Analysis of cannabinoids content was performed using Waters 2695 (Milford, MA, USA) separation module equipped with auto injector, sample cooler, vacuum degasser and column heater units.
Separation of all cannabinoids was accomplished on YMC PRO C18 (150 x 4 mm I.D., S-3μm) RP column coupled with C18 precolumn maintained at 30 °C by a CTO-20AC column oven. Isocratic elution consisted of acetonitrile:water (FA 0.1%) (4:1) was done in 30 min.
The flow rate was maintained at 0.8 ml/min. The cannabinoid CBD and CBD-A were monitored at 225 and 306 nm wave length respectively using dual absorbance detector Waters 2487 (Milford, MA, USA). The injection volume of 20 μl was injected using auto sampler at 10 °C. Data evaluation was performed using Empower 2 software.