Quantitative Analysis of 16 Cannabinoids with Complete Baseline Separation of Δ8 and Δ9 THC Utilizing the Triple Quad LCMS 8050

Significance of the topic

Analysis of cannabinoids in cannabis and hemp products demands high sensitivity and selectivity to ensure regulatory compliance and product quality. Clear differentiation of isomeric compounds such as Δ8-THC and Δ9-THC is vital due to their distinct pharmacological and legal implications.

Objectives and study overview

This study aimed to establish a rapid UHPLC-MS/MS method capable of quantifying sixteen cannabinoid analytes with complete baseline separation of Δ8- and Δ9-THC. The goal was to achieve a sub-five-minute run time while preserving analytical performance for routine high-throughput testing.

Methodology and instrumentation

Cannabinoid standards were prepared from an eleven-component Shimadzu mixture and five individual compounds from Sigma Aldrich. Calibration curves covered 1–2000 ng/mL in a 25:75 water/methanol solvent. Cannabis extracts were diluted up to 1:999 with ethanol. Separation utilized a Restek Raptor ARC-18 column under a mini-gradient protocol, promoting rapid elution and sharp peaks.

Instrumentation

• UHPLC: Shimadzu Nexera X2 with 40 °C column oven and 5 µL injection volume
• Column: Restek Raptor ARC-18, 1.8 µm particle size
• Mass spectrometer: Shimadzu LCMS-8050 triple quadrupole with heated ESI
• Gases: Nebulizing (3 L/min), Drying (10 L/min), Heating (10 L/min)
• Temperatures: Interface 370 °C, Desolvation Line 250 °C, Heat Block 400 °C

Main results and discussion

The method achieved baseline resolution of all sixteen cannabinoids, including the critical Δ8-THC/Δ9-THC pair. Twelve-point calibration yielded linearity (R² ≥ 0.9924) across 1–2000 ng/mL. The limit of quantitation was 1 ng/mL (S/N ≥10). Precision (%RSD) remained below 9.9% and accuracy ranged from 90.9% to 107.0%.

Benefits and practical applications

• Under-five-minute analysis enables high-throughput quantification of major and minor cannabinoids
• Accurate separation of Δ8-THC and Δ9-THC supports regulatory and quality control needs
• High sensitivity allows detection down to 1 ng/mL without matrix interference
• Suitable for raw materials and finished product testing in pharmaceutical, nutraceutical, and hemp industries

Future trends and possibilities

Incorporation of isotopically labeled internal standards can address matrix effects and native analyte levels. Expanding the analyte panel to include additional phytocannabinoids and metabolites, integrating high-resolution mass spectrometry for untargeted profiling, and automating sample preparation will further enhance throughput and analytical depth.

Conclusion

This application demonstrates that a sub-five-minute UHPLC-MS/MS workflow on the Shimadzu Nexera LCMS-8050 platform delivers a rapid, robust, and selective method for quantifying sixteen cannabinoids with reliable baseline separation of Δ8- and Δ9-THC, meeting critical demands of cannabis analytics.

Reference

1. Shimadzu Scientific Instruments. The Determination of CBD and General Cannabinoid Content in Hemp Oils Using HPLC with UV Detection. 2016.
2. Restek Corporation. 16 Cannabinoids on Raptor ARC-18 1.8 µm by LC-UV. 2017.