Studies On Structure Of Roots And Active Ingredients And HPLC Fingerprints In Gentiana Dahurica Fisch And Gentiana Macrophylla Pall

Gentiana dahurica Fisch and Gentiana macrophylla Pall,which belong to Gentianaceae Gentiana L Sect,as original gentian spices,were included in the "Pharmacopoeia of the People's Republic of China".The recent years' researches are focus on biological,chemical and pharmacological studies,but only few studies on Gentiana dahurica Fisch.In this study, we systematically investigated the development and different structure of roots,the storage site of effective secoiridoid glycoside,as well as the content of effective ingredient of different gentian species in different regions and HPLC fingerprint by using anatomy, histochemistry and phytochemistry methods.For deep studies on the structural development, the relationship between the structure and the effective ingredient in the Gentiana dahurica Fisch and Gentiana macrophylla Pall to provide the theoretical foundation,as well as the better gentiana herb selection,the relationship between the effective ingredient content and geography and other fields to provide scientific basis.1 The primary structure of Gentiana dahurica Fisch was same to other dicotyledons.It's consisted of epidermises,cortex and stele.The endodermis had clear casparian strips on its radial direction and the primary xylem were diarch.The secondary structure include the normal and the abnormal structure of the end root.The normal secondary structure was caused by the activities of vascular cadmium and cork cadmium.During the later secondary development stage,because of the abnormal activiton of the vascular cadmium and xylem parenchymatous.,The original dimensional string was divided into several parts,and then enclosed by the the cork-cells,at last the root splits into several branches.2 The root development of Gentiana macrophylla Pall was also same to the other dicotyledons.It could be divided into four stages,promeristem,primary meristem,primary structure and secondary growth stage.The primary structure and the early secondary progress were same as Gentiana dahurica Fisch.The later secondary progress was similar as Gentiana dahurica Fisch,but after the splitting of the dimension string,it would form the complete cambium ring.In the root of the third years gentiana,existing an abnormal structure which enclosed by cambium ring.In the root of the fourth years,emerging three rounds catheter, xylem and phloem alternate arrangement.3 The present histochemical result indicated that the secoiridoid glycosides were mainly exist in the parenchyma cells of xylem and phloem in gentiana's root.4 By using the HPLC and HPLC-ESI-MS~n connected technology,to study the methanol extraction of gentiana's root.According to the results,we speculated that the peak4 is 6'-O-β-Dglucosylgentiopicroside,the peak5 is swertiamarine and peak 6 is gentiopicroside. Through contrasting with the standard substance,to definite the peak 6 is gentiopicroside.5 Through HPLC methord,we measured 16 samples' gentiopicroside content.The results were all excess of the standards which provide by "Pharmacopoeia of the People's Republic of China".And between different species the content difference is obvious.Gentiana macrophylla Pall＞G.straminea.Maxim＞G.dahurica Fisch.Moreover,there's a relevance between gentiopicroside content and geographical distribution:to consider Shanxi Taibai and Sichuan Chuanzhusi as the center,the gentiopicroside content would reduce along with the northwest, southwest and northeast,the decline tendency along the northwest is the most obvious.The Gdahurica Fisch showed the similar regulation.Besides,the contents of 6'-O-β-Dglucosylgentiopicroside and swertiamarine showed a relationship,which is one loss and another gain.6 To measure and analyse the HPLC fingerprints of Gentiana macrophylla Pall from 9 regions in 7 provinces,the results showed that the basic chemical constituents of different regions are similar,but the contents different,the average similarity is 0.990.Cluster analysis showed that when connection coefficients is 2.25962,9 Gentiana macrophylla Pall samples were divided into 3 groups.The HPLC fingerprints of G.dahurica Fisch is similar with Gentiana macrophylla Pall',but the consistency is not as good as Gentiana macrophylla Pall, the average similarity is 0.971.When connection coefficients is 1.476995,G.dahurica Fisch samples were divided into 2 groups.Comparative and analyse the HPLC fingerprint of Gentiana macrophylla Pall,G.dahurica Fisch.and G straminea.Maxim,the average similarity of Gentiana maerophylla Pall is 0.988,G.straminea.Maxim is 0.979 and G dahurica Fisch is 0.941,which showed that G.straminea.Maxim is more suitable than G.dahurica Fisch as Gentiana macrophylla Pail's alternatives.Cluster analysis showed that when connection coefficients is 1.151655,16 samples were divided into 9 groups.Gentiana original medicals from different areas would cluster on the basic of species.