Direct LC–ESI–MS/MS analysis of plant glucosylceramide and ceramide species with 8E and 8Z isomers of the long chain base

ABSTRACT Glucosylceramides and ceramides with 8E and 8Z isomers of the long chain base are found in plants. These isomers have been difficult to quantify separately using liquid chromatography–tandem mass spectrometry (LC–MS/MS) because the isomers have the same retention time, their precursor and product ions have the same m/z values, and plant ceramide standards are not commercially available. Here we tested trial separations using various ODS columns and prepared plant ceramide standards generated by human glucocerebrosidase (imiglucerase) using commercially available plant glucosylceramide standards as the substrates. Consequently, we were able to quantify the isomers based on differences in retention times in a TSKgel ODS-120A column (Tosoh, Tokyo Japan) using LC–electrospray ionization–MS/MS (LC–ESI–MS/MS).