Abstract: In order to study the composition of bitter substances in beer, high performance liquid chromatography was applied to quantitatively determine humuliones, hulupones, and iso-α-acid in beer. By oxidizing hop pellets at 60 ℃, humuliones and hulupones as major products were separated using Alltima C18 column, and 0.05% phosphoric acid and acetonitrile were used for mobile phase gradient elution. C18 solid-phase extraction (SPE) tubes were used for sample pretreatment. The relative standard deviations (RSDs) were 2.18%-3.59%. The minimum detection limits for humuliones, hulupones, and iso-α-acid were 0.03, 0.03, and 0.05 mg/L, respectively, and their recoveries were in a range of 95%-115%. It was found that most commercial beers, such as lager beer, pilsner, wheat beer and stout, contained less humuliones and hulupones (＜2 mg/L), however, India Pale Ale (IPA) had higher levels and their composition varied significantly among different brands. In conclusion, this method is effective for conducting further studies on bitter senses of humuliones and hulupones, and also for regulating bitter substances in IPA.
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YANG Zhaoxia,LI Mei,SHENG Xianbin,et al. Simultaneous determination of humuliones, hulupones, and iso-α-acids in beer by high performance liquid chromatography[J]. Food and Fermentation Industries, 2019, 45(10): 182-187.
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