Five Principal Cannabinoids Separated and Detected in Nine Minutes Using the Axcend Focus LC®

Overview

Five cannabinoids* were separated and detect-ed in ~nine minutes using the Axcend FocusLC® High-Performance Liquid Chromatography system (HPLC): Cannabidiolic Acid, Cannabidi-ol, Cannabinol, Delta(9)-Tetrahydrocannabinol,and Delta(9)-Tetrahydrocannabinolic Acid. Due to its small size, low-weight, hand-portability,and lower solvent usage, the Axcend Focus LC is uniquely suited for field and laboratory analysis of cannabinoid compounds.

Key Points

This brief application note demonstrates the ability of the Axcend Focus LC to detect and separate five principal cannabinoids — Cannabid olic Acid (CBDA), Cannabidiol (CBD), Cannabi- nol (CBN), Delta(9)-Tetrahydrocannabinol (Δ9-THC); and Delta(9)-Tetrahydrocannabinolic acid A (THCA-A) — at 100 ppm in ~nine minutes with a 50:50water:methanol injection standard. UV detection occurred at 255 nm** on a 10 cm, 150 µm internal diameter nano-flow capillary column packed with 1.8 µm C18.

Introduction

United States cultivation, harvesting, processing,marketing and sales of “Industrial Hemp” has been legal since December 20, 2018. In addition, increased acceptance and legalization of medicinal and recreational marijuana in many U.S. states (as well as abroad), has created a significant and growing demand for lab and field cannabinoid analysis.An Axcend Focus LC was used in this experiment: a shoebox-sized, lightweight (17.25 lbs /7.8 kg), and hand-portable HPLC ideal for cannabinoid detection and analysis in field and lab settings. In addition, as a nano-flow capillary HPLC, the Axcend Focus LC consumes approximately 1/500th of the solvents of traditional HPLCs, dramatically reducing the Total Cost of Ownership (TCO) per instrument.

UV detection of five cannabinoids at 255 nm using the Axcend Focus LC. Elution Order — 3.3 min: CBDA; 3.6 min: CBD; 5 min: CBN; 5.8 min: Δ9-THC; and 7 min: THCA-A. Flow rate: 3 µL/min; Gradient: 65% - 85% B, 10 min; Equilibration Time: 2 min; Mobile Phase A (3:97:0.1 acetonitrile [ACN]:water [H2O]:trifluroacetic acid [TFA]); Mobile Phase B (97:3:0.1 ACN:H2O:TFA); Column: 10 cm x 150 µm i.d.; 1.8 µm C18.

Target Cannabanoids

• CBDA: Cannabidiolic Acid
• CBD: Cannabidiol
• CBN: Cannabinol
• Δ9-THC: Delta(9)-Tetrahydrocannabinol
• Injection Volume: 40 nL
• HCA-A: Delta(9)-Tetrahydrocannabino-lic acid A

Run Conditions

• Detection: UV at 255 nm
• Column Length: 10 cm
• Internal Diameter: 150 µm
• Packing: 1.8 µm C18
• Injection Volume: 40 nL
• Flow Rate: 3 µL/min

Summary

Five primary cannabinoids were separated and detected using the Axcend Focus LC. In addition,with only 36 µL of solvents consumed during this analysis, the Axcend Focus LC offers a significant reduction in the Total Cost of Ownership for organizations conducting cannabinoid testing using HPLCs.