Studies On The Chemical Constituents And Quantitative Determination Of Glucosyloxybenzyl 2-isobutylmalates In The Tubers Of Gymnadenia Conopsea

Gymnadenia conopsea ( L ) R. Br ( family orchidaceae ) is a rare traditional Chinese medicinal herb with highly clinical values. There have been more studies on chemical constituents of the tubers of G. conopsea in recent years, but only one literature has been reported on the research of quality standard, which is chemical fingerprint analysis of the tubers of G. conopsea. In order to evaluate the quality of this medicinal plant, and provide references for interspecies identification and cultivation, a systematic photochemical study was carded out at first, and then a high performance liquid chromatogramphy (HPLC) method was developed for the quantitative determination of the known active compounds in the tubers of G. conopsea.Twenty compounds were isolated from the ethanol extracts of the tubers of G. conopsea by various column chromatographies, nineteen of which were determined on the basis of physical properties and spectroscopic analysis as follows: hexadecanoic triglyceride (1), octadecylic triglyceride (2), stearic acid (3),β-sitosterol(4 ), daucosterol( 5 ), p-hydroxybenzaldehyde( 6 ), succinic acid( 7 ), di-(p-hydroxybenzyl) disulfide (8), 4,4′-dihydroxybenzyl sulfoxide (9), adenosine ( 10 ), coelovirins B ( 11 ), gynmadesideⅠ( 12), dactylorhin E ( 13), militarine ( 14), loroglossin ( 15), dactylorhin B ( 16), dactylorhin A ( 17), coelovirins F ( 18 ) and coelovirins G (19) . Compound 8 was obtained from nature products for the first time, eight compounds were obtained from the plants of Gymnadenia R.Br. for the first time.Five glucosyloxybenzyl 2-isobutylmalates, four of which were the known active compounds, were isolated from the tubers of G. conopsea and selected as the marker compounds, and a HPLC method was firstly developed for simultaneous quantitative determination of these five compounds in the tubers of G. conopsea. The assay had been successfully applied to determinate the contents of these compounds in G. conopsea and other seven plants of orchidaceae collected from different regions of China.Based on the above researches, several issues have been discussed covering the extraction and isolation methods of the chemical constituents of the tubers of G. conopsea, and the spectroscopic features of glucosyloxybenzyl 2-isobutylmalates, et al. Furthermore, this dissertation reviewed the advances on the study of chemical constituents, pharmacological activities and clinical applications of the plants of Gymnadenia R.Br., and summarized distribution of glucosyloxybenzyl 2-isobutylmalates in the plants of Orchidaceae. All these findings are helpful for further research on the plants of Gymnadenia R.Br.