An LC method for the analysis of phosphatidylcholine hydrolysis products and its application to the monitoring of the acyl migration process

An assay for quantitative analysis of phosphatidylcholine (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) and its hydrolysis products: 1-hydroxy-2-palmitoyl-sn-glycero-3-phosphocholine and 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine, sn-glycero-3-phosphocholine and palmitic acid using high-performance liquid chromatography with charge aerosol detector (CAD) was developed. The separation of the compounds of interest was achieved on a reversed-phase/hydrophilic interaction stationary phase with a mobile phase consisting of acetonitrile:methanol:10 mM ammonium acetate solution. The method was applied to control the acyl migration process of LPC regioisomers in the most common solvents used in the synthesis or modification of PC.

► Phosphatidylcholine (PC) hydrolysis product formation is a serious problem especially in the enzyme-catalyzed acyl exchange of PC.
► Quantitative HPLC analysis of PC and its hydrolysis products is presented.
► The method was applied to examine the stability of the lysophosphatidylcholine (LPC) isomers in 16 different solvents.
► 1-Palmitoyl LPC isomer is more stable than 2-palmitoyl LPC isomer.
► The acyl migration and hydrolysis processes are prefer at alkaline pH.