Study On The Material Basis Of Anti-dermatophytes Of Euphorbiae Humifusae Herba

Euphorbiae humifusae herba is the dry whole plant of Euphorbia humiusa Willd.and Euphorbia macuLata L..Euphorbiae humifusae herba has been widely used in traditional Chinese medicine,Mongolian medicine and Tibetan medicine.It has been used for the treatment of skin tinea diseases in clinical and exhibited antidermatophytes in pharmacological actions.However,the chemical constituents of antidermatophytes in Euphorbiae humifusae herba have not been investigated extensively.The present thesis studied on the material basis of the antidermatophytes of Euphorbia humiusa Willd.,combined various methods including activity screening,chemical isolation and identification and UPLC-PDA-QTOF-MSn.Firstly,different fractions were prepared from the ethanol extract of Euphorbia humiusa Willd..According to the results of activity screening and UPLC analysis,the antidermatophyte fractions were found.Then,various chromatography,including silica gel,MCI GEL CHP 20P,polyamide,ODS and Sephadex LH-20 column and preparative HPLC chromatographies were used to isolate and purify the constituents from active fractions.20 compounds were elucidated the structures,which included 13 flavonols,5 phenolic acids,2 three terpenes and sterol compounds.Flavonols included kaempferol(1),quercetin(2),kaempferol-3-O-β-D-glucopyrano-side(3),quercetin-3-O-α-L-arabinofuranoside(4),quercetin-3-O-(6"-O-galloyl)-β-glucopyranoside(5),kaempferol-3-O-α-D-arabinopyranoside(6),kaempferol-3-O-β-D-xylopyranoside(7),kaempferol-3-O-α-L-arabinofuranoside(8),Kaempferol-3-O-(6"-O-galloyl)-β-D-galactoside(12),quercetin-3-O-{rha(1→6)-[xyl(1→2)]-galactoside}(14),quercetin-3-rutinoside(15),quercetin-3-O-glucoside(16),quercetin-3-O-β-D-xyl(1→2)-glucoside(18).Phenolic acids include ethyl gallate(9),trans-ferulic acid(10),ethyl protocatechuate(11),3,3’-di-O-methyl ellagic acid-4-O-β-D-glucopyranoside(13),methyl gallate(17).Three terpenes and sterol compounds include cycloart-23-ene-3 β,25-diol(19),β-sitosterol(20).Compounds 5、7、10、11、12、14、18 were obtained from this plant for the first time.Moreover,the main components of the active fractions were characterized by using UPLC-PDA-QTOF-MSn technique,and 59 components were identified,including 44 phenolic acids and 15 flavonoids.Finally,the potential antidermatophytes constituents were probed by using grey relational analysis,and the result showed that the correlation coefficient between 29 constituents and activity is all greater than 0.6,of which the ellagic acid,kaempferol,quercetin have the highest correlation degree with anti-dermatophytes activity.Additionally,the development of the chemical constituents and pharmacological activities of Euphorbiae humifusae herba were also reviewed in this paper.