Direct mass spectrometric analysis of glycosphingolipid transferred to a polyvinylidene difluoride membrane by thin-layer chromatography blotting.
A simple, rapid method for the analysis of glycosphingolipid that combines "thin-layer chromatography (TLC) blotting" and mass spectrometry is reported. Glycosphingolipids developed by TLC were transferred to a polyvinylidene difluoride membrane by TLC blotting, after which the glycosphingolipid band on the membrane was excised and placed on a mass spectrometer probe tip, and a few microliters of triethanolamine was added as the matrix. The sample was analyzed by secondary ion mass spectrometry. About 1 microgram of glycosphingolipid subjected to TLC can be analyzed. The major advantage of this method is that glycosphingolipid can be analyzed structurally without purification by repeated column chromatography.
Taki
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Department of Biochemistry, Faculty of Medicine, Tokyo Medical and Dental University, Japan.
Ishikawa
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Handa
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Kasama
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D000818:Animals; D002240:Carbohydrate Sequence; D002855:Chromatography, Thin Layer / Q000379:methods; D004912:Erythrocytes / Q000737:chemistry; D005776:Gaucher Disease / Q000378:metabolism; D006028:Glycosphingolipids / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008567:Membranes, Artificial; D008969:Molecular Sequence Data; D011145:Polyvinyls; D013154:Spleen / Q000737:chemistry; D013552:Swine; D005465:Fluorocarbon Polymers
D006028:Glycosphingolipids; D008567:Membranes, Artificial; D011145:Polyvinyls; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers