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 using a fully automated gas chromatography – mass spectrometry methodology allowing both identification and accurate quantification of VOCs produced by roots of a monocotyledonous plant species at five selected developmental stages from germination to the end of tillering. 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 and linolenic acid derivatives produced via the lipoxygenase/hydroperoxide lyase pathway of higher plants. Our findings contrast with analyses documented on aboveground barley tissues that mainly emit C6 aldehydes, alcohols and their corresponding esters. Multivariate statistical analyses performed on individual VOC concentrations indicate quantitative changes in the volatile profile produced by barley roots according to plant age. Barley roots produced higher total and individual VOC concentrations when young seminal roots emerged from the coleorhizae compared to older phenological stages. Moreover, results also show that the C6/C9 volatile aldehyde ratio was the lowest at the end of tillering while the maximum mean value of this ratio was reached in seven day-old barley roots.