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Observation of different ceramide species from crude cellular extracts by normal‐phase high‐performance liquid chromatography coupled to atmospheric pressure chemical ionization mass spectrometry
Author(s) -
Pettus Benjamin J.,
Bielawska Alicia,
Kroesen BartJan,
Moeller Peter D. R.,
Szulc Zdzislaw M.,
Hannun Yusuf A.,
Busman Mark
Publication year - 2003
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1043
Subject(s) - chemistry , chromatography , atmospheric pressure chemical ionization , ceramide , mass spectrometry , high performance liquid chromatography , chemical ionization , sphingolipid , lipidomics , analytical chemistry (journal) , ionization , ion , biochemistry , organic chemistry , apoptosis
Normal‐phase high‐performance liquid chromatography (NP‐HPLC) coupled to atmospheric pressure chemical ionization mass spectrometry (APCI‐MS) allows qualitative analysis of endogenous ceramide and dihydroceramide species from crude lipid extracts utilizing chromatographic methods readily adaptable from commonly used thin layer chromatography (TLC) conditions. Qualitative information for the species comes from observation of differences in chromatographic and mass spectrometric behavior between species. Application to the analysis of ceramide and dihydroceramide from various cell lines is demonstrated. The results show the species profile in each cell line to be unique despite growth under identical conditions. The results from APCI‐MS analysis corroborate and enhance information acquired from use of the diacylglycerol kinase assay for total ceramide measurement. This technique readily allows the previously difficult distinction between ceramide and dihydroceramide species. Copyright © 2003 John Wiley & Sons, Ltd.