Chemical Constituents,quality Control And Diuretic Activity Screening Of The Water Extract Of Tetrapanax Papyrifer

Objectives:Tetrapanax papyrifer(Hook.)K.Koch is the dried stem pith of the plant of Araliaceae,which is used as medicine in the dry stem pith.It tastes sweet and mild,and has the effects of diuresis,anti-inflammatory,antipyretic,and lactation.At present,there are few studies on the chemical components of the stem and pulp of the plant.In this study,the chemical components of the plant were extracted,isolated,and identified,and the antioxidant activity was studied;The water extracts of Tetrapanax papyrifer were extracted by different organic reagents,and the diuretic activity of the extracted parts was screened;The UPLC characteristic atlas with obvious diuretic effect was established,and its quality evaluation and difference analysis were carried out through chemometrics,providing scientific basis for the quality evaluation of Tetrapanax papyrifer medicinal materials.Methods:1.The chemical components of Tetrapanax papyrifer were extracted by water decoction,and then separated and purified by macroporous resin D101 column chromatography,ODS column chromatography,Sephadex LH-20 column chromatography,and finally the monomer components were prepared by HPLC.The structure of the monomer was analyzed and identified by 1D-NMR,2D-NMR,CD,UV,IR and ESI-MS.The antioxidant activity was studied by DPPH spectrophotometry.2.The water extract of Tetrapanax papyrifer was extracted by petroleum ether,trichloromethane,ethyl acetate and water-saturated n-butanol reagent.The diuretic activity was screened by using water-loaded rat model.The urine of normal rats and water-loaded rats at 6 and24 hours after administration of different extraction sites were collected and the volume of urine was measured;Compare the contents of Na~+,K~+and Cl~-in the urine of normal rats and water load rats within 6 hours;The contents of AQP2and AQP4 in kidney homogenate were measured.3.The UPLC fingerprint of the chloroform part of the water extract of Tetrapanax papyrifer was established to evaluate the similarity.With the common peak area as the index,the quality evaluation and difference analysis of 16 batches of Tetrapanax papyrifer herbs were carried out through cluster analysis,principal component analysis and orthogonal partial least squares method.Results:1.In this study,three new compounds were obtained from the water extract of Tetrapanax papyrifer,which were named Tetrapanax papyrifer phenol A,Papyridinone A and Papyridinone B according to their structure;1 new natural product Papyridinone;Five compounds were first isolated from Tetrapanax papyrifer:2,3-dihydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)propan-1-one,dibutyl phthalate,(Z)-icos-9-enamide,9(Z)-octadecenamide,1-bromine-(8Z)-heptadene.2.The contents of Na~+,K~+and Cl~-in the urine of normal rats and water loaded rats within 6 hours after administration of different extraction sites were not significantly different from those of the negative control group;The contents of AQP2 and AQP4 in the kidney homogenate of normal rats and water-loaded rats also have no significant difference,but the urine volume of water-loaded rats after the administration of chloroform site has significant difference,so it is speculated that the chloroform site is the active site with diuretic effect.3.The UPLC fingerprint spectrum at the trichloromethane site of the aqueous extract of comfrey was established,and a total of 17consensus peaks were calibrated,referring to peak 4 as syringaldehyde and peak9 as benralactone;The results of the composite score of principal components indicated differences among samples from different origins,and seven differential components with VIP values>1 were selected by orthogonal partial least squares method to be useful as markers for the quality control of medicinal herbs of comfrey.Conclusions:In this study,nine compounds were obtained from comfrey,and it was found that Tetrapanax papyrifer A of comfrey has better antioxidant capacity and has the potential to become a novel natural plant antioxidant.Screening out the trichloromethane site has a more obvious diuretic effect,aiming at the site with diuretic activity,establishing that UPLC fingerprint of trichloromethane site of aqueous extract of comfrey can characterize its chemical quality characteristics more comprehensively,and the results of chemometric analysis of the fingerprint provide a scientific basis for the development of quality standards for comfrey.