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<p>The composition of hop oil contributes to the aroma of beer and the essential oil profile of hop samples contains valuable information for brewers. The aim of this study was to characterize the Hüller Bitterer hop variety, during the development of hop cones, by analysis the composition of volatile oil using in-tube extraction gas chromatography–mass spectrometry (ITEX-GC–MS). The obtained results show that the ITEX-GC/MS method is suitable for the determination of volatile compounds from hop samples. A number of 60 compounds were separated and 50 of them were identified. The most important volatile compounds found in Hüller Bitterer hop variety belonging to the monoterpenes and sesquiterpenes classes are represented by: β-myrcene, β-caryophyllene and α-humulene.</p><p> </p>