Premium
Lipid methodology — Chromatography and beyond. Part III. Analyses of natural and 2 H‐labeled glycerolipids by GC/MS and LC/MS with specific enzymic hydrolyses
Author(s) -
Kuksis A.,
Myher J. J.,
Marai L.
Publication year - 1985
Publication title -
journal of the american oil chemists' society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.512
H-Index - 117
eISSN - 1558-9331
pISSN - 0003-021X
DOI - 10.1007/bf03028751
Subject(s) - chemistry , chromatography , metabolic pathway , metabolism , biochemistry
Abstract In the concluding part of this series, the authors review recent attempts to adopt combinations of chromatography with other complementary analytical techniques to the study of stable isotope‐labeled molecules as tracers of glycerolipid metabolism. It is shown that LC/MS in combination with specific enzymic hydrolyses has special advantages for this purpose. Using deuterium‐labeled non‐lipid precursors, effective labeling of both newly synthesized fatty acids and glycerol has been obtained and their molecular association and positional distribution (fatty acids) in the newly formed glycerolipid molecules has been determined as an indicator of the metabolic pathways involved. The above experimental routines extend the analytical lipid methodology beyond the capabilities of chromatography and radio‐chromatography with or without complementary enzymatic analyses. The studies reviewed in the present part and in the previous two parts of this series provide outlines for a potential practical assessment of the various metabolic pathways of glycerolipids, including the identification and quantitation of the true precursor and product pools involved in specific biosynthetic or degradative transformations. Such investigations have not been possible in the past by chromatographic or radio‐chromatographic means. Some of the problems that remain may be subject to solution by means of GC/MS/MS and LC/MS/MS. It is hoped that the new and improved methodology will be matched in the future by comparable advances in the sampling of plasma and cellular components, in selected isotope labeling of the de novo products, and by improvements in the overall design of the metabolic experiments. Although the extension of lipid methodology beyond chromatography has greatly simplified the demands of the experimental design, it is obvious that improved experimental design and sampling techniques will result in further advances in the quality of the observations and in the understanding of lipid metabolism.