Extraction, Isolation And Identification Of The Metabolites From Plants Of Elsholtzia Splendens And Sedum Alfredii Hance Post-phytoremediation

In this paper, chemical composition in the 95% ethanol extract from the plants of Elsholtzia splendens has been studied, using ultrasonic extraction with 95% ethanol, together with column chromatography separation and purification methods (macro-porous resin, polyamide resin, silica gel column chromatography, etc.). Two phenyl-propanoid compounds (caffeic acid and rosmarinic acid) and three flavonoids (apige-nin, apigenin-7-O-β-D-glucoside and luteolin) have been isolated and purified as well as chemically identified by the spectroscopic means of IR,1H NMR/13C NMR, LC-MS, NOSEY, etc.). The purities of the compounds were more than 98% as de-tected by HPLC. Caffeic acid was an important metabolite in Elsholtzia splendens, with its content about 0.1305% based on the dry weight. A drastically negative corre-lation was found for copper concentration and caffeic acid content in the stems and leaves of Elsholtzia splendens when exposed to different copper levels. MTT cyto-toxicity was assayed at 50μmol concentration for these compounds on the lung ade-nocarcinoma cells A549, human epidermoid carcinoma A431, human breast cancer Bcap37, and their inhibition rate (IR) indicated that similar activity was found for apigenin (IR 77.07%) and luteolin (IR 71.94%) on the lung adenocarcinoma cell line A549, as close to the positive control with gefitinib (IR 80%). In addition, inhibition rate (IR) of luteolin on human epidermoid cancer cells A431 and breast cancer Bcap37 were the highest of 37.41% and 46.78%, respectively. Luteolin with mul-ti-target activity can be used as an important compound for the drug development of anti-lung adenocarcinoma cell line A549, which can produce the added value for plants of Elsholtzia splendens post-phytoremediation.Sedum alfredii Hance is a Chinese native zinc (Zn) hyperaccumulator. In this study, chemical composition in the 40% ethanol extract from the hyperaccumulator S.alfredii biomass (stems and leaves) has been analyzed, from which the substituted indole compound was isolated and purified. The separation via macroporous resin column has been optimized for the substituted indole compound from the stems and leaves of hyperaccumulator, and then further purified using preparative HPLC, the purity of the substituted indole compound detected by high performance liquid chro-matography (HPLC) was achieved to be 99.90%. Three-month growth was carried on both S. alfredii species in the hydroponic experiment, at two Zn treatments (0.5μmol/L and 50μmol/L). At high zinc (50μmol/L), hyperaccumulator grew nor-mally but its non-hyperaccumulator suffered from strongly Zn-induced toxicity. With Zn treatment elevated from 0.5 to 50μmol/L, both Zn concentration and the substi-tuted indole compound content were significantly increased in both S. alfredii species, and significantly higher in the stems than the leaves, more interestingly, stem Zn and stem substituted indole compound in hyperaccumulator were significantly higher than that in non-hyperaccumulator. The substituted indole compound is possibly an impor-tant metabolite, especially in the stems of the hyperaccumulator S. alfredii.