Chemical Constituents Of Artemisia Mongolica And Its Multi Index Quality Evaluation System

Objective:Through the comprehensive study on the chemical constituents of petroleum ether,dichloromethane and ethyl acetate extracts from Artemisia mongolica,the chemical information of Artemisia mongolica were found out to solve the problem that its chemical composition is not clear.At the same time,the multi-component simultaneous determination method and physical and chemical identification were studied,and the routine inspection methods were supplemented to establish the multi index quality evaluation system Artemisia mongolica.Methods:Artemisia mongolica(5.0 kg)were refluxed successively with petroleum ether,dichloromethane and ethyl acetate for 2 times,each time for 4 h.The petroleum ether,dichloromethane and ethyl acetate extracts were obtained by combining respectively the extracts and recovering the solvent under reduced pressure.The extracts were separated and purified by conventional silica gel and Sephadex LH-20 column chromatography to obtain a single compound.The structures of the compounds were determined by the determination and analysis of various spectral data of the single compound.According to the content requirements of impurity inspection in current Chinese Pharmacopoeia(2020),the heavy metal inspection,arsenic salt inspection,moisture determination,ash determination and extract inspection were carried out.The flavonoid compounds in Artemisia mongolica were identified by chemical method,while the TLC method was used to identify the aconol,1-(3-hydroxyphenyl)-2-(5-hydroxy-3-methoxyphenyl)ethane.The content of Artemisia mongolica in different batches were determined by HPLC.HPLC conditions were as follows:inertsil ods-sp(250 mm × 4.6 mm,5 μm);mobile phase:acetonitrile(a)-water(b),gradient elution,0-5 min,15%→25%a;5-15 min,25%→35%a;15-30 min,35%→65%a;30-40 min,65%→55%a;detection wavelength:261 nm;flow rate:1.0 ml.min-1 Results:Twenty compounds were isolated from different solvent extracts of Artemisia mongolica,namely chamazulene(1),mongolin A(2),ursolic acid(3),3-hydroxy-24(25)endamantane(4),ursanol(5),3-keto-24(25)endamantane(6),ursolic acid(7),dehydrofalcarindiol(12),benzyl salicylate(13),o-hydroxycapillene(14),1-(3-hydroxyphenyl)-2-(5-hydroxy-3-methoxyphenyl)ethane(15),1-(3-hydroxyphenyl)-2-(3,5-dimethoxyphenyl)ethane(16),1-(3-hydroxyphenyl)-2-(3,5-dimethoxyphenyl)ethane(17),artemonosid A(18),panalaside A(19),panalaside B(20).The results of TLC identification showed that the petroleum ether-ethyl acetate(10:1)was used as developing agent for ursolic alcohol and cyclohexane-acetone(7:1)for 1-(3-hydroxyphenyl)-2-(5-hydroxy-3-methoxy phenyl),the spots were clear and reproducible,and Rf values met the requirements.The results of general inspection showed that the contents of heavy metals,arsenic and water in 12 batches of Artemisia mongolica were in accordance with the standard of Chinese Pharmacopoeia.From the results of ash determination,the ash content and acid insoluble ash content in samples(A5,A6,A8 and A9)were higher,which may be due to unreasonable processing after the collection of medicinal materials and failure to dispose of some soil and sand particles.The average contents of water-soluble extracts and alcohol soluble extracts in 12 batches of Artemisia mongolica were 25.6%and 16.9%,respectively.Under the condition of selecting experimental chromatography,the calibration curves of 1-(3-hydroxyphenyl)-2-(5-hydroxy-3-methoxyphenyl)ethane(1),1-(3-hydroxyphenyl)-2-(3,5-dimethoxyphenyl)ethane(2),1-(3-hydroxyhenyl)-2-(3,5-dihydroxyphenyl)ethane(3)and artemonosid a(4)showed good linearity.The average recovery rates of 1,2,3 and 4 were 97.53%,96.96%,97.03%and 97.99%,respectively,and RSD was 1.30%,1.42%,1.44%and 1.22%,respectively.Conclusion:Through the study of chemical constituents of Artemisia mongolica,seven compounds(No.1-7)from the petroleum ether extracts,10 compounds(No.8-17)from the dichloromethane extracts and 3 compounds(No.18-20)from the ethyl acetate extracts were isolated and identified.Among them,compounds 2 and 15-20 are new compounds which have not been reported in the literature.Other compounds are isolated from this plant for the first time.Seven monoterpenes,sesquiterpenes and triterpenes(No.1-7),six alkynes(No.8-12,14),four diphenylethanes(No.15-18)and three other compounds(No.13,19,20)were isolated by structure type.In addition,the chemical,TLC identification and HPLC method for the determination of 4 diphenylethane compounds in Artemisia mongolica were established by qualitative and quantitative analysis.At the same time,the contents of heavy metal examination,arsenic salt examination,moisture determination,ash determination and extract examination were supplemented,which provided a strong basis for the quality control of the medicinal material.