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Quantitation of Aflatoxins B1, B2, G1, G2 and Ochratoxin A in Cannabis Using Integrated HPLC with a Fluorescence Detector

Quantitation of Aflatoxins B1, B2, G1, G2 and Ochratoxin A in Cannabis Using Integrated HPLC with a Fluorescence Detector

Introduction

Studies have found that many mycotoxins, including Aflatoxins G1, G2, B1, and B2,  and Ochratoxin A, are immunosuppressive, carcinogenic, neurologically toxic, and hepatotoxic, and that the mold itself can cause diseases such as lung infection and aspergillosis (1)(2)(3). Due to the high level of matrix complexity of cannabis products, mycotoxin testing in cannabis is very challenging. This study focuses on developing a powerful detection method that allows cannabis testing labs to confidently report concentrations below regulatory limits – Aflatoxins B1, B2, G1 and G2 combined to 20 ppb and 20 ppb for Ochratoxin A alone.

Experimental

Standards were run in triplicate at 2, 5, 10, 14, 20, and 50 ppb for the Aflatoxins and 10, 14, 20, and 50 ppb for Ochratoxin A. All standards were spiked in matrix and extracted so the actual concentrations of the injected material were 0.2, 0.5, 1.0, 1.4, 2, and 5 ppb for the Aflatoxins. The Ochratoxin A does not need as low of a detection limit because the limit is 20 ppb by itself; the Aflatoxins is 20 ppb for the sum of the four (4).

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  1. A Hazenkamp, “An evaluation of the quality of medicinal grade cannabis in the Netherlands”, Cannabinoids, 1, 1-9 (2006).
  2. GC Llewellyn, CE O’Rear, “Examination of fungal growth and aflatoxin production on marihuana”, Mycopathologia, 62, 109-112 (1977).
  3. M Sedmikova et al., “Potential hazard of simultaneous occurrence of aflatoxin B1 and ochratoxin A”, Veterinary Medicine - Czech, 46, 169-174 (2001).
  4. Full sample preparation and analytical conditions are available in Shimadzu’s document: SSI-HPLC-019, Quantitation of Aflatoxins B1, B2, G1, G2 and Ochratoxin A in Cannabis by LC Using Prominence-i and the RF-20Axs Fluorescence Detector
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