Studies On Quality Standards Of Acanthopanacis Cortex And Its Prepared Slices

Acanthopanacis cortex (Wu jiapi),a traditional Chinese medicine,is the dried root and root-bark of Acanthopanax gracilistylus W.W. Smith. In the clinical application, Acanthopanacis cortex and Periplocae cortex are often mixed because of the similar character in alias, medicinal part, product properties and the effect Periplocae cortex contains glucoperiplocymarin, which has a severe cardiotonic effect and if misused or overdosed may cause death as toxic component. On the other hand, little component was reported from Acanthopanacis cortex, the pharmacological effects also has not been clarified. The Chinese pharmacopoeia (Volume I,2010version) have only included description, microscopic identification, moisture, total ash,yet lack of TLC identification and HPLC determination and other items.Considering such a mess of clinical medication, we researched the Acanthopanacis cortex and its prepared slices of quality standard.Firstly, characteristic components from Acanthopanacis cortex is searched for distinguishing Periplocae cortex and three components are used as reference standard. By column chromatography, half preparation HPLC, recrystallization, we got three compounds, which all are diterpene. By IR, UV,MS,NMR,X-ray diffraction, the structure is identified, respectively ent-kaur-16-en-19-oic acid (WJP-1), ent-16β H,17-isovalerate-kauran-19-oic acid (WJP-2) and ent-16β H,17-methyl butanoate-kauran-19-oic acid (WJP-3). The cytotoxic activity was researched, and the results show that all of them have moderate intensity of cytotoxic activity. Therefore, whether from the characteristic, active, and content, the three compounds are suitable for the requirements of the reference standard.A new specific TLC identification method for Acanthopanacis cortex was established with WJP-1, WJP-2, WJP-3and control medicine the first time, with petroleum ether:acetone:isopropanol:formic acid=12:2:0.5:0.1for mobile phase, with10%vitriol ethanol solvent as chromoginic reagent, developed at105℃until spots are clear. The results found that11samples in25Acanthopanacis cortex samples and control medicine were corresponded in postion and color to the spot in chromatogram obtained with the WJP-1,2,3, and no WJP-1,2,3spots was observed in the Periplocae cortex chromatogram.The results show that the method is specific for identification of Acanthopanacis cortex. The method of determination WJP-1,2,3in Acanthopanacis cortex by HPLC-ELSD is firstly developed. Was set up for the first time are detected by HPLC WJP-1, WJP-2, WJP-3levels of method. Chromatographic conditions for WJP-1: chromatographic column:Phenomenex C182.6μm (100×4.6mm); mobile phase: Acetonitrile:water=50.2:49.8; The column temperature:20℃. There was a good linearity (R2=0.9998)with the range of2.006μg～40.12μg.The average recovery was101.21%with RSD of1.46%(n=6).The content is0.7008%2.1662%.Chromatographic conditions for WJP-2and WJP-3:chromatographic column:Agela promosil C185μm (250×4.6mm); mobile phase:0-20min acetonitrile:water=77:23;20to60min acetonitrile:water=77:23to acetonitrile: water=70:30; The column temperature:30℃. For WJP-2:there was a good linearity (R2=0.9999))with the range of1.154μg～11.54μg.The average recovery was99.11%with RSD of2.76%(n=6).The content is0.1634%～0.3388%.For WJP-3:there was a good linearity (R2=0.9999))with the range of2.076μg20.76μg.The average recovery was98.78%with RSD of1.75%(n=6).The content is0.1644%～0.5108%. It is proposed to contain not less than0.9%of WJP-1calculated with reference to the dried drug.We researched the11samples above items according to the method in The Chinese pharmacopoeia (Volume I,2010version).According to the experimental results,it is proposed to contain less than12%of moisture,11.5%total ash,3.5%acid insoluble ash, not less than10.5%of extraction.In the Chinese pharmacopoeia (Volume1,2010version),there is only processing method of Acanthopanacis cortex prepared slices, yet lack quality standard.The developed TLC identification and determination method of Acanthopanacis cortex is also suitable for prepared slices. It is proposed to contain not less than1.0%of WJP-1calculated with reference to the dried drug. It is proposed to contain less than11%of moisture,11.5%total ash,3.5%acid insoluble ash, not less than10.5%of extraction.