Document Detail

Separation and purification of sphingomyelin diastereomers by high-performance liquid chromatography.
MedLine Citation:
PMID:  10873280     Owner:  NLM     Status:  MEDLINE    
All naturally occurring sphingomyelins have the d-erythro-(2S,3R) configuration of the sphingoid base. We have developed a normal-phase HPLC method for the separation of this natural stereoisomer from the l-threo-sphingomyelin, which is the other stereoisomer commonly present in semisynthetic preparations of acyl-chain defined sphingomyelins. The chromatographic method was developed by modification of a previously reported method for phospholipid separation on a normal-phase diol column. The separation was accomplished by a binary gradient of solvent mixtures (A) hexane:isopropanol:acetic acid (82:17:1.0 by vol) and (B) isopropanol:water:acetic acid (85:14:1.0 by vol) with 0.08 vol% triethylamine added to both solvent mixtures. The program of gradient elution was optimized for maximal separation of sphingomyelin diastereomers. For detection of the lipids, a light-scattering detector was used. This analytical scale HPLC method was also used for purification of the stereoisomers (up to 0.5 mg of N-oleoyl-sphingomyelin in a single injection). The purified stereoisomers were at least 99% pure according to high-performance thin-layer chromatography and analytical HPLC.
B Ramstedt; J P Slotte
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical biochemistry     Volume:  282     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  2000 Jul 
Date Detail:
Created Date:  2000-08-21     Completed Date:  2000-08-21     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  245-9     Citation Subset:  IM    
Copyright Information:
Copyright 2000 Academic Press.
Department of Biochemistry and Pharmacy, Abo Akademi University, Turku, FIN 20521, Finland.
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MeSH Terms
Chromatography, High Pressure Liquid / methods*
Chromatography, Thin Layer
Sphingomyelins / isolation & purification*
Reg. No./Substance:
0/Solvents; 0/Sphingomyelins

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