Extraction, Purification And Pilot Process Study Of Glycyrrhizae From Swertia

Fish bile grass(Swertia davidii), the official said in Swertia davidii, it is an ethnic tujia folk medicine,which mainly contains iridoid class ingredients saponins and flavonoids, it is the main point of swertiamarine efficacy plants, which swertiamarine content accounted for about 3% of the original medicine.swertiamarine as seco iridoid components is a major component in Gentianaceae. It has the effect of inhibiting the release of acid phosphatase, can prevent liver injury caused by hypoxia in rats When the concentration of 50?g?m L-1 has the inhibitory effect on the liver cancer cells [1], at the same time it also has antioxidant, sedation, analgesia, eliminate edema, anti-inf lammatory and free radical scavenging effects.But now the literature of high purity swertiamarine prepared were less reported, according to the existing literature: using HP-300 purif ied from gentian ir idoid, column eluted swertiamarine mass fraction 2.44%, Zhou Dacheng etc. Studies have shown that HPD-100 macroporous resin type has good purification effectof swertiamarine, dried powder sample content is 52.29%.Xiao-bing fan in S-8 macroporous resin purification swertiamarine content can reach more than 50%.Marina using HPD-300 resin,in 20% ethanol elution mass fractionof the swertiamarine in the sample is 50% or higher.Existing literature on purification, the purified swertiamarine content is not high, this artic le will develope a preparation process of high-purity swertiamarine( mass fraction> 90%)and pass the pilot study by extraction, macroporous resin chromatography, activated charcoal column chromatography, at the same time test the activities of samples which after a macroporous resin chromatography and activated carbon chromatography. the main test content is as follows:(1) the optimization of extraction process conditionsin swertiamarine extraction rateas the index, based on the extraction tim e, extraction solvent, extraction temperature, extraction and liquid ratio conditions make single factor and orthogonal test to determine: the best extraction process of swertiamarineis solid- liquid ratio 1:10, temperature 60 ?, 10% ethanol concentration, extraction time is 10 min, according to this method, the extraction yield reached 88%(2) the purification process researchscreeningsix kinds of various macroporous resin, AB-8 macroporous resin was selected to separateswertiamarine.The diameter height ratio is 1: 8 loading at 80% of the adsorption capacity.first eluted with water to remove water soluble impurities, then eluted with 20% ethanol and finally collect fluid and rotary evaporated to dryness.the yellow powder extract's mass fraction is 60-80% during the process that Macroporous resin adsorption and elutionthe recovery is 85-95%. Put Good pretreatment of activated carbon and diatomite mixture into the glass column, take sample dissolved on 5 times the amount of water, than loading,standing overlight,with 2 bv water removing impur ity, then use methanol eluted14.7 bv, recovered solvents to dry, get solids, aftercrystallized with anhydrous ethanol,sample's content > 90%.(3) the pilot process debuggingcall pilot equipment in biochemical engine ering center in guizhou.first feeding 50 kg medicinal materials extractand concentrate, resin?the dosage of activated carbon, solvent dosage scaling up, the extract equipment should be washing with ethanol?waterbefore use.in the process of extracting temperature should not be warming too fast. with rubber gloves to avoid moisture absorption dry sample transfer.In this process sample's contentis more than 88%, the total recovery is about 58%.(4) biological activity testBy enzyme-linked immunosorbent assay(ELISA) to detect macrophage tumor necrosis factor(TNF alpha) secretion expression. add the sample under test, incubation develop after a certain time, cells that determine the content of TNF alpha supernatant fluid, determine the sample activation inhibition of cell. After LPS was added to macrophage training after a certain time, and then the sample was added, incubated for a certain time, taking the supernatant fluid, TNF-? determination, determine the sample activation or inhibition function to cells.In summary paper completed the Swertia bitter glycosides extraction conditions, macroporous resin and activated carbon filter packing, explored a cheap filler material was purified by high purity Swertia bitter glycosides way through commissioning and pilot materials costing determine the feasibility of this process. And the study found that high-purity extract has anti- inflammatory activity. These results will be fish guts grass Swertia bitter glycosides lay the foundation for comprehensive development and utilization.