Stability Of Soybean Lecithin And Enzymatic Preparation Of Glycerylphosphorylcholine

In this study,the stability of soybean lecithin of soybean phospholipid soft capsule by investigating the quality changes and reasons during storage was explored.The high purity glycerylphosphorylcholine(GPC)with medicinal value was prepared by phospholipase A1 hydrolyzed phosphatidylcholine(PC)of soybean lecithin PC35 and purification.Firstly,the stability of soybean lecithin of soybean phospholipid soft capsule were investigated by explore the reason of the quality changes of capsules during the normal storage and accelerated hydrolysis storage conditions.Results showed that PC content decreased and acid value increased.Lysophosphatidylcholine content increased along with the decrease of PC content.Water in capsule shell migrated to the PC during storage,caused PC hydrolyzation into LPC.Filler soybean oil also hydrolyzed,released free fatty acids.Both hydrolyzation products contributed to the increased acid value.Secondly,GPC was produced by phospholipase A1 hydrolysis of PC in an water-in-oil emulsion medium formed by soybean oil,water and soybean lecithin PC35.Influence factors on GPC yield of the hydrolysis,such as soybean oil ratio(soybean oil: PC35 weight),reaction temperature,water dosage(weight of PC35),enzyme dosage(weight of PC35)and reaction time,were discussed.After the single-factor experiment and optimized by the response design,the optimum reaction conditions were as follows: soybean oil ratio 4:1,reaction temperature 55oC,reaction time 3 hour,water dosage 30 wt%,enzyme dosage 5 wt%.And finally,the yield of GPC was obtained by 95.53%.Thirdly,under the optimum reaction conditions,the reaction system weight was amplified to ten fold in order to exam the possibility the reaction amplification.The results showed that good catalytic activity and the amplification reaction had good reproducibility.Then,the methods of purification combined with methanol extraction,centrifugation,deoil in acetone,silica gel column chromatography,decolorization of activated carbon,were used to purified enzyme hydrolysis products.And the main influencing factors elution flow rate and sample concentration on purification by silica gel column were optimized.The purified GPC products were quantitative analyzed by high performance liquid chromatography-evaporative light scattering detection(HPLC-ELSD)and qualitative analyzed by using liquid chromatography-mass spectrometry(LC-MS),31 P nuclear magnetic resonance(31P – NMR)and attenuated total reflection fourier transform infrared spectroscopy(ATR-FTIR).Finally,the results certified the product is GPC.