6460 jet stream triple quadrupole qqq mass spectrometer

An Agilent 1200 HPLC apparatus coupled with an Agilent 6460 Jet Stream triple quadrupole (QQQ) mass spectrometer was used in this study. Using a chromatographic Agilent Poroshell EC-C18 column with 2.7 μm particles and dimensions of 100 × 2.1 mm ID after an Agilent Poroshell EC-C18 precolumn with 4.6 μm particles, separation of cannabinoids was achieved with a mobile phase of water containing 0.1% formic acid (A) and acetonitrile containing 0.1% formic acid (B). The initial conditions were 34% of B held for 8 min; then, B was increased to 95% over 4 min and maintained for 1 min; then, B was reduced to 34% over 1 min and maintained for 6 min with an additional 3 min post-run. The flow rate was 0.2 mL/min, and the column temperature was maintained at 35 °C. Detection was performed in negative ion mode, and analytes were ionised by electrospray and monitored in multiple reaction monitoring (MRM) mode. Optimised mass spectrometer parameters were: gas temperature 300 °C, gas flow 5 L/min, nebuliser voltage 35 V, sheath gas temperature 250 °C at flow 11 L/min, and capillary and nozzle voltage 4000 V and 500 V, respectively. The MRM transition ions are shown in Table 1.

LC-MS/MS analysis was performed using Agilent 1200 HPLC instrument in conjunction with an Agilent 6460 JetStream Triple Quadrupole (QqQ) mass spectrometer. (The column: Phenomenex Kinetex C18; 150 × 2.1 mm ID, 2.6 μm particles). The gas temperature was 150 °C, and the gas flow was 5 L/min. The nebulizer voltage was optimized to 30 V, and the sheath gas temperature was 300 °C with a flow of 11 L/min. The capillary and nozzle voltages were 3500 V and 1000 V, respectively. The optimized gradient program of the mobile phase (A: 0.1% formic acid in water, B: 0.1% formic acid in acetonitrile) was used: 0 min 50% B, 3 min 90% B, 6 min 90% B, 7 min 50% B and held for 5 min. The mobile phase flow was 0.5 mL/min at 30 °C, and 5 µL of the sample was injected onto the column.