Study On The Extraction, Separation And Determination Of Sequoyitol From Taxus Media

Taxus media, a natural hybrid between Taxus cuspidata (female parent) and Taxus baccata (male parent), belongs to taxaceae taxus. Paclitaxel and docetaxel (synthesized by10-Deacetyl Baccatin Ⅲ,10-DAB) become the primary anti-cancer drugs widely used in the clinic, which leads to the economic value of taxaceae increased sharply.It can not meet the need of the social demand for Paclitaxel and10-DAB extracted from wild taxus. In addition,the wild yew endangered plants limits the source of Paclitaxel and10-DAB. In1996, China State Forestry Administration succeed to introduce and cultivate Taxus media for many years’trials. At present, it supplies ample resource for medicinal and research because of a number of Taxus media planting base. In recent years, sequoyitol found in taxaceae becomes another research focus, because it can lowers blood sugar and has potential development into a new treatment of diabetes drugs. The methods of isolation, purification and detection for taxanes are more mature. Nevertheless, there are less reports on sequoyitol to the public and many outcomes form patents to be protected. The study on the extraction,purification and detection of sequoyitol from Taxus media has not been reported yet in literature. It makes the study more meaningful to introduce and cultivate Taxus media successful in China. Therefore, the research is influential and significant. This paper carries on systematic experimental on sequoyitol involved in extraction and purification from Taxus media. Extraction conditions were optimized. In this study, both qualitative analysis and quantitative analysis of sequoyitol are established. The main results as follows:(1) Orthogonal design was used to optimize the extraction conditions of sequoyitol from Taxus media. The extracting test of orthogonal design L9(34) with branches and leaves of Taxus media for the extraction materials, three times extraction, considering solvent concentration (ethanol-Water), temperature and time as three factors and extract of sequoyitol charge for evaluation indicators established the optimization selection in extraction conditions of sequoyitol.It was follows:30%ethanol as solvent, extraction time2.5h-2.5h-2.5h, extraction temperature of65℃, three times extraction. The sequoyitol rate of extract is0.150%.(2) In the process of separation and purification, the branches and leaves of Taxus media extraction contains a lot of wax, pigments and other compounds, which lead to difficult to separate and purify. With that in mind,the research was designed with80%ethanol extraction(to product taxane) to remove most of the impurities of the pigment, wax, etc., then preferably obtained through orthogonal test process of extraction.The extraction solution by activated carbon chromatography is further processed to reduce pigment, reducing the adverse effects of recrystallization steps.(3) From the experimental result (by inositol instead of sequoyitol), it was suggested that the conductance of the sequoyitol aqueous solution is conjectured to be too lower to analysis.The result has important significance to monitor impurity such as amino acid, protein and other large conductance in step of chromatographic separation.(4) Established quantitative analysis of sequoyitol,named quantitative analysis method for "SPE-spectrophotometry-Zeisel determination method". By the method for determinating methoxyl content, we can calculate the content of sequoyitol. It has the advantage of good accuracy, high accuracy, stable, reliable and so on.(5)"Iodine-starch complex spectrophotometeric quantitative analysis" in "SPE-Spectrophotometry-Zeisel determination method"materially belongs to"Indirect iodine-starch spectrophotometric quantitative analysis".And we explain it by the dominant theory, which is a basic research results and has a more extensive application value.(6) Established qualitative analysis of sequoyitol to fill a gap in the method qualitative analysis of sequoyitol.(7) By Separation and Purification processes, the purity of sequoyitol sample reaches99.5%through detection of HPLC-PDA and MS result corresponds to CAS. The product can be used as quantitative reference standard and bring convenience for the research work.