Use of on-flow LC/1H NMR for the study of an antioxidant fraction from Orophea enneandra and isolation of a polyacetylene, lignans, and a tocopherol derivative

The CH2Cl2 extract of the leaves of Orophea enneandra displayed antifungal, antioxidant, and radical scavenging properties in bioautographic TLC assays. To obtain rapid information on the active compounds, on-flow LC/H-1 NMR and LC/UV/MS analyses of the antioxidant fraction were performed. The on-line information led rapidly to the partial identification of three closely related lignans, one tocopherol derivative, and one polyacetylene. This approach necessitated, however, large quantities to be injected to obtain satisfactory on-flow LC/H-1 NMR spectra, and isolation of the compounds was necessary to obtain complete NMR data. These compounds were isolated and identified as (-)-phylligenin (1), (-)-eudesmin (2), (-)-epieudesmin (3), polycerasoidol (4), and oropheic acid (5), a new polyacetylene. Their activities against the 2,2-diphenyl-1-picrylhydrazyl radical and the fungus Cladosporium cucumerinum were investigated. This paper indicates the possibilities and limits of on-flow LC/H-1 NMR in this type of study.