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A Validated Chiral HPLC Method for Resolution of Δ8 and Δ9 – Tetrahydrocannabinol Enantiomers

Authors: Tamara Tarbox, Isil Dilek PhD, Uma Sreenivasan PhD, Kenan Yaser
Presented at EAS, November 2009

Abstract
A chiral HPLC method to quantitatively determine the enantiomeric excess of Δ-tetrahydrocannabinol (Δ9-THC and Δ8-THC) was developed and validated. Baseline separation of all four enantiomers Δ9-THC and Δ8-THC were achieved within 25 minutes. Resolution of (+) and (-) enantiomers was =12 for Δ9-THC and =3 for Δ8-THC. The analysis was performed on a ChiralPak ADH column using Isopropanol:Methanol:n-Heptane (31:9:950) as mobile phase at a flow rate of 0.7 ml/min at 40ºC and monitored with a photodiode array detector at 225 nm. Validation steps involved measurement of accuracy, precision, linearity and range, robustness, system suitability, determination of limit of detection (LOD) and limit of quantitation (LOQ). The limits of detection (LOD) and quantitation (LOQ) were 0.54, 0.23 and 0.93, 0.60 µg/ml for (+) enantiomers of Δ8-THC and Δ9 -THC, respectively. The effects of deliberate changes to flow rate, temperature and injection volume and various mobile phase compositions on resolution of enantiomers were evaluated. The validated method has been demonstrated to be reliable for the determination of enantiomeric excess of tetrahydrocannabinols in pharmaceutical formulations.

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