Research Of Tibetan Medicine Rhamnella Gilgitica Mansfeld. Et Melch By HPLC

Rhamnella gilgitica Mansfeld et Melch. (Shengdeng) is traditional Tibetan herbal medicine which has several actions for cooling the blood, removing rheumatism, alleviating swelling and relieving pains. It's used for treatment of heat from blood, high altitude polyemia, hansen's disease ect. In this research, a new separating and determining method has been developed for the Tibetan medicine Rhamnella gilgitica Mansfeld. et Melch. At first, the refluxing (75% ethanol for 4h) and concentrating methods have been used to extract the effective components of the Tibetan medicine. Then, the optimal HPLC conditions were developed for HPLC fingerprint analysis. The samples were separated with DuPont ZORBAX ODS C₁₈ Column (4.6mm×25cm) by linear gradient elution using methanol(A:0～10min, 20%～25%, 10～20min,25%～40%, 20～55min, 40%～55%, 55～75min, 55%～75%, 75min～85min, 75%～90%)and ammonium formate (B, pH=3.5) as mobile phase at a flow rate of 1.0 mL/min and detected wavelength at 290nm. The accurate and feasible digitized chromatographic fingerprint spectrum (DFPS) of the Shengdeng was established, and analyzed with chemical model discriminating method, which can reflect the real constitution of samples. The above method can been used to distinguish the producing area of the medicine and to control the quality of Shengdeng samples. Moreover, a method of solvent extraction separated by HPLC and column chromatograph separation of Shengdeng was also studied to observe the efficiency of purification and separation, which provide the basis to identify several components. The detail content as follows in each chapter:In chapterl, the research status of quality control of Tibetan medicine(TM) with chromatography techniques was summarized. Including origin and characteristic of TM, the status standard for quality control of TM, the factors for quality of it, the concept and meaning of Chinese fingerprint spectrum, the status of analysis for TM with chromatography fingerprint spectrum, the principle and procedure of digitized chromatography fingerprint spectrum, the methods for analysis of fingerprint spectrum, the methods of extraction, purification, separation from Chinese herbal medicine ect.In chapter 2, a method for extraction, purification, separation was established. Compare the efficiency of extraction and purification with petroleum ether, ether, ethyl acetate, butyl alcohol by observing the behavior of separation and peak area when it was separated by HPLC. Concentration of extract was separated by Polyamide chromatograph and spotted on silica gel plate to examine the phenomenon of separation and color development by developing different solvent. This method can offer the basis for identify the existence of flavones.In chapter 3, an accurate and feasible HPLC fingerprint spectrum of Shengdeng was established. Compare the extract method between thermal reflux, soxhlet and super sonic at first, then compare the efficiency of extraction by different solvent and different extract time. Furthermore, optimal HPLC conditions were developed for HPLC fingerprint.In chapter 4, the methods for analyze the fingerprint spectrum were adopted. The feasible digitized chromatographic fingerprint spectrum (DFPS) for collected information from the chromatographic spectrum of 6 samples was established. Evaluate the quality of this herbal medicine by overlapping rate and difference rate. Calculate the similarity by similar software between the fingerprint spectrum which reflect the similarity of retention time. Classify the different quantitative samples with component and cluster analytical methods, excluding the inferior medicine, the common model can be used to identify of any Shengdeng medicine.