Development and Method Validation of Cannabinoid Contents In Cannabis sativa L. Leaves by Ultra High Performance Liquid Chromatography

Abstract

         Cannabis (Cannabis sativa L.) has been used to treat several diseases since the ancient time. Tetrahydrocannabinol (THC) is the major psychoactive compound and the other non-psychoactive ingredients are Cannabidiol (CBD) and Cannabinol (CBN). THC has been used for pain relief, reducing nausea from chemotherapy and muscle contraction treatment. CBD has antiepileptic mechanism, relieve pain and neuroprotective property for Parkinson's disease, while CBN has a potential use for anti-cancer therapy. Cannabis have been used as active ingredient of several drugs and products in the markets. However, the appropriate analytical methods for quality control of those compounds are still limited. This study aimed to develop analytical methods for quantification assessment of CBD, CBN and THC in cannabis leaf samples using the Ultra High Performance Liquid Chromatography (UHPLC) technique. Samples were prepared by refluxing in ethanol, filtering and evaporating until dry. The residue was redissolved and adjusted with methanol and injected into UHPLC system. Linearity was established for CBD, CBN and THC at concentration range of 1.07-106.89, 1.00-99.60 and 1.00-99.90 μg/ml, respectively, with a determination correlation coefficient (r) of 0.999. The %recovery was in the range of 96.83-100.89% and HORRAT was in the range of 0.30-1.96. The limit of detection (LOD) of CBD, CBN and THC were 0.006, 0.009, and 0.003 %w/w, respectively, while limit of quantitation (LOQ) was 0.020, 0.032, and 0.011 %w/w, respectively. It has been shown that the developed method is suitable and can be used to analyze the content of CBD, CBN and THC in cannabis leaves. Therefore, the appropriate specification of cannabinoid contents (CBD, CBN and THC) in cannabis leaf extract can be established.