The OSU Barley Flavor Project
Dustin Herb
Ph.D. Candidate
The OSU Barley Project
Oregon State University
January 17th, 2016
Goals
• Determine if flavor differences within modern barley
varieties exist.
• Identify factors that influence flavor difference.
• Develop methods to select for barley flavor.
• Genetically map gene(s) controlling the flavor
expression within barley.
Willamette Flavor
• 3 varieties (Full Pint; Copeland; Klages)
grown in Lebanon, OR in yield strips.
• Malted at the CMBTC.
• Brewed at New Glarus Brewing Co. and
Sierra Nevada Brewing Co.
• In house sensory, BIC sensory, and OSU
sensory.
• Difference test – revealed that flavor
differences were present among the
three varieties.
Farm
Malted
New Glarus Sierra Nevada
Sensory
Sensory
BIC Sensory
OSU Sensory
3 Varieties x 3 Locations
• 3 varieties (Full Pint; Metcalfe;
Klages) grown in 3 locations
(Oregon; Minnesota;
Saskatchewan).
• Malted at Rahr Malting Co.
• Wort Sensory and analytics at
Sierra Nevada Brewing Co.
• Genotype x Environment
interaction flavor effect.
Flavor
Variety Environment G X E Rep
Total
Taste
X
Malt
X
X
X
Fruity
X
X
X
Chocolate X
X
Toffee
X
X
Vegetable X
X
Dairy
X
Woody
Chemical
X
X
X
X
X
AROMA COMPOUNDS
Flavor
Buttery
Cheesy
Waxy
Nutty
Nutty
Nutty
Bready
Roasted
Chocolate
Spicy
Green
Floral
Fruity
Ethereal
Aldehyde
Chemical
Sulferous
Aroma Compound
2, 3-butanedione
Butyric acid
Nonanoic acid
2-ethyl-3,5-dimethyl pyrazine
2,6-diethyl pyrazine
2-acetyl furan
Furfuryl alcohol
2-formyl-1-methyl pyrrole
2-methyl butanal
2-methyl propanal
phenyl acetaldehyde
phenyl ethyl alcohol
ethyl hexanoate
ethyl acetate
3-methyl butanal
2-ethyl furan
dimethyl sulfide
Top Variety
Full Pint
Metcalfe
Full Pint
Full Pint
Metcalfe
Klages
Full Pint
Klages
Full Pint
Metcalfe
Metcalfe
Metcalfe
Metcalfe
Full Pint
Full Pint
Full Pint
Full Pint
SENSORY
Top Environment
Oregon
Minnesota
Oregon
Minnesota
Oregon
Oregon
Minnesota
Minnesota
Oregon
Minnesota
Oregon
Minnesota
Minnesota
Oregon
Oregon
Oregon
Minnesota
Flavor
Taste
Dairy
Malty
Toffee
Roasted
Chocolate
Fruity
Vegetable
Chemical
Dirty
Top Variety
Klages
Full Pint
Full Pint
Full Pint
Full Pint
Full Pint
Metcalfe
Metcalfe
Klages
Metcalfe
Top Environment
Saskatchewan
Oregon
Oregon
Oregon
Oregon
Oregon
Minnesota
Oregon
Oregon
Minnesota
Enzyme Brewing – Assessing the Malting Effect
• Determine what flavor
compounds are present in barley.
• Determine a flavor baseline for
barley.
• Evaluate the persistence of barley
and malt flavor compounds
throughout the brewing process.
• Same source barley, ½ malted, ½
brewed with Ondea Pro®.
• Identical brewing recipe.
Brewing parameters for the barley grain and barley malt brews at Oregon State
University pilot brewery in Corvallis, OR
Target
Gravity
Mashing Regime
Kettle
Boil
Bitterness
Rest I: Rest II: Rest III:
32 g / hl Iso15˚P 30 min, 60 min, 15 min, 60 min
Alpha-Acids
54C
64C
78C
Yeast/rate
1056 Ale
yeast
(%) Ferment
ABV Length
5.5
14 days
Finished beer measurements for barley grain and barley malt samples at the Oregon
State University pilot brewery in Corvallis, OR
Beers
Real
Color Turbidity Apparent
RDF
Extract
(˚SRM) (NTU) Extract (˚P)
(%)
(˚P)
pH
ABV Bitterness
(%)
(BU)
Barley
3.91
0.038
3.2
5.3
66.8
4.85
5.22
16.4
Malt
4.43
0.042
1.7
4.5
72.2
5.13
5.96
15.6
Barley vs. Malt beer flavor
Malty
8
Bitter
Nutty
7
6
5
Sweet
Bread
4
Barley
3
2
Body
Toffee
1
0
Malt
Mouthfeel
Grainy
Boozy
Fruity
Grassy
Floral
Oregon Promise Pop’n
• Golden Promise x Full Pint
• 202 double haploid population.
• Golden Promise is an iconic variety
for malting, brewing, and distilling.
• Full Pint is a contributor to the craft
brew Renaissance.
• The Oregon Promise will provide a
valuable resource for extending
current knowledge of malting and
brewing genes to the frontiers of
sensory assessment.
Testing Environments
Corvallis, OR
Madras, OR
Lebanon, OR
• 37 agronomic selection + parents
grown in all three locations.
• 14 random selection grown only
in Corvallis, OR.
Micro-malting & Malt analysis
Malt Quality
Sample
Full Pint
Golden
Promise
Control
Parents & Control
Color DP
Ext
Plump Moist pH
AA
BG
Prot
Turb
Viscos
AVG
83.50
381.33 1.46
156.00 78.41
10.30
4.95
5.99
40.78
5.27
1.12
STDEV
7.99
17.93
0.25
12.17
0.93
2.17
1.16
0.07
1.65
0.91
0.97
%CV
9.57
4.70
16.96
7.80
1.19
21.03
23.48
1.11
4.04
17.23
86.65
AVG
42.67
461.00 1.50
119.00 76.65
33.47
6.20
5.83
31.80
19.03
1.09
STDEV
4.38
155.19
0.26
38.20
1.70
11.27
1.80
0.02
3.61
7.60
0.94
%CV
10.25
33.66
17.17
32.10
2.22
33.68
29.06
0.34
11.35
39.94
86.61
AVG
63.53
97.33
1.35
139.00 79.15
16.60
4.70
5.92
40.29
4.30
1.01
STDEV
11.46
120.56
0.33
17.06
1.24
9.37
0.96
0.07
5.38
1.23
0.87
%CV
18.03
61.09
24.39
14.34
1.57
56.46
20.36
1.12
13.36
28.58
86.79
Malt Quality
Malt quality of agronomic Oregon Promise selections from all environments (Corvallis, OR; Lebanon, OR; Madras, OR)
analyzed by Rahr Malting Co. in 2015.
Agronomic Selections
Sample
COR
LEB
MAD
AA
BG
Color
DP
EXT
Plump
Moist
pH
Prot
Turb
Viscos
AVG
57.52
281.76
1.52
109.05
77.40
21.31
4.95
5.88
35.01
9.39
1.63
STDEV
%CV
15.80
27.47
198.93
70.60
0.23
15.47
26.47
24.27
1.49
1.92
8.07
37.88
1.78
35.99
0.07
1.12
8.33
23.78
8.11
86.38
0.12
7.63
AVG
50.02
383.95
1.47
114.05
77.92
14.15
5.41
5.95
34.05
12.26
1.31
STDEV
CV%
12.57
25.13
227.90
59.36
0.22
15.19
27.06
23.72
1.76
2.26
6.38
45.06
2.04
37.67
0.06
1.05
7.12
20.91
10.02
81.70
0.71
54.31
AVG
47.49
524.05
1.31
136.19
75.17
25.48
4.81
5.87
33.54
15.43
1.67
STDEV
CV%
12.35
26.01
279.69
53.37
0.17
13.26
25.57
18.77
1.51
2.01
15.51
60.89
1.32
27.44
0.05
0.81
3.99
11.88
16.74
108.49
0.47
27.91
Malt quality of random Oregon Promise selections from Corvallis, OR environment analyzed by Rahr Malting Co. in 2015.
Random Selections
Sample
COR
AA
BG
Color
DP
EXT
Plump
Moist
pH
Prot
Turb
Viscos
AVG
70.83
127.57
1.61
118.93
76.70
32.98
5.22
5.84
39.84
5.54
1.55
STDEV
CV%
19.62
27.70
95.64
74.97
0.23
14.40
25.48
21.43
1.33
1.74
8.40
25.48
1.49
28.52
0.06
0.95
8.05
20.22
1.77
32.01
0.08
5.06
Nano-brewing
1. Malt samples were milled.
2. 75g of malt grist and ~400
mL of water was placed in
mash bath cans.
3. Mash cans are brought to
standardized volume of
450 mL.
4. Congress mash is poured
over Ahlstrom filters.
5. Wort was filtered three
times for clarity.
6. Final wort was collected.
2
4
3
5
6
Nano-brewing
7. IsoHop® is added to the
congress wort and boiled
for 35 min.
8. Residual sediment and
trub filtered out with
Ahlstrom filters.
9. WhiteLabs Czech yeast
pitched and wort ferments
in tube at 9°C.
10. Beer is centrifuged,
bottled, and carbonated
then conditioned.
7
9
8
10
Sensory
Comparison to reference
• Difference testing
• Descriptive analysis
~13 experimental beers
2 parental beers - FP & GP
1 internal standard - Rahr Pils
1 embedded reference - Miller High Life
17 beer sample/session
10 tasting sessions
X
~170 beer samples tasted
Full Pint
0.2
0.18
0.16
0.14
0.12
0.1
0.08
0.06
0.04
0.02
0
FRUIT
Golden Promise
Full Pint
0.25
0.2
0.15
0.1
0.05
0
0
1
2
3
MALT
4
5
6
2
3
Full Pint
Relative frequency
0.3
Golden
Promise
0.25
0.2
0.15
0.1
0.05
0
0
1
2
3
4
TOFFEE
5
FLORAL
0.4
0.35
4
6
7
FRUIT
TOFFEE
Relative frequency
Golden Promise
0.3
Relative frequency
Relative frequency
MALT
5
6
7
0.2
0.18
0.16
0.14
0.12
0.1
0.08
0.06
0.04
0.02
0
Golden Promise
Full Pint
3
3.5
4
4.5
FLORAL
5
5.5
Parents & Reference
3
2
SWEET
Reference ROASTED
FLORAL
Reference
Reference
Reference
FRUITGolden PromiseGolden Promise
Reference
Golden
Promise
Reference
BITTER
Full Pint
Reference
Golden Promise
Reference CEREAL
Golden Promise
Reference Full Pint
Golden Promise
TOFFEE
Full Pint
Golden Promise
BREAD
Full Pint
Golden Promise
MALT
GRASS
Full Pint
Full Pint Full Pint
ASTRINGENT
GRAIN
Full Pint
Golden Promise
Full Pint
Golden Promise
F2 (18.01 %)
1
0
-1
-2
Full Pint
-3
-4
-3
-2
-1
0
F1 (42.84 %)
1
2
3
4
Environment Biplot
3
COR
COR
2
COR
COR
COR
LEB CORASTRINGENT
COR
BITTER
COR
LEB
LEB
GRAINCOR
COR COR
MAD
COR LEB
COR
LEB
COR
MAD
COR
MAD
COR
GRASS
COR CEREAL
LEB
MAD
LEB
COR
COR
LEB
MAD
COR
LEB
CORCOR MAD
LEB
MAD
MAD
LEB
LEB LEB
ROASTED
COR
LEB
LEB
COR
MAD
LEB
COR
COR
COR
FRUIT
LEB COR
MAD
COR LEBCOR
LEB
MAD MALT
MAD
BREAD
TOFFEE
LEB
MAD
FLORAL LEB COR LEB
LEB
SWEET
COR
LEB
COR
LEB MAD
LEB MADMADMAD
LEBMAD
MAD
COR
LEBLEB
MADMAD
MADLEBMAD
MAD LEBLEB
LEB
LEB
LEB
COR
COR MAD
MAD
MAD
MAD
LEB
MAD
LEB MAD
MAD
MAD
MAD
MAD
MAD
1
F2 (14.89 %)
COR
0
-1
LEB
MAD
-2
COR
MAD
MAD
-3
-4
-3
-2
-1
0
F1 (38.54 %)
1
2
3
4
Flavor Comparison
FRUIT
ASTRINGENT
FLORAL
BITTER
GRASS
SWEET
GRAIN
ROASTED
CEREAL
TOFFEE
MALT
BREAD
120145a
120156a
FULL PINTp
GOLDEN PROMISEp
120031
New Glarus Brewing Co.
Full, Malty, Smooth, Rich, Nice! - Best "Euro" Type
120058
Nice Foam, Clean, Smooth - Best "American" Type
120090
Best Foam, Sweet, Clean, Mild
120145
Balanced Malty and Hoppy, Clean, Sweet
120285
Sweet, full, very clean, bread, malt
Malty, Toffee, Caramel, Vanilla, Papery
Malty, Nutty, Floral, Phenolic, DMS
120322
Malty, sweet / dry, morish, low color
Malty, Toffee, Caramel, DMS
120331
Light, clean, malty, sulfur, Euro, juicy, nice
Diacetyl, DMS, Worty
120341
Sweet, full, biter after taste, malty, clean
120359
Wow, malty, biscuit, sweet, candy, floral
Acidic, Sour, Tart, Rubber
Fruity, Floral, Sweet
120373
Coffee! Roasted bitter, "White Stout"?, very unique
Malty, Worty
120744
Malty, bread, European, very nice
Nutty, Bananas, Floral
Full Pint
Full, malty, European
Malty, Sweet, Popcorn, Grain, Grass
Golden Promise Harsh bitterness
Rahr Malting Co.
Malty, Toasted/Bread, Roasted/Toffee
Honey, Nutty, Floral
Herbaceouse, Earthy, Green Apple
Fruity, Floral, Grass/Green, Astringent
Gas Chromatography-Mass Spectrometry
• Identify aroma compound peaks.
• Determine if significant
compound(s) affect sample flavor.
• Track compound deviation from
parental genotypes.
• Identify compounds that will be
useful in flavor selection
programs.
GC-MS
1.6e+006
1.4e+006
1.2e+006
1e+006
800000
600000
400000
200000
0
Time (s)
1210
1220
1230
TIC 13-FP:1
1240
1250
TIC 13-GP:1
1260
TIC 13-13:1
1270
1280
TIC 13-11:1
1290
1300
GC-MS
COMPOUND
FLAVOR
1-Butanol
Bananas, Sweet
1-Hexanol
Fruity, Sweet, Oil, Green, Ethereal
1-Octanol
Fruity, Floral, Citrus
1-Propanol
Alcoholic, Ethanol, Fruity, Apple, Nutty
2,3-Butanediol
Fruity, Buttery
2-Heptanone
Fruity, Coconut, Sweet, Spicy, Green
Acetic acid
Fruity, Sour, Vinegar
Benzaldehyde
Sharp, Cherry, Sweet, Almond
Benzeneacetaldehyde
Honey, Sweet, Floral, Rose, Cocoa, Spicy, Green, Earthy
Benzothiazole
Sulfur, Vegetable, Nutty, Coffee, Meat, Rubbery
Isobutyl acetate
Sweet, Fruity, Bananas, Tropic, Ethereal
Isopentyl hexanoate
Fruity, Pineapple, Sour
Nonanal
Rose, Orange peel, Cucumber, Fatty
GC-MS
Environmental biplot of aroma compounds
2.5
Parental biplot of aroma compounds
2.5
COR
2
FULL PINT
2
REF
REFERENCE
Acetic acid
Acetic acid
1.5
1.5
MAD
1
F2 (19.34 %)
F2 (19.34 %)
1
IsobutylREF
acetate
Benzeneacetald
1-Propanol
ehyde
REF
0.5
REF
IsobutylREFERENCE
acetate
Benzeneacetalde
1-Propanol
hyde
FULL PINT
REFERENCE
0.5
REFERENCE
0
0
MAD
MAD
2-Heptanone
-0.5
Benzaldehyde
Isopentyl
hexanoate
MAD
COR
-1
-1.5
-2.5
-2
-1.5
LEB
LEB
REFREF
2,3-Butanediol
1-Octanol
1-Hexanol
Nonanal
LEB
COR
Benzothiazole
COR
-0.5
0
0.5
F1 (31.78 %)
1
1.5
-0.5
-1
LEB
1-Butanol
-1
GOLDEN
FULL PINT
PROMISE
2-Heptanone
Benzaldehyde
-1.5
2
2.5
-2.5
Isopentyl
hexanoate
GOLDEN
GOLDEN
PROMISE
PROMISE
-2
-1.5
-1
-0.5
GOLDEN
2,3-Butanediol
1-Octanol
FULL
PINT
PROMISE
1-Hexanol
Nonanal
REFERENCE
REFERENCE
GOLDEN
Benzothiazole
PROMISE
GOLDEN
FULL PINT
PROMISE
FULL PINT
1-Butanol
0
0.5
F1 (31.78 %)
1
1.5
2
2.5
11.0
12.0
18.2
11_11479
11_20415
12_30843
11_10451
26.7
47.0
55.3
55.7
56.0
56.2
61.2
67.5
68.2
83.6
12_31305
12_30880
11_10431
11_10153
11_11098
11_20975
11_11219
11_20771
11_10143
11_21448
12_31395
11_20092
109.5
117.3
122.3
123.9
130.9
12_20832
11_21160
11_20139
12_31325
11_20504
64.9
TKW_LOD:6.6_a:-1.3
12.9
H_LOD:5.0_a:-2.6
HD_LOD:9.4_a:1.2
BSR_LOD:4.3_a:4.1
DP_LOD:3.1_a:6.8
ME_LOD:5.9_a:-0.5
AA = Alpha
H = Height
WP = Wort Protein
Amylase
HD = Heading
BD = Braird
BC = Barley Color Date
BSR14 = 2014
BG = Beta-glucan ME = Malt Extract Barley Stripe Rust
BP = Barley
P = Plump
Protein
SL = Spike Length
BSR = Barley StripeFPD = sdw1
Rust
(Dwarf)
DP = Diastatic
HD14 = 2014
Power
Heading Date
GPD = ari-e (dwarf S/T = Soluble/Total
gene)
Nitrogen
LR = Leaf Rust
TKW = Thousand
FAN = Free Amino Kernel Weight
Nitrogen
WC = Wort Color
Flavor
QTL
FAN_LOD:46.3_a:-39.4
BG_LOD:6.8_a:64.2
AA_LOD:56.5_a:-18.6
S/T_LOD:5.3_a:-4.9
WC_LOD:20.9_a:-0.3
WP_LOD:34.5_a:-0.6
FLAVOR
12_30382
11_20132
P_LOD:10.0_a:-2.1
186.9
187.5
ME_LOD:23.9_a:-1.0
12_30524
11_21422
11_10805
12_30929
11_10783
12_30930
11_10095
12_30869
12_30635
11_10819
11_21355
11_20829
11_10869
BC_LOD:5.5_a:-2.5
BG_LOD:6.6_a:-60.7
S/T_LOD:3.9_a:0.8
WC_LOD:4.3_a:0.1
90.8
92.6
98.5
106.1
106.2
106.3
106.4
107.4
111.2
118.2
132.0
146.3
148.5
TKW_LOD:28.4_a:-3.2
S/T_LOD:6.0_a:1.3
DP_LOD:3.3_a:-7.7
BP_LOD:6.9_a:-0.5
ME_LOD:15.6_a:0.8
P_LOD:3.8_a:-1.1 H_LOD:4.5_a:-2.8
HD_LOD:10.1_a:2.4 TKW_LOD:7.6_a:-1.4 DP_LOD:3.6_a:8.5
BC_LOD:9.2_a:3.1
SL_LOD:30.3_a:-0.9
11_21324
12_30214
11_10671
11_11221
11_20441
11_20645
11_11355
12_31427
11_20526
11_10518
11_21168
11_21061
11_11273
HD_LOD:4.6_a:-2.3
5H
0.0
12.5
14.4
14.9
16.4
30.3
35.4
36.5
42.0
43.4
57.6
60.5
63.0
BG_LOD:3.5_a:-41.8
3.1
6.0
6.1
19.6
27.8
BC_LOD:9.2_a:-3.2
0.2
12_30430
12_30856
11_10817
11_10462
11_21339
12_31289
11_20468
11_21025
11_10202
AA_LOD:9.0_a:-6.5
0.0
11_20722
12_30530
BG_LOD:2.8_a:37.4
11_20232
7H
0.0
2.9
S/T_LOD:52.1_a:-0.9
0.0
H_LOD:20.3_a:-6.3
44.0
44.3
45.9
48.2
52.3
HD_LOD:4.1_a:-1.5
12_31324
11_20557
11_20411
11_11180
11_10262
11_10946
11_10668
11_20939
12_30994
11_10467
11_21504
P_LOD:6.9_a:1.5
4H
TKW_LOD:9.4_a:1.5
11_21212
H_LOD:9.9_a:3.8
51.8
43.8
SL_LOD:8.5_a:0.4
11_21187
11_21261
11_11073
11_10525
11_10399
11_10648
11_10342
11_10297
11_11054
11_21166
11_10265
11_11402
11_20667
11_20419
12_10859
11_21242
11_11100
11_11533
11_10876
11_10900
11_21340
11_11323
11_11480
12_20183
11_20590
11_10791
11_10710
12_30953
11_10335
12_30754
11_20778
12_31346
12_30663
12_31367
0.0
18.7
31.0
38.2
2H
0.0
4.7
23.2
23.7
24.2
24.7
25.2
30.4
31.3
36.3
39.3
42.8
43.6
45.8
51.3
55.0
57.3
64.4
77.8
78.8
82.3
85.7
86.8
109.4
124.8
146.6
6HA, 6HB
0.0
0.5
8.5
14.6
15.4
18.3
30.6
32.2
SL_LOD:4.8_a:0.3
3H
SL_LOD:4.0_a:0.3
12_30918
12_30877
11_20712
11_10744
12_30336
11_10764
11_10438
12_31208
11_10552
11_20095
11_10516
11_10890
11_20990
11_20434
12_31160
11_21373
11_10522
11_20169
11_10911
11_20908
11_10586
11_10722
11_20383
11_10443
11_20772
0.0
16.0
23.9
27.7
37.4
38.4
41.2
41.8
47.1
47.5
49.6
56.3
58.9
69.1
71.1
71.8
78.6
79.0
0.0
1.6
3.6
3.7
17.1
23.5
25.1
FLAVOR
1HA, 1HB
?
QTL Mapping Pop’n
• Full Oregon Promise Population
• Nano-brewing sensory
• Pilot brewing trials with OP
agronomic and flavor selections
• Maris Beaver population (Full
Pint x Maris Otter)
• Winters & Facultative
• Full Stein population (Full Pint x
Violetta)
• Winters & Facultative
Exotic Barley Flavor
• Screening 174 genetically
diverse lines from the USDA
Core Collection.
• Micro-malted at Rahr Malting
Co.
• Wort analysis at Sierra Nevada
Brewing Co.
• Nano-brewed at New Glarus
Brewing Co. and OSU.
• Sensory at OSU.
Modification Flavor
• Degree of modification
• Under-, Well-, Overmodification
• Beta-glucan structure
• High- & low-molecular
weights
• Environmental factors
• Barley spec profile
• Structural: Soluble
carbohydrates ratio
• Lipid and fat levels
Barley Brand Projects
Nitrogen – Protein Level
and Amino Acid Flavor
Barley “Terroir” Flavor
Organic vs Conventional
Flavor Effects
Summary
• Significant flavor differences detected between genotypes.
• Random selections were highest in fruit, grass, cereal, malt, bread, and
toffee flavors in Corvallis environment.
• Significant flavor differences detected between environments.
• Genotype x Environment effects detected.
• Barley Flavor ≠ Malt Flavor
• Distribution of sensory data indicates that exploratory sensory methods are
effective in nano-brew samples.
• Identification of aroma compounds segregation indicates genotypic
differences for select compounds.
• GC-MS is effective for high-throughput screening and characterization of
“potential” flavor contributions.
• Flavor QTLs discovered!!!
Impact
• Breeders
• More tools on the tool belt (nano-brewing, sensory, GC-MS).
• Marker Assisted Selection for flavor QTLs
• Growers
• Regional taste or “terroir” production.
• Maltsters
• Identification of production regions and/or farms with best agronomics and flavor.
• Barley flavor specific malting (Custom malting).
• Value added product and new markets.
• Brewers
• Product diversification.
• Increased single malt brews.
• Proprietary or exclusive varieties contracts.
• Consumers
• More choices.
THINK BARLEY
DRINK BARLEY