Studies On The HPLC Fingerprint, Simultaneous Determination And Extraction Progress Of Elephantopus Scaber

Elephantopus scaber Linn., mainly distributed in the southern part of China, isa commonly used Chinese medicine to the treatment of cold, pertussis, hepatitis,conjunctivitis, nephritis and eczema. Its medicinal used parts are its whole plantsand roots. Pharmacological studies have shown that E. scaber has antiviral,antibacterial, anti-inflammatory, and antitumor activity, and its main activecomponents are caffeoylquinic acids and the sesquiterpene lactones.The objectives of the present studies are to establish the quality control methodthe whole plant of E. scaber, and investigate the extraction progress of the antiviralcomponents of3,5-O-dicaffeoylquinic acid and3,4-O-dicaffeoylquinic acid in theplant.Firstly, we established the quality control method of the whole plant of E.scaber using HPLC fingerprint analysis in combination of the simultaneousquantifications of three active components in E. scaber. Analyses were carried outon an WELCH XB C18column (4.6mm×250mm,5μm) column eluted withacetonitrile-0.2%glacial acetic acid as the mobile phase, and the flow rate was1.0ml/min, and the column temperature was25℃, the detection wavelength was set at235nm. In the HPLC chromatogram, nine common peaks were detected, andthree of them were identified to be3,5-O-dicaffeoylquinic acid,3,4-O-dicaffeoylquinic acid, and deoxyelephantopin. In addition, this method wasused for simultaneous quantification of three active compounds in E. scaber L,such as3,5-O-dicaffeoylquinic acid,3,4-O-dicaffeoylquinic acid, and deoxyelephantopin. After the method validation, linearities of3,5-O-dicaffeoylquinic acid,3,4-O-dicaffeoylquinic acid, and deoxyelephantopinwere good between the range of0.1128-5.076μg (R2=0.9997),0.2328-10.476μg(R2=0.9994), and0.067-2.51μg (R2=0.9999), respectively. The averagerecovery rates were96.23%、98.61%and96.73%, respectively. The method hadgood precision, reproducibility, and stability (RSD <3.0%). The method set up inthe present was successfully applied to control the quality of E scaber.Secondly, we investigated the extraction progress of the antiviral componentsof3,5-dicaffeoylquinic acid and3,4-dicaffeoylquinic acid in the plant by means oforthogonal experimental design method. The contents of these two activecompounds were tested with the established HPLC method. Experimental resultsshowed that the best extraction conditions were that the powder plants wereextracted in70%ethanol under70C and pH value of7for1hour, and the ratio ofplant material and solvent was1:20. In this conditions, the extraction yield of both3,5-dicaffeoyl quinic acid and3,4-dicaffeoyl quinic acid could reach8.16mg/g and2.54mg/g.