Optimized Extraction Method for Neutral Cannabinoids Quantification Using UHPLC-HRMS/MS

Area of Science:

• Analytical Chemistry
• Pharmacognosy
• Forensic Science

Background:

• The global cannabis market is growing, increasing demand for cannabis herbal extracts (CHE) in clinical applications.
• Neutral cannabinoids like cannabidiol (CBD), tetrahydrocannabinol (THC), and cannabinol (CBN) are key active compounds in CHE.
• Non-standardized cannabinoid content in CHE presents challenges for pharmaceutical quality control and therapeutic efficacy.

Purpose of the Study:

• To develop and optimize a multivariate method for extracting CBD, THC, and CBN from commercial CHE.
• To validate a predictive model using Response Surface Methodology for extraction efficiency.
• To establish a rapid, cost-effective, and reliable analytical method for cannabinoid quantification in CHE.

Main Methods:

• Utilized Full Factorial and Box-Behnken designs for optimizing extraction parameters.
• Employed methanol/hexane (9:1 v/v) solvent system with minimal volume and short extraction times.
• Quantified cannabinoids using ultra-high-performance liquid chromatography coupled to high-resolution tandem mass spectrometry (UHPLC-HRMS/MS).

Main Results:

• Agitation and sonication times were identified as critical extraction parameters, followed by solvent volume.
• The optimized method demonstrated high precision (<15%), accuracy (69-98%), sensitivity (23-39 ng kg-1), and linearity.
• Successfully processed 16 commercial CHE samples within a 3-hour timeframe, proving practicality and reproducibility.

Conclusions:

• Developed a significantly faster, cheaper, and more efficient protocol for cannabinoid extraction compared to existing literature methods.
• Achieved state-of-the-art analytical sensitivity and specificity using UHPLC-HRMS/MS for reliable routine analysis.
• The validated method is suitable for quality control of commercial CHE in both human and veterinary pharmaceutical applications.