H NMR Spectroscopy for Determination of the Geographical Origin of Hazelnuts.

A total of 262 authentic samples was analyzed by H NMR spectroscopy for the geographical discrimination of hazelnuts ( Corylus avellana L.) covering samples from five countries (Germany, France, Georgia, Italy, and Turkey) and the harvest years 2013-2016. This article describes method development starting with an extraction protocol suitable for separation of polar and nonpolar metabolites in addition to reduction of macromolecular components. Using the polar fraction for data analysis, principle component analysis was applied and used to monitor sample preparation and measurement. Several machine learning algorithms were tested to build a classification model. The best results were obtained by a linear discrimination analysis applying a random subspace algorithm. The division of the samples in a trainings set and a test set yielded a cross validation accuracy of 91% for the training set and an accuracy of 96% for the test set. The identification of key features was carried out by Kruskal-Wallis test and t test. A feature assigned to betaine exhibits a significant level for the classification of all five countries and is considered a possible candidate for the development of targeted approaches. Further, the results were compared to a previously published study based on LC-MS analysis of nonpolar metabolites. In summary, this study shows the robustness and high accuracy of a discrimination model based on NMR analysis of polar metabolites.