Preparation and electrophoretic separation of Bodipy-Fl-labeled glycosphingolipids.
Several glycosphingolipids were labeled with the fluorphore Bodipy-Fl and analyzed using capillary electrophoresis with laser-induced fluorescence detection. GM1-, LacCer-, and Cer-Bodipy-Fl were prepared through acylation using the N-hydroxysuccinimide ester of Bodipy-Fl. Several other glycosphingolipids including GT1a-, GD1a-, GM2-, GM3-, GD3-, and GlcCer-Bodipy-Fl were enzymatically synthesized. Micellar electrokinetic capillary chromatography with a TRIS/CHES/SDS/α-cyclodextrin buffer produced better separation than an established borate/deoxycholate/methyl-β-cyclodextrin buffer. The nine Bodipy-Fl-labeled glycosphingolipid standards were separated in under 5 min, theoretical plate counts were between 640,000 and 740,000, and the limit of detection was approximately 3 pM or 240 ymol analyte injected onto the capillary.
The University of Notre Dame, Department of Chemistry and Biochemistry, Notre Dame, IN 46556, USA.
Scott A
SA
Sarver
Richard B
RB
Keithley
David C
DC
Essaka
Hidenori
H
Tanaka
Yayoi
Y
Yoshimura
Monica M
MM
Palcic
Ole
O
Hindsgaul
Norman J
NJ
Dovichi
D001881:Borates; D001896:Boron Compounds / Q000737:chemistry*; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D003840:Deoxycholic Acid; D019075:Electrophoresis, Capillary / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006028:Glycosphingolipids / Q000737:chemistry* / Q000302:isolation & purification; D057230:Limit of Detection; D012967:Sodium Dodecyl Sulfate; D013654:Taurine / Q000031:analogs & derivatives; D014325:Tromethamine; D047391:alpha-Cyclodextrins; D047392:beta-Cyclodextrins
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001881:Borates; D001896:Boron Compounds; D005456:Fluorescent Dyes; D006028:Glycosphingolipids; D047391:alpha-Cyclodextrins; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D003840:Deoxycholic Acid; D014325:Tromethamine; C050927:2-(N-cyclohexylamino)ethanesulfonic acid; D013654:Taurine; D012967:Sodium Dodecyl Sulfate