| It was shown that ruscogenin had excellent prospects in pharmaceutical field because of its effects in anti-inflammatory, anti-thrombotic, anti-tumor, etc. The 2010 version of “Chinese pharmacopoeia” included the ruscogenin as quality evaluation standard material for radix ophiopogonis. Recently, the biological activity and potential clinical value of ruscogenin had been recognized gradually, which rapidly increased the demand for ruscogenin.At present, the ruscogenin was prepared by separation and purification using medicinal herbs radix ophiopogonis as raw material. However, the problem was the high preparation cost for ruscoginin because of its low content in radix ophiopogonis and comprehensive utilization. On the other hand, radix ophiopogonis as a kind of “medicine food homology” contained various effective components group, and possessed individual development feasibility. Based on the comprehensive utilization of radix ophiopogonis, the paper created a line of industrialization for the preparation of ruscoginin, which could provide various products.Firstly, the extraction process of total saponins and polysaccharide was optimized. The parameters to be optimized were extraction by 6 times volume of the 80% ethanol for 3 times, and every time the extraction process for 1 hour, 0.44% total saponins and 64.46% total polysaccharide of radix ophiopogonis were obtained.The alcohol precipitation experiments of polysaccharide carried out for the separation of saponins and polysaccharide. The density of alcohol precipitation solution was 1.20 g/mL, adjusted the concentration of ethanol to 75% and rested at room temperature for 16 h. The total saponins metastasis rate of radix ophiopogonis was 91.20%, yield of dry extract reduced 76.97%, the total saponins increased from 0.25% to 1.27%, and 656 mg ophiopogon polysaccharide was obtained for every gram materials.The macroporous resin was used for the further purification of radix ophiopogonis. According to the test result, the D101 resin was chosen for the experiment, the height-diameter ratio was 12:1, flow rate was 2 BV/h, the pH value of injection solution was adjusted to 2.47 by HCl, sodium sulfate was added 3% and the sample injection condition was 7 BV as 500 μg/mL total saponins solution. The leak rate was less than 10% after repeated experiments. The desorption conditions were determined that washed 3 BV by water and then eluted 6 BV by 80% ethanol with flow rate of 3 BV/h. Experiments showed that the elution rate of total saponins was more than 90%. The recycle rate was much more than 80% when the purification process was scaled up 6, 10 and 20 times. 3.21 mg total saponins was prepared for every gram materials and the purity increased to 58%.Ruscogenin was prepared by acid hydrolysis, and the hydrogen ion concentration of the system was adjusted to 0.4 mol/L using sulphuric acid and reaction for 2.5 h at 105 ℃. Ruscogenin of 57.93 μg was obtained using the concentrated solution(836.15 μg/mL) of total saponins per milliliter. The column chromatographic technique was applied in the purification of Ruscogenin, and the parameters to be optimized were the height-diameter ratio of 12:1 and the binary eluent( V(n-hexane):V(ethyl acetate)=1:1) with 3-3.5 BV eluate. The purity of ruscogenin from column chromatographic technique improved to 40%. The chromatographic conditions were as follows: the determine wavelength of 205 nm, the flow rate of 60 mL/min, injection content of 10 mg and the mobile phase being neat methanol.In this paper, ruscogenin process with higher comprehensive utilization rate of materials and lower production cost was established. 200 g ruscogenin with the purity of 58% was obtained for every gram materials, when polysaccharide was obtained also. In addition, this process can meet different needs with the multiple products at different purity, and provide technical support for product development of Radix Ophiopogonis. |
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