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Determination of 11 Cannabinoids in Biomass and Extracts of Different Varieties of Cannabis Using High-Performance Liquid Chromatography

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An HPLC single-laboratory validation was performed for the detection and quantification of the 11 major cannabinoids in most cannabis varieties, namely, cannabidiolic acid (CBDA), cannabigerolic acid (CBGA), cannabigerol (CBG), cannabidiol (CBD), tetrahydrocannabivarin (THCV), cannabinol (CBN), Δ9-trans-tetrahydrocannabinol (Δ9-THC), Δ8- trans-tetrahydrocannabinol (Δ8-THC), cannabicyclol (CBL), cannabichromene (CBC), and Δ9-tetrahydrocannabinolic acid-A (THCAA). The analysis was carried out on the biomass and extracts of these varieties. Methanol–chloroform (9:1, v/v) was used for extraction, 4-androstene-3,17-dione was used as the internal standard, and separation was achieved in 22.2 min on a C18 column using a two- step gradient elution. The method was validated for the 11 cannabinoids. The concentration-response relationship of the method indicated a linear relationship between the concentration and peak area with r2 values of >0.99 for all 11 cannabinoids. Method accuracy was determined through a spike study, and recovery ranged from 89.7 to 105.5% with an RSD of 0.19 to 6.32% for CBDA, CBD, THCV, CBN, Δ9-THC, CBL, CBC, and THCAA; recovery was 84.7, 84.2, and 67.7% for the minor constituents, CBGA, CBG, and Δ8-THC, respectively, with an RSD of 2.58 to 4.96%. The validated method is simple, sensitive, and reproducible and is therefore suitable for the detection and quantification of these cannabinoids in different types of cannabis plant materials.
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Document Type: Research Article

Affiliations: 1: ElSohly Laboratories, Inc., 5 Industrial Park Dr, Oxford, MS 38655, USA 2: National Center for Natural Products Research, School of Pharmacy, the University of Mississippi, University, MS 38677, USA; Department of Pharmacognosy, Faculty of Pharmacy, University of Alexandria, Alexandria, Egypt 3: National Center for Natural Products Research, School of Pharmacy, the University of Mississippi, University, MS 38677, USA; Department of Pharmacognosy, Faculty of Pharmacy, Minia University, Minia, Egypt 4: National Center for Natural Products Research, School of Pharmacy, the University of Mississippi, University, MS 38677, USA 5: National Center for Natural Products Research, School of Pharmacy, the University of Mississippi, University, MS 38677, USA; Department of Bimolecular Sciences, School of Pharmacy, the University of Mississippi, University, MS 38677, USA 6: American Herbal Products Association, 8630 Fenton St, Suite 918, Silver Spring, MD 20910, USA 7: ElSohly Laboratories, Inc., 5 Industrial Park Dr, Oxford, MS 38655, USA; National Center for Natural Products Research and Department of Pharmaceutics and Drug Delivery, School of Pharmacy, the University of Mississippi, University, MS 38677, USA

Publication date: November 1, 2015

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  • The Journal of AOAC INTERNATIONAL publishes refereed papers and reviews in the fields of chemical, biological and toxicological analytical chemistry for the purpose of showcasing the most precise, accurate and sensitive methods for analysis of foods, food additives, supplements and contaminants, cosmetics, drugs, toxins, hazardous substances, pesticides, feeds, fertilizers and the environment available at that point in time. The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better methods of preparing samples for analysis; collaborative studies substantiating the performance of a given method; statistical techniques for evaluating data. The Journal will also publish other articles of general interest to its audience, e.g., technical communications; cautionary notes; comments on techniques, apparatus, and reagents.
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