Sensitivity enhancement and second-dimensional information from solid phase extraction-capillary electrophoresis of entire high-performance liquid chromatography fractions.
A novel separation technique is demonstrated for peptide analysis of entire 100 microL high-performance liquid chromatography (HPLC) fractions in a single, low pH, capillary zone electrophoresis (CZE) separation. The method employs a hybrid capillary consisting of a removable inlet section containing a reverse-phase packing and a standard CZE bare-fused silica separation capillary. The packed tip allows sample to be concentrated at flow rates greater than 20 microL/min and released in a controlled manner for CZE separation. Separations are comparable to standard CZE with minor modifications in selectivity. Using a mixture of model peptides, the hybrid capillary is shown to effect enhancement in sensitivity of > or = 100-fold. Five fractions from the HPLC separation of peptides obtained from the digestion of bovine serum albumin (BSA) were analyzed by the solid phase extraction-capillary electrophoresis (SPE-CE) method which was shown to provide rapid second-dimensional information. Practical concentration limits of detection for peptides are calculated to be 1-10 ng/mL with this technology.
Strausbauch
M A
MA
Department of Surgery, Mayo Clinic/Foundation, Rochester, MN 55905, USA.
Madden
B J
BJ
Wettstein
P J
PJ
Landers
J P
JP
D000818:Animals; D002138:Calibration; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods*; D010455:Peptides / Q000032:analysis*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
D010455:Peptides; D012710:Serum Albumin, Bovine; D014357:Trypsin