The effect of conditioning of fused-silica capillaries on their electrophoretic performance.
In CE practice, conditioning of the capillary tube with strong base, acid, or both in sequence, has been recognized as essential to obtain reasonable precision in electroosmotic mobility (EOM), and consequently, in the migration times of analytes. This report focuses on the comparative study of three different approaches for capillary conditioning: etching with HF, etching with NaOH, and leaching with HCl after NaOH treatment. EOM-based measurements, including the hysteresis effect that characterizes the dependence of EOM with pH, were used to evaluate the conditioned silica surface. Additionally, indirect inspection of the conditioned surface was carried out by examining the electrophoretic profile of the cationic probe Ru(bpy)(3) (2+), known to strongly interact with the ionized silica surface while displaying no significant acid-base activity. It was shown that, once conditioned and prior to any CE measurement, extensive rinse of the capillary with the running electrolyte at high flow rate was essential to attain relatively rapid re-equilibration of the tube with the fluid. Insufficient rinse with the running electrolyte would result in negatively biased EOM measurements and significant drift in migration times. It was also established that relatively high flow rates of 1 M NaOH conditioning solution (4-5 column volumes per minute) was required to attain capillaries with reproducible electrophoretic performance within a reasonable conditioning time (typically, 1 h). In addition to relatively more extensive rehydroxylation of the silica surface, evidenced by the highest EOM values obtained, the sequential use of NaOH-etching and HCl-leaching provided better precision than HF-etching or NaOH-etching alone.
Gómez
Jorge E
JE
Department of Chemistry, Universidad del Valle, Cali, Colombia.
Sandoval
Junior E
JE
D053841:Electroosmosis / Q000379:methods; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D006858:Hydrofluoric Acid / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D009942:Organometallic Compounds / Q000737:chemistry; D015203:Reproducibility of Results; D012822:Silicon Dioxide; D012972:Sodium Hydroxide / Q000737:chemistry
D009942:Organometallic Compounds; C415768:tris(2,2'-bipyridyl)ruthenium(II); D012972:Sodium Hydroxide; D012822:Silicon Dioxide; D006858:Hydrofluoric Acid