Purification Of Triterpene Saponins With Bioactivities From The Seeds Of Camellia Oleifera Abel

Camellia oleifera Abel (Oil tea), a plant of Camellia L., which originated in China, well known as an important source of edible oil obtained from its seeds. It is widely distributed in south China, of which the roots, stems, leaves, flowers and seeds were used as tranditional Chinese medicines. According to the National Camellia Industry Development Plan (2009-2020), issued by the National Development and Reform Commission and other ministries, the total size of the planting of Oil tea in China will reach 4.7 million hectares in 2020. Oil tea seeds after oil extraction(oil tea seed cakes) will exceed 700 million tons. Currently, oil tea seed cakes are used as pesticide and fertilizer in a small scale, lots of oil tea seed cakes were abandoned. Extract tea saponin from oil tea seed cakes is the way of the comprehensive utilization of waste. Tea saponin can be applied to many fields such as industry, agriculture, which in high purity also can be used in the pharmaceutical industry. It is not only increase farmers’income, but also has advantage to the environment, resulting in considerable economic and social benefits.Chemical constituents and bio-activities of Camellia L. have been reported numerously, and it is suggest that abundant of triterpenoid saponins as the main active ingredients in Camellia L, which have gastroprotective effect, inhibitory activity on alcohol absorption, anti-inflammatory activity, antiallergic activity, anti-hyperlipidemic activity, inhibitory effect on melanogenesis, etc.,131 tea saponins have been isolated from the Camellia L., and 13 saponins are reported from Camellia oleifera Abel,5 saponins were identified in its root and flower respectively. Only 2 new and 1 known saponins were isolated from oil tea seeds, and the activities of them were rarely reported. Therefore, in order to improve the comprehensive utilization of oil tea seeds, it is necessary to do purification and related activity research of saponins in oil tea seeds.Our research was mainly concentrated on the triterpenoid saponins in seeds of Camellia oleifera Abel, harvested from South of Anhui Province. This work will contribute significantly to the systematically understanding for the type and activity of saponins in seeds of Camellia oleifera Abel, and promote comprehensive utilization of seeds resources to develop new pesticides and new medicines.1. Oil tea seeds were defatted and extracted by 70% ethanol, then isolated by chromatograph of silica gel, ODS, Pre-HPLC, sephadex LH-20, etc. The structures of isolated compounds were characterized by spectroscopic analysis (ESI-MS, NMR, UV). One tea seed saponin called camelliasaponin B1 was identified, and three other saponins are purified and the structures of them are still in analysing.2. Column chromatographic conditions on separating and purifying saponins in oil tea seeds were established. According to the UV spectrums of oil tea seeds saponin monomers, 278 nm was selected as UV detection wavelength in HPLC method for identification and determination saponins. It will provid a reference for determination of total saponins in oil tea seeds, also laid the foundation for the research of Camellia seeds saponins.3. The killing activities of crude extract of seeds, macroporous resin fractions and total saponins in oil tea seeds to Meloidogyne spp. were studied. The results showed that crude extract of seeds and total saponins in oil tea seeds had strong killing activities to root-knot nematode,50% ethanol macroporous resin fraction had a weak activity, and no activities with the other components.4. The results of orthogonal experiments, on Bursaphelenchus xyluplilus catalase and carboxylesterase activities of saponin, showed that under the concentration of 1000 μg/mL total saponins in oil tea seeds had maximum activity for both enzymes and the other factors had less effects.5. Total saponins and camelliasaponin B1 both had strong cytotoxic and anti-tumor activity. The results showed that the inhibition rate of camelliasaponin B1 were good in 4 human tumor cells, the 50% inhibition concentration (IC50) of human nasopharyngeal carcinoma KB cells, human leukemia HL-60 cells, human gastric cancer BGC-823 cells and human hepatoma Bel-7402 cells were 16.54 μmol,18.07 umol,25.79 μmol and 17.65 μmol, respectively. Total saponins showed sensitivity against human gastric cancer BGC-823 cells with IC50 to be 5.98 mg/L, the IC50 of human leukemia HL-60 cells, human hepatoma Bel-7402 cells and human nasopharyngeal carcinoma KB cells were 12.54 mg/L,11.02 mg/L and 19.76 mg/L, respectively.6. Total saponins and camelliasaponin B1 under the concentrations of 10 mg/mL and 10 μmol showed significant protective effect on PC 12 cells injured by hydrogen peroxide respectively. The protective activity was positive correlated with the concentration of camelliasaponin B1. Also total saponin showed good protection at concentration of 0.1 mg/mL, but the protective effect had not showed the relationship to the concentration of saponins.The type and content of saponins are rich in tea seed oil, with varieties of bio-activities which are confirmed by the experiments. It was believed saponins have further study of the meaning and application value.