Enhanced detection of porphyrins by capillary electrophoresis-laser induced fluorescence.
A highly sensitive technique for the analysis of urinary porphyrins using capillary electrophoresis (CE) coupled with laser-induced fluorescence (LIF) detection is reported. Separation of mesoporphyrin IX, coproporphyrin, uroporphyrin and the penta-, hexa- and heptacarboxylic acid porphyrins was achieved in 11 min using a 10 mM 2-(N-cyclohexylamino)ethanesulfonic acid (CHES, pH 10) -75 mM sodium dodecyl sulfate (SDS) buffer. Migration time and peak area repeatability were less than 1 and 5% relative standard deviation (RSD), respectively. Limits of detection of 20 pM (2 x 10(-11) M) were achieved employing the recently introduced Nichia violet diode laser for excitation at 400 nm. This represents an enhancement in sensitivity of over two orders of magnitude compared to previous reports. This high sensitivity for urinary porphyrins was demonstrated through the quantification of coproporphyrin and uroporphyrin in urine samples after up to a 100-fold dilution.
Melanson
Jeremy E
JE
Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada.
Lucy
Charles A
CA
D002138:Calibration; D019075:Electrophoresis, Capillary / Q000379:methods* / Q000592:standards; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D006801:Humans; D007834:Lasers; D011166:Porphyrins / Q000032:analysis / Q000302:isolation & purification* / Q000652:urine; D012015:Reference Standards; D012680:Sensitivity and Specificity
D011166:Porphyrins