Study On The Preparation And Activity Evaluation Of Phospholipids With High Purity From Shrimp Head

Phospholipids are widely found in all kinds of organisms.The heads of shrimp are also rich in phospholipids.Marine phospholipids have the physiological functions of both phospholipids and omega-3 polyunsaturated fatty acids because they are rich in side chains of ?-3 polyunsaturated fatty acids.Therefore,choosing shrimp head as raw material for in-depth study of marine phospholipids and realizing the high-value utilization of shrimp by-products has important ecological significance and economic development value.In this paper,the phospholipid extracts of shrimp heads were prepared from four representative shrimp in Shandong Province,namely,Penaeus chinensis(Fenneropenaeuschinensis,FC),Penaeus vannamei(Penaeusvannamei,PV),Penaeus chinensis(Penaeusjaponicas,PJ)and Procambarus clarkii(Procambarusclarkia,PCC).The structure,distribution and content of phospholipids in four kinds of shrimp heads were quantitatively and qualitatively analyzed by liposome method based on UHPLC-MS/MS technique.A total of 8 phospholipid types containing 1519 structures(PC,PE,PS,PI,SM,CL,LPC,LPE)were detected and identified,of which the highest content was PC(86.46%),followed by LPC(6.95%)and PE(6.5%).Specifically,the content of PC in PCC(81.94%)was significantly lower than that in the heads of the other three shrimp species.PE(86.61%)in PV,PJ,and PCC is about twice as much as PE in FC.The content of LPC in FC and PCC is higher(9.31%),while the content of LPC in PV and PJ is lower(5.36%).The content of PG in PCC is much higher than that of the other three shrimp heads,which is close to 3 times of the PG content of other shrimp heads.In addition,compared with the other two shrimp heads,the contents of LPE and CL in PCC and PV were significantly higher,while the content of PS in PV was the highest(0.07%).Multivariate statistical analysis showed that there were significant differences in phospholipid composition among the four shrimp heads,and the phospholipid distribution in FC and PJ was relatively similar.The specific phospholipid components in all kinds of shrimp heads were determined by further analysis.The cardiovascular activities of phospholipids in four kinds of shrimp heads were evaluated and compared by zebrafish model,including anti-inflammatory,angiogenic,antithrombotic and cardioprotective effects.All shrimp head phospholipid groups had obvious anti-inflammatory,antithrombotic and cardioprotective activities.The angiogenic activity of phospholipids from PCC and PV was significantly stronger than that of the other two kinds of shrimp head phospholipids.Because the contents of LPE and CL in PCC and PV are much higher than those in the other two shrimp heads,it is inferred that LPE and CL may have angiogenic activity,which need to be verified by further experiments.In view of the comparison and analysis of the quality of shrimp head,we chose Penaeus japonicus to prepare and purify high-purity phospholipids from shrimp head.The preparation method of shrimp head phospholipid is alcohol solvent extraction-n-hexane extraction-acetone precipitation method.the yield of shrimp head phospholipid is 2.18%,the total phospholipid content is 91.96%,and the PC content is 76.21%.Phospholipids from the head of Penaeus japonicus were isolated and purified by column chromatography,and five types of phospholipids(PC,PE,PI,PS,SM)were obtained,of which PC was the most.UHPLC-Q-ExactiveHF/MS was used for analysis and structure identification,and ammonium thiocyanate spectrophotometry was used to determine the content.The purity of all kinds of phospholipids was between 83.35 and 95.05%.A series of zebrafish models were used to evaluate the cardiovascular activity of phospholipids from the heads of five species of Penaeus japonicus.It includes anti-inflammation,promoting angiogenesis,anti-thrombosis and heart protection.Studies have shown that five kinds of shrimp head phospholipids have therapeutic effects on four disease models.Among them,the anti-inflammatory activity of PC was the strongest among the five kinds of shrimp head phospholipids.The effect of high concentration group(80 ?g/mL)was even similar to that of ibuprofen,the vascular protective activity of PC and PI were the strongest among the five kinds of shrimp head phospholipids.There was no significant difference in the antithrombotic effect of all kinds of shrimp heads phospholipid in cardioprotective species,except for 20 ?g/mL.PE also had strong activity at other concentrations.The activity of phospholipids from five shrimp heads increased with the increase of concentration.Through this project,focusing on the by-products of shrimp head,this paper comprehensively compared the structure,distribution,content and cardiovascular protective activity of phospholipids from four common shrimp heads by using a series of techniques,such as UHPLC-MS/MS,liposome,zebrafish activity screening,column chromatography and so on.The head of Penaeus japonicus was selected to prepare high purity phospholipids,and five types of phospholipids were isolated,combined with the evaluation results of cardiovascular activity of all kinds of phospholipids.it laid a solid theoretical foundation for the high-value utilization of phospholipids and the targeted development of all kinds of phospholipids.