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Issue 9, 2005
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Single-step analysis of low abundance phosphoamino acidsvia on-line sample preconcentration with chemical derivatization by capillary electrophoresis

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Abstract

New strategies for rapid, sensitive and high-throughput analysis of low abundance metabolites in biological samples are required for future metabolomic research. In this report, a direct method for sub-micromolar analyses of phosphoamino acids was developed using on-line sample preconcentration with 9-fluorenylmethyloxycarbonyl chloride (FMOC) derivatization by capillary electrophoresis (CE) and UV detection. Analyte focusing by dynamic pH junction and FMOC labeling efficiency were influenced by several experimental factors including buffer pH, ionic strength, sample injection length and FMOC concentration. About a 200-fold enhancement in concentration sensitivity was achieved under optimal conditions relative to conventional off-line derivatization, as reflected by a detection limit (S/N ≈ 3) of 0.1 µM. In-capillary sample preconcentration with chemical labeling by CE offers a unique single-step analytical platform for high-throughput screening of low abundance metabolites without intrinsic chromophores.

Graphical abstract: Single-step analysis of low abundance phosphoamino acids via on-line sample preconcentration with chemical derivatization by capillary electrophoresis

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Article information


Submitted
01 Apr 2005
Accepted
26 May 2005
First published
20 Jul 2005

Analyst, 2005,130, 1263-1270
Article type
Paper

Single-step analysis of low abundance phosphoamino acids via on-line sample preconcentration with chemical derivatization by capillary electrophoresis

A. S. Ptolemy and P. Britz-McKibbin, Analyst, 2005, 130, 1263
DOI: 10.1039/B504480D

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