| Paris polyphylla Smith var.yunnanensis（Franch.）Hand.-Mazz.is a variant of the Paris polyphylla of Paris genus of Liliaceae.In recent years,a new varieties of P.polyphylla Smith var.yunnanensis which named YDCL03 was discovered.It is valued because of its high yield and high breeding rate,especially distributed in the Gaoligong Mountains of the southern section of the west,because compared with P.polyphylla Smith var.yunnanensis（Franch.）,no matter in the plant morphology characteristics,morphology and number of the cluster buds and other places of YDCL03 are quite different.Since 2013,it was investigated the resources,distribution,cultivation and utilization of P.polyphylla Smith var.yunnanensis（polygerm varieties）in Guizhou,Sichuan and Yunnan provinces,and researched their botany character.In addition,it was carried out a full range of botanical research on YDCL03,and successfully obtained the certification of new materials in Sichuan province and summary of scientific cultivation techniques.The main research topics were as follows:（1）Base on-the-spot investigation,literature review and the experience of demonstration base of P.polyphylla for six years,it was basically mastered the distribution and the situation of cultivation and utilization of P.polyphylla Smith var.yunnanensis（Franch.）and its polygerm varieties.It was carried out a full range of botanical research on P.polyphylla Smith var.yunnanensis（Franch.）and its polygerm varieties.And it was systemly summaried the scientific cultivation techniques of P.polyphylla.（2）To establish a method of determination seventh steroidal saponins content in P.polyphylla Smith var.yunnanensis（polygerm varieties）by UPLC-ELSD with a ACQUITY UPLC? BEH C18（2.1 mm×50 mm,1.7 μm）column,and gradually diluted with acetonitrile（A）-water（B）at a flow rate of 0.2 mL·min-1,elegant tube temperature 55℃,flow rate of nitrogen 40 psi and gain 500,gradient elution program:0-1 min 10%-25%A;1-2.5 min,25%-30%A;2.5-4 min,30%-35%A;4-6 min,35%-40%A;6-8 min,40%A-42%A;8-21 min,42%A-50%A;21-23 min,50%A-90%A.The results showed that: 1)The 23 samples of P.polyphylla var.yunnanensis（polygerm varieties）were collected from different places,were detected.Polyphyllin saponin H was detected in all of the samples of P.polyphylla var.yunnanensis（polygerm varieties）,but polyphyllin saponin Ⅵ was only detected in 4 samples.Herein,it was suggetted that polyphyllin saponin H should be as one of the standards of quality evaluation of PARIDIS RHIAOMA.2)It was used principal component analysis and cluster analysis to analyze 23 samples based on seven steroid saponins content.The results showed that there were no obvious differences about the chemical composition and contents among P.polyphylla var.yunnanensis（polygerm varieties）,the only differences was the differences of their botanical character.3)The method was convenient,accurate and suitable for the quantitative analysis of P.polyphylla var.yunnanensis（polygerm varieties）.（3）Based on the above instrument method,it was established the fingerprint of the YDCL03 for the first time,and was used the similarity analysis and cluster analysis to study on recognition of fingerprint pattern.Taken polyphyllin saponin II as the reference peak,it was calibrated eight common characteristic peaks,and was identified four common peaks which were polyphyllin saponin Ⅶ,polyphyllin saponin H,polyphyllin saponin II,and polyphyllin saponin I.The similarity of the fourteen samples was the range of 0.301-0.996,the mean value was 0.751,and it was indicated that there are many differences among YDCL03.In addition,the cluster analysis based on the relative peak area of common peaks was also showed the difference amongYDCL03.The method was convenient,accurate and suitable for the quantitative analysis of P.polyphylla var.yunnanensis（polygerm varieties）.（4）It was first time to study and evaluate the quality standard of YDCL03,included the original plant identification,medicinal properties and TLC,moisture and ash inspection,alcohol extract and content determination,and provided a scientific and effective method for the quality control of medicinal materials about YDCL03.（5）It was firstly based on our own base demonstration base of P.polyphylla to compare the germination of P.polyphylla var.yunnanensis and YDCL03,and artificial pollination and natural pollination seeds.The results showed that there was no difference among P.polyphylla var.yunnanensis（polygerm varieties）seeds,and the germination rate and emergence rate of artificial pollination were significantly higher than that of natural pollination.The peeled seeds were treated by 100 mg·mL-1 gibberellin,and cultured to sprout by wet sand at the temperature of 4-18℃,then sown in the field.It could shorten the seed emergence period one year and up to more than 80% of the rate of emergence.（6）It was the fist time to use wet digestion-ICP-OES method to detected As,Cd,Cu,Hg,Pb,Cr,Fe,Mn,Sr,Zn of P.polyphylla var.yunnanensis（polygerm varieties）.The results showed that under optimized conditions,it was simultanely differmined with non-interference each other.There was no requirement of limitation of heavy metal in 2015 edition of the ChinesePharmacopoeia（one section）.It was consulted the requirement of limitation of heavy metal of the green industry standard for the import and export of medicinal plants and preparations of standards of the foreign trade of the People’s Republic of China,as follow: Pb(≦5.0 μg·g-1),Cu(≦20.0 μg·g-1),As(≦2.0 μg·g-1),Cd(≦0.3 μg·g-1),and Hg(≦0.2 μg·g-1).The result showed that Pb,Cu,as and Cd of all samples were not exceeded the limitation except for Pb limitation of sample four.Hg was not detected in all samples.（7）A steroidal saponin was isolated from P.polyphylla var.yunnanensis（polygerm varieties）by HSCC for the first time,and chemical structural formula was identified by MS and NMR as:（25R）-26-[（-D-glucopyranosyl）oxy]-22-oxhydryl-5-en-3-ylO-D-glucopyranosyl-（1-4）-O-[-L-rhamnopyranosyl-（1-2）]-O-L-rhammnopyranosyl-（1-4）-O-L-rhammnopyranos ide.（8）The results showed that the new strain was based in the quality of ordinary P.polyphylla var.yunnanensis,but also with high yield,strong resistance,easy cultivation and other excellent characteristics.In a word,YDCL03 was a variety worth popularization and in-depth researches. |
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