A Validated GC-MS Method for Major Terpenes Quantification in Hydrodistilled Cannabis sativa Essential oil.

Introduction: Terpenes, which are found in high concentrations in the essential oil fraction of the Cannabis sativa flower, have demonstrated potential in many therapeutic and industrial applications.

Objectives: This work reports on a method developed for quantifying 18 terpenes in C. sativa essential oil obtained through hydrodistillation. The following method has been evaluated for specificity, selectivity, accuracy, linearity, precision, stability, limit of detection, and limit of quantification.

Materials and methods: Samples were prepared by separating the essential oil fraction through hydrodistillation and then diluting with ethyl acetate containing a 100 μg/mL solution of n-tridecane and octadecane as internal standards. Analysis was performed on a gas chromatograph mass spectrometer (GCMS) using selected ion monitoring (SIM).

Results: The developed method enabled quantification of isomers of nerolidol and ocimene and several coeluting compounds, with recoveries of 87.35%-116.61%. Two cultivars of C. sativa flower were evaluated, and the dominant terpene compounds in both cultivars were β-myrcene (5.85-8.62 mg/g dried plant) and β-caryophyllene (3.89-4.69 mg/g), followed by α-humulene (1.35-1.99 mg/g), limonene (0.91-1.33 mg/g), and α-bisabolol (0.66-0.68 mg/g).

Conclusion: This method provides an accurate and reliable procedure for separating and quantifying the major terpene compounds in C. sativa flower using hydrodistillation and GCMS with SIM. The simplicity and solvent-free nature of the hydrodistillation extraction, combined with the specificity and accuracy of using SIM and external standards, enables the determination of total and individual terpenes concentrations within plant material and supports numerous industrial and therapeutic applications.