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.
Highlights
Cannabis sativa is one of three generally recognized plant species of Cannabis [1]
The isomers: CBD and THC were monitored in positive ion mode using an identical multiple reaction monitoring (MRM) transition (315.0 ! 193.0) and the compounds were distinguished by retention time (Fig 1C)
A second transition was used for qualification of each analyte, and deuterated standards were used as internal standards
Summary
Cannabis sativa is one of three generally recognized plant species of Cannabis [1]. C. sativa has been used for industrial textiles, food production (hemp), medicinal, and illicit psychoactive properties (marihuana) for several thousand years [2]. Not until the mid-20th century were the cannabinoids responsible for the biological effects of C. sativa first identified [10,11]. C. sativa contains a family of approximately 60 structurally similar cannabinoids [12], the majority of research to date has focused upon the psychoactive Δ9-tetrahydrocannabinol (THC) and the structurally similar non-psychoactive cannabidiol (CBD). THC is a partial agonist of CB1 and CB2 whereas CBD is a negative allosteric modulator and so the overall physiological effect of C. sativa is often related to both THC and CBD content [15,16]. While THC and CBD are the most relevant cannabinoids to mammalian biology, C. sativa produces both in their inactive acidic forms [17,18]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
