Studies On Extraction, Analysis And Isolation Of Ceramides From Konjac

Ceramides, a group of important bioactive compounds, exist comprehensively in animal and plant.In recent years,ceramides have become an important functional ingredient in pharmaceutical health care products,food and cosmetic,especially their high moisturizing effects, which makes they possible to be widely used in cosmetic. With great value in the market and add value, it has attracted the interests of many researchers.This paper focused on the extraction,analysis, isolation and purification of konjac ceramide. The konjac ceramides were extracted by the refluxing method.Firstly,extracting effects of different solvents was compared,then according to the orthogonal experimental design,the optimum extracting conditions of konjac ceramides was showed as following:extracting solvent was 95% ethanol,extracting temperature was 80℃,solid-liquid ratio was 1:10,extracting time was 8 h.In this conditions ceramides were extracted from different sources of konjac,the conclusion demonstrated that there are more or less difference among cultivars, the different position of the same cultivar.The content of phospholipids in the extract was quantitated by molybdenum blue spectrophotometry,with ascorbic acid as the reductant and the content sphingomyelin in the extract was quantitated by HPLC-ELSD.The resluts showed that the content of phospholipids and sphingomyelin is highest in konjac fly flour.Thus the konjac fly flour has certain development and utilize potential.If konjac fly flour as raw material to fermente,we can obtain the ceramide by enzymolysis of Phospholipase C.The lack of a chromophore in the ceramide molecule makes it impossible to identify these molecules using ultraviolet(UV)detection,the previous analytical method need derive,the procedure cumbersome and time-consuming.An analytical method using HPLC-ELSD has been developed for the determination of ceramide extracted from konjac. After a serial of trial, the optimum elution process is methanol concentration starting from 60% to 90% in 5min, and then to 95% in 20 min. Under the optimized conditions, the content of konajc and konjac fly flour was analysed. cerII was found in konjac but not in Amorphophallus Konjac and Amorphophallus Konjac peel.Preliminary purification of ceramides was achieved by chromatography on a silica column. Most impurities were eliminated by elution with 10 bed-volume petroleum ether and ethyl acetate (95/5). Ceramides were eluted with petroleum ether and ethyl acetate (7/3,6/4).ceramide was isolated and prepared by semi-preparative High Performance Liquid Chromatography and validated by analytical column,then ceramide was studied by ESI mass spectrometry and gained its molecular 556.