Purification of l‐α‐Glycerylphosphorylcholine from the Enzyme Reaction Solutions by Resin Column Chromatography

Abstract

Colorless L-a-glycerylphosphorylcholine (L-a-GPC) was obtained at 98.8% purity, and with a specific rotation of -2.5°via a four-step procedure. L-a-GPC was first produced by phospholipase A 1 hydrolysis of phosphatidylcholine in an aqueous medium. Almost all the impurities were then removed using two successive D001 cation and D301-111 anion exchange resin column chromatography procedures, followed by decoloration with active carbon to remove minor pigments and remaining impurities. D001 resin and D301-111 resin showed remarkable ability for L-a-GPC isolation after being used eight times. Thus, this study presents a simple and cost-effective method for preparing L-a-GPC with high purity, low costs, and environmental friendliness, and encourages future investigation into its adaptation for industrial applications.

Chemical and Enzymatic Approaches to Esters ofsn‐Glycero‐3‐Phosphoric Acid

“…Lecitase Ultra is the most used PLA 1 as a biocatalyst to obtain GPC starting from PC under a wide range of experimental conditions. More in detail, Zhang et al [92] . reported the hydrolysis in aqueous medium starting from PC of food grade soy lecithin: GPC was obtained in 70–74 % yield and up to 98.8 % purity by silica or ion‐exchange chromatography [92,93] .…”

“…Bang et al [98] . prepared GPC via Lecitase Ultra‐catalyzed hydrolysis of soy PC in an n ‐hexane‐water system, thus achieving enhanced productivity of GPC compared to other published enzymatic reaction systems [91–93] …”

“…[91] Lecitase Ultra is the most used PLA 1 as a biocatalyst to obtain GPC starting from PC under a wide range of experimental conditions. More in detail, Zhang et al [92] reported the hydrolysis in aqueous medium starting from PC of food grade soy lecithin: GPC was obtained in 70-74 % yield and up to 98.8 % purity by silica or ion-exchange chromatography. [92,93] Hydrolysis of PC catalyzed by PLA 1 can lead to the formation of GPC as well as sn-1-LPC due to the migration of the acyl group from the sn-2 position to the sn-1 position in LPC.…”

“…More in detail, Zhang et al [92] reported the hydrolysis in aqueous medium starting from PC of food grade soy lecithin: GPC was obtained in 70-74 % yield and up to 98.8 % purity by silica or ion-exchange chromatography. [92,93] Hydrolysis of PC catalyzed by PLA 1 can lead to the formation of GPC as well as sn-1-LPC due to the migration of the acyl group from the sn-2 position to the sn-1 position in LPC. [94] Plückthun and Dennis [95] investigated acyl migration in LPC and reported that about 90 % of thermodynamically stable 1-acyl LPC and 10 % 2acyl LPC were present in the mixture at equilibrium under the experimental conditions used.…”

Enzymatic preparation of food‐grade l‐α‐glycerylphosphorylcholine from soy phosphatidylcholine or fractionated soy lecithin

“…In the former case, an extracting solvent, such as diethyl ether, is used to remove the free fatty acids (FFA) that form during the reaction . The chromatographic method commonly utilizes column chromatography with an ionic or polar stationary phase and organic solvents (e.g., methanol) as a mobile phase to separate l ‐α‐GPC from substances with lower polarities …”