Base Hydrolysis (or not) for Targeted Sphingolipid Quantitation

Abstract

Sphingolipids are a diverse class of lipids that exhibit various structures and functions. Their analysis presents several challenges due to dynamic range, structural diversity, and complex sample matrices. Addressing these challenges requires meticulous sample preparation and dedicated analysis methods. Liquid chromatography (LC) coupled with mass spectrometry (MS) is a commonly used technique for detecting and quantifying sphingolipids. An alkaline hydrolysis step is routinely used during sample preparation to enrich for sphingolipids. The added chemical step during sample preparation can impact lipid extraction efficiency and removes the opportunity to analyze other lipid classes simultaneously. We will present our findings comparing sphingolipid enrichment using mild base hydrolysis pre-treatment versus no hydrolysis in C18 and HILIC LC-MS/MS methods.