Rapid method of preparation of lysoglycosphingolipids and their confirmation by delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry.
We have recently developed a rapid method for the preparation of lysoglycosphingolipids on a small scale, in high yield. This procedure of microwave-mediated saponification of about 1 mg of glycosphingolipid with 0.5 ml of 0.1 M NaOH in methanol for two minutes can be easily repeated if larger amounts of lyso-compounds are needed. We have also found that the new methodology of delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry in the reflector mode is extremely effective for the confirmation of different lysoglycosphingolipids together with their long chain base components. The combined method of preparation and confirmation of lysoglycosphingolipids is also important for the identification of long chain bases of various sphingolipids, because the usual analytical method of long-chain bases of sphingolipids depends on acidic methanolysis, which results in the formation of by-products such as O-methylsphingosines and threo-sphingosines.
Department of Biochemistry, Shinshu University School of Medicine, Nagano. agingol@gipac.shinshu-u.ac.jp
T
T
Taketomi
A
A
Hara
K
K
Uemura
E
E
Sugiyama
D000818:Animals; D002240:Carbohydrate Sequence; D004912:Erythrocytes / Q000737:chemistry; D005776:Gaucher Disease; D006028:Glycosphingolipids / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification; D006041:Goats; D006801:Humans; D008872:Microwaves; D008969:Molecular Sequence Data; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D006028:Glycosphingolipids