Innovative Development and Validation of an HPLC/DAD Method for the Qualitative

Jacob Bell

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Innovative development and validation of an HPLC/DAD method for the qualitative and quantitative determination of major cannabinoids in cannabis plant material

Benjamin De Backera, Corresponding Author Contact Information, E-mail The Corresponding Author, Benjamin Debrusb, Pierre Lebrunb, Laetitia Theunisa, Nathalie Duboisa, Lies Decockc, Alain Verstraetec, Philippe Hubertb and Corinne Charliera

aLaboratory of Clinical, Forensic and Environmental Toxicology, CIRM, CHU Sart-Tilman, University of Liège, B-4000 Liège, Belgium

bLaboratory of Analytical Chemistry, CIRM, Department of Pharmacy, University of Liège, B-4000 Liège, Belgium

cDepartment of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium
Received 28 August 2009;
accepted 1 November 2009.
Available online 6 November 2009.

Abstract

GC is commonly used for the analysis of cannabis samples, e.g. in forensic chemistry. However, as this method is based on heating of the sample, acidic forms of cannabinoids are decarboxylated into their neutral counterparts. Conversely, HPLC permits the determination of the original composition of plant cannabinoids by direct analysis. Several HPLC methods have been described in the literature, but most of them failed to separate efficiently all the cannabinoids or were not validated according to general guidelines. By use of an innovative methodology for modelling chromatographic responses, a simple and accurate HPLC/DAD method was developed for the quantification of major neutral and acidic cannabinoids present in cannabis plant material: Δ9-tetrahydrocannabinol (THC), THC acid (THCA), cannabidiol (CBD), CBD acid (CBDA), cannabigerol (CBG), CBG acid (CBGA) and cannabinol (CBN). Δ8-Tetrahydrocannabinol (Δ8-THC) was determined qualitatively. Following the practice of design of experiments, predictive multilinear models were developed and used in order to find optimal chromatographic analytical conditions. The method was validated following an approach using accuracy profiles based on β-expectation tolerance intervals for the total error measurement, and assessing the measurements uncertainty. This analytical method can be used for diverse applications, e.g. plant phenotype determination, evaluation of psychoactive potency and control of material quality.

Keywords: HPLC; Cannabis sativa; Cannabinoids; THC; CBD; CBG


Source: Innovative development and validation of an HPLC/DAD method for the qualitative and quantitative determination of major cannabinoids in cannabis plant material
 
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