Expanding the Cannabinoid Target List for a Rugged HPLC Potency Method

Significance of the Topic

The quantification of non-psychotropic cannabinoids is critical due to their broad therapeutic potential including pain relief, anti-inflammatory effects, seizure suppression and metabolic regulation. A reliable and high-throughput analytical method is essential in research and quality control to ensure product safety and label accuracy.

Objectives and Study Overview

This study aims to extend a previously established HPLC potency method covering 11 cannabinoids to a high-resolution protocol detecting 18 analytes within a 30-minute run time. The core goals include method robustness, accuracy, and applicability to diverse cannabis matrices such as tincture oils and dried flower.

Methodology

An integrated HPLC approach with a UV-Vis detector was used. Calibration standards from 0.5 to 85 mg/L and QC levels at 2.5, 25 and 75 mg/L were prepared. Separation employed a phosphoric acid–water/methanol gradient (60% to 95% B) over 24 minutes. Sample injection volume was 5 µL and the column was maintained at 50 °C. Linear regression and QC metrics were evaluated using seven replicates.

Instrument Used

• System: Shimadzu Cannabis Analyzer for PotencyTM (220-94420-00)
• Detector: UV-Vis at 220 nm
• Column: Shim Pack XR-ODS II, 75 × 3.0 mm, 2.2 µm
• Guard: NexLeaf CBX Cartridge, 2.7 µm
• Mobile Phase A: 0.085% Phosphoric Acid in Water
• Mobile Phase B: 0.085% Phosphoric Acid in Methanol
• Flow Rate: 1.0 mL/min
• Run Time: 30 min per injection

Main Results and Discussion

All 18 cannabinoids achieved excellent linearity (R2 ≥ 0.9996) and QC accuracy between 98.8% and 103.7%. The high-resolution chromatogram demonstrated baseline separation within a 5 µL injection of 50 ppm mixture. Application to a commercial CBD tincture yielded 637.8 mg CBD vs. 500 mg label claim, while a flower sample showed total CBD of 94.4 wt.% and THC of 3.36 wt.%.

Benefits and Practical Applications

• High throughput quantitation of major and minor cannabinoids in oils and plant material
• Robust method suitable for routine QA/QC and regulatory testing
• Accurate potency verification aligning with product labeling

Future Trends and Possibilities

Future developments may include coupling with mass spectrometry for enhanced sensitivity and selectivity, faster gradient programs to increase sample throughput, expansion to emerging cannabinoids and integration into automated workflows and portable platforms for on-site analysis.

Conclusion

This work presents a rugged, high-resolution HPLC method extending cannabinoid profiling to 18 targets while maintaining accuracy and efficiency, supporting reliable potency assessment in research and commercial settings.

References

1. Pezeshk N, Young C. Expanding the Cannabinoid Target List for a Rugged HPLC Potency Method. Shimadzu Scientific Instruments. Pittcon Virtual Expo 2021.