2006 Elsevier B.V. All rights reserved.

引用

页码：323 / 328

页数：6

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