Improving detection in capillary electrophoresis with laser induced fluorescence via a bubble cell capillary and laser power adjustment.
Bubble cells have been frequently employed in capillary electrophoresis (CE) to increase the light path length with UV detection to provide an increase in the observed sensitivity of CE; however this approach has not been commonly used for laser-induced fluorescence detection (LIF) with CE. In this paper we study the influence of laser power on the sensitivity of detection in using conventional and enlarged fused silica capillaries for CE with LIF. When using the bubble cell capillary, the laser power must be decreased relative to use of the conventional capillary to reduce the effects of photodegradation of the species being illuminated by the laser. Even though the light intensity was decreased, an increase in sensitivity of detection was observed for most compounds when a bubble cell was used. This increase ranged from a factor of 8 for riboflavin (410 nm excitation) to 3.2 for most aromatic compounds (266 nm excitation), when using a 3x bubble cell compared with a conventional capillary. The bubble cell capillary was used for native detection of IgG by LIF at 266 nm. A limit of detection of 60 ng mL(-1) was obtained from a 20 pg injection, which was 40 times more sensitive than silver staining in conventional SDS/PAGE.
Rodat
Audrey
A
Laboratoire des IMRCP, Université de Toulouse, Université Paul Sabatier, 31062 Toulouse, Cedex, France.
Gavard
Pierre
P
Couderc
François
F
D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods*; D005453:Fluorescence; D007834:Lasers; D015203:Reproducibility of Results