Study On The Preparation Process And Quality Standards Of Senecio Cannabifolius Extract

Objective: To select the decoction extraction process of Senecio cannabifolius extractthrough the orthogonal design，and to establish the quality standard of Senecio cannabifoliusextract, also to provide the reference for the research on the standardization of qualitystandard and the industrial production.Methods: Using the extraction time （A）, the solvent multiples （B）, the number of extraction（C） and the extraction temperature （D） as four factors and three levels of the orthogonalexperimental design was made for the decoction extraction process. As an indicator, thecontent of chlorogenic acid and coffee acid were determined by using HPLC.Chromatographic conditions： acetonitrile-0.05%trifluoroacetic acid solution （12:88） asmobile phase，the chromatographic column was AlltimaTMC18（250mm×4.6mm,5μm）, thedetection wavelength was327nm, the column temperature was21±2℃, the flow rate was1.0ml·min^-1, the injection volume was20μl.The quality standard of Senecio cannabifolius extract was established by the thin layerchromatography and fingerprint analysis as qualitative methods, and the high performanceliquid chromatography as quantitative methods.The stationary phase of Thin layer chromatography was silica gel plate, The mobilephase（developing solvent） was the upper solution of butyl acetate-formic acid-water（7.5:2.5:1）.The fingerprint was established by using high performance liquid chromatography, thechromatographic conditions: the chromatographic column was Alltima TMC18（250mm×4.6mm,5μm）, the mobile phase was0.05%three fluorine acetic acid aqueous solution （A）-methanol （B）, the elution mode was gradient elution, elution program:0-5min95%-95%A，5-15min95%-90%A，15-25min90%-90%A，25-40min90%-88%A，40-60min88%-85%A，60-70min85%-84%A，70-80min84%-82%A，80-90min82%-80%A，90-100min80%-80%Aï¼›the detection wavelength was327nm, the column temperature was21±2℃,the flow rate was1.0ml· min^-1, the injection volume was20μl. Using the peaks ofchlorogenic acid and caffeic acid as the reference peak, the chromatographic fingerprint dataof the10batches of samples was determined, and then the analysis was made by the similarityevaluation system of chromatographic fingerprint （2004A）.The content of chlorogenic acid and coffee acid as reference was determined by usingHPLC, and quantified by external standard method. Chromatographic conditions： thechromatographic column was AlltimaTMC18（250mm×4.6mm,5μm）, acetonitrile-0.05%trifluoroacetic acid solution （12:88） as mobile phase，the detection wavelength was327nm,the column temperature was21±2℃, the flow rate was1.0ml·min^-1, the injection volumewas20μl.Results: The optimum extraction conditions of Senecio cannabifolius extract were16timesthe volume fraction of water at100℃for2.5h, extracted for two times, under this condition,it can obtain higher extraction rate of coffee acid and chlorogenic acid.The qualitative identification of TLC in the quality standards for Senecio cannabifoliusextract was the good separation, strong specificity. The test sample and the control in thecorresponding position showed the same spot.There were about20chromatographic peaks in the chromatographic fingerprint of Senecio cannabifolius extract, among them there are9common fingerprint peaks, chlorogenicacid used as reference peak （peak S1, peak3）, the relative retention time of the other8common fingerprint peaks were:0.384,1.090,1.576,2.127,2.238,2.434,2.520,2.751, andthe relative peak area were:0.358,0.311,1.326,1.351,1.693,0.958,0.671,0.574. Thesimilarity among common mode for the fingerprints data of the10batches of the samples wasmore than0.9.On the determination of the content of Senecio cannabifolius extract, the linearrelationship of chlorogenic acid in0.14μ g～1.7μ g was good, R=0.9972, and the averagerecovery was98.41%, RSD=2.1%（n=6）, the linear relationship of coffee acid in0.02μ g⁰.25μ g was good, R=0.9999, and the average recovery was95.49%, RSD=1.3%（n=6）. Thecontent of chlorogenic acid was2.75mg·g^-1, and coffee acid was0.15mg·g^-1Conclusion: The decoction extraction process of Senecio cannabifolius extract is simple,stable, and has high extraction rate. The qualitative identification of TLC in the qualitystandards for Senecio cannabifolius extract is simple, stable, specificity and reproducibility,and can be used as a qualitative method for its quality control. HPLC chromatographicfingerprint can be used for the identification and quality control of Senecio cannabifoliusextract. On the determination for the content of chlorogenic acid and coffee acid in Seneciocannabifolius extract, the result is accurate and reliable, reproducible, and can be used for thequality control of Senecio cannabifolius extract.