Evaluation of reversed-phase nano liquid chromatography conditions by using reversed-phase thin layer chromatography based on Hansen solubility parameters for the analysis of amphiphilic glycosylsphingolipid transformations

摘要

Highlights?Analysis conditions for amphiphilic glycosphingolipids were obtained by examining Hansen solubility parameters using RP-TLC.?Removal of phospholipids was achieved by using ZrO2–SiO2.?Transformation of fluorescently labeled lactosylsphingolipid at Golgi was investigated.?Glycan metabolism was analyzed by using nano-LC.AbstractPulse chase analysis is often used in investigating dynamics of cellular substances. Fluorescently labeled lactosyl sphingosine molecule is useful in chasing its transformation, however the analysis of such metabolites in attomole level is of extreme difficult due to the presence of large amount of endogenous amphiphilic molecules such as glycosphingolipids, sphingomyerin, and glycerophospholipids. Nano LC suites for analyzing the attomole scale metabolites, therefore removal of endogenous substances prior to nan