Sphingosine determination at the picomole level using dimethylaminoazobenzene sulphonyl chloride.
A sensitive method (lower detection limit 5 picomoles) is proposed for the determination of sphingoid bases liberated from mammalian glycosphingolipids by acid hydrolysis. The azo dye 4-dimethylaminoazobenzene-4'-sulfonyl chloride reacts with the primary amino group of the sphingosine bases, forming a stable derivative. Excess reagent, which is degraded during the derivatization, and free amino sugars as common hydrolysis products of glycosphingolipids are completely separated by reversed-phase high-performance liquid chromatography. This method was applied to the determination of the glycosphingolipid content of mouse spleen and thymus.
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006028:Glycosphingolipids / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D013110:Sphingosine / Q000302:isolation & purification*; D013154:Spleen / Q000378:metabolism; D013950:Thymus Gland / Q000378:metabolism; D004124:p-Dimethylaminoazobenzene / Q000031:analogs & derivatives
D006028:Glycosphingolipids; C010572:4-N,N-dimethylaminoazobenzene-4'-sulfonyl chloride; D004124:p-Dimethylaminoazobenzene; D013110:Sphingosine