Development, validation, and application of capillary zone electrophoresis method for determination of pyrimidine glucosides in seeds of Vicia faba L. var. minor. Issue 3 (2nd September 2020)

• Record Type: Journal Article
• Authors: Kowalczyk, Mariusz; Krzyżanowska‐Kowalczyk, Justyna; Stochmal, Anna
• Is Part Of: Phytochemical analysis. Volume 32:Issue 3(2021)
• Page Start: 375
• Page End: 381
• Publication Date: 2020-09-02
• Subjects: capillary electrophoresis -- convicine -- pyrimidine glucosides -- Vicia faba -- vicine; Plants -- Analysis -- Periodicals; Plants -- chemistry -- Periodicals; 572.2
• DOI: 10.1002/pca.2984
• Languages: English
• ISSNs: 0958-0344

Abstract

Abstract: Introduction: 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. Objectives: 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. Methodology: 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. Conclusion: 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. Abstract : The CZE‐UV method for monitoring pyrimidine glucosides was developed, fully validated, and used for quantitative analyses in fava beans. The 5‐bromouridine was applied as an internal standard to improve the reliability and robustness of the method. Obtained limits of detection and quantitation are sufficient for quantifying vicine and convicine in seeds and other plant‐derived materials.