| Malania oleifera Chun et S. Lee is an important and special oil tree cultivated in Guangxi, China. In this thesis, systematic research was carried out on different parts of Malania oleifera, such as seeds, shell, branches with leaves and outer part of pericarp (exocarp and mesocarp). Chemical component of fatty acids, essential oils, polysaccharides, flavonoids, lignins, protein and antioxidant were studied. The antitumor, antioxidation, antilipooxidant and antimicrobial activities of effective constituent were investigated. Research results obtained in this study will greatly enhance the comprehensive utilization rate of Malania oleifera and provide basic data for further development of Malania oleifera. The main research contents and results were showed as follows:(1) The extraction of fatty acids from the seeds was conducted using petroleum ether (90-120℃) in a soxhlet extractor and essential oils were extracted by hydrodistillation. Total fatty acids (TFAs) of lipids were transformed into their corresponding methyl esters and analyzed by gas chromatography-mass spectrometry (GC-MS). The yield of fatty acids extracted from the seeds was60.5±2.5%, which contains fourteen chromatographic components, of which10compounds were identified, representing98.98%of the lipids, the major components were nervonic acid (55.70%), octadecenoic acid (23.81%), docosenoic acid (13.13%); The yields of essential oils were0.1±0.02%,0.2±0.03%and1.1±0.12%, respectively from fresh seeds, branches with leaves and outer part of pericarp. Seventeen chromatographic components and fifteen compounds (99.90%) were identified in the essential oils of seeds with benzyl alcohol (58.42%) and benzaldehyde (29.66%) as main constituents. Ten components and eight compounds were identified, representing99.87%of the essential oils of branches with leaves, the major componts were benzaldehyde (90.68%), benzyl alcohol (2.95%), benzoic acid (4.17%) and mandelonitrile (0.07%). Seven compounds were identified, representing99.83%of the essential oils of outer part of pericarp, the major constituents include benzaldehyde (87.15%), benzyl alcohol (6.51%) and mandelonitrile (5.66%).(2) The analysis method of polysaccharides with pertinence to the residue of Malania oleifera was established. Based on the results of single factor experiment, response surface method (RSM) was used for optimizing ultrasound-assisted extraction of malania polysaccharides. Results showed that the optimized conditions of ultrasonic power490W, extraction time45min and ratio of solid to liquid as1:25, under such circumstances, the yield of polysaccharide was5.02%. The crude polysaccharides were purified by alcohol sedimenting technique, sevag method (deproteinization), macroporous-polyamide resin (decoloration), DEAE cellulose-52chromatography and sephadex G-100column chromatography in that order. Two main fractions, GP-1and GP-2, were obtained through the extraction and purification steps. High performance gel permeation chromatography (HPGPC) of the individual fractions showed that each fraction was represented by a narrow peak on the chromatograms. The average molecular weights of the two fractions were determined to be42.95kDa and2597kDa for fractions GP-1and GP-2, respectively. The GP-1was composed of glucose, rhamnose, and arabinose in a molar ratio of5.8:3.2:1while GP-2was composed of glucose, rhamnose, arabinose, mannose and fructose in a molar ratio of13:3.4:1.6:1:1. Furthermore, the influence of fractions GP-1and GP-2upon antitumor activities to the human nasopharyngeal carcinoma (CNE-1) cells and lung cancer (H460) cells in vitro were evaluated. The results indicated that GP-1and GP-2played an inhibited effect on the cancer cells and didn’t kill human normal liver cells (HL-7702). Growth inhibition rate of GP-2against CNE-1cells was higher than that of GP-1and the maximum inhibition rate reached64.49%.(3) The extraction of total flavonoids from Malania oleifera leaves and its residue after volatile oil extraction were studied with the methods of solvent extraction. Results showed that the yields of total flavonoids from M. oleifera leaves and its residue were16.78%and12.31%under the optimized conditions, respectively. The purity of flavones were79.80%and78.62%increased by39.19%and43.97%respectively from crude extract flavones of Malania oleifera leaves and its residue after purification by HPD400resin. Rutin has been firstly purified from its crude extracts from Malania oleifera by HPLC. The confirmation of chemical structure was performed by MP, IR, MS,’HNMR and UV spectroscopy. The purity of rutin reached95.59%by analytical HPLC. The antioxidant, antilipooxidant and antimicrobial activities of Malania oleifera flavonoids were investigated. The result showed that the scavenging activity of total flavonoid separated by HPD400on DPPH·were better than that of crude extract, rutin and tannic acid, the highest scavenging rates of purification of total flavonoids from leaves (residues) reached89.78%and91.69%in the cocentration of1000μg·mL-1, respectively. The purification of total flavonoids from malania leaves had very strong antioxidation on lard. Bacteriostasic experiments showed that purification of total flavonoids have bacteriostatic bioactivities against Monilia albican, Salmonella, Staphyloccocus aureus to some extent, the bacterial inhibition diameters of samples on Staphylococcus aureus were greater than12.0mm. Rutin palyed a significant role in antioxidation and antimicrobial activity.(4) Lignins were extracted from Malania oleifera shell and branch by traditional alkaline method, ultrasound-assisted method and microwave-assisted method. The yields and the contents of lignins in Malania oleifera shell and branch were determined. Meanwhile, the lignins obtained were structurally analyzed by infrared and (IR) and UV spectroscopy. The contents of lignin in M. oleifera shell and branch were44.41(±0.65)%and34.57(±0.52)%respectively by Klason method, the lignins extraction rates reached45.21(±0.59)%and63.78(±0.73)%respectively from Malania oleifera shell and branch under the conditions of ultrasound-assisted extraction at40℃water bath for1hour with 0.5mol·L-1KOH water as the solvent and the solvent/material ratio of30:1(mL·L-1). UV and IR spectral analyses showed that the lignin extracted by ultrasound-assisted method remained its original structure, there were many syringyl and guaiacyl groups in Malania oleifera shell and branch. And compared the relative molecular weight of lignins extracted by three different extraction methods with viscosity method.(5) The antioxidant components were extracted by reflux extraction from peel and flesh of Malania oleifera and its antioxidation were studied under two different situations. The results showed that the polyphenols from Malania oleifera flesh extraction rate reaches3.29%under the optimized conditions of extraction at70℃water bath for1hour twice with water (pH4) as the solvent and the material/solvent ratio of1:30(g·mL-1). The clearing capacity of polyphenols for·OH and DPPH·were very significant by scavenging test, were74.00%and76.54%respectively, in the order of clearing capacity for DPPH·were water extraction> ascorbic acid> tannic acid.(6) The content of protein was measured, and single factor experiment and orthogonal factor experiment were carried out to optimize the technology of extraction, which was used to extract the protein from Malania oleifera residue. Results showed that the content of protein in Malania oleifera residue was31.07%, the optimum conditions were as follows:The extraction temperature is room-temperature, the extraction solvent is phosphate buffer solution, and pH=7.5, the extraction time is3hours and the ratio of material to liquid is1:25. Under this circumstance, the extraction rate reaches86.01%.(7) It was revealed that abundant resources were hidden in Malania oleifera by the means of components analysis of fatty acids, essential oils, polysaccharides, flavonoids, proteins, lignin and antioxidant of polyphenols. Therefore, research carried on Malania oleifera has great significance. |
