Recent Advances in the Enantiomeric Separation of Tropane Alkaloids by Liquid Chromatography: Analytical Strategies and Applications in Pharmaceutical, Biological, and Food Samples

This review examines recent advances in the enantiomeric separation of tropane alkaloids (TAs), focusing on atropine, a racemic mixture of (+)- and (−)-hyoscyamine whose enantiomers differ significantly in pharmacological activity, with (−)-hyoscyamine being the more potent antimuscarinic agent. Enantiomeric purity and racemisation control are therefore essential in pharmaceutical formulations, biological samples, and food, where TAs are regulated contaminants due to their toxicological effects. The review covers analytical strategies reported from 2010 to 2026, with emphasis on high-performance liquid chromatography (HPLC), often coupled with tandem mass spectrometry (MS/MS) to improve sensitivity and selectivity. Polysaccharide-based chiral stationary phases operated in normal-phase or polar organic modes, usually with amine additives, represent the most widely employed approach. Protein-, cyclodextrin-, and antibiotic-based chiral selectors have also demonstrated successful enantiomeric resolution. Sample preparation is particularly challenging for food matrices, requiring efficient extraction and clean-up while preventing enantiomeric racemisation. Published strategies include solid-phase extraction (SPE), matrix solid-phase dispersion (MSPD), and QuEChERS protocols. Despite significant analytical advances, chiral analysis of TAs in foods remains underexplored. Recent studies have reported (−)-hyoscyamine concentrations exceeding regulatory limits in herbal infusions, underscoring the need for further research to support European Food Safety Authority (EFSA) recommendations and improve consumer risk assessment.