| Honeybee comb is a kind of natural products which has a variety of excellent physiological functions.Nowadays,there were not so many researches on activity ingredient identification and antioxidant properties of honeybee comb.This article was the first time to use multiple solvents to extract the active ingredient from honeybee comb,and used a variety of antioxidant methods and cell model of in vitro anti-inflammatory,to conduct a comprehensive evaluation.This rearche designed to provide a theoretical basis for the excellent physiological activity of the honeybee comb,and to provide a realistic basis for further development and utilization of the potential function of the honeybee comb.1.Honeybee comb was the object of study in this article,water,different concentrations of ethanol,acetone,petroleum ether,ethyl ether,ethyl acetate were used as the extraction solvent,and then we got 10 kinds of extracts of honeybee comb.Total polyphenols and total flavonoids of hydrophilic extracts were determined.HPLC,HPLC-ESI-MS/MS and GC-MS methods were used to analysis and identify the compositions of hydrophilic extracts.The different hydrophilic extracts of honeybee combs,had different contents of total polyphenols and flavonoids,50%alcohol extract contained the highest total polyphenol content in the material,was 9.75 ± 0.16 mg gallic acid/g honeybee comb,95%ethanol extract contained the highest total flavonoid content,was 14.2 ± 0.26mg rutin/g honeybee comb.For the identification of the honeybee comb,the HPLC method was used firstly,and the external standard method was used to analysis the aqueous extract and ethanol extracts preliminarily and qualitativly,then we got 4 kinds of compounds.To identify the above 4 compounds accurately,the HPLC-ESI-MS/MS method was used.Ultimately three compounds had been identified,they were 4-hydroxybenzoic acid,kaempferol,and chrysin.Among these three kinds of compounds,chrysin is one kind of flavonoids for the first time identified in the extract of honeybee comb.Analyzed the lipophilic extract components qualitatively,Volatile components were identified by GC-MS.There were that 15 kinds of components were identified in the petroleum ether extracts.Among these,19 kinds of components were identified in acetone extracts and ether extract;12 kinds of components were identified in ethyl acetate extracts.In the composition of the lipophilic extracts identified,alkanes contained the most,accounting for more than 50%of the identified components.Four lipophilic extracts have all identified the presence of octacosanol,the content was more than 8%,the relative contents of octacosanol was the highest in acetone extract,which was 32.644%.2.In this dissertation,antioxidant properties of honeybee comb was determined by seven kinds of methods,such as the trolox equivalent antioxidant capacity(TEAC),reducing power,DPPH radical scavenging activity,anion radical scavenginga activity,hydroxyl free radical scanvenging activity,?-carotene bleaching assay,and ORAC methods.And the comprehensive antioxidant capacities of the hydrophilic and lipophilic extracts of honeybee comb have been evaluated.The comprehensive antioxidant properties of hydrophilic extracts listed from strong to weak as follows:75%ethanol extract,50%The ethanol extract>25%ethanol extract,aqueous extract,95%ethanol extract>100%ethanol extract.The comprehensive antioxidant properties of four kinds of lipophilic extracts listed from strong to weak as follows:acetone extract>diethyl ether extract,ethyl acetate extract>petroleum ether extract.3.The anti-inflammatory activities in vitro of hydrophilic extracts from honeybee comb were determinated.The four kinds of inflammatory cytokines,such as COX-2,PGE2,IL-8 and TNF-a,were producted by lipopolysaccharide(LPS)-stimulated RAW264.7 cells.The results shows that the hydrophilic extracts inhibieted all of these four kinds of inflammatory cytokines,especially the COX-2 and PGE2 were singnificantly inhibieted by these extacts.The comprehensive anti-inflammatory activities of these six kinds of hydrophilic extracts were evaluated,and the results showed that the 50%ethanol extract had the stongest comprehensive anti-inflammatory property,meanwhile the 95%ethanol extact showed the weakest comprehensive anti-inflammatory property. |
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