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Enhanced detection of porphyrins by capillary electrophoresis‐laser induced fluorescence
Author(s) -
Melanson Jeremy E.,
Lucy Charles A.
Publication year - 2002
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200206)23:11<1689::aid-elps1689>3.0.co;2-l
Subject(s) - chemistry , capillary electrophoresis , repeatability , chromatography , laser induced fluorescence , fluorescence , sodium dodecyl sulfate , detection limit , electrophoresis , dilution , analytical chemistry (journal) , optics , physics , thermodynamics
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 m M 2‐( N ‐cyclohexylamino)ethanesulfonic acid (CHES, pH 10) – 75 m M 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 p M (2×10 –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.