In chemical ecology, the roles played by root-emitted volatile organic compounds (VOCs) in biotic interactions and the quantitative analysis of such chemicals in root tissues remain poorly documented. In this context, this study aims at developing a fully automated analytical methodology allowing both identification and accurate quantification of VOCs produced by roots of a monocotyledon plant species. Briefly, VOC emitted by crushed barley roots are successively trapped by dynamic headspace sampling on Tenax TA adsorbents, thermally desorbed and cryofocused, separated by gas chromatography (GC) and finally analysed by mass spectrometry (MS) in both SCAN and selected ion monitoring modes. Results show that barley roots mainly produce four volatile aldehydes, namely hexanal, (E)-hex-2-enal, (E)-non-2-enal and (E,Z)-nona-2,6-dienal. These molecules are well-known linoleic (C18:2) and linolenic (C18:3) acid derivatives produced via the lipoxygenase and the hydroperoxide lyase pathways of higher plants. Our findings contrast with analyses documented on aboveground barley tissues that mainly emit C6 aldehydes, alcohols and their derivative esters. Moreover, preliminary results indicate quantitative changes in the volatile profile contained in barley roots according to plant age. Multivariate statistical analyses are currently underway to quantitatively assess these changes using plants at five selected developmental stages ranging from germination to the end of tillering.