| Sea cucumbers contain a lot of nutrients.With the improvement of people's living standards,the demand for sea cucumbers is gradually increasing.Sea cucumbers contain a large amount of autolysing enzymes.When the sea cucumbers are salvaged and brought ashore,the released autolysing enzymes will dissolve the sea cucumbers.In order to prevent the sea cucumbers from autolysing,the sea cucumbers need to be cooked immediately after fishing,resulting in a large amount of cooking liquid.Sea cucumber is rich in polysaccharides and has a variety of biological activities,and nutrients will be lost during the cooking process.Therefore,it is of great guiding significance to study the extraction method of sea cucumber cooking liquid polysaccharides and make full use of sea cucumber polysaccharides.In this paper,three different extraction methods were used to obtain polysaccharides from sea cucumber boiled liquid,and further separation and purification were carried out,and the composition,structure and properties of polysaccharides were analyzed.The main research results are as follows:Using ethanol/sodium carbonate two-phase system to extract polysaccharides from sea cucumber cooking liquid,it was determined that the optimal extraction conditions of salting-out extraction method were:ethanol was 30%,sodium carbonate was 15%,and the recovery rate of sea cucumber cooking liquid polysaccharide was83.88%.Three different methods were used to extract polysaccharides from sea cucumber boiled liquid.The total sugar content,protein content,sulfate content and uronic acid content of the polysaccharides in the salting-out extraction method were 32.81±0.61%,27.66±0.62%,7.81±0.50%,8.18±0.66%;CPC method was 14.25±0.15%,17.31±0.69%,23.86±0.76%,7.81±1.34%;alcohol precipitation method was7.38±0.86%,32.91±0.49%,31.48±0.78%,0.17±0.24%.The monosaccharide composition was analyzed by high performance liquid chromatography.The monosaccharide composition of polysaccharide by salting out extraction method was mannose,glucosamine,glucuronic acid,galacturonic acid,galactosamine,glucose,galactose,arabinose,Fucose,itsmolarratiois3.44:0.90:0.07:0.01:0.08:1.00:0.31:0.05:0.46;the composition of CPC method polysaccharide includes mannose,glucosamine,glucuronic acid,galacturonic acid,aminosaccharide Lactose,glucose,galactose,arabinose,fucose,the corresponding molar ratio is 4.94:10.54:5.50:0.24:9.05:1.00:21.99:0.32:191.32;alcohol precipitation polysaccharide is composed of mannose,glucosamine,glucose It is composed of uronic acid,galacturonic acid,glucose,galactose,arabinose,and fucose,and the molar ratio is2.99:1.28:0.04:0.62:1.00:0.65:0.05:2.68.The properties of the three polysaccharides were analyzed,and the infrared spectra showed that the three polysaccharides all contained fucose methyl and acetylamino groups.The thermodynamic properties of the polysaccharides were analyzed by DSC,and the three sea cucumber cooking liquid polysaccharides all had an endothermic peak.and an exothermic peak,the endothermic peaks of the three polysaccharides are similar,but the exothermic peak of the polysaccharide by the salting-out extraction method is 279.19?,and the exothermic peak of the polysaccharide by the alcohol precipitation method is 267.80?,which are significantly higher than those by the CPC method,indicating that the salting-out extraction method The thermal stability of the polysaccharide by the method of salting out extraction was higher than that of the other two methods.Polysaccharides from sea cucumber boiled liquid were separated and purified by DEAE-sepharose fast flow ion chromatography.The salting-out extraction method polysaccharide was separated and purified to obtain 4 components,and component 1and component 3 were recovered and named as Y1 and Y3.The CPC method polysaccharide was separated and purified to obtain 4 components,and components 3and 4 were recovered and named C3 and C4.By high performance liquid chromatography analysis,Y1 is composed of mannose,glucosamine,glucuronic acid,glucose,galactose,arabinose and fucose,and its molar ratio is1:0.201:0.013:0.303:0.098:0.042:0.016;Y3 is mainly composed of mannose,glucosamine,glucuronic acid,galactosamine,galactose and fucose,and the molar ratio is 1.00:1.73:1.13:1.71:1.55:6.65;C3 monosaccharide is composed of mannose,glucosamine,glucuronic acid,galacturonic acid,galactosamine,galactose and fucose,the molar ratio is 1.00:5.56:0.78:2.31:8.08:10.01:107.74;C4 contains mannose,glucosamine,Galactosamine,galactose,fucose,the molar ratio is1.00:3.22:1.94:6.00:76.81.The sulfate content of Y1 was 1.29%,26.48%,16.17%and 8.50%,respectively.The weight-average molecular weights were 4.249?10~4,8.256?10~4,2.062?10~4 and 3.474?10~4 Da,respectively.The infrared spectra showed that the four components Y1,Y3,C3 and C4 all had vibration peaks caused by fucose methyl groups.In the Raw264.7 cell viability experiment,the sample Y1 and C3 had no significant effect on the Raw264.7 cell viability when the dose was lower than 800?g/m L.When the sample C4 concentration was 800?g/m L,the Raw264.7 cell viability was significantly decline.According to the review,the polysaccharide extraction rate,chemical composition and monosaccharide composition obtained by different extraction methods of sea cucumber cooking liquid are different,which provides a theoretical basis for the further development and utilization of sea cucumber cooking liquid polysaccharide. |
PDF Full Text Request