Varian 500 ion trap mass spectrometer

Extraction of artemisinin and its derivatives from shoots of 4 months old transformed and wild type plants was carried out using the reported methodology [28 (link)]. HPLC-MS analysis was performed with a Varian 500 Ion Trap Mass spectrometer (Varian, Spain) coupled to a 212-HPLC system equipped with an auto sampler 410 (Varian, Spain). The analytes were separated by a C18 (5μm) column (150mmx 4.6mm) (AKDAY Chromatografica, Spain) using a mobile phase consisting of water with 0.1% formic acid (A) and acetonitrile (B) in a gradient as follows: (t (min), % B) (0, 50) (10, 50) (15, 99) (23, 50) (26, 50). The flow rate was 1ml/min. Electro spray ionization (ESI) in positive mode was applied with a capillary voltage of 4000V and plate voltage of 600V. Other parameters include: spray chamber temperature 50 C, nebulizing gas (N₂) pressure: 35psi, dry gas (N₂) 10 l/min at 35psi and 400C. Mass analyzer scanned from 66–500 u. Fragmentation amplitude was 1.0 V. Three injections per sample were applied.

Extraction of artemisinin and its derivatives from leaves of 4–5 month old transformed and wild type plants was carried out with a previously published method [25 (link)]. HPLC–MS analysis was performed with Varian 500 Ion Trap Mass spectrometer (Varian, Spain) coupled to a HPLC system equipped with auto sampler (Varian, Spain). Separation of the analytes was achieved by C 18 (5 µm) column (150 mm × 4.6 mm) (AKDAY Chromatografica, Spain) using mobile phase consisting of water with 0.1 % formic acid (A) and acetonitrile (B) in a gradient as follows: [t (min), % B] (0, 50) (10, 50) (15, 99) (23, 50) (26, 50). The flow rate was 1 ml/min. Electrospray ionization (ESI) in positive mode was applied with a capillary voltage of 4000 V and plate voltage of 600 V. Other parameters include: spray chamber temperature 50 °C, nebulizing gas (N₂) pressure: 35 psi, dry gas (N₂) 10 L/min at 35 psi and 400 °C. The mass analyzer scanned from 66 to 500 u. Fragmentation amplitude was 1.0 V.