The Separation And Purificition Of Stigam Maydis And Abandoned Tobacco By High-Speed Countercurrent Chromatography

Stigma maydis is an important Chinese medicinal material, which is often used to treat Diabetes Mellitus, jaundice and nephritis. One of the significant characteristics of the flavonoids is clearance the free radicals, and also can inhibit the activities of large quantities enzymes. Flavonoids as the main active constituent of many herbs, has been studied for long time.At present, Al(NO₂)₃ coloration method rutin as the standard is one of the ordinary method for deteminating flavonoids in food and herb. The common separation technologies are column chromatography, centrifugal TLC, thin layer chromatography and paper chromatography.Massive abandoned tobacco leaves and dust have been wasted. Solanesol is an important pharmaceutical intermediate which has the strong antitumor bioactivity. It is also the important material of drug synthesis and the irreplaceable pharmaceutical intermediate for ubiquinone. Solanesol only can be obtained from plant leaves.In this thesis, the extraction and systematic isolation of Stigma maydis and abandoned tobacco by high-speed Countercurrent Chromatography have been described.By arthogonal method, the optimized extraction condition of Isorhamnetin from Stigma maydis is: 80% ethanol solution, 1:8 solid-liquid ratio, 3 hours for 2 times. The total flavonoids content of the extraction is 9.07%, the isorhamnetin content is 1.53%.Preparative high-speed countercurrent chromatography (HSCCC) has been used successfully for the isolation and purification of Isorhamnetin from stigma maydis. This was achieved in two stages: firstly HSCCC protocol with a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (HEMW) at volume ratio of 5:5:5:5, v/v was used and Isorhamnetin of 65.64% was obtained, secondly HEMW 5:5:6:4, v/v was used to resolve the components from the first stage. 700 mg amount of the crude extract was separated, yielding 11.8 mg at a high purity of 98%.Preparative HSCCC was used for Solanesol separation from crude extract of the waste tobacco leaf with a nonaqueous solvent system composed of n-hexane-chloroform-acetonitrile at an optimum volume ratio of 10:3:7. The solanesol purity is over 96% as determined by HPLC analysis.