Journal:Enzyme immunoassay for measuring aflatoxin B1 in legal cannabis

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Full article title Enzyme immunoassay for measuring aflatoxin B1 in legal cannabis
Journal Toxins
Author(s) Di Nardo, Fabio; Cavalera, Simone; Baggiani, Claudio; Ciarello, Matteo; Pazzi, Marco; Anfossi, Laura
Author affiliation(s) University of Turin
Primary contact Email: laura dot anfossi at unito dot it
Year published 2020
Volume and issue 12(4)
Article # 265
DOI 10.3390/toxins12040265
ISSN 2072-6651
Distribution license Creative Commons Attribution 4.0 International
Website https://www.mdpi.com/2072-6651/12/4/265/htm
Download https://www.mdpi.com/2072-6651/12/4/265/pdf (PDF)

Abstract

The diffusion of the legalization of cannabis for recreational, medicinal, and nutraceutical uses requires the development of adequate analytical methods to assure the safety and security of such products. In particular, aflatoxins are considered to pose a major risk for the health of cannabis consumers. Among analytical methods that allow for adequate monitoring of food safety, immunoassays play a major role thanks to their cost-effectiveness, high-throughput capacity, simplicity, and limited requirement for equipment and skilled operators. Therefore, a rapid and sensitive enzyme immunoassay has been adapted to measure the most hazardous aflatoxin B1 in cannabis products. The assay was acceptably accurate (recovery rate: 78–136%), reproducible (intra- and inter-assay means coefficients of variation 11.8% and 13.8%, respectively), and sensitive (limit of detection and range of quantification: 0.35 ng mL−1 and 0.4–2 ng mL−1, respectively corresponding to 7 ng g−1 and 8–40 ng g−1 in the plant), while providing results which agreed with a high-performance liquid chromatographytandem mass spectrometry (HPLC-MS/MS) method for the direct analysis of aflatoxin B1 in cannabis inflorescence and leaves. In addition, the carcinogenic aflatoxin B1 was detected in 50% of the cannabis products analyzed (14 samples collected from small retails) at levels exceeding those admitted by the European Union in commodities intended for direct human consumption, thus envisaging the need for effective surveillance of aflatoxin contamination in legal cannabis.

Keywords: mycotoxins, food safety, medicinal herbs, competitive immunoassay

Introduction

Cannabis sativa is a plant of the Cannabaceae family and is well-known for its content of biologically active chemical compounds, among which are the major compounds Δ9-tetrahydrocannabinol (Δ9-THC) and cannabidiol (CBD). The flowering or fruiting tops of the Cannabis plant have been controlled in the United States under the Controlled Substances Act since 1970 under the drug class “Marihuana.”[1]

Cannabis products can be used for medicinal purposes (whether using the psychoactive constituent THC or the non-psychoactive constituent CBD, generally referred to as "medical cannabis"), in manufacturing ("industrial hemp"), and for non-medical intoxication ("recreational or psychoactive cannabis").[2] The number of active constituents found in cannabis and the variety of their effects have also suggested cannabis' potential use as a dietary supplement and nutraceutical.[1][3] According to the World Health Organization (WHO), recreational cannabis is the most widely used illicit drug and the most largely cultivated and trafficked worldwide.[4]

The therapeutic application of cannabis is increasing around the world.[5] For example, a medicine based on cannabis extract has been approved by the European Medicines Agency.[6] THC can be medically administered as capsules, mouth spray, or as flowers for making tea. And the U.S. Food and Drug Administration (FDA) has approved one cannabis-derived and three cannabis-related drug products.[7]

The cultivation and supply of cannabis for industrial use has been legal in the European Union since 2013, provided the cannabis' THC content does not exceed 0.2%.[8] In 2018, the U.S. legalized the production and marketing of hemp, provided that its THC content is below 0.3% on a dry weight basis.[1]

As cannabis increasingly becomes legalized for recreational purposes, dietary supplements, and various medical applications, growth of the global legal market of such products looks favorable in the coming years. However, the toxicity of common cannabis contaminants to humans is largely unknown. Due to the ambiguity between legal and illicit production and supply of cannabis products, there is a significant lacking in the literature regarding the prevalence of cannabis contaminants and of their harmfulness to humans. Contemporarily, the expanded use of cannabis products demands further research in this area, especially for therapeutic uses.[9]

Several classes of contaminants can be present in cannabis, including heavy metals, which are able to bioaccumulate in Cannabis plants[10]; pesticides, (including illegal pesticides; given how long cannabis has been illegal, pesticide guidelines or maximal limits for pesticide residues have not been set for this substrate); microbiological contaminants; and toxins from microbial overloads, such as ochratoxins and aflatoxins.[11][12]


References

  1. 1.0 1.1 1.2 U.S. Food and Drug Administration. "FDA Regulation of Cannabis and Cannabis-Derived Products, Including Cannabidiol (CBD)". U.S. Food and Drug Administration. https://www.fda.gov/news-events/public-health-focus/fda-regulation-cannabis-and-cannabis-derived-products-including-cannabidiol-cbd. Retrieved 10 July 2019. 
  2. Mead, A. (2019). "Legal and Regulatory Issues Governing Cannabis and Cannabis-Derived Products in the United States". Frontiers in Plant Science 10: 697. doi:10.3389/fpls.2019.00697. PMC PMC6590107. PMID 31263468. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6590107. 
  3. Hartsel, J.A.; Eades, J.; Hickory, B.; Makriyannis, A. (2016). "Chapter 53: Cannabis sativa and Hemp". In Gupta, R.C.. Nutraceuticals: Efficacy, Safety and Toxicity. Academic Press. pp. 735–754. ISBN 9780128021477. 
  4. World Health Organization. "Cannabis". Management of substance abuse. World Health Organization. https://www.who.int/substance_abuse/facts/cannabis/en/. Retrieved 20 April 2020. 
  5. Bridgeman, M.B.; Abazia, D.T. (2017). "Medicinal Cannabis: History, Pharmacology, And Implications for the Acute Care Setting". P & T 42 (3): 180–88. PMC PMC5312634. PMID 28250701. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5312634. 
  6. European Monitoring Centre for Drugs and Drug Addiction (December 2018). "Medical use of cannabis and cannabinoids: Questions and answers for policymaking". EMCDDA. doi:0.2810/979004. https://www.emcdda.europa.eu/publications/rapid-communications/medical-use-of-cannabis-and-cannabinoids-questions-and-answers-for-policymaking_en. Retrieved 04 November 2019. 
  7. Corroon, J.; Kight, R. (2018). "Regulatory Status of Cannabidiol in the United States: A Perspective". Cannabis and Cannabinoid Research 3 (1): 190-194. doi:10.1089/can.2018.0030. PMC PMC6154432. PMID 30283822. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6154432. 
  8. "Regulation (EU) No 1307/2013 of the European Parliament and of the Council". Official Journal of the European Union. 20 December 2013. pp. 608–70. https://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2013:347:0608:0670:EN:PDF. 
  9. Dryburgh, L.M.; Bolan, N.S.; Grof, C.P.L. et al. (2018). "Cannabis Contaminants: Sources, Distribution, Human Toxicity and Pharmacologic Effects". British Journal of Clinical Pharmacology 84 (11): 2468-2476. doi:10.1111/bcp.13695. PMC PMC6177718. PMID 29953631. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6177718. 
  10. Zerihun, A. Chandravanshi, B.S.; Debebe, A. et al. (2015). "Levels of Selected Metals in Leaves of Cannabis Sativa L. Cultivated in Ethiopia". SpringerPlus 4: 359. doi:10.1186/s40064-015-1145-x. PMC PMC4503701. PMID 26191486. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4503701. 
  11. Llewellyn, G.C.; O'Rear, C.E. (1977). "Examination of Fungal Growth and Aflatoxin Production on Marihuana". Mycopathologia 62 (2): 109–12. doi:10.1007/BF01259400. PMID 414138. 
  12. Wilcox, J.; Pazdanska, M.; Milligan, C. (2020). "Analysis of Aflatoxins and Ochratoxin A in Cannabis and Cannabis Products by LC-Fluorescence Detection Using Cleanup With Either Multiantibody Immunoaffinity Columns or an Automated System With In-Line Reusable Immunoaffinity Cartridges". Journal of AOAC International 103 (2): 494–503. doi:10.5740/jaoacint.19-0176. PMID 31558181. 

Notes

This presentation is faithful to the original, with only a few minor changes to presentation. Some grammar and punctuation was cleaned up to improve readability. In some cases important information was missing from the references, and that information was added. In the original article, citations 1 and 4 are duplicates; that duplication was removed for this version.