Single-step analysis of low abundance phosphoamino acids via on-line sample preconcentration with chemical derivatization by capillary electrophoresis.
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 approximately 3) of 0.1 microM. 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.
Ptolemy
Adam S
AS
Department of Chemistry, McMaster University, Hamilton, ON, L8S 4M1, Canada.
Britz-McKibbin
Philip
P
D000818:Animals; D019075:Electrophoresis, Capillary; D005449:Fluorenes / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D009862:Online Systems; D019606:Phosphoamino Acids / Q000032:analysis* / Q000737:chemistry; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
C496789:9-fluorenylmethoxycarbonyl; D005449:Fluorenes; D019606:Phosphoamino Acids