Preparation,Identification And Antioxidation Of Astaxanthin From Different Sources

Astaxanthin is widely found in certain algae,fungi,crabs,shrimps,flamingos,and ornamental fish,and has strong biological activities,such as anti-oxidation,anti-aging,anti-cancer,anti-cancer,and UV damage.It has a wide range of applications in medicine,food,and cosmetics.In this paper,P.rhodozyma and H.pluvialis were used as raw materials to purify and prepare astaxanthin.A general quantitative analysis model first established in this study,in order to use the general formula to solve the source of the same substance with stereoisomers in the mixed sample from natural products.Then the model was verified by a mixture of P.rhodozyma,H.pluvialis and chemically synthesized astaxanthin;The protective effect of astaxanthin,β-carotene echinenoneon and a mixture of the three components and crude extract of P.rhodozymon UVA oxidative damage of HepG2 and HSF cells were studied.The main results of the experiment were as follows:(1)P.rhodozyma astaxanthin with a purity of 92.8%was prepared by using P.rhodozyma as a raw material.(2)The conditions for the separation and preparation of H.pluvialis astaxanthin by HSCCC were optimized,and the optimal conditions were as follows:two-phase solvent system:n-hexane:ethyl acetate:ethanol:water(6.5:5:6.5:3,v/v/v/v),flow rate of 3 mL/min,temperature of 25℃,rotate speed of 850 r/min,sample concentration of 10 mg/mL and sample volume of 10 mL.Purified by HSCCC.H.pluvialis astaxanthin with a purity of 88.6%was prepared.(3)A general quantitative analysis model was established to directly determine the source and concentration of each astaxanthin in the mixed astaxanthin sample,and a mixture of P.rhodozyma astaxanthin,H.pluvialis astaxanthin and chemically synthesized astaxanthin was used.This example was validated against this model.The results showed that the deviation between the concentration and the actual concentration in the actual total peak area was in the range of 0-7 μg/mL,and the recovery was between 88.90-103.56%.The deviation between the concentration and the actual concentration was calculated from the theoretical total peak area in the range of 0-3 μg/mL,and the recovery was between 96.20-107.20%.It was indicated that the quantitative analysis model of astaxanthin was feasible and reliable.(4)The protective effect of astaxanthin(P.rhudozyma),β-carotene echinenoneon and a mixture of the three components and crude extract of P.rhodozyma on UVA oxidative damage of HepG2 cells were detected.The results showed that they all could enhance the activity of GSH and T-AOC,which demonstrated that they all had protective effect on UVA oxidative damage of HepG2 cells.(5)The protective effect of astaxanthin(P.rhodozyma),β-carotene echinenoneon and a mixture of the three components and crude extract of P.rhodozyma on UVA oxidative damage of HSF cells were detected.The results showed that compared with the negative control group they all could reduce the activity of LDH and MDA,which demonstrated that they all had protective effect on UVA oxidative damage of HSF cells.The results of this thesis provide a reference for the preparation of astaxanthin from P.rhodozyma and H.pluvialis,and also provide a basis for quantitative analysis of mixtures of stereoisomers from different sources and the anti-oxidation studies of carotenoids.