Study The Simultaneous Extraction Technology Of The Main Flavonoids And Essential Oil From Seabuckthorn (Hippophae Rhamnoids L.) Leaves

In the present study,simultaneous determination of rutin,quercetin,kaempferol and isorhamnetin was established in the extract of seabuckthorn(Hippophae thamnoides L.)leaves by HPLC.Ionic-liquid-based ultrasound/microwave-assisted hydrodistillation(ILUMSE-HD)simultaneous extraction of the main flavonoids and essential oil from seabuckthorn leaves.Extraction yields of rutin,quercetin,kaempferol,isorhamnetin and essential oil were used as indexes,the extraction conditions of them were optimized.Enrichment and separation of four flavonoids in the ionic-liquid extraction solutions by macroporous adsorption resins was studied and their antioxidant activities were preliminary evaluated.The optimal conditions for extration and separation of four flavonoids were builded.Furthermore,essential oils from seabuckthorn leaves were qualitative analysed by the GC-MS,which provides the theoretical basis for the rational and efficient use of seabuckthorn leaves resources.1.The method on simultaneous determination of rutin,quercetin,kaempferol and isorhamnetin in the extract of seabuckthorn leaves by HPLC was established as follows:Chromatographic conditions:chromatographic column:HIQ SIL C18(4.6mmΦ×250mm.x5μm),the flow rate was ImL/min.The injection volume was 10 uL and the column temperature was set at 30℃.The UV detection wavelength applied was 368 nm.The mobile phase was composed by methanol-acetonitrile-water(40:15:45,v/v/v)adding 1.0%acetic acid.2.The technology on simultaneous extraction of rutin,quercetin,kaempferol,isorhamnetin and essential oil from seabuckthorn leaves by ILUMSE-HD was builded for the first time and essential oils in seabuckthorn leaves were analysed by gas chromatographic method.1-butyl-3-methyl imidazole bromine salt([C4mim])was selected as the most suitable ionic liquids and its suitable molarity is 1 M.The BBD experiment design for the novel and efficient extraction method were utilized for parameter optimization.The optimal parameters of ILUMSE-HD are as follows:liquid/solid ratio:12:1extraction time:34 minmicrowave power:600WUnder the best extration conditions of ILUMSE-HD,the extraction yields of four compounds and essential oil were respectively:RU(9.18±0.35 mg/g),QU(5.52±0.23mg/g),KA(3.03±0.11mg/g),IS(5.64±0.24 mg/g)and EO(0.095%%±0.004%).Compared with the ethanol ultrasonic assisted extraction(EUAE),ionic-liquid-based ultrasounic assisted extraction(ILUAE),ionic-liquid-based microwave assisted extraction(ILMAE),the extraction yields of four flavnoids obtained by ILUMSE-HD had significantly increased and the extaction yields of four flavnoids obtained by ILUMSE-HD had significantly increased and the extaction time was reduced significantly.Therefore,ILUMSE-HD was remarkable superiority.In addition,the results of GC-MS indicated that essential oil components were unobvious change.At the same time,the GC-MS chromatograms of essential oil from seabuckthorn leaves obtained by ILUMSE-HD and HD methods were compared and the results showed that the former extraction was confirmed to extract higher contents of essential oil.Therefore,the novel extraction technology had the advantages of fast and efficient,green environmental protection,saving energy and time,which would be widely applied to simultaneous extract and analysis of active ingredients and essential oil in plant materials.3.Rapidly enrichment and efficiently separation of rutin,quercetin,kaempferol and isorhamnetin from seabuckthorn leaves by using macroporous adsorption resin were studied.Six different types macroporous adsorption resin including AB-8、SA-3、NAK-9、D101、HPD100B and HPD400 were used and the results showed that six different type macroporouses adsorption resin were suitable to enrich and efficiently separate the above four flavonoids.In six different types macroporous adsorption resin,the AB-8 macroporous adsorption resin was the best in enrichment and separation consequence with the recovery yields of rutin,quercetin,kaempferol and isorhamnetin respectively were 91.43%,81.11%,85.57%,73.22%.4.The antioxidant activities of extracts enriched by different type of resins were preliminary valuated and the results showed that the extracts enriched by AB-8 resins have the strongest antioxidant activities with the DPPH radical scavenging abiliity of 0.061 mg CAE/g,FRAP experimental value was 1.585 mmol FeSO4/g and the TEAC value was 0.533 mmol Trolox/g in ABTS test.In conclusion,in this paper,on the one hand,the main flavonoids and essential oil in seabuckthorn leaves were efficient simultanoues extracted,on the other hand,essential oil in seabuckthorn leaves were qualitative analyzed,and four kinds of flavonoids(rutin,quercetin,kaempferol,isorhamnetin)were enrichly separated and furthermore,their antioxidant activities were preliminary evaluated.This paper would provide a theoretical basis for the development and efficient utilization of seabuckthorn leaves resources.ILUMSE-HD technology was bulided,which would provide an important reference for the efficient and simultaneous extraction of active ingredient and essential oil in the plant materials.