Pharmacognosy Studies On Rhizoma Bergeniae Scopulosae

Rhizoma Bergeniae Scopulosae is the Rhizoma of Bergenia scopulosa T. P. Wang which belongs to the family of Saxifragaceae, was used as the test material. The main contents of this paper include the following several aspects of research work:(1) The pharmacognosy study of the Rhizoma Bergeniae Scopulosae; (2) The content of bergenin,arbutin and gallic acid in the Rhizoma Bergeniae Scopulosae from different regions were determinated by HPLC; (3)To study the quality criteria of herbs and optimize the extraction process of bergenin by the method of ultrasonic assisted extraction and response surface. The main research results were as follows:1. Study on the pharmacognosy characteristics of Rhizoma Bergeniae Scopulosae.The medicinal material of Rhizoma Bergeniae Scopulosae shows irregular cylindrical, thick in shape. The surface color is brown. There are many links with some epibiotic leaf and leaf base. Hard texture, easy to break and farinaceous. In the edge of the transverse section, a ring of vascular bundle can be seen with naked eye. In microscope, there are some remained epidermises outside the rhizome, which were consisted of a layer rectangular or quasi-square epidermal cell. The cork cells formed outside the cortex and consist of more than 10 layers cork cells. The cortex is thicker in which the parenchyma cells contain abundant starch grains and crystal. The vascular bundle is collateral vascular bundle and the pericycle fibers can be seen occasionally. The cambium existed between phloem and xylem. The interfascicular cambium existed between the two adjacent vascular bundles. The pith is broad and the parenchyma cells of the pith contain a lot of starch grains, calcium oxalate crystal cluster and brown inclusions.The color of medicinal material powder is tan to brownish red and the smell is light and tastes a little bitter. Microscopic identification of the medicinal powder, there are a lot of calcium oxalate cluster of crystal and the change size changed biggish, the diameter range from 15μm to 75μm. There are many starch grains in the medicinal powder which is single grain type, elliptic or pear-shaped, usually slightly pointed at both ends and minority is quasi-circular, the diameter is from 4μm tol8μm, the length is about 20μm. The sclereid is not rich, but most of them are quasi-circular, few of them are circular. The vessel elements belong to spiral duct, scariform duct and reticulate duct, the diameter is from 10μm to 25μm. The cork cell is quasi-rectangle and the diameter is 20μm to 25μm. They overlap multilayer offenly and the colour is light tan.2.The content analysis of bergenin on Rhizoma Bergeniae Scopulosae.This study establishes an HPLC method for simultaneous determination of bergenin in Rhizoma Bergeniae Scopulosae from different areas. The chromatographic separation was carried out using Inertsil ODS-3C18 column (4.6mm×150mm,5μm), the mobile phase was methanol:0.2% phosphoric acid (9:91), the detection wavelength was set at 275nm, the temperature was 30℃, and the flow rate was 1.0mL/min. The calibration curve was linear in the range of 15 to 75μg/mL and the regression equation for bergenin was is quasi-circular, the diameter is from 4μm tol8μm, the length is about 20μm. The sclereid is not rich, but most of them are quasi-circular, few of them are circular. The vessel elements belong to spiral duct, scariform duct and reticulate duct, the diameter is from 10μm to 25μm. The cork cell is quasi-rectangle and the diameter is 20μm to 25μm. They overlap multilayer offenly and the colour is light tan.2.The content analysis of bergenin on Rhizoma Bergeniae Scopulosae.This study establishes an HPLC method for simultaneous determination of bergenin in Rhizoma Bergeniae Scopulosae from different areas. The chromatographic separation was carried out using Inertsil ODS-3C18 column (4.6mm×150mm,5μm), the mobile phase was methanol:0.2% phosphoric acid (9:91), the detection wavelength was set at 275nm, the temperature was 30℃, and the flow rate was 1.0mL/min. The calibration curve was linear in the range of 15 to 75μg/mL and the regression equation for bergenin wasY=14942.0X-428.6 (R2=0.9998), the average recovery was 100.58% with RSD of 1.77% (n=6). The result showed that there were some differences in the content of bergenin among different origins. The contents of bergenin were range between 14.68 to 70.19 mg/g. The study showed that this method was simple, efficient and accurate, which can be used for the analysis of bergenin in Rhizoma Bergeniae Scopulosae.3. The research on bergenin extraction processTechnology of ultrasonic assisted extract Bergenin in Rhizoma Bergeniae Scopulosaes was optimized by using response surface method. The optimized extraction process conditions as follows:ultrasonic power was 270W, extraction time was 40 minutes, ethanol concentration was 75%, and extraction temperature was 59 ℃. Under the above conditions the bergenin yield was 6.43% and the extraction rate can reach 91.61%.4. The content analysis of arbutin on Rhizoma Bergeniae Scopulosae.The contents of arbutin in Rhizoma Bergeniae Scopulosae from different areas were determined using HPLC. The chromatographic separation was carried out using Inertsil ODS-3 C18 column(4.6mm×150mm,5μm), the mobile phase was methanol:water (8:92), the detection wavelength was set at 282nm, the temperature was 30℃, and the flow rate was 1.0mL/min. The calibration curve was linear in the range of 10.3 to 103.0μg/mL and the rearession eauation for arbutin was Y=4468.8X-999.1 (R2=0.9999). the average recovery was 99.58% with RSD of 1.79%(n=6). The result showed that there were some different in the content of arbutin among different origins, the contents of arbutin were range between 5.34 to 18.32 mg/g. This method is simple, efficient and accurate, which can be used for the analysis of arbutin in Rhizoma Bergeniae Scopulosae.5. The content analysis of gallic acid on Rhizoma Bergeniae Scopulosae.This study establishes a method for determination of gallic acid in Rhizoma Bergeniae Scopulosae that from different areas. The HPLC separation was performed on an Inertsil ODS-3C18 column (4.6mm×150mm,5μm), the mobile phase was a solution of methanol-0.4% phosphoric acid (7:93) with a flow rate of 1.0 mL/min, detection was monitored at 271nm, and column temperature of 30℃, There was well linear relationship for gallic acid in range of 5.3 to 53μg/mL and the regression equation for it was Y=31446X-15550 (R2=0.9999), the average recovery was 99.995% with RSD of 1.704% (n=9). The result showed an obvious difference in the contents of gallic acid among 14 different origins. The contents of gallic acid were range between 1.7044 to 8.6393mg/g. This study showed that the established method is simple, accurate, and repeatable, which can be used for the quality control of Rhizoma Bergeniae Scopulosae. The Rhizoma Bergeniae Scopulosae from Wenxian county has further development and utilization value for its richer gallic acid.6. The research on the quality standards of medicinal materials.According to the Chinese pharmacopoeia (2010 edition) method, moisture, ash content and extract content in Rhizoma Bergeniae Scopulosaes were analyzed. The moisture shall not be over 10.0% and the ash content may not be over 10.0%. The extracted material by hot dipping of dilute ethanol method shall not be less than 40.0%. Bergenin content not less than 1.32%, arbutin content not less than 0.48% and gallic acid content is not less than 0.15%.