| Objective: Tibetan medicine of Herpetospermum caudigerumhas has an efficity treatment on hepatitis b and its ethyl acetate extracts,fatty oil and polysaccharides are the function effective parts to protect liver.This topic used the effective parts as the research object,respectively,studied its quality control method,in order to provide reference to control and evaluate the consistency and stability of the quality from the effective parts.At the same time,preliminary pharmacodynamics studied on the ethyl acetate extracts and oil,and lay the foundation for further effectiveness of material research.Methods:(1)The HPLC method was adopted to establish the chemical fingerprint of ethyl acetate extracts from Herpetospermum caudigerum,and established the active ingredient content determination method.of Herpetoilde A and herpetone.(2)The GC method was adopted to establish content determination method of main fatty acid in oil(palmitic acid,linoleic acid,oleic acid,stearic acid)from Herpetospermum caudigerum.With reference to the national standards to study the physical and chemical properties of Herpetospermum caudigerum oil,determined the saponification value,acid value,iodine value,peroxide value,etc.(3)The UV method was adopted to establish the content determination method of polysaccharides from Herpetospermum caudigerum.Various extraction methods(reflux extraction,ultrasonic assisted extraction)were studied to extract the polysaccharides from Herpetospermum caudigerum to determine the optimal extraction process.(4)The mouse were injected ConA through tail vein to establish the model of immunological liver injury,and gives oil and ethyl acetate extracts and both doses of compatibility of fatty oil and ethyl acetate extracts of Herpetospermum caudigerum.to explore the protective effects of active parts of ethyl acetate extracts and oil of Herpetospermum caudigerum by ConA induced immunological liver injury in mice.Results and Conclusion:(1)Establish different sources chemical fingerprint analysis method of ethyl acetate extracts from Herpetospermum caudigerum.calibration of 19 common peak,adopted the reference substance to identify three peaks,respectively is Herpetoilde A,balanophonin and herpetone.At the same time measured 10 batches content of herpetone from ethyl acetate extracts with different origin.10 batches ethyl acetate extracts from Herpetospermum caudigerum from different origin similarity is between 0.848 ~ 0.994.The results showed that different regions of Herpetospermum caudigerum.ethyl acetate effective extracts of fingerprint similarity is higher,Herpetoilde A content exists certain differences.In addition,HPLC method was adopted to establish the herpetone content determination method of the ethyl acetate extracts,and showed a good liner relationship in ranger of 0.1804~ 0.6012 μg(r = 0.9999),The method is simple feasible and reproducible.(2)GC method was adopted to establish the content determination method of fatty acid is feasible.palmitic acid in rang of 28.87 ~ 28.87 μg(r = 0.9998);Linoleic acid in rang of 9.53 ~ 54.48 μg(r = 0.9998);Oleic acid in rang of 206.20 ~ 206.20 μg(r =0.9998);Stearic acid in rang of 24.55 ~ 24.55 μg(r = 0.9998),All of them showed a good liner relationship.oleic acid content is the highest in fatty oil indifferent sources.The method is accurate,reliable,reproducible and could be used as quality control of palmitic acid,oleic acid,linoleic acid,stearic acid inoil acid value,saponification value,iodine value,peroxide value also could reflect the quality of Herpetospermum caudigerum.(3)The content of the polysaecharides were determined by Spectrophotometric method(phenol-sulfuirc)acid colorimetrictitration method.the content of polysaecharides of Herpetospermum caudigerum from various cultivation locations varied from 1.15% ~ 2.16%.The method established was accurate,simple and economical.It can be used to determine the content of polysaccharides of Herpetospermum caudigerum.By means of two extraction methods(refluxing extraction,ultrasonic assisted extraction)process of polysaccharides of Herpetospermum caudigerum.By the comparison of two methods,the reflux extraction yield increase nearly 11%,reflux extraction is better than that of ultrasonicassisted extraction.But there was no significant difference.The research of polysaccharides from Herpetospermum caudigerum lay the foundation of research further of pharmacodynamics.(4)Ethyl acetate extracts and oil and their combination of Herpetospermum caudigerum could decrease liver index,spleen index,the contents of ALT,AST,NO,MDA and prevented the reduction of SOD activities.Meanwhile,pathological damage in liver was relieved.Compared with the model group,Effect fraction of HSO and HE in 2:1 ratio group showed the best effect on immunological liver injury in mice.Ethyl acetate extracts compatibility with oil can protect immunological liver injury in mice.The mechanism may be associated with anti-inflammatory,anti-oxidation and scavenging oxygen free radical. |
