A reliable and validated LC-MS/MS method for the simultaneous quantification of 4 cannabinoids in 40 consumer products

Abstract

In the past 50 years, Cannabis sativa (C. sativa) has gone from a substance essentially prohibited worldwide to one that is gaining acceptance both culturally and legally in many countries for medicinal and recreational use. As additional jurisdictions legalize Cannabis products and the variety and complexity of these products surpass the classical dried plant material, appropriate methods for measuring the biologically active constituents is paramount to ensure safety and regulatory compliance. While there are numerous active compounds in C. sativa the primary cannabinoids of regulatory and safety concern are (-)-Δ⁹-tetrahydrocannabinol (THC), cannabidiol (CBD), and their respective acidic forms THCA-A and CBDA. Using the US Food and Drug Administration (FDA) bioanalytical method validation guidelines we developed a sensitive, selective, and accurate method for the simultaneous analysis CBD, CBDA, THC, and THCA-A in oils and THC & CBD in more complex matrices. This HPLC-MS/MS method was simple and reliable using standard sample dilution and homogenization, an isocratic chromatographic separation, and a triple quadrupole mass spectrometer. The lower limit of quantification (LLOQ) for analytes was 0.195 ng/mL over a 0.195–50.0 ng/mL range of quantification with a coefficient of correlation of >0.99. Average intra-day and inter-day accuracies were 94.2–112.7% and 97.2–110.9%, respectively. This method was used to quantify CBD, CBDA, THC, and THCA-A in 40 commercial hemp products representing a variety of matrices including oils, plant materials, and creams/cosmetics. All products tested met the federal regulatory restrictions on THC content in Canada (<10 μg/g) except two, with concentrations of 337 and 10.01 μg/g. With respect to CBD, the majority of analyzed products contained low CBD levels and a CBD: CBDA ratio of <1.0. In contrast, one product contained 8,410 μg/g CBD and a CBD: CBDA ratio of >1,000 (an oil-based product). Overall, the method proved amenable to the analysis of various commercial products including oils, creams, and plant material and may be diagnostically indicative of adulteration with non-hemp C. sativa, specialized hemp cultivars, or unique manufacturing methods.

<p>Cannabinoid, Terpene, and Heavy Metal Analysis of 29 Over-the-Counter Commercial Veterinary Hemp Supplements</p>

“…Prior work for human products has examined a smaller profile of cannabinoids in over-the-counter hemp products, showing numerous discrepancies with what was tested versus labeling concentrations. [12][13][14] Notes: a COA results that were based on dry materials used in preparation of final product; these products were not used in final calculations of percentage of company COA. Company reported COA bolded numbers represent COA analysis from the same company we utilized in this study (#11, 18,25).…”

“…Two recent publications examining selected cannabinoid concentrations in human over-the-counter products showed a tremendous disparity between labeling claims and analysis of the products, with over 40% having less than the labeled amount and over 40% having more than the labeled amount. 12,13 The THC concentrations in a Canadian study were less than 0.01% for all products showing compliance with the Canadian standard for Cannabis CBD products. 12 In a study examining 14 European products, all with THC concentrations being below the 0.2% allowable limit, CBD concentrations varied from either total cannabinoids or specific CBD concentrations as labeled.…”

“…12,13 The THC concentrations in a Canadian study were less than 0.01% for all products showing compliance with the Canadian standard for Cannabis CBD products. 12 In a study examining 14 European products, all with THC concentrations being below the 0.2% allowable limit, CBD concentrations varied from either total cannabinoids or specific CBD concentrations as labeled. 14 This disparity in products can be from a range of issues including batch to batch variation, intentional improper labeling, degradation over time, poor extraction techniques and lack of certification of the laboratories being used to measure cannabinoids.…”

Quantification of Cannabinoids in Cannabis Oil Using GC/MS: Method Development, Validation, and Application to Commercially Available Preparations in Argentina

“…This process was optimized for dilution solvent and sonication time. Dilution solvents had been selected according to the existing literature [23,25,32,33] and to the physical-chemical properties of the studied analytes. The solvents evaluated were methanol, ethanol, diethyl ether, and petroleum ether.…”

“…Several methods, based on GC [21][22][23][24] or LC [24][25][26] have been published for the determination of Δ 9 -THC, CBD, and other cannabinoids in cannabis oil. GC, one of the most used techniques for the quantitative analysis of cannabinoids in plant materials, has been in use for a long time.…”