Isolation,Identification And Antioxidant Activity Of Astaxanthin From Antarctic Krill

Objective: Antarctic krill is one of the important biological resources that can be developed and utilized in the Antarctic waters,and is rich in active substances such as astaxanthin.The aim of this study was to establish a high performance liquid chromatography-high resolution mass spectrometry(HPLC-HRMS)qualitative and quantitative analysis method to study the difference in structure and composition of astaxanthin from different sources,and to separate the antarctic krill astaxanthin by column chromatography.And the antioxidant activity of senescent mice was evaluated.Methods: A high-throughput identification and quantification method for high performance liquid chromatography-high resolution mass spectrometry(HPLC-HRMS)of Antarctic krill astaxanthin ester was established.The high purity astaxanthin was prepared by silica gel column chromatography and thin layer chromatography using Antarctic krill as raw material.The three forms of astaxanthin were qualitatively and quantitatively determined by high performance liquid phase method.The anti-aging experiment of D-galactose induced aging mice was carried out by preparing purified astaxanthin ester,and the difference of antioxidant activity of astaxanthin from synthetic astaxanthin and Antarctic krill was compared.Results: A total of 27 astaxanthin esters were detected by HPLC-HRMS in this study.Among them,the higher content of astaxanthin diesters were: Asta-C14:0/C16:0,AstaC14:0/C14:0,Asta-C16:0/C16:0;the higher content of astaxanthin monoesters were Asta-C14:0,Asta-C16:0 and Asta-C18:1.Silica gel column chromatography with different ratios of acetone-petroleum ether solution could be used to separate and prepare astaxanthin diester,monoester and free astaxanthin;V(n-hexane): V(acetone): V(acetic acid)= 8:2:0.2 was the best developer for thin layer chromatography.The proportion of3 R,3’R in astaxanthin diester and monoester in Antarctic krill was more than70%,and the ratio of 3S,3’S was the lowest.This study found that astaxanthin had a significant anti-aging effect on D-galactose-induced aging mice.Compared with the model group,the activities of SOD,GSH-Px,CAT and other enzymes in serum,liver and brain tissues of the astaxanthin intervention group were significantly higher than those in the model group(P<0.05),and the enhancement of T-AOC was also obvious(P<0.05).The content of MDA was significantly decreased(P<0.05).And the antarctic krill source astaxanthin had stronger activity in SOD,GSH-Px and T-AOC than synthetic astaxanthin.Conclusion: A high-resolution,high-throughput mass spectrometry method for astaxanthin in Antarctic krill was established,which had the advantages of high sensitivity and high accuracy.It could analyze the structure and composition of astaxanthin esters and provide technology for the development of Antarctic krill resources.support.The separation of different forms of astaxanthin was achieved by silica gel column chromatography.It was found that the astaxanthin from krill had obvious anti-aging effect on D-galactose-induced aging mice,which could improve the total antioxidant capacity of the body.It reduced the production of lipid peroxidation products,reduced liver pathological damage,and achieved the purpose of delaying aging.The antioxidant effect of Antarctic krill astaxanthin was better than that of synthetic astaxanthin.