BACKGROUND Consumption of fava beans seeds can be harmful to both humans and animals due to the presence of pyrimidine glucosides: vicine and convicine. Therefore, seeds, feed supplements, and protein extracts derived from fava beans should be monitored for the contents of pyrimidine derivatives. OBJECTIVE The main objective of this work was the development and validation of an inexpensive, uncomplicated, quick, and robust method for the quantitation of vicine and convicine in seeds of fava beans. METHODS Plant material was extracted using a dual-phase extraction system to remove lipophilic contaminants and deactivate residual enzymes. Pyrimidine glucosides together with the internal standard: 5-bromouridine were separated using capillary zone electrophoresis (CZE) on an uncoated fused-silica capillary with alkaline, tetraborate-based electrolyte. Linearity, limits of detection and quantitation, precision, accuracy, recovery, and robustness of the method were investigated. The method was applied for quantitation of pyrimidine glucosides in seeds. RESULTS Validation results suggest that the method is suitable for quantitation of pyrimidine glucosides, although it may not be sensitive enough for low-concentrated samples. Application of the method showed that seeds of the investigated variety of fava beans contain both vicine at 10-15 mg/g of dry weight and convicine at 3-5 mg/g of dry weight. CONCLUSIONS Developed and validated CZE-ultraviolet (UV) method is suitable for analysis of pyrimidine glucosides in seeds of Vicia faba L. var. minor. It can be used as an inexpensive, greener alternative to more advanced methods while providing equally reliable and accurate results.