Nigella sativa subsp. leaves

Elyasi, Rizan, Majdi, Mohammad, Krause, Sandra T., Kücükay, Nagihan, Azizi, Abdolbaset, Jorg & Degenhardt, 2022, Identification and functional characterization of a γ-terpinene synthase in Nigella sativa L (black cumin), Phytochemistry (113290) 202, pp. 1-7 : 5

5.2. Extraction of terpenes from N. sativa leaves , flowers and seeds

Fine powder (80–100 mg) of three replicates of N. sativa samples ground in liquid nitrogen was used for metabolite analysis. 880 μl of nhexane was added to each sample and 20 ng nonyl acetate was used as an internal standard. After short vortexing, samples were shaken for 2 h at room temperature and subsequently centrifuged for 1 min at 20,000 g. The supernatant was transferred to a clean micro-tube fitted for GC vials. Terpene analysis was carried out with a gas chromatograph (GC- QP2010 Plus, Shimadzu, Duisburg, Germany). Alternatively, volatile terpenes were extracted by a solid phase microextraction fiber (SPME) (100 μm polydimethylsiloxane; Supelco, Belafonte, PA, USA). The fiber was exposed in the headspace of 100 mg fine plant powder for 45 min at 35 ◦ C in a water bath. The adsorbed products were analyzed by GC-MS.