Development and validation of reverse phase high performance liquid chromatography method for the determination of delta-9-tetrahydrocannabinol and cannabidiol in oromucosal spray from cannabis extract
2018
Abstract A simple, fast, precise, accurate and responsive high performance liquid chromatography method was developed and validated carefully for determining an amount of delta-9-tetrahydrocannabinol and cannabidiol. A reverse phase Zorbax C-18 column 4.6 mm × 100 mm, 3.5 μm was eluted by using a mixture (85:15) of methanol and water as the mobile phase in an isocratic system with a flow rate of 1.0 ml/min and the injection volume was 10 μl, at a wavelength of 220 nm. The method was developed and validated through linearity, accuracy, precision and detection and quantitation limits studies. A good linear relationship ( R 2 ) of delta-9-tetrahydrocannabinol and cannabidiol were 0.9998 and 0.9995, respectively. The obtained % recoveries were found to be 98.3% and 96.5%. The relative standard deviations values of peak areas were found to range from 0.20% to 2.55% and 0.30% to 3.28% for delta-9-tetrahydrocannabinol and cannabidiol, respectively. Delta-9-tetrahydrocannabinol and cannabidiol presented limits of detection of 0.12, 0.23 μg/ml and limits of quantitation of 0.40, 0.76 μg/ml. The developed method could be employed for quantitative analysis of cannabis extract and oromucosal spray formulation.
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