Development of the UHPLC-DAD method for simultaneous determination of four active pharmacological ingredients in cannabinoid plant material
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A stability-indicating ultra-high-performance liquid chromatography (UHPLC) with DAD detector was developed and validated for the simultaneously determination of Δ9-tetahydrocannabitol (THC), cannabidiol (CBD), Δ9-tetahydrocannabinolic acid (THC-COOH) and cannabidiolic acid (CBD-COOH) in extracts of Cannabis sativa L.. The separation was achieved on a Kinetex, C18, 100 x 2.1 mm, 2.6 µm. The gradient separation was started with mixture of methanol and 12mM ammonium acetate in ratio 75:25 (%/%). The methanol was increased to 100% from 0.0 min to 10.0 min, then maintained 100% for the next 1.0 min, then decreased to 75% from 11.0 min to 15.0 min and held at 75% from 15.0 min to 20.0 min. The injection volume was 5.0 µL and the wavelength of detection was controlled at 228 nm.
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