Dry Hopping

Myths versus Reality

1
Major volatile compounds
after main fermentation
(at start of dry hopping !)
2
 Compounds present in beer after main fermentation
IR CPSil5 Compounds Flavour Samples
YEAST GLYCOSILES PRECURSORS T 0 Day
HOP CYSTEINES BY PRODUCTS C (ppm)
ACTION ON FATTY ACIDS bière
FD

BACKGROUND INFORMATIPON
730 isobutylacetate banana, fruity 0.037 128
774 ethylbutyrate banana, fruity 0.058 512
812 3-methylbutanoic acid cheese 0.101 512
852 isoamylacetate Banana, apple 0.843 128
921 4-methylpentanoic acid cheese 0.009 <128
Dry leaves, goaty,
957 hexanoic acid 0.094 512
fatty acids
974 2-methyl-2-pentanoic acid grassy, acid 0.056 <128
982 β-myrcene lemon, sweet 0.014 <128
1001 hexylacetate + sulfitic grassy, hop 0.002 512
1018 phenylacetaldehyde sweet, hony 0.010 <128
1088 linalool lemon 0.009 <128
1101 β-phenylethanol rose 6.672 512
1226 citronellol lemon 0.037 <128
1260 4-ethylguaiacol spicy 0.011 ND
1294 4-vinylguaiacol cloves 0.70 <128
3
Compounds present in beer after main fermentation

IR CPSil5 Compounds Flavour Samples

YEAST GLYCOSILES PRECURSORS T 0J

HOP CYSTEINES BY PRODUCTS
ACTION ON FATTY ACIDS C (ppm) bière FD

808 3-methyl-2-buten-1-thiol (MBT) hop, cheese ND 512

850 3-mercaptopropanol potatoes ND 512

890 2-mercaptoethylacetate grilled meat ND 512 Analysis performed

at the lab of Prof Sonia Collin
UCL
905 4-mercapto-4-methyl-2-pentanone catty ND <128

1374 beta -damascenone cooked apple ND <128

1432 gamma-decalactone peach ND <128

1465 delta-decalactone coconut ND <128

1441 4-vinylsyringol Cloves ND <128

1578 ethyldodecalactone grassy ND <128

1687 delta-dodecalactone fruity, olive ND <128

4
EBC HOP SYMPOSIUM GDX/JMR/SDK/2009
5
ALCOHOLS
ALDEHYDES
LACTONES
TERPENES
Monoterpenes
Sesquiterpenes

MCF-ACIDS (Medium chain fatty acids)

LCF-ACIDS
GLUCOSIDE OF TERPENES
Monoterpenes
Sesquiterpenes

GLUCOSIDE OF PHENOLS
Phenolic acids
Polyphenols
6
ACIDS
ALCOHOLS
TERPENES
Monoterpenes
Sesquiterpenes
VOLATILE PHENOLS
CYSTEINE DERIVATIVES (THIOLS)

MCF-ACIDS
LACTONES
GLUCOSIDE OF TERPENES
Monoterpenes
Sesquiterpenes
GLUCOSIDE OF PHENOLS
Phenolic acids
Polyphenols

7
Fate of major beer volatiles
by dry hopping (maturation) in
presence of S.cerevisiae
8
MAJOR BEER VOLATILES

MONOTERPENES

CITRUS FLAVORS

9
YEAST GLYCOSILES PRECURSORS

HOP

IR CPSil5 Compounds Flavour Samples

T 0J T 14J T 21J
TERPENES C (ppm) bière FD C (ppm) bière FD C (ppm) bière FD

982 β-myrcene Citrus, sweet 0.014 <128 0.014 <128 0.040 <128

1088 linalool Citrus 0.009 <128 0.05 >128 0.06 >1024

1226 citronellol Citrus, fresh 0.037 <128 0.020 <128 0.118 512

10
CITRUS FLAVORS
IR CPSil5 Compounds Flavour Samples
T 0J T 14J T 21J
TERPENES C (ppm) bière FD C (ppm) bière FD
C (ppm)
bière
FD

982 β-myrcene Citrus, sweet 0.014 <128 0.014 <128 0.040 <128

>1024
1088 linalool Citrus 0.01 <128 0.05 >128 0.06 = dil 1024x

1226 citronellol Citrus, fresh 0.037 <128 0.020 <128 0.118 512

11
DRY HOPPING: evolution of linalool
Influenced by :
- Time
- Temperature
- Hop variety (oil content)
- Hop quantity added
- Hop dispersion methods
(static /dynamic)
- Beer type and yeast type

Linalool

12
CITRUS FLAVOURS

IR CPSil5 Compounds Flavour Samples

T 0J T 14J T 21J
TERPENES C (ppm) bière FD C (ppm) bière FD C (ppm) bière FD

982 β-myrcene Citrus, sweet 0.014 <128 0.014 <128 0.040 <128

1088 linalool Citrus 0.009 <128 0.034 >128 1.192 >1024

512
1226 citronellol Citrus, fresh 0.037 <128 0.02 <128 0.12 = dil 512x

13
DRY HOPPING: evolution of citronellol
Influenced by :
- Time
- Temperature
- Hop variety
- Hop quantity
- Hop dispersion methods
(static /dynamic)
- Beer type and yeast type

14
PRESENCE OF CITRONELLOL

METABOLISM CASCADE OF MONOTERPENE ALCOHOLS BY LAGER

AND ALE YEASTS PROPOSED BY KING AND DICKENSON. THE ASTERIC
INDICATES A CHIRAL CENTER.
15
IN LATE HOPPED WORT
(10 min before end of boiling)

Citation :

HAPPOSHU
16
G

17
Fate of the other volatile compounds
as the …
• Acids
• Esters
• Alcohols
• Aldehydes
• Lactones
• Thiols
• Others

18
 CHEESY FLAVOURS
HOP

YEAST

IR
Compounds Flavour Samples
CPSil5
T 0J T 14J T 21J
SHORT CHAIN FATTY ACIDS
C (ppm) bière FD C (ppm) bière FD C (ppm) bière FD

3-methylbutanoic
acid
cheesy
812 0.101 512 0.102 512 0.179 >1024

4-methylpentanoic
acid
cheesy
921 0.009 <128 0.003 <128 0.018 512

957 hexanoic acid mild cheesy 0.094 512 0.102 512 0.179 >1024

2-methyl-2-
pentanoic acid
grassy, acid
974 0.056 <128 0.037 <128 0.160 512
19
HOP CONDITIONNING AND STORAGE CONDITIONS

CHEESY : linked to bad storage of hop bales

and/or pellets and to the use of aged hop
(presence of degradation products as 2-methyl-
propionic acid/ isobutyric acid and of 2-methyl-
butyric acid/ isovaleric acid)

20
FRUITY, FLORAL FLAVOURS

YEAST

HOP
IR CPSil5 Compounds Flavour Samples
T 0J T 14J T 21J
ESTERS C (ppm) bière FD C (ppm) bière FD C (ppm) bière FD

730 isobutylacetate banana, fruity 0.037 128 0.065 512 0.096 >1024

banana,
774 ethylbutyrate fruity, flowery
0.058 512 0.113 >1024 0.156 >1024

banana,
852 isoamylacetate sweet
0.843 128 1.086 128 1.821 >1024

1001 hexylacetate grassy, hop 0.002 512 0.002 512 0.011 512

21
 FRUITY, FLORAL FLAVOURS

YEAST GLYCOSILES PRECURSORS ACTION ON FATTY ACIDS

HOP CYSTEINES BY PRODUCTS

IR CPSil5 Compouds Flavour Samples

T 0J T 14J T 21J
HIGHER ALCOHOLS C (ppm) bière FD C (ppm) bière FD C (ppm) bière FD

1101 β-phenylethanol rose 6.672 512 6.532 512 11.685 >1024

IR CPSil5 Compouds Flavour Samples

T 0J T 14J T 21J
ALDEHYDES C (ppm) bière FD C (ppm) bière FD C (ppm) bière FD

phenylacetaldehyde sweetened,
1018 0.010 <128 0.015 <128 0.022 512
honey
22
23
 YEAST GLYCOSILES PRECURSORS ACTION ON FATTY ACIDS
HOP CYSTEINES BY PRODUCTS

FRUITY, FLORAL FLAVOURS

IR CPSil5 Compounds Flavour Samples

T 0J T 14J T 21J
LACTONES C (ppm) bière FD C (ppm) bière FD C (ppm) bière FD
gamma-
1432 peach ND <128 ND <128 ND 512
decalactone
delta-
1465 coconut ND <128 ND <128 ND 512
decalactone
Ethyl-
1578 grassy ND <128 ND <128 ND 512
dodecalactone
delta- fruity,
1687 ND <128 ND <128 ND 512
dodecalactone olive 24
WHY LACTONES ?
FRUITY FLAVORS

25
FRUITY FLAVORS

BIOSYNTHESIS
YEAST PEROXISOME

26
FRUITY FLAVORS

YEAST PEROXISOME

BIOSYNTHESIS

27
SULFITIC FLAVOURS
CYSTEINES BY PRODUCTS
YEAST

IR CPSil5 Compounds Flavour Samples

T 0J T 14J T 21J

C (ppm) bière FD C (ppm) bière FD C (ppm) bière FD

3-methyl-2-buten-1-thiol hop,
808 ND 512 ND >1024 ND >1024
(MBT) cheese

850 3-mercapto-propanol potatoes ND 512 ND >1024 ND >1024

grilled
890 2-mercapto-ethylacetate meat
ND 512 ND >1024 ND >1024

4-mercapto-4-methyl-
905 catty ND <128 ND <128 ND 512
2-pentanone
28
Concentration of
major non-volatile compounds
after maturation in presence of raw hops
• Polyphenols
• Glucosides
29
Non volatile compounds extracted
POLYPHENOLS
Year 2008

30
POLYPHENOLS
MOST INTERESTING PHENOLIC COMPOUNDS

• Phenolic acids
• Flavonoids
Flavanoids
Proanthocyanidin = anthocyanogen
• Resveratrol

31
 POLYPHENOLS PHENOLIC ACIDS
HOP
YEAST
ONLY IF PRESENCE OF
BRETTANOMYCES sp
ACTION ON PHENOLIC ACIDS
(Vinyl reductase activity)

PRESENCE OF “POF or PAD YEASTS”

(Phenolic acid decarboxylase activity)
Several TOP fermenting yeasts
FERULIC ACID = Phenolic acid
(Barley, Hops)
CLOVES AND SPICY FLAVORS

IR CPSil5 Compounds Flavour Samples

T 0J T 14J T 21J
PHENOLS C (ppm) bière FD C (ppm) bière FD C (ppm) bière FD

1260 4-ethylguaiacol spicy 0.011 ND 0.000 ND 0.067 >1024

1294 4-vinylguaiacol Cloves, dentist 0.70 <128 1.32 <128 3.36 >1024

32
Formation of vinylguaiacol vinylphenol and styrene
from phenylalanine by S. cerevisiae !

33
POLYPHENOLS

FLAVANOIDS

WHY ARE “FRESH” FLAVANOIDS INTERESTING !

BECAUSE….
34
Non volatile compounds extracted
- POLYPHENOLS

- GLUCOSIDES
35
NON VOLATILE COMPOUNDS
EXTRACTED
POLYPHENOLS
PHENOLIC ACIDS
FLAVANOIDS

GLUCOSIDES
GLYCOSIDES

• IMPORTANCE OF THE HOPS

• INFLUENCE OF THE YEAST STRAINS

• CULTURE YEASTS
• WILD YEASTS
Brettanomyces sp

36
 Non volatile compounds extracted
- GLUCOSIDES
A few theory…
Glucosidic bound can be hydrolysed by :
- High temperature
- Enzymatic activity
- endogenous presence (yeast by
maturation, bottle refermentation)
- (addition of exogenous beta-
glucosidase during the process) –

37
- GLyCOSIDES

Non volatile compounds extracted

In beer, the aglycon part is liberated by two
different types of activities

- by β -1-3-glucanase activity in case of S. cerevisiae

strains
- by β -glucosidase activity in case of Brettanomyces sp.
strains

38
 Glycosidic bounds hydrolysed by endogenous

Non volatile compounds extracted

presence (by yeast during maturation and/or during bottle fermentation)

Preferential links detected in hops

Terpenes
• TERPENIC SUB – UNIT + SUGAR : FLAVOUR PRECURSOR

linalool - β-D- glucoside

α – terpineol – β – D - glucoside
1- octen-3-ol- β – D – glucoside
3 – hydroxy – β – damascenone – D – glucoside

• PHENOLIC SUB – UNIT + SUGAR : SHELF – LIFE PROTECTOR

PRECURSOR
Flavonoids
Kaempferol and Quercetin – 3 – glycosides
glycosidic sub-unit : mono or di- glycosides – glucose , galactose, rhamnose

Flavanoids
As yet, flavanol dimers or multimers glucoside aren’t detected nor described 39
Glucosidic bound hydrolyses by endogenous presence

Non volatile compounds extracted

(yeast by maturation, bottle refermentation)
• Importance of the yeast strain and
yeast species

40
Influence of hop quantity and of yeast strain
TYPICAL YEASTS β-glucanase and β glucosidase activities
of different yeast strains and yeast species
LEGEND :
SC : S.CEREVISIAE (ALE YEAST)
BB : BRETTANOMYCES BRUXELLENSIS
BC : BRETTANOMYCES CUSTERSII
*SC + D : S. CEREVISIAE + CARBOHYDRATE
SOURCE (GLUCOSE)
AR : AR 2000 GLYCOSIDASIC CONCENTRATE OF
ASPERGILLUS NIGER (COMMERCIALLY AVAILABLE)

41
 Glucosidic bound hydrolyses by endogenous presence

Non volatile compounds extracted

(yeast by maturation, bottle refermentation)
• Importance of the aglycon moiety
(glucoside of higher alcohols)
cis-3-hexen-1-ol

1-octen-3-ol

42
HOP
GLYCOSILES PRECURSORS
YEAST

FRUITY, FLORAL FLAVOURS

IR CPSil5 Compounds Flavour Samples

T 0J T 14J T 21J
OTHERS
C (ppm) bière FD C (ppm) bière FD C (ppm) bière FD

cooked
beta -
1374
damascenone
apple, ND <128 ND <128 ND >1024
rose

43
BEER DRY HOPPING IS A CONFLICTUAL ART

WHEN RAW HOP EXTRACTION TAKES PLACE IN PRESENCE OF ACTIVE YEAST

ACTIVE YEAST BY DRY HOPPING IMPROVES THE BEER VALUE BY :

- PROTECTING BEER FROM OXIDATION BY CAPTURING MOLECULAR OXYGEN

- PROTECTING BEER BY EXERTING A REFRESHING EFFECT

- IMPROVING (IN A FIRST STEP) THE BEER FLAVOUR BY :

• increasing the content of citronellol

• synthesizing nice flavours as lactones
• liberating aglycons from glucosides :
- Linalool, Geraniol,…

44
BEER DRY HOPPING IS A CONFLICTUAL ART

WHEN RAW HOP EXTRACTION TAKES PLACE IN PRESENCE

OF ACTIVE YEAST

ACTIVE YEAST by dry hopping

DECREASES THE BEER VALUE BY :

- LIBERATING ESTERASES THAT DECREASE THE AROMA

- METABOLIZING H+ ACCEPTORS AS SOME ALDEHYDES
AND OTHER POTENT SUBTILE FLAVOURS AS LACTONES

45
 BEER DRY HOPPING IS A CONFLICTUAL ART
WHEN BOTTLING TAKES PLACE IN PRESENCE OF ACTIVE YEASTs

46
 TRADITIONNAL EUROPEAN METHODS
Orval
C Methods by batch dry hopping
A
B D

A : Parts of 4 KG of dried hop cones are compacted

under inert gas (N2) in onion bags that are sealed in
aluminium packets by the hop supplier. of 4 KG.
Sierra Nevada
These are stored in the fridge of the brewery at + 2 /+ 4°C.
B : Just before use, the aluminium packets are
opened and the onion bags containing the
hop cones are decompressed.
C : Before introduction in the empty tank, E
the hop cones are decompacted too
D : In order to improve the contact between
the beer and the hop bells, the hop bag can be
attached to the tank. E : Hop plugs of 5 Oz ( 140 Gr) are also available
47
by hop suppliers and can be added to beer casks directly.
 Methods by continuous
extraction in separated vessels
1.1. by hop cones
Sierra Nevada Brewery
1.2. by hop pellets
Stone Brewery (designed plant)
NEW AMERICAN METHODS New Belgium Brewery

Pneumatic Method
= dry method
Out- Indoor hop
cellar canoon
Lagunitas Brewery
48
With authorization of Steven Pauwels, CBC Dry hopping subcommittee
HOP and BEER

49