Study On The Preparation Of The Anemones Raddeanae Rhizoma Cataplasm And Its Quality Standard

Object:To lay a foundation for the development of a kind of new Chinese drugs for treating rheumatoid arthritis by the pharmaceutical study of the Anemones Raddeanae Rhizomacataplasm.Methods:1.Study on the extraction processThe yield rate of the dry extract and the content of the raddeanin A that was determined by HPLC were used as the evaluation index, three extration methods were compared to optimize the extraction process of the Anemones Raddeanae Rhizoma, that is, water-extracting technology, refluxing extraction technology with different concentrations alcohol(optimization by L9(34)orthogonal test) and the percolation process (optimization by L9(34)orthogonal test).2.Study on the purification processThe yield rate and the content of the raddeanin A that was determined by HPLC were used as the evaluation index to investigate the purification technological parameter of5kinds of macroporous adsorption resin (MAR) that were used to purify total saponins of Anemones Raddeanae Rhizoma, and the purification ability of the5kinds of MAR were compared to choose the best method.3.Study on the preraration process for Anemones Raddeanae Rhizoma cataplasmL18(3’)orthogonal test was used to optimize the preraration process for Anemones Raddeanae Rhizoma cataplasm and the viscosity, smear and expansion ability, appearance and peeling strength of the Anemones Raddeanae Rhizoma cataplasm samples were used as the evaluation index.4.Study on the quality standard and stability of the Anemones RaddeanaeRhizoma cataplasmTLC was used to identify the raddeanin A in Anemones Raddeanae Rhizoma cataplasm. HPLC was used to determine the content of the raddeanin A, and the chromatographic conditions were:The Shim-pack VP-ODS(4.6mm×150mm) column by a gradient elution using acetonitrile (A) and0.1%phosphoric acid solution(B) as the mobile phase and flow rate of1.0mL/min, the elution process was presented in the table below; the detection wavelength was at206nm; column temperature was35℃. To examine the content of ointment, adhesion, formability and the weight diversity. The preliminary study on stability of the preparation was finished with the characteristics of the preparation, qualitative identification, content of ointment, adhesion, content of the raddeanin A as the evaluation index.5.Study on the release rate and percutaneous rateThe release rate and percutaneous rate of raddeanin A in Anemones Raddeanae Rhizomacataplasm were determined by HPLC.Results:1.The optimised extraction and purification process were:Coarse powder,10times of80%ethanol, refuxing and extracting for2times,1.5h for every turn, the extracts were collected together and concentrated to4.83μl/mL, and then added to the resin column with the flow rate was2times colunm volume per hour, the ratio of the diameter and the height of the resin bed wasl:8, eluted the colunm with7times70%ethanol and the elution rate was2times colunm volume per hour. The content of the total saponins in the eluate could reach81.21%, and the recovery rate was82.65%.2.The optimised preraration process for Anemones Raddeanae Rhizoma cataplasm were:PANA:PVPK-30:glycerin:poval:gelatine:MCC:azone:extract powder=3:2:4:1:1:0.2:1:4.3.The results of the quality standard were:(1)The TLC method which was used to identify raddeanin A was simple and convenient, and the chromatogram was clear. (2)The results of the content determine:the linear range of raddeanin A was2.3μg～11.5μg.(3)The results of the precision experiment:the precision of raddeanin A (RSD) was1.86%,(4)The results of the recovery rate:the average recovery rate of raddeanin A was99.68%, RSD=1.77%.(5)The results of the reproducibility:the average content of raddeanin A was0.65%, RSD=1.84%. The content of raddeanin A in the preparation was used as the quality control index, and we suggested that its content in the preparation should temporarily be limited to8mg per sheet of the Pasqueflower. The content of the ointment, adhesion, weight diversity, formability were all conformed to the stipulation.4.The results of release rate and percutaneous rate study:the release rate of rddeanin A of the preparation in water was98.13%, and the accumulated percutaneous rate in vitro (rats) during36h was0.68%.Conclusion:The research contents of this study contained preparation research of Anemones Raddeanae Rhizomacataplasm, release rate and percutaneous rate study, quality standard study, preliminary stability investigation and preliminary clinic observation. Our study laid a solid foundation for the further development of the Anemones Raddeanae Rhizomacataplasm.