Study On The Extracting Technology Of Acehytisine Hydrochloride From The Hairy Roots Of Aconitum Coreanum

Aconitum Coreanum（Levl.） Rapaics species belongs to Aconitum genus in the Ranuculaceae family,as well as has the effect of Qufeng desiccant, warming meridians and relieve pain. A series ofpharmacological studies demonstrated that Guanfu base A （GFA）, a naturally occurring diterpenealkaloid originally isolated from the root of Aconitum coreanum, has been proven to exhibit significantanti-arrhythmic activity. Hydrochloric GFA received the certificate of State Category I New Drug issuedby the Chinese Health Ministry. In this paper, the hairy roots of Aconitum coreanum were systematicallyresearched, including extraction, separation, qualitative and quantitative analysis of GFA. Pulsedelectric field （PEF）, percolation extraction, ultrasonic-assisted extraction, heat reflux extraction, organicsolvent extraction, macroporous resin extraction, cation exchange resin extraction, aci dye colorimetry,LC-MS, which are used in experiments. Meanwhile, the pilot production process of acehytisinehydrochloride was also tested. The results are as follows:1. Compared to the other three extraction methods consisting of percolation extraction,ultrasonic-assisted extraction and heat reflux extraction, it indicated that the highest yield of GFA andextract yield was up to3.91‰and39.32%by PEF with the optimal formulation conditions of20kV·cm-1electric field intensity,8pulse,1:14solid-to-solvent ratio and90%ethanol-water solution.Meanwhile, the extraction time of PEF was less than1min, without exposure to high temperatureswhich could effectively avoid changing target material into other substances.2. The total alkaloids were determined at410nm by the method of improved acidic dyecolorimetry from materials of the hairy roots of Aconitum coreanum, and the optimum extractionconditions were confirmed by single factor tests. The results showed that the best test conditions wasthat bromothymol blue as chromogenic acid dye was2mL, pH of buffer solution was8.0,5mL. Thecalibration curve was linear in the range of8.7μg·mL^-1-43.3μg·mL^-1, and the regression equation beingy=0.0459x-0.0126（r=0.9911）. The average recovery was98.8%（RSD=0.516%）. The content of GFAwas determined by HPLC, the calibration curve was linear in the range of0.10μg-1.00μg, and theregression equation being y=15.97x-1.7241（r=0.9924）. The average recovery was100.86%（RSD=0.96%）.3. Separation and purification of GFA were studied in the hairy roots of Aconitum coreanum, theresults showed that organic solvent extraction, using methyl tert-butyl ether （MTBE） as the mainextraction solvent, had a large advantages over macroporous resin extraction and cation exchange resinextraction, such as simple, low cost, less dosage of the acid and alkali, short productive cycle and goodeconomical benefit. In addition, the yield of GFA was2‰by organic solvent extraction which could beapplied in the industrial production of GFA from Aconitum coreanum.4. Using the tuberous root of Aconitum coreanum in place of the hair root for the pilot test, whichshown that based on the PEF extraction for total alkaloid, MTBE as the main extraction solvent, theextraction technology was stable, feasible and then to determine the feasibility of industrialization.