Medicinal Ingredients In Hippophae Rhamnoides L. And The Preliminary Study Of Activity

Seabuckthorn(Hippophae rhanmoides L.),synonyms for the vinegar,acid prickly,black thorn,is a deciduous shrubs in the family Elaeagnaceae(Elaeagnaceae).It contains a variety of bioactive substances.Flavonoids is one of the major bioactive substances,which has been confirmed with a variety of medicinal functions such as anti-inflammatory,anti-oxidation,hypoglycemic and other.The current pharmacological study is only for the total flavonoids of seabuckthorn,but the total flavonoid composition is not clear,restricting the mechanism of seabuckthorn flavonoids in-depth study.Flavonoids extracted from three parts of seabuckthorn with high purity were obtained.A new HPLC and LC-MS/MS method for the determination of flavonoids was established.The separation and identification of 6 flavonoids were realized.Effect of flavonoids extract on blood glucose in mice provided a reference for the further study of flavonoids in different parts of seabuckthorn.The quality control of seabuckthorn fruit pods,seeds and leaves was carried out.Main components such as water,sugar,fat and flavonoids were determined.The results of extraction of flavonoids from different parts of seabuckthorn showed that the extraction rate of flavonoids in seabuckthorn was 5.04% of seabuckthorn pomace,3.67% of seabuckthorn seeds,6.85% of seabuckthorn leaves.Macroporous adsorption resin and polyamide resin were compared for the purification step.After purification by D101 macroporous resin,the purity of seabuckthorn flavonoids reached 73.3% of seabuckthorn pomace,66.7% of seabuckthorn seeds and 36.7% of seabuckthorn leaves.The extraction of seabuckthorn leaves was optimized by single factor experiment.Volume fraction of ethanol,ratio of material to liquid,extraction time and extraction times were optimized.The best extraction method of crude flavonoids from seabuckthorn leaves: ethanol volume fraction 70%,extraction temperature 80 oC,ratio of material to liquid 1:16,extraction time 2 h,extraction times 3 times,the extraction rate of flavonoids reached 7.10%.The HPLC method and the LC-MS/MS method for 6 flavonoids(rutin,catechins,isorhamnetin,myricetin,quercetin and kaempferol)were established,and the methods were validated,including the LOD,LOQ,precision,stability.The method covered the main aglycones of seabuckthorn flavon,and it achieved a complete analysis of seabuckthorn flavonoids.It also reached the detection limit to the ng level,which provides a new idea for the trace analysis of flavonoids.The differences of flavonoids in different parts of seabuckthorn were systematically analyzed.The results of LC-MS/MS showed that: the main flavonoids in seabuckthorn fruit pomacea were isorhamnetin 45.42 mg/g,quercetin 23.41 mg/g,rutin 20.51 mg/g,catechins 4.12 mg/g;the main flavonoids in seabuckthorn seeds were isorhamnetin 52.46 mg/g,quercetin 44.15 mg/g,kaempferol 32.48 mg/g;the main flavonoids in seabuckthorn leaves were kaempferol 132.08 mg/g,isorhamnetin 42.36 mg/g and quercetin 27.78 mg/g.The results of hypoglycemic test in mice showed that the flavonoids of seabuckthorn seeds and seabuckthorn leaves had a rapid hypoglycemic activity.According to the results of LC-MS/MS,the rapid hypoglycemic action of seabuckthorn flavonoids may be related to the content of isorhamnetin,quercetin and kaempferol,which provides a reference for the development and utilization of seabuckthorn resources and the medicinal function of seabuckthorn.