School of Food and Nutritional Sciences

University College Cork

Ireland

Critical review of the understanding

of chill haze formation in beer
Jean Titze, Antonie Herrmann and Vladimír Ilberg

EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Haze – A critical quality factor [5]

Physico-chemical Sensorical
stability stability

Quality

Microbiological stability

1 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Haze development (acc. to relevant literature) [2]

protein + polyphenol

(protein)m + (polyphenol)n
precipitation
low temperatures, 0 °C (e.g. due to catalysis by
reversible at >20 °C metal ions or oxygen)

(protein)a + (polyphenol)b (protein)x + (polyphenol)y

reversible chill haze permanent haze

2 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Characteristics of beverages [9]
Considering the characteristics of a filtered beer,
following physico-chemical definition can be given:

Definition
Filtered beer can be defined as a water-
ethanol-mixture in which colloids (e.g.
proteins), besides real dissolved substances
(e.g. salt), are dispersed.

The haze behavior of a dispersion is mainly

determined by the interaction between these colloids.

3 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Colloid Science [9]
Link between molecular effects and solid-state characteristics

Molecular Colloid Science Solid-state

Chemistry/ Physics/
Physics Chemistry
Big
molecules 1 µm

„small“
1 nm particles

Particle Analysis:
Particle size
Maillard reaction Rheology
while wort boiling Surface charge while mashing

4 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Importance of surface – Example of the fauna

Arctic fox The arctic fox has a good relation

(Vulpes or Alopex lagopus) between surface and volume,
to keep its body temperature.

The small ears show

the surface optimization.
Fennec
(Vulpes zerda)

In comparison to that, the desert fox

can afford to have big ears.
5 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Surface charge of a particle [9]

Surface with unsaturated groups (e.g., carboxyl or amino):

Surface
COO

SO 42

+
NH3

6 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Physical stability of dispersions [9]

7 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Forcier test [4,6]
Start Turbidimetry (90°); +20 °C Start value = a

Incubator
+40 °C or +60 °C, 24 h
Cool down to
One „Warm day“ room temperature
= 48 h (2 days) Fridge or ice water bath
0 °C, 24 h
Warm up to
room temperature
Turbidimetry (90°); +20 °C Result = b

No
b − a ≥ 2 EBC

Yes
End
8 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Hazing curve of a filtered beer (real time test) [3,7]
2,5
Lag Phase Increase Phase Stationary Phase

2
Haze 90° [EBC]

1,5

0,5

0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14

Storage time at 20 °C [month(s)]

9 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Haze curves during a long-term forcing test [7-9]
24
after
nach 24 h warm phase
Wärmebehandlung
22
nach Kältebehandlung
after 24 h cold phase
20

18

16
Haze 90° [EBC]

14

12

10

3.3
2.1
6.1

3.2
7.2

7.3

4.4
1.12
5.12
9.12

10.1
14.1
18.1
22.1
26.1
30.1

11.2
15.2
19.2
23.2
27.2

11.3
15.3
19.3
23.3
27.3
31.3
25.12
27.11

13.12
17.12
21.12

29.12

2009 2010
10 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Mathematical fitment of the haze curves [8,10]
25 5,5
after 24 h warm phase
after 24 h cold phase 5
haze difference
20 4,5
relative haze difference
4
R2 = 0.9453

Haze difference [EBC]

3,5
15
Haze 90° [EBC]

R2 = 0.9508 2,5
10
2

1,5
5 1

0,5

0 0
0 4 8 12 16 20 24 28 32 36 40 44 48 52 56 60 64

Time [d]

11 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Water release due to cold temperatures (synaeresis) [1,9]

−H2O

Cooling

Amorphous area
with fringed
polypeptide chains

Inclusions of water molecules

preferred in the amorphous area
12 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Particle size distribution of an artificial aged beer [8,10]
artifical aged beer (3 weeks at 60 °C)

artifical aged beer after 24 h at 0 °C (chill haze)

100
artifical aged beer after 24 h at 0 °C
and after that 24 h at 20 °C
Intensity sum distribution [%]

80

60

40

20

0
0,1 1 10 100 1000 10000
Particle size [nm]

13 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Particle size distribution of a whey protein solution
Whey proteins at 20 °C

Whey proteins after 24 h at 0 °C

100
Whey proteins after 24 h at 0 °C and 24 h at 20 °C

Whey proteins after 24 h at 0 °C and 48 h at 20 °C

Intensity sum distribution [%]

80

60

40

20

0
0,1 1 10 100 1000 10000
Particle size [nm]

14 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Conclusion 1/2 [8,10]
1. Colloidal haze (formation of permanent haze)
The colloidal haze formation of beer is purely a matter of a thermal effect,
which is the reason for the following two mechanisms:
1.1 Already denatured proteins in beer agglomerate according to collision
(DLVO-theory).
1.2 Native proteins start unfolding. After that, they are able to
agglomerate.
Hence, the basic requirement for haze formation is thermal energy. With
every warmday the amount of denatured proteins, which can agglomerate
right away, as well as the number of native proteins, which first unfold and
then can agglomerate, increase.

15 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Conclusion 2/2 [8,10]
2. Chill haze (formation)
Due to cold temperatures the water gets out of the molecule of the not
denatured proteins with a change in the properties of the hydration hull.
Two following activities are possible:
2.1 According to the diversification of the protein hydration the index of
refraction of the molecule changes and the particle gets visible.
2.2 Due to the modification of the hydration, hydrophobic groups of the
protein can interact with themselves or with already denatured
proteins and reversibly accumulate with them.
Both phenomena are of reversible nature. Independent of the beer aging
status, they lead to an increase of haze due to the cold temperature.

16 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Acknowledgement

The authors would like to thank

Mr. Markus Schmidt

from the Research Center Weihenstephan for Brewing and

Food Quality for the performance of the long-term forcing test.

17 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 References 1/2
1 DRITTES PHYSIKALISCHES INSTITUT (DPI), 2011: Lehre der Biophysik und der Physik komplexer Systeme
– Computergestützte Biophysik I. Fakultät für Physik. Georg-August-Universität, Göttingen. URL:
http://www.dpi.physik.uni-goettingen.de/Lehre/Bachelor/ images/ comp_biophys1.jpg, letzter
Zugriff am 05.11.2011.
2 GUZMAN, J. E., MCKEOWN, I. P., GLEAVES, M., STEWART, G. G., DOYLE, A., 1999: Selective protein
adsorption with silica gel. – MBAA Technical Quarterly 36, Nr. 2, S. 227-230.
3 MÉLOTTE, L., 2008: Measurements of Beer Colloidal Stability. XIIIth J. De Clerck Chair, Sept. 2008,
URL: http://www.uclouvain.be/cps/ucl/doc/inbr/documents/DeClerckChair Melotte.pdf, letzter
Zugriff am 05.11.2011.
4 TITZE, J., ILBERG, V., FRIESS, A., JACOB, F., PARLAR, H., 2007: Enhanced long term stability measurement
using a charge analyzing system. – In: Proceedings of the 31th EBC Congress, Venice, European
Brewery Convention, S. 718-735.
5 TITZE, J., 2010: Qualitätssiegel oder Medaille? – Güte und Sicherheit von Produkten richtig
kommunizieren. – Getränkeindustrie 64, Nr. 10, S. 52-55.
6 TITZE, J., CHRISTIAN, M., ILBERG, V., JACOB, F., 2010a: Particle analysis – A combined method to
analyze the colloidal characteristics of particles. – BrewingScience 63, Nr. 5/6, S. 62-71.
7 TITZE, J., CHRISTIAN, M., JACOB, F., PARLAR, H., ILBERG, V., 2010b: The Possibilities of Particle Analysis
Demonstrated by the Measurement of the Colloidal Stability of Filtered Beer. – Journal of the
Institute of Brewing 116, Nr. 4, S. 405-412.
8 TITZE, J., HERRMANN, A., ILBERG, V., 2012: Ein chemisch-physikalischer Erklärungsansatz – Neue
Erkenntnisse zum Mechanismus der Kältetrübung. – BrauIndustrie 97, no. 4, pp. 12-15.
18 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 References 2/2
9 TITZE, J., 2012: Theoretische und praktische Aspekte bei der Anwendung der
Ladungstitrationsmethode zur schnelleren Vorhersagbarkeit der kolloidalen Stabilität
untergäriger Biere, Technische Universität München, Fakultät Wissenschaftszentrum
Weihenstephan für Ernährung, Landnutzung und Umwelt, Dissertation, submitted.
10 TITZE, J., HERRMANN, A., ILBERG, V., 2012: Recent Findings on the Mechanism of Chill Haze – a
physico-chemical Explanatory Approach. – World Brewing Congress. Portland (Oregon, USA),
24.07.-01.08.2012.

19 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Thank you for your attention!

20 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen
 Contact

Jean Titze National University of Ireland

Senior Research Scientist
University College Cork
School of Food and Nutritional Science
College Road
Cork
IRELAND
Phone +353 21 490 3435
Mobile +353 83 114 4342
E-mail j.titze@ucc.ie

Prof. Dr. Vladimir Ilberg

Gartenbau und Lebensmitteltechnologie

Lebensmitteltechnologie/Verfahrenstechnik

Am Staudengarten 11 | 85350 Freising/Germany

Tel +49 (0)8161 - 71 5030 | Fax +49 (0)8161 - 71 5251
vladimir.ilberg@hswt.de | www.hswt.de

21 TITZE, J., HERRMANN, A. and ILBERG, V.: Critical review of the understanding of chill haze formation in beer.
EBC Symposium "From Chiller To Filler“  11 September 2012  Carlsberg Research Centre  Copenhagen