Analysis On The Chemical Constituents From The Rhizome Of 15 Paris Species

Objective: Rhizoma Pariclis is a rare and valuable traditional Chinese medicine,with many funticons including clearing away heat and toxic material,relieving swelling,relieving pain,cool liver and so on.It is mainly used for treating sore throat,snakebite,traumatic injury,epilepsy disease.In the 2015 edition of the "People's Republic of China Pharmacopoeia",only two legal species are listed: Paris polyphylla Smith var.yunnanensis(Franch.)Hand.–Mazz.and Paris polyphylla Smith var.chinensis(Franch.)Hara.There are 38 species(including variants)of the genus,of which 18species(including variants)are commonly used as medicinal in Paris L.In order to investigate the similarities and differences of chemical composition among different species of Rhizoma Paridis and the feasibility of using other species as medicinal for replanting,we collected and identified 15 species of Paris L.and carried out analysis and identification of their chemical composition and quality evaluation.Methods:(1)Depending on recent references,we summarized and established the database of Paris saponins,mass spectrometry fragmentation patterns,and the retention behaviours on reversed-phase chromatography of Paris steroidal saponins.(2)We used the UPLC-Q-TOF MS technology to analysize the extracts from rhizome of 15 species in genus Paris.Metabonomic analysis and utilized multivariate statistical analysis methods were processed to find biomarkers between different species of Paris.(3)Utilizing the Unify software and the database of Rhizoma paridis,structural analysis and identification of the chemical composition of the P.forrestii,P.vietnamensis and P.polyphylla Smith var.Yunnanensis.(4)Based on the semi-quantitative analysis of UPLC-Q-TOF MS,further focused on the 8 main active components(polyphyllins VII,H,VI,II,I,and V,dioscin,and gracillin)in Paridis Rhizoma have established HPLC-UV content determination methods,and 183 samples were assayed.Results:(1)We established the database of Paris L.and summarized mass spectrometry fragmentation mechanism and the retention behavior on reversed-phasechromatography of Paris steroidal saponins.(2)An method for metabolite profiling of Paris was established,and 15 species in Paris were analysis by UPLC-Q-TOF MS.Finally,171 compounds were identified or tentatively identified in P.forrestii,P.vietnamensis and P.polyphylla Smith var.yunnanensis,inccuding 56 compounds were found in Paris for the first time.(3)Through multivariate statistical analysis of the data of UPLC-Q-TOF MS on 15 Paris species,the results showed that PCA score plot showed P.daliensis,P.axialis,P.forrestii and P.polyphylla Smith var.yunnanensis can be clustered into one category;P.polyphylla var.stenophylla,P.mairei,and P.delavayi can be clustered into one category;P.vietnamensis solely generated a class;while P.polyphylla Smith var.chinensis in different origins showed divergent.P.polyphylla var.pseudothibetica is between the P.polyphylla Smith var.yunnanensis and P.polyphylla Smith var.chinensis;however,it's tended to P.polyphylla Smith var.yunnanensis.After analyzing the P.forrestii with P.polyphylla Smith var.yunnanensis,and P.vietnamensis with P.polyphylla Smith var.yunnanensis by OPLS-DA,each of 15 metabolites as biomarkers were found,which can distinguish them.(4)A method for simultaneous determination of8 main active steroidal saponins(polyphyllins VII,H,VI,II,I,and V,dioscin,and gracillin)in rhizoma of genus Paris was established to evaluate the quality of the medicinal materials of the Rhizoma Pariclis.There are significant differences in the content of steroidal saponins from different species.The P.mairei,P.polyphylla var.stenophylla,and P.delavayi are not suitable for medicinal usage because of their low content of steroidal saponins.The contents of steroidal saponins in P.polyphylla var.chinensis from different places are quite different.The P.polyphylla var.yunnanensis,P.forrestii,P.daliensis,and P.axialis have a higher content of steroidal saponins which can reach to 5.0%.Although the contents of four saponins qualified in pharmacopeia are lower in the P.vietnamensis(Takht.)H.Li.,the contents of 8 total saponins are higher.Thus,it suggest that P.forrestii,P.daliensis,and P.axialis have the potential pharmic value and may be used in the future.Conclusion: In this study,we systematically studied the qualitation and quantitationanalysis of chemical composition for 15 species of the genus Paris,and the potential alternative sources of medicinal materials for Rhizoma Pariclis were discovered.The key difference markers between different medicinal ingredients were found and identified,and the result of using chemical composition to provenance identification is of great significance.There are some limitations for the determination method of rhizoma Pariclis in “China Pharmacopoeia”.We proposed that the quality control method together with the fingerprint could be more reasonable,and qualifing polyphyllin H is more better than polyphyllin VI,which is more suitable to be used as an indicator of content determination.The results of this study will lay the material foundation for the comprehensive protection,utilization and development of genus Paris.It also provides scientific data for accurate poverty alleviation through the use of Paris planting in border areas.