Studies On The Anti-ultraviolet Activity Of Laver Extracts

Objectives:1. Preparation of phycobiliprotein, polysaccharide and mycosporine-like amino acids（MAAs） from laver.2. Investigation the protective effect of the phycobiliprotein, polysaccharide and MAAs onNIH3T3exposed to ultraviolet rays. Discussion the application of the extracts in cosmeticprotecting against UV rays.Methods:1. Simultaneity extract phycobiliprotein and polysaccharide from Laver by ultrasonic. Andthen extract MAAs by water.2.The phycobiliprotein was purified by salting-out and column chromatographic method. Itwas purified by DEAE-cellulose chromatography and HAP chromatography respectively, andmake a comparison between the two stuffing.3. Culture NIH3T3to build the in vitro model of cell lesion caused by UVA. Test theextracts proliferating ability on NIH3T3cell and protection against UV rays by MTT way.Results:1. On the basis of orthogonal array design experiments, four crucial extraction parameterswere optimized, the results was as follows: the ratio of the weight of raw material to the volumeof water was15mg/ml, ultrasonic run time in a circle was4s, ultrasonic interim time in a circlewas3s, extractive time was90min. Under such conditions, the extraction ratio ofphycobiliprotein was14.4%, the extraction ratio of polysaccharide was33.2%.2. Using UV spectrum analysis, the maximum absorption peak of extractive was332nm.Using liqud chromatography and LC-MS analysis, the extractive was MAAs.3. Phycobiliprotein was purified by salting-out and column chromatographic method. First,make the saturation of ammonium sulfate to25%to get rid of the impurity protein, and thenadding the ammonium sulfate to60%to get phycobiliprotein, The crude phycobiliprotein wasdesalted by Sephadex G-25. Then the crude phycobiliprotein was purified by DEAE-cellulosechromatography and HAP chromatography respectively, the purity (A_max\A₂₈₀) of phycobiliprotein was1.0～1.5by DEAE-cellulose chromatography, and the purity （Amax\A280） ofphycobiliprotein was2.15by HAP chromatography. The samples were purified by HAchromatography again and the purity would be improved.4. The phycobiliprotein, polysaccharide and mycosporine-like amino acids （MAAs） have noobvious cytotoxicity on NIH3T3, and they have proliferation effect. In the test of NIH3T3lesion by UVA, the survival rate had significant difference in the group of NIH3T3withtreatment of the phycobiliprotein, polysaccharide and MAAs, compared to the model group.Conclusion:The NIH3T3cells can be damaged an inhibited by UV rays. The phycobiliprotein,polysaccharide and mycosporine-like amino acids have great proliferated effect on NIH3T3andprotect the cells from damaging under ultraviolet rays exposing.