Preparation And Analysis Of Diosgenin From Dioscorea Zingiberensis

Diosgenin is an important precursor for synthetic production of steroid drugs. Inthis paper, the detection, extraction and preparation of diosgenin were studied, mainconclusion is as flow:1. The thin layer chromatogram(TLC) were developed. The optimal conditionwere as flow: silica gel G as stationary phase,chloroform- acetone(97:3,V/V) as themobile phase, coloration by 4% molybdenum phosphate in ethanol. Under thesecondition, a good separation was obtained, and the Rf of diosgenin was 0.43.2. A method of the reversed phase high performance liquid chromatographicdetermination of diosgenin in plant was developed. The optimal HPLC conditionswere as follow: Kromasil C18 column as the stationary phase, methanol as the mobilephase, 1mL/min of flow rate, UV detector(208nm). Under these condition, a goodseparation was obtained, and the retention time of diosgenin is 9.3 minnutes. Thecalibration equation is y=258440x+46992, the related coefficient is 0.9993. The linearrange is 3.876-19.38μg.the RSD of system is 1.75. The average recovery is 96.21%.The method is accurate and reproducible to determine the sample.3. A method of the reversed phase high performance liquid chromatographicdetermination of diosgenin in cultivated cell was developed. The optimal HPLCconditions were as follow: Kromasil C18 column as the stationary phase,methanol-water(95:5,V/V) as the mobile phase, 1mL/min of flow rate, UVdetector(208nm). Under these condition, a good separation was obtained, and theretention time of diosgenin is 21.3 minnutes. The calibration equation isy=354944x-39538, the related coefficient is 0.9996. The linear range is 3.876-19.38μg.the RSD of system is 1.46. The average recovery is 97.83%. The method is accurateand reproducible to determine the sample.4. Ternary phase(sulfuric acid isopropanol and water) was studied by conditionof single factor methodology. On the basis, Central Compsite Design and ResponseSurface Methodology were employed to optimize sulfuric acid concentration, iisopropanol concentration, hydrolyzing time and hydrolysis liquid dosage.The optimalcondition were as flow: sulfuric acid concentration was 2.5N, isopropanolconcentration was 75%, refluxing time was 8 hours, hydrolysis liquid was 25mL. Theextraction rate was over 68% than traditional hydrochloric acid hydrolysismethodology.5. By combination of silica column chromatography and preparative highperformance liqiuid chromatography, diosgenin purity was 97.12%. The product wasidentified as diosgenin by melting point ,IR, MS and HNMR.