A Rugged High-Resolution Potency Determination of 18 Cannabinoids in Hemp Plant Material and Finished Tinctures using the Cannabis Analyzer for Potency

Importance of the Topic

Non-psychotropic and psychoactive cannabinoids are gaining recognition for their therapeutic benefits, ranging from pain and inflammation relief to seizure suppression and metabolic regulation.
Accurate quantification of these compounds is critical for research, product labeling, regulatory compliance and quality control in the rapidly expanding cannabis and hemp industries.

Objectives and Study Overview

This study aimed to develop and validate a high-resolution, rugged HPLC-UV method capable of separating and quantifying 18 major cannabinoids in hemp plant material and finished tincture oils within a single 30-minute run.
The method builds on a previous 11-component analysis, expanding the target list while retaining throughput and robustness.

Instrumentation Used

• Shimadzu Cannabis Analyzer for Potency™ integrated HPLC system with built-in UV-Vis detector operating at 220 nm
• Shim-pack XR-ODS II column (75 × 3.0 mm, 2.2 μm) with NexLeaf CBX guard column cartridge
• Mobile phases: 0.085 % phosphoric acid in water (A) and 0.085 % phosphoric acid in methanol (B)
• Gradient elution: 60 % B (5 min) → 72 % B (over 11 min) → 95 % B (6 min) → hold (2 min) → re-equilibration (1 min) → re-equilibration (60 % B, 5 min)
• Flow rate 1 mL/min, column temperature 50 °C, injection volume 5 μL
• LabSolutions software (v5.99) for data acquisition, calibration and custom potency calculations

Methodology

Calibration standards for 18 cannabinoids were prepared at six concentration levels (0.5–85 mg/L) with QC mixtures at 2.5, 25 and 75 mg/L. A 1/C weighting was applied in linear regression and all curves achieved R² ≥ 0.999.
Dry flower samples were homogenized using a Geno/Grinder (100 mg sample with steel balls), extracted in methanol, diluted 100× and filtered through 0.45 μm syringe filters before injection.
Oil-based tinctures were diluted 1000× by dissolving 10 μL in isopropanol, adding methanol, and filtering prior to analysis.

Results and Discussion

The method achieved baseline resolution of 18 cannabinoids within 30 min and demonstrated excellent precision (RSD < 0.4 %) and accuracy (98.8–103 %) across QC levels.
System sensitivity and dynamic range supported quantification of both major (CBD, THC) and minor cannabinoids (THCV, CBDA, CBGA, CBCV, etc.).
Analysis of two dry flower samples revealed total CBD levels of 96.3 % (flower-1, non-hemp) and 87.2 % (flower-2, compliant hemp), with corresponding total THC of 3.6 % (exceeding the 0.3 % limit) and negligible THC, respectively.
Tincture profiling quantified CBD and THC contents consistent with product labels, demonstrating the method’s applicability to finished formulations. Custom equations in LabSolutions enabled rapid conversion to potency metrics (mg/g, wt %) within the same workflow.

Benefits and Practical Applications of the Method

• High throughput analysis supporting 30-minute runtime with integrated wash and re-equilibration steps
• Accurate quantitation of a broad cannabinoid panel for R&D, QC and regulatory compliance
• Simple sample preparation protocols adaptable to both plant and oil matrices
• Turnkey solution with intuitive software for calibration, QC monitoring and potency reporting

Future Trends and Potential Applications

Further expansion to emerging cannabinoids (e.g., CBT, CBDP) and conjugated forms via advanced detectors (MS, MS/MS) can enhance profiling capabilities.
Integration with automation platforms or benchtop systems for on-site testing and real-time QC monitoring promises faster decision-making in cultivation, processing and product release.
Advanced data analytics, machine learning and LIMS connectivity will streamline compliance and R&D efforts in evolving regulatory landscapes.

Conclusion

A robust, validated HPLC-UV method was established for the separation and quantification of 18 cannabinoids in hemp biomass and tinctures within a 30-minute run time.
The approach demonstrated excellent linearity, precision and accuracy, making it a practical tool for laboratories requiring comprehensive potency analysis.

References

• Sifferlin A. Can Medical Marijuana Help End the Opioid Epidemic? Time Magazine. July 2016.
• Bachhuber MA, Saloner B, Cunningham CO, Barry CL. Medical Cannabis Laws and Opioid Analgesic Overdose Mortality in the United States, 1999–2010. JAMA Internal Medicine. 2014;174(10):1668.