Silica gel 60

Silica gel 60 and Sephadex^® LH-20 (GE Healthcare, Uppsala, Sweden) were used for column chromatography, while Silica gel 60 F₂₅₄ aluminum sheets (Silicycle^®, Ville de Québec, QC, Canada) were used for analytical thin layer chromatography (TLC). Compounds were visualized by exposure under UV_254/365 light and by spraying with p-anisaldehyde reagent followed by heating. All solvents for chromatographic procedures were of analytical grade, while those for MS analysis were of spectrometric grade. NMR experiments were acquired at 303 K in CDCl₃ and acetone-d₆ on a Bruker (Billerica, MA, USA) AVANCE III NMR spectrometer and on an Agilent (Santa Clara, CA, USA) DD2 600 spectrometer, operating at 14.1 Tesla, observing ¹H and ¹³C at 600 and 150 MHz, respectively. High-resolution MS and MS/MS analysis of the pure compounds were performed on a Bruker MicrOTOF-QII ESI-QTOF mass spectrometer coupled to a Dionex (Sunnyvale, CA, USA) Ultimate 3000 RS UHPLC. LC-MS and MS analyses of CHCl₃ extract and the plant material were carried on a Waters (Milford, MA, USA) Xevo G2-S QTOF mass spectrometer equipped with exchangeable ESI and ASAP sources and coupled to an Acquity UPLC (Waters) chromatograph. A CD spectrum of 3, recorded in MeOH on a Jasco (Groß-Umstadt, Germany) J815 spectropolarimeter, did not show any cotton effects.

Optical rotation was measured on a JASCO P-1020 digital polarimeter (Tokyo, Japan). UV data were recorded on a JASCO V-530 UV/VIS Spectrophotometer (Tokyo, Japan). High-resolution ESIMS data were obtained on a Bruker APEX II spectrometer (Billerica, MA, USA)). The IR spectrum was measured on a Perkin Elmer system 2000 FT-IR spectrophotometer (Waltham, MA, USA). The NMR spectra were obtained by JEOL JNM-ECS 400 MHz NMR (Akishima, Japan). Merck (Darmstadt, Germany) silica gel 60 and GE Healthcare (Chicago, IL, USA) Sephadex LH-20 were used for column chromatography. The instrumentation for HPLC was composed of a Shimadzu LC-10AD pump (Kyoto, Japan) and a Shimadzu SPD-M10A PDA detector.

Perkin Elmer (Waltham, MA, USA) system 2000 FT-IR spectrophotometer was used for IR spectrum measurement. JASCO (Tokyo, Japan) P-1020 digital polarimeter was utilized for optical rotation measurement. UV and ECD spectra were recorded on JASCO V-530 UV/VIS spectrophotometer and JASCO J-815 CD spectrometer, respectively. Electrospray ionization (ESI) mass data were obtained from Waters (Milford, MA, USA) 2695 separations module and Bruker (Billerica, MA, USA) APEX II spectrometer (high resolution). NMR spectra were obtained by Bruker AVIII HD 700 MHz FT-NMR and AVAN CEIII 600 MHz FT-NMR. Merck (Darmstadt, Germany) silica gel 60 and GE Healthcare (Chicago, IL, USA) Sephadex LH-20 were used for column chromatography. The instrumentation for HPLC was composed of a Shimadzu (Kyoto, Japan) LC-20AD pump and a Shimadzu SPD-M20A PDA detector.