Materials and Reagents

1. Rauvolfia nukuhivensis (Voucher JFB 2808): In our example, the bark of Rauvolfia nukuhivensis was collected at Maauu in “Terre Déserte” area on Nuku Hiva Island, Marquesas archipelago, French Polynesia, at an altitude of 477 m and identified by Dr Jean-François Butaud. A voucher specimen (JFB 2808) has been deposited at the Herbarium of French Polynesia. The sample was dried after collection at 30 °C using an air dryer device and stored in the room until grinding and extraction.
   
2. Ethanol (95%) (Thermo Fisher Scientific, Fisher chemical)
   
3. Methanol (HPLC grade from Fisher chemical)
   
4. Dichloromethane (HPLC grade from Fisher chemical)
   
5. Cyclohexane (HPLC grade from Fisher chemical)
   
6. Ethyl acetate (HPLC grade from Fisher chemical)
   

Equipment

1. SEB Prep'line grinder (SEB)
   
2. Semi-preparative HPLC column Luna 5 μm C18 (50 mm x 10 mm) (Phenomenex, model: RP-C18 )
   
3. Vacuum liquid chromatography (Buchner filter from Buchi, chromatography performed with vacuum pump) (BÜCHI Labortechnik AG)
   
4. High Pressure Liquid Chromatography (HPLC) Waters 600 system equipped with a Waters 717 Plus autosampler, a Waters 998 photodiode array detector (WATERS, model: Waters 998 photodiode array detector ), and a Sedex 75 evaporative light-scattering detector (SEDERE, model: Sedex 75)
   
5. NMR spectrometer (Bruker Corporation, model: Bruker Avance 500 MHz )
   
6. Polarimeter (jascoinc, model: Jasco P-2000 )
   
7. Spectropolarimeter (jascoinc, model: Jasco J-810 )
   
8. Mass spectrometer (Bruker Corporation, model: Bruker ESQUIRE 3000 plus )
   
9. Thermo Finnigan LTQ Orbitrap mass spectrometer (Thermo Fisher Scientific)
   

Procedure