Preparation Of Main Ingredients Of Grape Seed And Oligomeric Proanthocyanidins

To provide a reference for the realization of grape seed utilization and deepprocessing. Author uses winemaking grape seed as raw materials, systematically studythe process of extraction of the main ingredients of grape seed, The experimental resultswere as follows:Useing petroleum ether（boiling range60-90℃） as solvent to extract grape seedoil,the optimal parameters are: extraction time3h, extraction temperature84℃,extraction liquid-solid ratio9:1. Under this condition, the extract rate of grape seed oilis14.5%. GC-MS analysis results indicate that: wool grape seed oil contains sevenkinds of fatty acid, and main fatty acids content of linoleic acid53.50%, palmitic acid25.94%, stearic acid13.86%.The optimal parameters to extract proanthocyanidins from skimmed grape seed are asfollows: ethanol65%, liquid-solid ratio6:1, each extraction3h, extraction two times.Under optimal parameters, proanthocyanidins extraction rate is5.7%, reach the purityof33.1%. Using biuret method,the extracted grape seed protein content was measured,the value is8.92%,The best parameters of the polyamide resin isolated proanthocyanidins crude extractswere determined. The value of pH is7, the flow rate of sampling was2mL/min, thesample solution concentration was1.1mg/mL, using60%acetone-water as eluent, theelution volume is three times column volume.Optimum parameters for analytical HPLC purified dimer is: packing ODS-AQ;diameter of5μm; flow rate1mL/min; injection volume was20μL; column size4.6×250mm; detect split ratio of5:1; detection wavelength of280nm; columntemperature25℃; mobile phase A: acetone, mobile phase B: ultra-pure water; processtime0-10min: A10%-10%;10-60min: A10%-25%;60-120min: A25%-35%.The optimal parameters preparative liquid chromatography dimer is as follows:packing ODS-AQ; diameter40μm; flow rate of500mL/min; injection volume was400mL; column size20×150cm; detect split ratio of5:1; detection wavelength280nm;column temperature25℃; mobile phase A: acetone, mobile phase B: ultra-pure water;process time0-10min: A10%-10%;10-60min: A10%-25%;60-120min: A25%-35%. Finally, using LC-MS and13C,1H nuclear magnetic resonance spectra for high purityproanthocyanidins structures were identified to grape seed proanthocyanidin dimers B2.