## 2 Materials and Methods
### Obtention of Vacuum Distillation Fractions and Pure \(\alpha\)-copaene  
A vacuum-sealed, silver-plated column with dimensions of 40 cm long \(\times\) 3 cm \(\O\) i. and packed with Dixon rings was used to obtain the distilled fractions. An angled Bertrand collector-separator was used to maintain the vacuum during fraction collection. The vacuum was maintained at 5 mm Hg throughout the whole distillation. The chemical composition of distilled fractions was studied by gas chromatography (GC). A Fisons 8000 GC equipped with a Hewlett Packard HP-1 (crosslinked methyl silicone) column (25 m \(\times\) 0.32 mm; film thickness = 0.52 \(\upmu\)m) was used. Compounds were identified by their retention times and characteristic fragmentation patterns in mass spectra obtained by gas chromatography-mass spectrometry (GC-MS) and literary references [51]. The mass spectra were obtained by the electronic impact technique in a Varian Saturn II spectrometer, coupled with GC.  
Extraction of \(\alpha\)-copaene from DCO and IF fractions was performed by two methods: (1) _Open column_. The \(\alpha\)-copaene separation was carried out with silica gel (63-200 \(\upmu\)m) in proportion to 50 g g\({}^{-1}\) of DCO and using hexane as eluent. Column monitoring was made by analytical thin-layer chromatography (silica gel 60 F254 chromatographs), using as revealing I\({}_{2}\); (2) _Preparative HPLC_. A Varian chromatograph was used, with a model 9065 pump and a model 9065 PolyChrom UV detector. The column used was LiChrosorb Si 60 of 7 \(\upmu\)m (25 \(\times\) 2.5 cm\({}^{2}\)).  
Purity confirmation of the isolated compounds was carried out in analytical HPLC [column Spherisorb, 5 \(\upmu\)m (25 \(\times\) 0.7 cm\({}^{2}\))] by studying them with eluents of different polarities. In parallel, a UV detector-photo diode array was used to record chromatograms at different wavelengths. UV spectra were obtained at three different times corresponding to the rising, apex, and falling of each chromatographic peak. Thus, a compound was considered pure when a single peak was recorded at the different wavelengths and the UV spectrum overlapped with the internal standard.  
Infrared spectroscopy (IR) spectra were obtained as a liquid film on a Nicolet 710 FT-IR spectrophotometer, spanning the 4000-600 cm\({}^{-1}\) region. V\({}_{\rm max}\) is indicated for the main absorption bands. One-dimensional nuclear magnetic resonance (NMR) (\({}^{1}\)H, \({}^{13}\)C, and DEPT) and the correlation spectroscopy (COSY) spectra were recorded on a Varian Gemini 300 MHz spectrometer, in CDCl\({}_{3}\) as a solvent.  
The enantiomeric identity of the \(\alpha\)-copaene isolated from DCO was determined by enantio-selective gas chromatography. For this, a 30 m long \(\times\) 0.25 mm \(\O\) i. trifluoroacetyl gamma-cyclodextrintrin (G-TA) column was used. The chromatograph conditions were an initial temperature of 80 \({}^{\circ}\)C for 60 min, rising 1 \({}^{\circ}\)C min\({}^{-1}\) up to 170 \({}^{\circ}\)C, and using He as carrier gas at a pressure of 35 kPa. The optical rotation values were determined in a PerkinElmer polarimeter using light with a wavelength corresponding to line D of the sodium emission spectrum, using CHCl\({}_{3}\) as a solvent. The concentrations were expressed in g 100 mL\({}^{-1}\).