Design and performance of a light-emitting diode detector compatible with a commercial capillary electrophoresis instrument.
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 mM Orange G as the indirect detection probe, 10 mM 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 microM (excluding chloride 0.56 microM), with separation efficiencies in the range of 154,000-274,000 theoretical plates.
Australian Centre for Research on Separation Science, School of Chemistry, University of Tasmania, Hobart, Tasmania, Australia.
Cameron
C
Johns
Miroslav
M
Macka
Paul R
PR
Haddad
D019075:Electrophoresis, Capillary / Q000379:methods*; D006639:Histidine / Q000737:chemistry; D007202:Indicators and Reagents; D012680:Sensitivity and Specificity
D007202:Indicators and Reagents; D006639:Histidine