WORLD BREWING CONGRESS 2016 World Brewing Congress August 13-17, 2016 Comparison of characteristics of taste and Sheraton Downtown Denver Denver, CO 80202, U.S.A. 129 in using different hop varieties Takamasa Hasegawa, Tomoyuki Nakahama, and Takako Inui / Suntory Beer Ltd., Osaka, Japan Objective Results are essential components that contribute to the hoppy aroma and taste-related characteristics such as fullness, sweetness, bitterness and astringency of beer. Among ■Results of hops ■Results of beer ■Results of hop vs beer the different hop-derived compounds, polyphenols are specifically considered to play an important role in imparting taste, and their profiles are thought to vary between Fig 1. Comparison of profile in each hop variety harvested Fig 3. Influence of hop varieties on taste characteristics of beer Fig 5. Correlation between content of each polyphenol in hop hop varieties, resulting in different characteristics with regard to taste as well as over 3 years and beer Hallertauer Mfr. N=7 A high correlation (more than 0.7) is pink if positive and blue if negative aroma in beer. 56 56 This study investigated the polyphenol profiles of 6 hop varieties, and the polyphenol 2013 2014 2015 Hallertauer Tradition Sweetness Bitterness 4-hydroxybenzoic acid Chlorogenic acid p-Coumaric acid trans-Ferulic acid Sinapic acid F 4-Hydroxybenzaldehyde 2013 2014 2015 54 54 profiles and taste characteristics of beer prepared using each hop variety. Procyanidin B2 Chlorogenic acid A A:Monophenol Procyanidin B1 p-Coumaric acid 52 * 52 B:Flavonol Procyanidin C1 trans-Ferulic acid E 50 50 In beer Materials and methods C: (+)- 4-Hydroxybenzoic acid D: 48 48 In hop ■ (−)-Epigallocatechin Sinapic acid Hop raw materials E:Flavanol 46 46 Xanthohumol 8-Prenylnaringenin Gentisic acid

(−)-Epicatechin Syringaldehyde score Normalized F: Varieties Parentage Origin Xanthohumol Quercetin 44 44 B Hallertauer Mittelfrüh landrace Germany Isoxanthohumol Gentisic acid D 8-Prenylnaringenin 56 Fullness 56 Astringency Saazer landrace Czech Republic Naringenin C 54 54 * Tettnanger landrace Germany Saazer Tettnanger Perle Cascade 52 52 (−)-Epicatechin (−)-Epigallocatechin (+)-Catechin Procyanidin B1 Procyanidin B2 Procyanidin C1 a Hallertauer Tradition Hallertauer Gold × 75/15/106M Germany 2013 2014 2015 2013 2014 2015 2013 2014 2015 2013 2014 2015 50 50 Perle Northern Brewer × 63/5/27Mb Germany Cascade (Fuggle × [Serebrianca × Fuggle USA 48 48 - seedling]) × open-pollinated 46 46 44 44 0.45 Distance 1.0 a47% Hallertauer, 15% Saazer, 9% Spalter, 28% wild hops, 1% Northern Brewer The content of polyphenols in hop showed a correlation with that in beer, with the bHallertauer Mfr, Spalter, Saazer. The percentage of mixed breed are not known. exception of isoxanthohumol and (−)-epigallocatechin, which were generated from Cf. “Genetic diversity and phylogenetic relationships among accessions of hop, , as xanthohumol and proanthocyanidin during the boiling of hop. determined by amplified fragment length polymorphism fingerprinting compared with pedigree data” Seefelder S, et al. Plant Breeding. 119: 257–263 (2000) * : Significant difference (P<0.05) Hallertauer Mfr., Saazer, and Tettnanger have more proanthocyanidin (F). Conclusion ■Extraction and quantification of hop polyphenols Perle, Cascade, and Tradition have more flavanone (C) and prenylflavonoid (D). ■Each hop variety exhibited a different polyphenol profile. • Sample preparation • LC-MS/MS conditions Fig 2. Principal component analysis of polyphenol profiles in each hop ■Fresh beer samples using 6 hop varieties showed different taste Hop Beer Fig 4. Principal component analysis of taste score of beer LC-MS/MS : SHIMADZU LCMS-8040 characteristics, especially in sweetness and astringency. Dichloromethane variety harvested over 3 years Column : Luna® 5 µm C18(2) 100 Å, ■ Wash Filtration LC Column 150 x 4.6 mm Sweet Astringent It was observed that the differences in polyphenol profiles as well as taste 70% Acetone/water Mobile phase : A: water containing 0.1% formic acid characteristics between hop varieties were strongly linked to the genetic B: acetonitrile containing 0.1% formic acid Fullness similarities. Extraction Clean up（SPE) Tettnanger Flow rate : 0.7 mL/min Hallertauer ■It was suggested that the contents of proanthocyanidin were correlated Gradient (min/%B) : 0/5, 5/5, 35/98,40/98,40/5,50/5 Mfr. Saazer Perle Filtration Concentration Injection : 10 μL with the intensity of fullness while the contents of flavanone and Tradition Tettnanger prenylflavonoid were correlated with the intensity of astringency.

.1%) Saazer 30 LC-MS/MS LC-MS/MS ( ■

(29.6%) The content of each polyphenol in beer was similar to that in hop. PC2 ■Brewing trial ■ PC2 Sensory evaluation Hallertauer Mfr. Tradition Futurework • Brewing size: 100 L • Seven trained panelists Cascade • Malt ratio: 100% • Four sensory descriptors: Cascade Perle 1. To verify how much each polyphenol in hop contributed to the taste in beer, Sweetness, Bitterness, by the addition of extracted polyphenols from hop or hopped beer or by the • Conditions of hop addition: Lightness addition of individual chemicals to beer 145 g of each variety of hop was added at the start of boiling. Fullness, Astringency PC1 (53.6%) Bitterness was adjusted using CO2 extract. • Scoring: 2. To elucidate factors other than hop varieties that affected polyphenol profile • Fermentation conditions：Lager yeast, 10℃ Scored from 0 to 3 and normalized PC1 (35.5%) in hop, such as cultivation and brewing conditions