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Preparation and electrophoretic separation of Bodipy-Fl-labeled glycosphingolipids.

Journal of chromatography. A (2012-02-11)
Scott A Sarver, Richard B Keithley, David C Essaka, Hidenori Tanaka, Yayoi Yoshimura, Monica M Palcic, Ole Hindsgaul, Norman J Dovichi
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
CHES, ≥99.0% (titration)
Sigma-Aldrich
CHES, BioXtra, ≥99.0% (titration)
Sigma-Aldrich
CHES, BioUltra, ≥99.5% (T)

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