Selective extraction and effective separation of galactosylsphingosine (psychosine) and glucosylsphingosine from other glycosphingolipids in pathological tissue samples.
To facilitate the study of the chemical pathology of galactosylsphingosine (psychosine, GalSph) in Krabbe disease and glucosylsphingosine (GlcSph) in Gaucher disease, we have devised a facile method for the effective separation of these two glycosylsphingosines from other glycosphingolipids (GSLs) in Krabbe brain and Gaucher spleen samples. The procedure involves the use of acetone to selectively extract GalSph and GlcSph, respectively, from Krabbe brain and Gaucher spleen samples. Since acetone does not extract other GSLs except modest amounts of galactosylceramide, sulfatide, and glucosylceramide, the positively charged GalSph or GlcSph in the acetone extract can be readily separated from other GSLs by batchwise cation-exchange chromatography using a Waters Accell Plus CM Cartridge. GalSph or GlcSph enriched by this simple procedure can be readily analyzed by thin-layer chromatography or high-performance liquid chromatography.
Department of Biochemistry, Tulane University School of Medicine, 1430 Tulane Avenue, SL-43, New Orleans, LA 70112, USA. yli1@tulane.edu
Yu-Teh
YT
Li
Su-Chen
SC
Li
Wayne R
WR
Buck
Mark E
ME
Haskins
Sz-Wei
SW
Wu
Kay-Hooi
KH
Khoo
Ellen
E
Sidransky
Bruce A
BA
Bunnell
D000293:Adolescent; D000328:Adult; D000818:Animals; D001923:Brain Chemistry; D002648:Child; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002855:Chromatography, Thin Layer / Q000379:methods; D004285:Dogs; D005776:Gaucher Disease / Q000473:pathology*; D006028:Glycosphingolipids / Q000737:chemistry*; D006801:Humans; D007223:Infant; D007965:Leukodystrophy, Globoid Cell / Q000473:pathology*; D008253:Macaca mulatta; D051379:Mice; D008817:Mice, Mutant Strains; D011609:Psychosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013154:Spleen / Q000737:chemistry
D006028:Glycosphingolipids; D011609:Psychosine; C035742:sphingosyl beta-glucoside