Study On Extraction And Inhibitory Effect Of Pulsatilla Saponins From

Traditional Chinese medicine of Pulsatilla is the dried root of Pulsatilla chinensis(Bge.) Regel, contained in the "Shen Nong’s Herbal Classic".Its bitter taste,quantity of cold, go through the stomach, large intestine.It has the effect of detoxification, diarrhea, dampness insecticidal. Modern pharmacological studies have shown that it has enhanced immune function, anti-inflammatory, anti-oxidation,anti-tumor activity against pathogenic microorganisms and other pharmacological effects. In this paper, we regard Pulsatilla chinensis as raw materials, content of pulchinenoside B4 as the index.The ultrasonic assisted solvent and microwave assisted solvent method were used to extract, using the response surface methodology to optimize methods. Then vitro antibacterial activity of pulchinenoside was studied.Specific results are as follows:(1) Set up a method for determination of the content of pulchinenoside B4.Performed by HPLC assay to select mobile phase of acetonitrile- water- phosphoric acid(27: 73: 0.1, v / v), under the conditions of column temperature 25℃, eluted with a flow rate of 1.0 m L ? min-1, detected wavelength 201 nm, each injection volume of10 μ L, In this liquid phase conditions to determine the content of pulchinenoside B4,The results showed that the relationship is good from 0.025 to 0.6 mg m L-1.Correlation coefficient R2= 0.9996, The accuracy and the precision of this method are in line with the requirements of pulchinenoside.(2) 25 parts from different origins of Pulsatilla were purchased.There are 16 copies of Pulsatilla chinensis by micro structural identification. By using the way above HPLC, we determine the content of pulchinenoside B4.The results showed that the highest content of it is from the Northeast region. So take it as further experimental material.(3) The extraction of pulchinenoside by ultrasonic assisted solvent extraction and microwave assisted solvent method, effects of solid-liquid ratio, ethanol concentration,extraction power and other factors affecting the extraction rate are inspected, and the response surface to each method was optimized. The extraction rate of pulchinenoside is 5.45% and 5.33% with the microwave and ultrasonic method. For the first time, the new scheme of ultrasonic microwave assisted solvent extraction of pulchinenoside is proposed and experimentally. The conditions are confirmed as follows: the solid-liquid ratio 1:8, ethanol concentration 60%, ultrasonic power 240 W, microwave power of 300 W, firstly ultrasonic extraction within 15 minutes, secondly microwave extraction within 2 minutes, then repeating once. Under these conditions, the yield of pulchinenoside is 6.77 %.This method can significantly improve the extraction rate.(4) Extract the pulchinenoside by the best method in this paper and bacteriotasis experiment in vitro is carried out.Then deal with Escherichia coli, enteric bacilli,achromobacter, pseudomonas aeruginosa, klebsiella pneumoniae by the filter paper method. The results showed that these inhibition zone diameter are 13.66±0.21 mm、19.25±0.31 mm、16.79±0.26 mm、15.69±0.17 mm、14.87±0.23 mm.