【作者】 魏静；

【导师】 钟耕；

【作者基本信息】 西南大学 ， 农产品加工与贮藏， 2008， 硕士

【摘要】 神经酰胺（ceramide）是由神经鞘氨醇（sphingosine）和长链脂肪酸缩合而成的酰胺,主要存在于角质层的细胞膜和细胞间基质中。神经酰胺最早是从动物的神经鞘中分离得到,由此而得名,后来,人们又陆续从众多的海洋生物如海星、海葵、海绵、珊瑚中分离得到,也从被囊类动物和某些高等植物如大豆、洋葱中得到。由于神经酰胺在化妆品中具有的高效保湿、美白和屏障作用,以及在医学、分子生物学等领域日益广泛的应用,使得神经酰胺的提取和纯化分离更加广泛。而随着疯牛病等动物性疾病的发生,动物源性神经酰胺的安全性受到了人们的质疑,同时工业上化学合成的神经酰胺的安全性也对人体有较大的危害。因而,从上世纪末期开始,科学家们转而向更安全更低廉的植物源性神经酰胺展开了研究。现阶段,已经有很多植物中的神经酰胺的分子结构被鉴定出来,从而为神经酰胺单体在医药、化妆品和食品等方面的广泛应用提供了前提。现有的研究表明,魔芋的表皮中含有大量的神经酰胺,远远高于已经进行研究过的植物,但是在加工成魔芋精粉的过程中,这些表皮组分却作为废弃物而留在了魔芋飞粉中。因而,对魔芋飞粉中神经酰胺的提取和鉴定尤为重要。本试验首次对魔芋飞粉中神经酰胺的提取作了初步探索,由于神经酰胺结构的特殊性,本试验主要以神经酰胺粗提物作为各项指标的参考标准。试验首先比较了两种不同提取溶剂（氯仿和乙醇）的提取效果,以及振荡浸提和超声波浸提的提取效果,从而确定提取溶剂和所采用的提取方法。然后通过单因素试验,确定提取温度、时间、料液比和提取溶剂对提取效果的影响,通过正交试验确定提取的最佳工艺参数,并与柱层析（LSC）和薄层层析（TLC）等手段相结合,提取分离魔芋飞粉中的神经酰胺成分,用紫外光谱、红外光谱、质谱和核磁共振等方法,结合文献鉴定了其中神经酰胺单体化合物的分子结构。其结果如下:1.采用单因素试验与正交实验相结合的方法,分析了提取溶剂浓度（D）、料液比（B）、温度（C）和时间（A）对魔芋飞粉中神经酰胺得率的影响。其影响程度由大到小依次为C＞B＞A＞D,即温度＞料液比＞时间＞溶剂浓度,影响最小的为溶剂的浓度。通过实验得到优化的神经酰胺提取工艺参数为:提取温度60℃,提取时间35min,提取料液比1:4,提取溶剂为90%乙醇。神经酰胺粗提物的最高提取率可以高达2.89%。2.采用柱层析（LSC）、薄层层析（TLC）和溶剂结晶相结合的方法分离得到一个神经酰胺单体化合物,通过本实验得到了一条分离纯化该化合物的详细的工艺路线。并利用红外光谱、紫外光谱、质谱和核磁共振等方法,结合文献鉴定了该单体化合物分子式为C₃₈H₇₅NO₅（N-（1′-羟甲基-2′-羟基）-3-十八烯烃-2,4-二羟基十九脂肪酰胺）,分子量为625。本试验对魔芋飞粉中神经酰胺的分离纯化和结构进行了研究和分析,为神经酰胺的研究提供了基础资料,将有助于神经酰胺的进一步开发利用。更多还原

【Abstract】 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.更多还原