Olivaro Cristina, Núñez Nerea, Basile Patricia, Mederos América, Reyno Rafael, Saurina Javier, Núñez Oscar
Espacio de Ciencia y Tecnología Química, CENUR Noreste, Universidad de la República, Tacuarembó, Uruguay.
Department of Chemical Engineering and Analytical Chemistry, University of Barcelona, Barcelona, Spain.
Front Mol Biosci. 2025 Aug 29;12:1646758. doi: 10.3389/fmolb.2025.1646758. eCollection 2025.
The genus, part of the legume family, comprises over 180 species distributed across diverse ecosystems worldwide. Its broad genetic diversity enables adaptation to various environmental conditions and represents a valuable resource for breeding programs targeting key agronomic traits. One of the most attractive features of species is the presence of condensed tannins in the forage, which, in ruminants, help prevent bloat, exhibit antiparasitic properties, enhance the absorption of non-ammonia nitrogen compounds, and reduce greenhouse gas emissions.
This study aimed to develop a UHPLC-HRMS method for classifying ten cultivars produced in Uruguay using a non-targeted metabolomic fingerprinting approach. Five cultivars belong to , three to , and two are interspecific hybrids. The analysis focused on phenolic compound-rich fingerprints. Principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) were used for data exploration and classification, and to identify key phenolic compounds with high discriminant potential. Finally, cultivar-specific polyphenolic compounds were tentatively identified based on chromatographic and high-resolution mass spectrometry (HRMS/MS) data obtained from all cultivars.
When defining four classes (, , and the two hybrids), the optimal PLS-DA model required six latent variables and achieved 100% classification accuracy, with both sensitivity and specificity reaching 100%. Additional PLS-DA models were developed to assess intra-species discrimination among the 3 and 5 cultivars, with varying degrees of separation observed. In each PLS-DA model, VIP loadings scores allowed the selection of the most discriminant phenolic compounds for each class under study. A total of 105 compounds, including phenolic acids, flavonols, flavan-3-ols, proanthocyanidins, and organic acids, were tentatively identified by analyzing all cultivars.
该属属于豆科，包含180多个物种，分布于全球不同的生态系统。其广泛的遗传多样性使其能够适应各种环境条件，是针对关键农艺性状的育种计划的宝贵资源。该属物种最吸引人的特征之一是其牧草中含有缩合单宁，在反刍动物中，这些缩合单宁有助于预防瘤胃臌胀、具有抗寄生虫特性、增强非氨氮化合物的吸收并减少温室气体排放。
本研究旨在开发一种超高效液相色谱-高分辨质谱（UHPLC-HRMS）方法，采用非靶向代谢组学指纹图谱方法对乌拉圭生产的10个品种进行分类。其中5个品种属于该属某一种，3个属于另一种，2个是种间杂交品种。分析聚焦于富含酚类化合物的指纹图谱。主成分分析（PCA）和偏最小二乘判别分析（PLS-DA）用于数据探索和分类，并识别具有高判别潜力的关键酚类化合物。最后，根据从所有品种获得的色谱和高分辨质谱（HRMS/MS）数据，初步鉴定出各品种特有的多酚类化合物。
在定义四个类别（两个种和两个杂交品种）时，最优的PLS-DA模型需要六个潜在变量，分类准确率达到100%，灵敏度和特异性均达到100%。还开发了其他PLS-DA模型来评估3个某一种和5个另一种品种内的种内判别情况，观察到了不同程度的分离。在每个PLS-DA模型中，变量重要性投影（VIP）载荷得分允许选择所研究的每个类别中最具判别力的酚类化合物。通过分析所有品种，共初步鉴定出105种化合物，包括酚酸、黄酮醇、黄烷-3-醇、原花青素和有机酸。
