The Potency Determination of 15 Cannabinoids using the Hemp Analyzer

Importance of Topic

Industrial hemp and its derived products have experienced rapid market growth driven by applications in pharmaceuticals, nutraceuticals, cosmetics and textiles. Reliable potency testing of cannabinoids is critical to ensure product quality, consumer safety and regulatory compliance. High performance liquid chromatography (HPLC) remains a key analytical tool for quantifying multiple cannabinoids in complex hemp matrices.

Study Objectives and Overview

This work aimed to develop and validate a fast, robust HPLC method for simultaneous quantitation of 15 major cannabinoids. The method leverages the Shimadzu Hemp Analyzer, an integrated HPLC system with UV detection, to achieve high sensitivity, accuracy and reproducibility for routine potency analysis in research, manufacturing and quality control laboratories.

Methodology and Instrumentation

Sample preparation involved mixing certified reference materials and supplementing with cannabinoid standards to achieve a 100 µg/mL 15-component mixture. Calibration employed six concentration levels (0.5–100 µg/mL) with three QC levels (2.5, 25 and 75 µg/mL). Key instrument parameters:

• System: Shimadzu Hemp Analyzer with UV‐Vis detector at 220 nm
• Mobile phases: 0.085% phosphoric acid in water (A) and in acetonitrile (B)
• Gradient: 70% B → 95% B over 9 min, return to 70% B, total run ~15 min
• Column: NexLeaf CBX 2.7 µm, 4.6 × 150 mm with guard cartridge
• Flow rate: 1.6 mL/min; Oven temperature: 35 °C; Injection: 5 µL

Main Results and Discussion

All calibration curves for the 15 cannabinoids exhibited excellent linearity (R² ≥ 0.9993) using 1/C weighting. QC recoveries ranged from 91% to 108% with relative standard deviations (RSDs) below 5% across low, medium and high levels. Limits of detection and quantitation were determined by ASTM signal‐to‐noise criteria (S/N 3 and 10 respectively), yielding LODs between 0.10 and 0.23 µg/mL and LOQs between 0.31 and 0.70 µg/mL. Representative chromatograms confirmed baseline separation and consistent retention times across replicate injections.

Benefits and Practical Applications

This method offers:

• High throughput: under 15 min per run
• Quantitative accuracy: recoveries within 90–110%
• Reproducibility: RSDs <5% for retention time and peak area
• Wide dynamic range: 0.5–100 µg/mL calibration
• Compliance: meets regulatory and research requirements for hemp potency analysis

Future Trends and Opportunities

Advancements may include:

• Transition to UHPLC for further speed and resolution gains
• Integration of mass spectrometric detection for enhanced specificity
• Automation and online sample preparation to streamline workflows
• Expansion to minor cannabinoids and metabolites profiling
• Application in stability testing and product shelf-life studies

Conclusion

The developed HPLC method on the Shimadzu Hemp Analyzer demonstrates rapid, accurate and sensitive quantitation of 15 cannabinoids. Its robust performance supports diverse applications in hemp research, product development and quality control, addressing industry needs for reliable potency testing.

Reference

• Grand View Research. Industrial Hemp Market Size, Share & Trends Analysis Report By Product, By Application, By Region, And Segment Forecasts, 2020–2027.
• Armbruster DA, Tillman MD, Hubbs LM. Limit of detection (LOD)/limit of quantitation (LOQ): comparison of empirical and statistical methods exemplified with GC-MS assays of abused drugs. Clinical Chemistry. 1994;40(1):1233–8.
• Shrivastava A, Gupta VB. Methods for the determination of limit of detection and limit of quantitation of the analytical methods. Journal of Pharmacy & Bioallied Sciences. 2011;2(1):21–5.