Reversed-phase liquid chromatographic method for the determination of 7-nitrobenz-2-oxa-1,3-diazol-4-yl-labelled lipid analogues.
This paper reports the development of a dual column system for the simultaneous separation of fluorescent short-chain ceramide, 6-[(7-nitrobenz-2-oxa-1,3,-diazol-4-yl[NBD])amino]hexanoyl-sphingo sine and its metabolites, C6-NBD-sphingomyelin and C6-NBD-glucosylceramide, as well as the fluorescent derivatives of choline and serine phosphatides. The method enables the separation of these lipids in a single run on the basis of the polarity of their headgroups and hydrophobicity of their acyl backbone. The fluorescent properties of the NBD-label make it possible to quantitate small amounts of NBD-lipid analogues. The sensitivity of the presented method thus permits the use of small sample volumes and the determination of NBD-lipid analogues secreted into mouse bile directly, without prior extraction or concentration steps.
Frijters
C M
CM
Department of Gastrointestinal and Liver Diseases, Academic Medical Center, The Netherlands.
Tuijn
C J
CJ
Hoek
F J
FJ
Groen
A K
AK
Oude Elferink
R P
RP
Zegers
B N
BN
D000818:Animals; D001646:Bile / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods*; D005456:Fluorescent Dyes; D005963:Glucosylceramides / Q000302:isolation & purification* / Q000378:metabolism; D051379:Mice; D010069:Oxadiazoles / Q000302:isolation & purification* / Q000378:metabolism; D013109:Sphingomyelins / Q000302:isolation & purification / Q000378:metabolism
C095088:6-(N-(7-nitro-2,1,3-benzoxadiazol-4-yl)amino)hexanoyl-glucosylceramide; D005456:Fluorescent Dyes; D005963:Glucosylceramides; C104907:N-6-(7-nitro-2,1,3-benzoxadiazol-4-yl)aminohexanoysphingomyelin; D010069:Oxadiazoles; D013109:Sphingomyelins