Alkaloid-enriched extracts from the tree Neltuma flexuosa (Fabaceae): chemical characterization, fractionation, and cytotoxic effects
DOI:
https://doi.org/10.15517/3jcswj50Keywords:
cytotoxicity, food chemistry, piperidine alkaloids, rainfall, toxicologyAbstract
Introduction: Neltuma flexuosa is a key species in arid regions, offering nutritional benefits but also containing alkaloids that can pose toxicological risks. Objective: To analyze the chemical composition and alkaloid content of N. flexuosa pods collected over four years (2019-2022) in Cafayate, Argentina, considering the influence of climatic variations, and to evaluate their potential cytotoxic effects on C6 glioma cells. Methods: Dried and ground pods were analyzed for chemical composition using standard methods. Alkaloid-enriched extracts (AEE1-AEE4) were obtained through acid/base extraction and analyzed by HPLC-HRMS for juliprosine and juliprosopine quantification. AEE4 was fractionated using flash chromatography, yielding fractions (F1-F5), which were also analyzed for alkaloid content. Cytotoxicity on C6 glioma cell lines was evaluated using crystal violet staining assay, and apoptosis induction was confirmed in C6 cells treated with AEE4 using the TUNEL assay. Results: Chemical analysis revealed interannual variations in pod composition, particularly in fiber and protein content. HPLC-HRMS identified juliprosine and juliprosopine, with the highest concentrations in AEE4. Fractionation of AEE4 by flash chromatography yielded an alkaloid-rich fraction (F4). Cytotoxicity assays demonstrated dose-dependent reductions in cell viability, with AEE4 exhibiting the lowest 50 % cytotoxic concentration (CC50) values. Furthermore, F4 displayed the highest cytotoxicity against C6 cells. TUNEL assays confirmed the induction of apoptosis by AEE4. Conclusions: These findings provide valuable insights into the chemical composition, alkaloid content, and cytotoxic effects of N. flexuosa extracts, contributing to the development of guidelines for their safe and sustainable use as a feed resource.
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