Study Of Procyanidins From Sea Buckthorn Seed On Anti-myocardial Ischemia Activity

Sea Buckthorn which belongs to Hippophae Plants of Elaeagnus Branch is also called Hippophe or acid gill. In the world not only our country is the largest on the distribution of Hippophae Plants, and it is also the largest on the reserves. In 1977, Seabuckthorn was writen in the book of "People's Republic of China Pharmacopoeia" as a kind of traditional Chinese medicine. The Procyanidine of Sea Buckthorn Seed was taken as the object of study in this topic.Find out the part of anti-myocardial ischemia activity and then isolate the effective part.At last the structure of component from the effective part was identified.These studies may set a foundation for the further development of Sea Buckthorn Seed resources.First,Procyanidine were extract from ethanol extracts of Sea Buckthorn Seed by ethyl acetate. Then four parts were abtained from the ethyl acetate extracts through Macroporous Resin AB-8 chromatography column. Afterwards different patrs were given to ischemic myocyte of neonatal rat respectively for drug intervention and then detect the survival rate of ischemic myocyte by MTT.The experiments showed that the ethyl acetate extracts and the 10% ethanol eluates from Macroporous Resin AB-8 could significantly increase the survival rate of ischemic myocyte and had the pretective effects on myocardial ischemia injury.Second,four compounds were abtained from 10% ethanol eluates by Sephadex LH-20, HW-40 and ODS chromatography column and the elution process was detected by TLC. Their structures of five compounds were determined by means of NMR analysis. The experiments of anti-myocardial ischemia activity determination showed that epigallocatechin and catechin are the main active component.Third, taking yield rate of the Procyanidine as index, the single factor and the orthogonal experiment were carried on to obtaine the optimization extraction craft parameter.Finally using water as extractant,the extraction plan was summed up as followed: temperature 50℃, solid-liquid ratio 1:10(W/V), extraction time 1h,extraction times 3 times.Finally macroporous resin purification craft of Procyanidine in Sea Buckthorn Seed was studyed. The experiments showed that AB-8 has good adsorption and desorption capacity.It is helpfull for separation that the concentration of the samples arrive at about 30mg/mL.The Procyanidine with good purity was gained through Macroporous Resin AB-8 chromatography column by 1 BV water and 2.8 BV 10% ethanol.After useing 4 times Macroporous Resin AB-8 need regeneration treatment.