Study On Isolation And Identification Of Essential Oils And Polysaccharides From Magnolia Officinalis And Their Bioactivities

Magnolia officinalis Rehd.et Wils.is one of the species of the genus magnoliaceae Magnolia,Magnolia officinalis flower is Magnolia's dry flower bud.This project systematically explores both the extaction and composition analysis for the volatile oils and the extraction,separation,purification and structure identification for the polysaccharides isolated from magnolia and magnolia officinalis flowers,as well as antioxidation,antitumor,anti-inflammatory,antibacterial,fall blood sugar and light aging resistance and so on many kinds of biological activity was explored,the main results ar as follows:The essential oils of magnolia and magnolia officinalis flowers were extracted by two methods of steam distillation?SD?and simultaneous distillation-extraction?SDE?.The chemical composition analysis conducted by gas chromatography-mass spectrometry?GC-MS?.The results showed four samples are different from components and categories.The SD method to extract volatile oil contained 21 kinds of compounds in magnolia,magnolia officinalis flowers's volatile oil contains 25 kinds of compounds;SDE method to extract of magnoliae volatile oil contains 27 kinds of compounds,magnolia officinalis flower essential oil contains 31 compounds.The main ingredient of magnolia officinalis flower essential oil is d-limonene,magnolia essential oil main ingredient are eucalyptus alcohols and d-limonene.Antioxidation,antitumor,anti-inflammatory and anti-bacterial activities of four essention oils were investigated.The results showed that the total reducing power of value of the volatile oil samples are very low,the ability to remove DPPH free radicals and ABTS+·is in general,the ability to reduced Fe³⁺and remove hydroxyl free radical ability are low,there has certain antioxidant capacity.Magnolia and magnolia officinalis flower essential oil have inhibitory effect on Hep G2 liver cancer cells,MDA-MB-231 breast cancer cells and H1299lung cancer,and have the concentration dependence.In anti-inflammatory experiments,LPS was used to induce RAW264.7 mice macrophage cell for building anti-inflammator model.The result shows that four kinds of volatile oil can inhibit inflammation factor such as NO,IL6 and TNF-?to release.The further research found that it could also effectively restrain the IL-6?i NOS and TNF-?m RNA expression and cytokines secretion.The optimum extracted by hot water of magnolia and magnolia officinalis flower polysaccharide are determined by single factor experiments and response surface analysis.The potimum extraction for magnolia is as follows:91?,liquid ratio of 32.00m L/g,extracting time 3.00h,the yield of magnolia officinalis polysaccharides is 4.12%;The potimum extraction for magnolia officinalis flower is as follows:94?,liquid ratio of 25.50m L/g,extracting time 3.40h,the yield of magnolia officinalis flower polysaccharide is 5.50%.After the extraction of polysaccharide by hot water,then decolorization,alcohol precipitation,proteins removal,freeze drying were carried out.Four components?MO-??MO-??MO-?I?MO-?V?were gained from magnolia officinalis by using DEAE-52 anion exchange column chromatography,magnolia officinalis flower coarse polysaccharide obtained MOF-?,MOF-?I,MOF-?II three components.From the GPC analysis,MO-?,MO-?,MOF-? are homogeneous purified polysaccharide,the molecular weight were:5886da,39195da,6756da.And physical and chemical properties of three kinds of homogeneous polysaccharide were analyzed?Structure analysis of MO-?,MO-?,MOF-? polysaccharides:MO-?,MO-?,MOF-? mainly consists of rhamnose,arabinose,mannose,glucose,galactose,and molar ratio were:1:1.14:1.34:14.29:2.22,1.02:2.27:1:12.99:4.14,1:51.38:1.09:42.04:12.43.Nuclear magnetic resonance?NMR?results showed that MO-? and MO-? are acid polysaccharide,are composed of five kinds of monosaccharide,and contain two kinds of pyranoside?????.MOF-? was acid polysaccharide containing sulfuric acid group,which composed of five kinds of monosaccharide,containing alpha and beta two main pyranoside,and main chain type is 1-6glycosidic bond connection.Congo red test showed that MO-?,MO-? did not exist three spiral space conformation,MOF-? exist three spiral space conformation.The hypoglycemic activity of MO-?,MO-? and MOF-? polysaccharide results showed that the three kinds of polysaccharide has certain inhibitory effect to alpha glycosidase enzymes,MO-? was anticompetitive inhibitor,MO-? and MOF-? were competitive inhibitor.Hep G2 cells were used as a receptor model to study the effects of polysaccharides on glucose consumption in the insulin resistance cells.The result showed polysaccharide can promote the glucose consumption in certain concentration range.The most significant effect appeared in three kinds of polysaccharide at the concentration of 800?g/m L.Anti-aging experiment results showed that MO-?,MO-? and MOF-? polysaccharide can improve Ha Ca T cells proliferation rate after irradiation,and has protective effect on the irradiation damage cells.By determining content of polysaccharide inflammation factor in photoinjurd cells?NO,MMP1,IL-6,TNF-??,in the certain concentration range,polysaccharide can reduce the release of inflammation factors.