Authentication Of Germplasm Resource Of Rhizoma Pinelliae And The Effect Of Processing Method On Its Quality

Rhizoma Pinelliae,is the dried tuber of plant Araceae Pinellia Pinellia ternata(Thunb.) Breit.with the roles of drying dampness to resolve phlegm, downbearing the adverse flow of Qi to stop vomiting,dissipating palpable and nonpalpable masses.According to statistics in the 558 kinds of Chinese medicine prescriptions,Rhizoma Pinelliae ranks frequency of use of 22.As an important traditional Chinese medicine,Rhizoma Pinellia pharmacological effects range widely.However,the content of the composition of Rhizoma Pinelliae is fairly low and the active ingredient is also not clear.For Rhizoma Pinelliae by different processing methods caused by the enormous differences in quality have not been sufficient attention,there was no scientific processing standard for a long time.Because of quality of different resources,processed in different methods and harvested in different time,the quality of Rhizoma Pinelliae vary significantly and seriously affect its clinical efficacy.We collected dried and fresh samples of Rhizoma Pinelliae from different habitat as object.With isoelectric focusing electrophoresis,gas chromatography,and HPLC determination,these samples were analyzed to explore identification methods of their germplasm resources and different processing methods of the impact of their chemical composition,which lay the foundation for the germplasm research to identify and quality standards study.With isoelectrofocusing electrophoresis technology,we compared and analyzed the electrophoresis action of protein in fresh Rhizoma Pinelliae from different places,different period of maturity and germination germplasm materials.This technology can form smooth PH gradient by adding ampholyte and make protein stop electrophoresis at the isoelectric points.The samples of Chinese traditional medicine can be analyzed and identified with protein isoelectrofocusing electrophoresis technology.In order to reduce expensive reagent consumption and finish IEF technology with the ordinary electrophoresis equipment,we have used improved IEF technology which is effectual and the reappearing is good.The experimental results show that: Although the IEF electrophoresis atlas of the protein in Rhizoma Pinelliae from different habitat in the mature stage is the same,but there are significant difference among the ones of the germination period of the cultivation of resources;each of Rhizoma Pinelliae in the germination period from the same area(such as wild growing pinellia tuber Jingzhou) has the same electrophoresis atlas of the protein.Therefore,we can identify different areas or different varieties of germplasm resources with Rhizoma Pinelliae in the germination period.In this paper,the samples from different habitat processed in different methods are analyzed by GC method.Results showed that:there are five specific components of a very rich steroid in the whole samples of Rhizoma Pinelliae from the west of Hubei province which dried 2h by 70℃then bake with low temperature,and the peaks of the five element in each sample have a high similarity.But the samples of Rhizoma Pinelliae without proper heat treatment,drying or directly naturally dried were detected in peak height due steroid barely detectable component or components of the corresponding.We also tested the samples of Rhizoma Pinelliae in the germination,did not detect the steroid-specific components.In our analysis,some enzymes in Rhizoma Pinelliae involved in steroid ingredients' metabolism,germination period, which are particularly active,rapid metabolism.So it is hard for steroid component to be detected.Enzymes such as polyphenol oxidase which contained in low-temperature drying and direct dried samples,have not been destroyed.Just because of the drying process inhibited some of the activities. Once the temperature increases,the influx of air change,the amount of enzymes become active.Rhizoma Pinelliae without proper temperature treatment to kill enzymes,in the roles of polyphenol oxidase,will occur browning even decay.For preventing this phenomenon,someone joined Rhizoma Pinelliae with one kind of antiputrefactive called the preservative sodium sulfate which is easy to cause cancer.It has seriously affected the quality of Rhizoma Pinelliae.In this paper,we used the conventional methods in the Chinese Pharmacopoeia,found that some samples of Rhizoma Pinelliae contain sodium pyrosulfite indeed.The component of Rhizoma Pinelliae with low-temperature and naturally dried easily changes,which also easily decompose.That's because of the damage of polyphenol oxidase in it.It is revealed that the processing method to Rhizoma Pinelliae has a very close relationship to its quality and storage.In order to confirming the above experimental result,we analysised the water-soluble components contained in Rhizoma Pinelliae.With HPLC chromatography,the samples of Rhizoma Pinelliae processed by different methods have been determined.The mobile phase is methanol-water system, examination wave-length 210 nm,the column temperature 30℃,flow velocity 1.0 ml/min.The experimental results showed that:Compared with the samples of Rhizoma Pinelliae which dried 2h by 70℃then bake with low temperature and fresh Rhizoma Pinelliae just excavated,the water-soluble chemical constituents of the samples of Rhizoma Pinelliae without proper heat treatment to kill the enzyme and fresh Rhizoma Pinelliae in the germination decreased significantly.And when there are correct sample and reference substance,we can also analysis the fingerprint of water-soluble chemical in Rhizoma Pinelliae furthermore with the HPLC chromatography established in this paper.Finally,using the method inducing dormancy of the mice with subliminal and suprathreshold dose of sodium pentobarbital adding ethyl acetate or n-butanol part of Rhizoma Pinelliae(at high temperatures to kill enzyme) ethanol extract,we compared the sedative and hypnotic act of these two part. The results showed that:the ethyl alcohol part of the total ethanol extract, compared with the n-butanol part,has better sedative and hypnotic effects of sedative and hypnotic.The active ingredients should be in its ethyl acetate part of the total ethanol extract.