Atropisomeric scaffolds are a common design element found in pharmaceuticals, many deriving from an N–C axis of chirality. The handedness associated with atropisomeric drugs is oftentimes crucial for their efficacy and/or safety. With the increased use of high-throughput screening (HTS) for drug discovery, the need for rapid enantiomeric excess (ee) analysis is needed to keep up with the fast workflow. Here, we describe a circular dichroism (CD) based assay that could be applied to theeedetermination of N–C axially chiral triazole derivatives. Analytical samples for CD were prepared from crude mixtures by three sequential steps: liquid–liquid extraction (LLE), a wash-elute, and complexation with Cu(ii) triflate. The initialeemeasurement of five samples of atropisomer 2 was conducted by the use of a CD spectropolarimeter with a 6-position cell changer, resulting in errors of less than 1%ee. High-throughput ee determination was performed on a CD plate reader using a 96-well plate. A total of 28 atropisomeric samples (14 for 2 and 14 for 3) were screened foree. The CD readings were completed in 60 seconds with average absolute errors of ±7.2% and 5.7%eefor 2 and 3, respectively.