Elaboration and Validation of HPLC/DAD Method for Quality Control of Products Containing Cannabidiol

Abstract

Background: Cannabidiol (CBD) is the main, physiologically active, but psycho-inactive constituent of the glandular hairs of Cannabis sativa. CBD exhibits diverse pharmacological activities and it is used as anticonvulsant, sedative and hypnotic agent. Other biological properties shown by CBD are antipsychotic, anti-inflammatory and neuroprotective. Objective: The aim of this study is to develop, validate and apply high performance liquid chromatography (HPLC/DAD) method with diode array detection for identification and assay of CBD in food supplements obtained by different types of extraction and purification. Method: HPLC method with isocratic elution using column C18 ODS, 250 x 4.6, 100 A, 5 μm, mobile phase - mixture of acetonitrile and water in ratio 80:20v/v, flow rate of 1.0 ml/min and DAD detection at 220 nm. Results: The method was developed and validated according to European Pharmacopoeia and International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH). For the purposes analytical parameters repeatability, linearity, accuracy, LOD, LOQ, specificity and system suitability test were studied. The developed method was successfully applied on a series of food supplements containing CBD. Conclusion: The developed method is of practical importance for determination of purity, identification and assay tests of food supplements containing CBD extract in accordance with EU regulation concerning food authorizations because it is useful for analytical tests for identification, purity determination and assay of products containing CBD extract and could serve as part of the testing process in the pharmaceutical industry.

Progress in the analysis of phytocannabinoids by HPLC and UPLC (or UHPLC) during 2020–2023

“…Cannabis seeds have long been used as a food source, and there are several food products available in the marker that contain purified cannabinoids or cannabis extracts. [175][176][177][178][179][180][181] Therefore, the qualitative and quantitative analysis of cannabinoids in food and food supplements and beverages is essential for quality assurance and the control of dietary intake of cannabinoidcontaining food items. 2 An HPLC-DAD method, employing a simple isocratic elution with 80% ACN in water (both containing 0.1% HCOOH) at 1 ml/min, has recently been reported for the analysis of cannabinoids, particularly CBD (5) and THC (14) in cannabis-infused ice cream 103 (Table 9).…”

“…This study also looked at the stability of these cannabinoids in ice cream over period of as long as 270 days. A similar isocratic HPLC-DAD method was used for the determination of CBD (5) in various food supplements that contain CBD ( 5) 178 (Table 9). Dawson and Martin 177 investigated chocolate matrix interference on cannabinoid analytes, for example, CBD (5), CBG (8), CBN (10), and THC (14), using an HPLC-DAD method with a much shorter run time (7 min), using an ACN/water gradient and at a slightly higher temperature (40 C) than the room temperature (Table 9).…”

“…Since the publication of the review article 2 on the application of HPLC and UPLC (or UHPLC) methods in the analysis of phytocannabinoids covering the period of 2010–2019, several publications have appeared in the literature on this topic 12–185 . All these publications highlight the importance of HPLC and UPLC (or UHPLC) techniques as well as showcase remarkable advancements in sensitivity and versatility associated with these techniques in relation to phytocannabinoid analysis 12 .…”

“…Cannabinoids may be present in food and beverages, either as contaminants or as food additives 2,175,176 (Table 9). Cannabis seeds have long been used as a food source, and there are several food products available in the marker that contain purified cannabinoids or cannabis extracts 175–181 . Therefore, the qualitative and quantitative analysis of cannabinoids in food and food supplements and beverages is essential for quality assurance and the control of dietary intake of cannabinoid‐containing food items 2 .…”