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Design and performance of a light‐emitting diode detector compatible with a commercial capillary electrophoresis instrument
Author(s) -
Johns Cameron,
Macka Miroslav,
Haddad Paul R.
Publication year - 2004
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/elps.200405913
Subject(s) - detection limit , detector , absorbance , capillary electrophoresis , analytical chemistry (journal) , light emitting diode , diode , capillary action , materials science , chemistry , optoelectronics , isoelectric focusing , optics , chromatography , physics , biochemistry , composite material , enzyme
Indirect photometric detection in capillary electrophoresis (CE) has been predominantly performed in the UV region, in part due to a lack of suitable high‐intensity and low‐noise light sources in the visible spectral region. A new photometric detector based on light‐emitting diodes (LEDs) as light sources and compatible with a commercially available CE instrument has been designed and constructed and its performance evaluated. The utility of this detector was successfully demonstrated by the indirect photometric detection of anions using a dye as probe and absorbance measured in the visible region. The detector exhibited very low baseline noise (around 0.03 mAU), stable output, and improved upper limit of detection linearity (502 mAU) compared with previously used LED detectors. The detector was tested for indirect detection of anions separated with an electrolyte containing 4 m M Orange G as the indirect detection probe, 10 m M histidine as an isoelectric buffer, and 0.05% hydroxypropylmethylcellulose to suppress the electroosmotic flow. Extremely low detection limits were obtained ranging from 0.16–0.36 μ M (excluding chloride 0.56 μ M ), with separation efficiencies in the range of 154 000–274 000 theoretical plates.