Determination of 5 Hemp-Based Cannabinoids & 12 Standard Cannabinoids by Hand-Portable HPLC

Overview

This application note demonstrates the ability of the Axcend Focus LC® portable High-Performance Liquid Chromatography system (HPLC) to detect and separate 5 principal cannabinoids present in 50 mg of a hemp-variant of cannabis flowers (dry weight) , namely Cannabidiolic Acid (CBDA), Cannabigerol Acid (CGBA), Tetrahydrocannabinolic Acid (THCA), Cannabidiol (CBD), and Cannabichromene (CBC). For comparison, 12 additional cannbinoids were also determined in a Restek* standard, with run-times for both analyses of ~18 minutes with a 50:50mixture of ACN 0.3 M NH4Ac buffer pH 4.6 (Mobile Phase A) and MeOH 50% ACN (Mobile Phase B).

Hemp Sample Preparation

• 50 mg of cut Hemp flower inserted into a 5 mL glass tube
• 1 mL of Methanol added
• Shaken for 2 min
• Filtered through a 0.2 um Filter

Run Conditions

• Detection: UV at 235 nm
• Column Length: 10 cm
• Internal Diameter: 150 μm
• Packing: 1.7 μm C8
• Injection Volume: 40 nL
• Flow Rate: 0.97 μL/min

Hemp Cannabinoid Detection

Analysis of 12 Cannabinoids Standard

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. As shown below, we also detected and identified 12 separate cannabinoids in a Restek standard.

UV detection of 12 cannabinoids in a Restek standard at 235 nm using the portable Axcend Focus LC. Elution Order shown above and below. Flow rate: 0.97 μL/min; Gradient %B: 0 min—3%, 1 min—25%, 10 min—25%, 17 min—60%, 18 min—95%, 20 min—95%; Equilibration Time: 4 min; Mobile Phase A (50% H2O 50% ACN 0.3 M NH4Ac buffer pH 4.6); Mobile Phase B (50% MeOH 50% ACN); Column: 10 cm x 150 μm i.d.; Phenomenex Kinetex 1.7 μm C8. (See table below for more details.)