FOSC 4081: Brewing Science

Lab 3: Pilot-Scale Mashing and Fermentation

Lab Completed: October 29, 2015
For
Dr. Andrew Macintosh
Submitted: November 13, 2015

_____________________________
Jonathan Totten
B00450823

_____________________________
Matthew Zwicker
B00582641

Lab 3: Pilot-Scale Mashing and Fermentation

Executive Summary

Lab 3: Pilot-Scale Mashing and Fermentation

Table of Contents
Executive Summary.............................................................................................................ii
Table of Contents..................................................................................................................i
List of Figures.......................................................................................................................i
List of Tables.......................................................................................................................ii
1

Introduction................................................................................................................3

Material and Methods.................................................................................................3

Results and Discussion...............................................................................................6

References............................................................................................................................9

List of Figures
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Lab 3: Pilot-Scale Mashing and Fermentation

Figure 1: Kettle on the right and the mash tun with the added malt

Figure 2: Plate heat exchanger used for cooling and fermentation vessel

Figure 3: Calibration curve of absorbance vs yeast cell count

Lab 3: Pilot-Scale Mashing and Fermentation

List of Tables

Lab 3: Pilot-Scale Mashing and Fermentation

Introduction

Creating a well-balanced beer that will satisfy the palate of the consumer begins with the
selection of quality ingredients including: hops, malt, yeast and water. Totten & Zwicker
craft breweries is considering purchasing a major portion of malt for the 2015/16 fiscal
year to brew their popular chocolate porter ale. To determine if the malt passes Totten &
Zwickers standards of excellence, a pilot-scale mash and fermentation was completed.
The objectives of the pilot-scale mashing and fermentation are:

To analyze various mash parameters in a pilot-scale mash and become familiar

with the mashing process.

To analyze various fermentation parameters in a pilot-scale fermentation using

standard procedures.
To contrast fermentation data with a control fermentation for which you will be

given the data.

To model and compare the key fermentation parameters (density and absorbance)
of both the experimental and control fermentations, using appropriate equations.

It is expected that the pilot-scale mash and fermentation will have similar results to the
control

Material and Methods

The lab consisted of two main parts: the mashing step and the fermentation step.

To begin the mashing step the brewing water was pre-heated in the steam kettle. The
EBC congress mash was used to determine the ratio of malt to water (1:4). The water
from the kettle was transferred to the mash tun and kept at a constant temperature of
70oC. The malt was then added to the mash tun that consisted of 80% 2-row barley
(11.76kg), 10% crystal malt (1.47kg), and 10% chocolate malt (1.47kg) for a total of 14.7
kg. During the mashing procedure frequent samples were collected to perform iodine
tests, color and density tests. The mash was stirred with an oar before taking samples.
Samples were taken at the beginning, end and sparging steps. Using the ESC-4.5.1
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Lab 3: Pilot-Scale Mashing and Fermentation

(Section 8.1.6) method, the saccharification (solution turns dark purple to yellow
indicating sugar) of the mash was tested by taking samples every five minute until
complete. The samples for the color and density were prepared by transferring to 50 mL
of mash to centrifuge tubes and then the malt was crashed cooled on ice to 20 oC to slow
hydrolytic activity of the enzymes. The samples were then centrifuged for two minutes at
5300 rpm and then filtered through Whatman #4 paper. Using the ASBC Beer-10 method,
the color of the mash was measured by measuring the absorbance at 430 nm wavelength
with a spectrophotometer. The density of the malt was measured using the ASBC Beer-3
method and an Anton Parr densitometer. Figure 1 shows the kettle boiler on the right used
to boil the water for the mash and the picture on the left shows the mash tun with the
water and ground malt.

Figure 1: Kettle on the right and the mash tun with the added malt
After one hour the mash was filtered through a strainer and the malt was added
back to the mash and the wort was added to the empty kettle. The wort was heat until it
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Lab 3: Pilot-Scale Mashing and Fermentation

reached 100oC and then 50% of the fresh hops from Malagash, Nova Scotia and two cups
of cascade hops were added. The wort was boiled for 25 minutes and the remainder of the
fresh hops was added. Finally, the wort was boiled for a further five minutes. The hot
wort was then transferred through a (type) plate heat exchanger to the fermentation vessel
by transfer hoses. The heat exchanger was fitted with a thermometer, sight glass and
oxygen insert. The plate heat exchanger cooled the hot wort from 100 oC to 30oC and also
injected medical grade oxygen from a compressed cylinder into the wort to oxygenate it
before it entered the fermenter. It should be noted that during the transfer from the kettle
to the fermentation vessel, the plate heat exchanger was clogged with hop debris from the
kettle and a significant amount of wort was lost. Figure 2 shows the plate heat exchanger
used for cooling on the left and the fermentation vessel on the right.

Lab 3: Pilot-Scale Mashing and Fermentation

Figure 2: Plate heat exchanger used for cooling and fermentation vessel
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Lab 3: Pilot-Scale Mashing and Fermentation

An ale Smack pack brewing yeast was used for the fermentation. To count the
yeast cells in suspension the ASBC Yeast-4 method was used. The fermentation
continued over the weekend after the lab was completed (insert length of fermentation).
The post analysis crew took samples at 9:00am, 1:30pm and 5:00pm the day after mash
and fermentation to measure the density. Appendix A contains the tasks allocation for the
mash crew, the boil fermentation crew, post analysis crew, and the clean-up crew.

Results and Discussion

Mashing is a process by which wort is prepared. To begin the mashing process

pre-heated water (or liquor) was transferred to a mash tun and then the malt was added.
There are key physical differences between the wort and the mash. The mash is the
combination of ground malted grains, water and sometimes supplementary enzymes and
salts. On the other hand, the wort is the liquid extracted from the mashing process. The
extraction process leaves few to no grain husks left in the wort. The wort contains the
starch from the malted grains that is enzymatically broken down to sugars. There are
typically two types of mashes; infusion mashing and decoction mashing. Infusion
mashing is the process where the mash is heated to different temperatures at which
specific enzymes work optimally. A single rest at one stable saccharification temperature
is used in this technique. Decotion mashing is no longer a widely used technique due to
the introduction of automated temperature control. The basic principle of this technique is
to extract part of the mash from the system, boil it, and return it back to the rest of the
mash. This process allows the use of physical pulping to help break down the cell walls in
the malt in addition to raising the mash to a second temperature by mixing the two parts
of the mash.
During the mashing process a beta-glucanase rest is typically done at 40 degrees
Celsius (oC) to break down the grains cell walls and make starches more available.
Highly polymerized beta-glucans are located within the cell walls of the starchy
endosperm of the malt. Beta glucans are polysaccharides such as cellulose which are
made of glucose molecules connected by beta glycosidic bonds. At 70 oC an alpha and
beta amylase rest are typically down simultaneously. The beta-amylase enzyme becomes
active first and begins by breaking down the starch chains from the end of the molecules,
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Lab 3: Pilot-Scale Mashing and Fermentation

forming two chain glucose molecules (maltose). Alpha-amylase becomes active shortly
after the beta-amylase where the starches are broken down from the inside of the starch
molecule creating chains of one to four glucose molecules in length (dextrins or
maltodextrins). It should be noted that barley does not contain alpha amylase; however, it
does develop in the grain during the malting process.
Calcium salts are sometimes added to the mashing process as the mash liquid
(wort) can contain large amounts of phosphates that were derived from the ground malted
grains. The phosphate ions can have a buffering effect where the hydrogen ions from the
water are reacted with the phosphate, resulting in a pH increase. The added calcium will
react with the phosphate by the following reaction:
+
+2 H
2 Ca 3(PO 4 )2
2++2 HPO4
3 Ca

The calcium phosphate is insoluble in the liquid and precipitates, releasing hydrogen ions
and lowering pH. The lower pH improves beta-amylase activity thus increasing sugar
extract content. The calcium ions also protect and stabilizes the alpha-amylase enzyme
from inhibition by heat. The lowered pH will also provide a greater resistance to
microbiological infection. Calcium is not added during our test mashing procedure
because the outcome of the test mash is to determine the properties of the ground malt.
Therefore, it is crucial to understand how the malt affects pH of the wort without the
addition of other compounds such as calcium salts.
Brewers generally manage pH values mostly for enhancement of enzyme
performance in the mash. Beta and alpha amylase perform optimally between ranges of
5.4 to 5.6 and 5.6 to 5.8 respectively. pH is sometimes measured during mashing because
pH is difficult to predict in advance even if a recipe is being followed. Enzyme activity
during the mash is crucial and as mentioned before, a pH that is too high or too low will
affect the activity of the enzyme and how much starch is converted to sugars.

Lab 3: Pilot-Scale Mashing and Fermentation

The presence of starch in the mash is measured by the iodine test. The iodine is
usually added to the starch in the form of potassium iodide. This will usually start as a
blue-black colour. Due to the structure of amylopectin in the starch, the combination of
iodine and starch will form a yellow colour. As the starches become hydrolysed into
smaller particles, the reaction which forms the yellow colour will stop occurring.
A w /w =(OE AE)0.419
Where,
Aw/w = % Alcohol (weight/weight)
OE = Original Density (or Extract - oP at the beginning of fermentation)
AE = Apparent Density (or Extract - oP at the end of fermentation)
A w /w =(9.67 P3.6 P)0.419
A w /w =2.54 3
Beers are reported in some parts of the world on a weight per weight basis;
however, beer in Canada is reported in a volume per volume basis. Therefore the actual
alcohol percentage in volume per volume is calculated by the formulas below:
SG=1+

P
P
258.6
227.1
258.2

((

Where,
o

P = Degree Plato

SG = Specific Gravity
SG=1+

3.6
3.6
258.6
227.1
258.2

((

SG=1.0144
A V /V =

A W /W SG
0.7907
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Lab 3: Pilot-Scale Mashing and Fermentation

Where,
Av/v = % Alcohol (volume/volume)
A V /V =

2.4531.0144
0.7907

A V /V =3.26
An alcohol level of 3.26% volume per volume does not seem reasonable for a
porter. Porters are dark, top fermented beers made with generous amounts of hops. The
typical porter alcohol level is in the range of 4.5% to 6% by volume. Therefore, the
brewed chocolate porter has a significantly lower alcohol level than a theoretical porter.

2.5
2
1.5

Absorbance

f(x) = 0.58 ln(x) - 8.52

R = 0.99

1
0.5
0
0.00E+00

5.00E+07

1.00E+08

Cell Count

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Lab 3: Pilot-Scale Mashing and Fermentation

Figure 3: Calibration curve of absorbance vs yeast cell count

References
American Society of Brewing Chemists. (1975). Apparent extract. Retrieved from
http://methods.asbcnet.org/methods/Beer-3.pdf
American Society of Brewing Chemists. (2002). Color.
Retrieved from http://methods.asbcnet.org/methods/Beer-10.pdf
American Society of Brewing Chemists. (2011). Miniature fermentation assay.
Retrieved from http://methods.asbcnet.org/methods/Yeast-14.pdf
American Society of Brewing Chemists. (2010). Wort color and sample preparation.
Retrieved from http://methods.asbcnet.org/methods/Wort-9.pdf
Briggs, D. E., Boulton, C. A., Brookes, P. A. & Stevens, R. (2004). Brewing science and
practice. Cambridge, EN: Woodhead Publishing Ltd.

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Lab 3: Pilot-Scale Mashing and Fermentation

Cutaia, A. J., Reid, A. J. & Speers, R. A. (2009). Examination of the relationships
between
original, real and apparent extracts, and alcohol in pilot plant and commercially
produced beers. Journal of the Institute of Brewing, 115(4), 318-327.
European Brewery Convention. (2000). Extract of malt: Congress mash. Retrieved from
http://analytica-ebc.com/index.php?mod=contents&method=297
Oliver, G. (2012). The Oxford Companion to Beer. New York, NY: Oxford University
Press, Inc.

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Lab 3: Pilot-Scale Mashing and Fermentation

Appendix A Task Allocations

Analysis

Mash Crew
Jake Steinburg and Matt Gillis

Boil Fermentation Crew

Dylan Walker and Riley Giffen

Transfer

Matt M and Andrea Neyens

Jonathan Totten and Matt Zwicker

Clean Up Crew
Po-Lin Hsu and Tianlin Zhao
Post Analysis Crew
Parker McNeil and Bonnie Moore
Annmarie Hessian
Sai Kalyanaraman and Vitor Motta

9:00 am
1:30 pm
5:00 pm

Appendix B Raw Data

Test

Mash first half

Mash End
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Lab 3: Pilot-Scale Mashing and Fermentation

Iodine

Black (No
change)

Test
Mash first half
Colour (Abs430)
2.438
2.725
2.742

Test
Density (Plato)

Yellow
(Change)
Mash End
2.261*
2.221*
2.19*

Mash first half

Mash End
5.7
9.1
5.4
9.3
5.4
9.2

Date/Time
Density (Plato)

Cell counts
(cells/mL)

(4:40 pm 29-10-15)
(9:00 am 30-10-15)
(1:30 pm 30-10-15)
(5:00 pm 30-10-15)
(10:45 am 2-11-15)

Ferm Time
0
16.333
20.833
24.333
90.083

Date/Time

Ferm Time

(4:40 pm 29-10-15)
(9:00 am 30-10-15)
(1:30 pm 30-10-15)
(5:00 pm 30-10-15)
(10:45 am 2-11-15)

0 hr
16.333 hr
20.833 hr
24.333 hr
90.083 hr

Readings
(Triplicate)
9.5
5.2
3.8
3.7
3.6

9.3
5.3
3.6
3.5
3.6

10.2
5.2
3.6
3.4
3.6

Cell count
(cells/mL)
N/A
1.39 x 10^7
3.6 x 10^7
6.2 x 10^7
1.4 x 10^7

15