Abstract
A method is described for the direct quantitation of phosphatidylcholine molecular species by reverse phase high-performance liquid chromatography employing flame ionization detection. The method is shown to be applicable to plan phosphatidylcholine. The molecular species are separated with a C18 column eluted in an isocratic mode. Detection by a commercially available flame ionization detector overcomes the problems of detecting underivatized naturally occurring lipids using ultraviolet detectors, and allows direct and rapid mass determination of the resolved molecular species. Detection limits for quantitation are defined.
Highlights
In this report we describe the application of this flame ionization detection (FID) with reverse phase high-performance liquid chromatography (HPLC) for the determination of plant PC molecular species
Commercial PC standards were used to develop this optimal mobile phase by modification of the mobile phase used for separation of Dunaliella PG molecular species [8]
In addition to identification by retention times, the identity of molecular species was confirmed by fatty acid analysis of individual components recovered after HPLC separation
Summary
MaterialsPC standards were obtained in 99% purity from Sigma Chemical Company (St. Louis, MO). The leaves were pulselabeled for 1 hr with [14C]oleic acid (sp act 52.6 mCi/mmol; New England Nuclear, Boston, MA) [10]
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