Study On The Chemical Constituents And Quality Control Of Anoectochilus Roxburghii

In this study,Anoectochilus roxburghii was studied in terms of chemical constituents and HPLC fingerprint and content analysis.Based on the results of on-line HPLC-FRAP antioxidant activity detection and on-line Sephadex LH-20×RPLC analysis,alcohol elution of macroporous resin column with strong antioxidant activity were selected from crude extract of A.roxburghii,utilize various chromatographic methods(including Sephadex LH-20 column chromatography,ODS column chromatography,preparative HPLC),26 compounds were obtained;the water elution of macroporous resin column were separated by HILIC column chromatography,preparative HPLC,and 2 compounds were obtained.Structures of 24 compounds were identified by physicochemical constants,NMR and MS.Three of them were new compounds.A preliminary analysis of the antioxidant activity of the compounds was performed.Results show that flavonoids are major antioxidant active components in A.roxburghii.The fingerprints of common cultivars and wild species of A.roxburghii were established.24 common peaks were identified.The relative retention time RSD values of the common peaks were between 0.034%and 1.377%and RSD values of relative peak area were between 27.33%and 129.59%.6 common peaks were identified by comparison with the retention time of the compounds isolated.Similarity analysis was performed with the software "Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine" published by GPC(Version 2004A).The similarity was good in 14 batchs of samples.The fingerprints were used to identify the confusing species of A.roxburghii,the similarity of three kinds of A.formosanus samples were lower than that of A.roxburghii,confirming that the HPLC fingerprint can be applied to identify two speciesA HPLC method for the determination of flavonoids after acid hydrolysis and determination of trigonelline in A.roxburghii were established.The contents of quercetin,kaempferol,isorhamnetin aglycone and trigonelline in common cultivars and wild varieties were determined and results show that there is a big difference in the content of the four substances.The methods were also used to determine and compare the contents of four components in A.rmosanus,the content of trigonelline in A.formosanus is much lower than that of A.roxburghii,indicating that the content of trigonelline can be used as a criterion for distinguishing two easily confused varieties.