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A method for the quantitative analysis of molecular species of alkylacylglycerol and diacylglycerol
Author(s) -
Warne Thomas R.,
Robinson Mitchell
Publication year - 1990
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02544045
Subject(s) - lipidology , diacylglycerol kinase , clinical chemistry , chemistry , chromatography , computational biology , biology , biochemistry , enzyme , protein kinase c
We describe a method for the quantitative analysis of molecular species of diacylglycerol and alkylacylglycerol as their diradylglycerobenzoate derivatives. Synthetic internal standards were used to provide quantitative determinations of the low levels of diacylglycerol and alkyl‐acylglycerol and their individual molecular species in cultured cells. Diradylglycerols were isolated by thin‐layer chromatography (TLC), converted to their benzoate derivatives and separated into subclasses by TLC. The molecular species of each subclass were analyzed by reversed‐phase high performance liquid chromatography. Thirty‐six species of diglyceride‐type molecules were identified in Madin‐Darby canine kidney cells. These cells were shown to contain 7.88 nmoles of diacylglycerol and 3.97 nmoles of alkylacylglycerol per μmole of phospholipid. Both subclasses contain predominantly monoenoic and saturated species. This technique should be valuable for studies examining the origin and metabolism of these important intracellular mediators.