Mass Confirmation of Cannabinoids in Cannabis sativa by using HPTLC-QDa

Importance of the Topic

The accurate identification of the main cannabinoids—cannabinol (CBN), cannabidiol (CBD), and Δ9-tetrahydrocannabinol (Δ9-THC)—in Cannabis sativa is critical for quality control, regulatory compliance, and ensuring safe and effective therapeutic use. Variations in cannabinoid content arise from strain differences and cultivation conditions, so a robust analytical approach is essential for consistent patient dosing and product standardization.

Objectives and Study Overview

This study aimed to develop and validate a high-performance thin layer chromatography (HPTLC) method coupled with mass detection (QDa) to confirm the presence of CBN, CBD, and Δ9-THC in various cannabis matrices, including oils, tinctures, and plant material.

Methodology

Standards of CBN and CBD (1 mg/mL in methanol) and Δ9-THC (1 mg/mL in ethanol) were diluted 1:10 in methanol. Four cannabis products (two plant samples, one oil, one tincture) were extracted in methanol (500 mg sample, sonication 10 min, centrifugation). Sample volumes applied: 5 μL standards, 8 μL plant extracts, 5 μL oil, 2 μL tincture. HPTLC separation used silica 60 F254 plates developed in cyclohexane/di-isopropyl ether/diethylamine (52:40:8). Plates were derivatized in Fast Blue salt B and visualized under white light.

Used Instrumentation

• Automatic TLC Sampler 4 for precise sample application
• Chromatogram Immersion Device for plate derivatization
• TLC-MS Interface 2 with oval elution head
• ACQUITY QDa Detector in negative electrospray ionization mode
• Empower Chromatography Data Software for data processing

Main Results and Discussion

Mass spectra acquired (m/z 50–650 at 10 points/sec) showed clear molecular ions for CBN, CBD, and Δ9-THC in both standards and cannabis oil extracts. The QDa detector provided selective confirmation of each cannabinoid, overcoming limitations of UV detection and eliminating the need for additional derivatization for low-UV-absorbing compounds. HPTLC separation combined with mass detection allowed unambiguous identification even in complex matrices.

Benefits and Practical Applications

• Rapid screening and confirmation of major cannabinoids in diverse cannabis products
• Improved selectivity compared with UV-only methods
• Streamlined workflow by direct mass detection without extra derivatization
• Applicable to quality control in pharmaceutical, nutraceutical, and regulatory laboratories

Future Trends and Potential Applications

Advancements may include expansion to minor and novel cannabinoids, integration with high-resolution mass spectrometry for structural elucidation, further automation of HPTLC-MS workflows, and development of portable platforms for in-field analysis. Quantitative HPTLC-MS and comprehensive fingerprinting approaches could enhance standardization across the cannabis supply chain.

Conclusion

The HPTLC-QDa method delivers a robust, selective, and efficient tool for confirming key cannabinoids in Cannabis sativa products. By combining high-resolution planar separation with direct mass detection, this approach addresses critical quality control needs and streamlines analytical workflows.

References

1. United Nations. Recommended methods for the identification and analysis of cannabis and cannabis products; 2009.