The Extraction, Separation And Identification Of Ceramide From Fly Konjac Flour

Ceramide is an amide which integrates by sphingosine and long chain fatty acid.It mainly consists in cutin of cell velum and radicel substrate between the cells,and that was the name coming. The ceramide was firstly separated from animal Sphingosine,and lately,it was found in numerous halobios,such as starfish,actinia,sponge and coral,and also found in some plants such as soy and onion.The researches on extracting and purifying of ceramide are doing extensively,because the ceramide is of highly humid-protecting,whitening and barrierring,and is also used more and more widely in the fields of medicine and molecule-biology.As the occuring of animal diseases such as mad-cow illess,the safety of zoo oriented ceramide was oppugned,and the chemosynthesis ceramide maight be harmful to human body.Since the last century,scientists turned to study the safer and cheaper vegetal-ceramide.A lot of molecular structures of ceramides from vegetables had been identified,which promise the abroadly uses of ceramide monocase in medicine,cosmetic and food.Some studies showed that there were so many ceramides in the coat of konjac,and the ceramide contains were highly then all the vegetables that had been learned,but the coat of konjac were abandoned with the fly konjac flour in processing,so it was important to extract and identify the ceramide from fly konjac flour.The extraction of ceramide from fly konjac flour was elementary studied,and because of the especial structure of the ceramide,we chose the crude extracts as the various standards.Firstly, extracting effects of two solvents(chloroform and ethanol)was compared,and so was the surging and ultrasonic extracting,finaly the extracting method was fixed on.Through the single factor experiment,the influence of the temperature,time,the material to solvent ratio,and solvent concentrationwas studied,and the optimized parameter was confirmed by the orthogonal experiment, LSC and TLC.The molecule structure of ceramide monocase was identified by ultraviolet spectrum, infrared spectrum,mass spectrum and nuclear magnetic resonance and so on.The results were as follows:1.The single factor and orthogonal experiment were combined to analysis the influence of the solvent,the amount of solvent,temperature and time to the ceramide gain.The influence degree was C＞B＞A＞D,wich means temperature＞the material to solvent ratio＞time＞solvent concentration,the least influence was the concentration of the solvent.So the best technics parameter was:temperature 60℃,time 35min,the material to solvent ratio was 1:4,and the solvent was 90%ethanol.The best extracting rate was highly to 2.89%.2.We got a ceramide monocase by combining the LSC,TLC and crystal technology,and a detailed technics route for separating and purifying compound.Though the use of the ultraviolet spectrum, infrared spectrum,mass spectrum and nuclear magnetic resonance,and combined the literature,we identified the molecular formula was C₃₈H₇₅NO₅(N-(1'- hydro-methyl -2'-hydroxide)-3-octadeolefin -2, 4- dihydroxyl nonade ceramide),molecular weight was 625.The separation,purifying and the structure of the ceramide from fly konjac flour were Studied and analyzed,which would provide the data to the study of the ceramide,and might help extensive exploitation to the ceramide.