Vol.

58, 1952] seed: pure yeast culture 129

and also as to whether it may be contaminated
with wild yeast, or infected with bacteria.
Biologically pure pitching yeast.—By this
is meant culture yeast that contains no wild
yeast and no bacteria. When the old yeast
is replaced by a pure culture yeast, it becomes
possible to clean out the whole plant, and to
be assured that the new fermentation will
proceed for a considerable period free from
infection.
Uniformity.—When a suitable strain of
culture yeast has been selected, the fer
mentations will proceed with a regularity
and uniformity unobtainable by other means.
Acknowledgement.—Thanks are due to
John Davenport & Sons Brewery, Ltd., for
permission to use the results of the work
done at their brewery, and to Messrs. A. P.
Grant and Gordon Leek, B.Sc, for their
practical assistance in carrying out the tests
described.

COMMUNICATIONS

EFFECT OF STORAGE ON BEERS CONCENTRATED BY FREEZING,

AND ON BEERS KEPT IN THE FROZEN STATE;
SOLUBILITY OF CARBON DIOXIDE IN ICE

By R. E. Essery, B.Sc, Ph.D., F.R.I.C.

(Institute of Brewing Research Laboratories, The University, Edgbaston, Birmingham)

and R. Gane, M.A., M.Sc, Ph.D., A.R.I.C.

(Low Temperature Research Station, Cambridge)
Received \2ih October, 1951

Freezing and thawing do not of themselves appear to influence haze potentialities

or flavour of beer, though there is some loss of gas. Storage of concentrates may lead
to the production of haze, sediment and a vinous flavour, though the diluted products
fine satisfactorily and loss of body can be compensated for by addition of dextrin
and peptone, whilst addition of appropriate amounts of hop-resin concentrate and of
hop oil may also be useful. In general, storage of concentrates at 0-15° C. gives
best results. With whole beers maintained in the frozen state, storage at — 10-
— 20° C. is satisfactory at least up to 18 months, though overfoaming may occur from
rapidly-thawed containers. Since the solubilities of carbon dioxide in ice at — 5°
and at — 20° C are, respectively, 1/20 and 1/100 that in water at 0° C, high pressure
may develop in frozen containers, re-establishment of equilibrium on thawing being
slow*

In a previous paper, Essery, Gane & T. N. Experimental

Morris (this Journal, 1947, 204) described
some experiments on the concentration of
beer by partial freezing and removal of ice,
and some preliminary results on the effect of
storage on concentrates so obtained. The
present communication reports the results of
a more extended study of storage effects, not
only on concentrates, but also on beers
frozen and held at low temperatures.
Beer Concentrates
Analysis of samples.—Specific gravity,
original gravity, and alcohol concentration
were determined on beers and concentrates
by standard methods. The apparent con
centration of the concentrates was calculated
(a) from the ratio of the O.G. values; (b) from
the ratio of the alcohol concentrations; (c)
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130 ESSERY AND GAME: STORAGE OF FROZEN BEER [J. Inst. Brew.

the ratio of the refractive indices. It will be
seen from Table I that the three methods give
comparable results. It will also be seen,
from the ratio (O.G.—1,000)/alcohol, that
there is a slight loss of alcohol during con
centration in Series I and II. This is
probably due to evaporation in the centrifuge,
since a similar effect was shown in Tables III
and IV of the previous paper.
TABLE I Analysis of Beers and of Concentrates prepared by Freezing
for the ales 00053% of hop-resin concentrate
(1 oz. per brl. of diluted liquid) and 0000027%
of essential oil of hops (I fl. oz. per 653 brl.)
gave good results, but stout required double
the amount of hop concentrate. These addi
tions were made after dilution but before
fining. Care is required in determining the
amount of hop oil to be added, as excess gives
a vague aromatic odour which was not

Apparent degTce of con-

concentration

1.000 From
Present Original Alcohol From From refract
Series Sample gravity gravity % (v/v) Alcohol O.G. alcohol ive index

I I.P.A.:
Control 1000*50 1032-94 3*30 9-80
Concentrate 1033*13 112910 12*02 10*23 3-92 3*70 4*20

Oatmeal stout:
Concentrate .. ., 1042-05 1135*20 1214 11-14 — — —

II Burton:
Control 1003*30 103000 3*45 8*87
Concentrate 1020*50 1107*15 11-73 9*13 8*50 340 3*45

III Pale ale:

Control 1002*85 1028-23 319 8-83
Concentrate 101401 1110*52 13*78 8*07 . 4*23 4*30 413

IV Brewery A:
LP.A:
Cans 100902 1044*30 4*50

Brewery B:
I.P.A.:
Bottles 1010*23 1043*04 4*38 — — — —

Cans 1009-87 1043*13 4-27

Pale ale:
Bottles 1008*93 1033*35 3*20
- z
— —-

Cans 1008*89 1032*84 3*12

Brown ale:
Bottles 1011-74 1032-71 2*04 — —

Cans 1012-47 1032*70 204 —

In general, dilution of concentrates back favoured. Moreover, the hop concentrate

to the concentration of the original beer gave
products which were thin and weak. Dilu
tion back to a concentration of 1*3 gave an
acceptable beer, which was, however, still
deficient in body and bitterness. Body could
be secured by addition to the diluted product
of 4% of dextrin and 0-1% of peptone.
Bitterness could be replaced with hop-resin
concentrate and aroma by essential oil of
hops. Precise amounts thought necessary
were largely a matter of personal taste, but
used must be hop-resin concentrate, as the
"complete" concentrate gives opalescence,
with very poor fining.
Freezing and thawing without storage.—Four
samples of the I.P.A. of Series I (see below)
were treated thus: [a) left on the bench over
night; (b) stored at 0° C. overnight and then
allowed to reach room temperature on the
bench; (c) frozen overnight in a glass vessel
and then allowed to reach room temperature
on the bench; (d) as (c) but frozen in a tin

Vol. 58, 1952] ESSERY AND GANEI STORAGE OF FROZEN BEER
vessel. Samples (b)t (c) and (d) showed
marked chill haze at low temperature, but
this cleared as the samples warmed up and,
finally, all were indistinguishable from the
control (a), except for loss of gas from (c) and
(i). There is therefore no evidence that the
mere processes of freezing and thawing
exercise any effect either on the flavour or
on the formation of irreversible haze.
Freezing and storage.—The I.P.A. of Series I
was a fined cask-beer, frozen at — 12° C.
(10-4° F.), which was centrifuged to remove
ice. A portion of the concentrate was
carbonated at room temperature under a
pressure of 15 lb. The two portions were
bottled and crowned, the crowns being sealed
with wax; storage thereafter was at 0°, 15°,
20° and 25° C. (32°, 59°, 68° and 77° F.).
Controls of the original beer were pasteurized
at 65-6° C. (150° F.) for 20 min., and stored
with the concentrates, while fresh samples of
the same type of beer were obtained from the
brewery at each sampling. Results may be
summarized as follows:
Haze and a settled deposit appeared in all
samples after 5 months (the first sampling),
but did not appear to increase thereafter.
The dilutions fined satisfactorily, and gave
bright beers. Vinous, "old-ale" flavour
appeared in all concentrates and in their
dilutions, more rapidly at the higher storage
temperatures, but differences lessened as the
storage time increased, less vinous samples
catching up on the more vinous ones so that,
after 15 months (final sampling), there was
little detectable difference. It was generally
agreed that carbonation of the concentrate
gave a smoother and more pleasant product,
both as regards the concentrate itself and
after it was diluted. After 15 months, all
samples gave a " reasonable " beer on dilution,
all having a distinct "old-ale" flavour. It
was agreed that storage at 15° C. gave the
best product, with 0° C. a close second and
20° and 25° C. not so satisfactory.
An oatmeal stout was also examined, being
a sample which had been concentrated at
— 12° C. and stored in bulk at that tempera
ture for 6 months. As the quantity was
small, it was examined only on bottling and
after 15 months.
The concentrate was rather harsh and
bitter, but again carbonation gave a smoother
product. On dilution, it gave a pleasant
drink which retained the stout flavour and,
while lacking body, was equivalent to a good
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131
bitter beer. After 15 months, it gave a
reasonable drink on dilution, improved by
restoration of bitterness with hop-concen
trate. The best storage temperatures were
found to be 0° C. and 15° C.
Fined cask-beer, described as "low-gravity
Burton," was used in Series II. This beer
was chilled in store at — 2-2° C. (28° F.) and
filtered cold before concentration at — 12° C.
Carbonation, storage, and dilution were as
for Series I.
Carbonation of the concentrate improved
the product. The results were similar to
those of Series I, but the storage temperature
effect was not so pronounced. Vinous
flavour was marked in all samples after 11
months, at which time the beers were begin
ning to deteriorate; after 13£ months they
were valueless. Chilling and filtration before
concentration had an unexplained adverse
effect on storage-life, but there was no
perceptible effect on the onset of haze and
deposit in the concentrates.
For Series III, a fined cask-beer of the
Pale Ale type was concentrated at — 10° C.
(14° F.), the beer being handled throughout in
an atmosphere of carbon dioxide, with careful
exclusion of air. Storage and dilution (in an
atmosphere of carbon dioxide) were other
wise as for Series I, except that carbonation
was not applied.
Vinous flavour was noted after 17 weeks,
but was then only faint in the dilutions. It
became marked after 32 weeks and strong at
72 weeks. The influence of storage-tempera-
ture was not so marked as in previous series,
and the expressed preferences were largely a
matter of personal taste, with a tendency for
tasters to select the beer stored at 15° C. as
the most satisfactory. No taster made
adverse comment on the vinous flavour.
Handling in carbon dioxide seemed to
increase storage-life very markedly. Apart
from the vinous flavour at 72 weeks, there was
no evident reason why the samples, at all
temperatures, should not have withstood
longer periods of storage.
Beers Maintained in the Frozen State
For Series IV, Brewery A provided beer of
I.P.A. type, in £-pint cans. The cans were
gold-lacquered inside, with soldered seams,
the interior lacquer being covered with a
coating of waxy material soft enough to be
scraped off with the thumb-nail. Controls
were stored at 5° C. (41° F.), the remainder of

132 ESSERY AND GANEI STORAGE OF FROZEN BEER
the cans being divided into three sets which
were frozen and stored at — 3°, — 10°, and
-20°C. (26-6°, 14°, and - 4° F.) respec
tively. All samples at — 3° C. remained
liquid and supercooled. While an occasional
leaker was found, there was no general
evidence of damage to seams, distortion of
cans, or damage to the interior coating on
freezing.
After 6 days, all samples were bright and
there were no obvious differences between
them. All possessed a slight after-taste,
rather unpleasant, which was referred to as
"can flavour." After 7$ weeks, all samples
showed an irreversible haze which increased
slightly with time of storage. After 62J
weeks, there was no evidence that storage had
interfered with flavour, except that the 5° and
— 3° C. samples were slightly unclean, per
haps representing the beginning of deteriora
tion. The — 10° and — 20° C. samples
were quite satisfactory, apart from the can-
flavour, and there was throughout a general
tendency to prefer these two. Condition
remained good, without overfoaming, and no
vinous flavour appeared.
Brewery B provided I.P.A., Pale Ale, and
Brown Ale, in bottles and cans. The beers
were treated as for Brewery A, except that
the bottles to be frozen were uncrowned,
60 ml. were removed from each to allow for
expansion, and the bottles were re-closed
with corks tied in. This necessarily involved
some loss of condition. The cans were of the
same type as those from Brewery A and again
no evidence was obtained of damage due to
freezing.
After 6 days, all samples remained bright,
and no differences could be observed other
than those due to loss of condition in the
bottles. After 14 weeks, samples remained
bright and no marked differences could be
observed between controls and frozen sam
ples. With the cans, however, condition
was very high and a rather large amount of
beer was lost by overfoaming when the cans
were opened. After 30 weeks, heavy sedi
ment and haze commenced to appear
irregularly, mainly in the bottles. Differences
in flavour were slight, but there was a
tendency for tasters to prefer cans to bottles
and the lower temperatures of storage to the
higher. Slight can-flavour was reported by
one observer. After 42 weeks, there were no
marked differences, but more decided can-
flavour was beginning to appear* No onset
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[J. Inst. Brew.
of vinous flavour was noted, and there was
again a general tendency to prefer the beers
stored at the lower temperatures.
After 18 months, the final samples were
returned to the brewery for report. The
general opinion was that the beers had kept
well and were commercially good when care
fully decanted from the sediment. The
brewery reported no overfoaming of cans,
probably because they thawed the samples
slowly over 5 days, thus allowing the carbon
dioxide to come into equilibrium, whereas the
experimental samples were thawed overnight.
The beers stored at — 20° C. tended to pour
in layers, pale fluid coming first, followed by a
darker, heavier layer, but these layers mixed
well in the glass, with little haze or sediment.
The cans generally contained more sediment
than the bottles but no remark was made as
to can-flavour or vinous taste.
Solubility of Carbon Dioxide in Ice
The overfoaming of the cans from Brewery
B, when thawing is fairly rapid (but not
apparently, when thawing is prolonged),
suggests liberation of CO2 as the beer partially
freezes, with relatively slow attainment of
equilibrium on thawing. This implies that
the pressure in frozen containers is likely to
be higher than in the unfrozen state.
Table II gives the observed pressures in
cans at various temperatures together with
the concentration of CO3 in the unfrozen
beer. It will be seen that at - 2-5° C. the
pressure is generally lower than at 5*0° C.
(which is to be expected if the beer remains
liquid and supercooled), but that when the
beer is actually partially frozen from — 11° C.
downwards pressure rises rapidly and, at
lowest temperatures, can rise as high as some
seven times that in the unfrozen cans. The
pressure in the cans of Brewery B is higher
than in those of Brewery A, since the beer
from B originally contained some 9% more
carbon dioxide than that from A. This
renders the solubility of carbon dioxide in ice
a matter of some interest. A search of the
literature disclosed no figures; hence, some
measurements were made, with the results
shown in Table III.
Two methods were employed. In the first,
distilled water, saturated with carbon dioxide,
was frozen in a constant-temperature room,
with slight agitation, in an atmosphere of
carbon dioxide. When frozen, the carbon
dioxide above the ice was swept out with
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Vol. 58, 1952] ESSERY AND GANE: STORAGE OF FROZEN BEER 133

nitrogen, and the flask was sealed and thawed; ice forms in a bottle would have some in
the carbon dioxide swept out with nitrogen fluence on the incidence of fracture. The
was then absorbed and weighed. Alterna pressure of carbon dioxide might have some
tively, the carbon dioxide may be swept out effect on loss of condition through springing Of
with air. and correction applied for the crowns, starting of staves and seams, and so
carbon dioxide content of the air; both pro on. However, while in general no such
ceedings give identical results. In the second effect was observed, one or two leaking cans
TABLE II
were found, and the probable explanation of
the complete loss of gas in some of the bottles
Pressure in Beer Cans maintained at Low
Temperature is that tied-in bungs could not hold the gas
as well as properly-applied crowns. The
overfoaming of the cans after fairly rapid
Carbon Pressure in
dioxide in can
thawing, but not apparently, after slow
beer (mg./ Tempera (Lb./sq. thawing, suggests that the expelled carbon
Sample 100 g.) ture (" C.) in.) dioxide may not rapidly return to equilibrium.
Brewery A: TABLE III
I.P.A. 305 + 5-0 4-6. 0, 0 Solubility of CO, in Ice
- 2-5 0, 6
- 110 25, 28
- 100 34, 40 Temperature Mg. CO, per
— 20-0 40 Method <*C.) 100 g. of ice

Brewery B: Water (control) 0 335

I.P.A. 320 + 5-0 7-5
-2-5 3 A - 2-2 10-5
- 12-0 40 B - 2-8 16
B -6-3 14
Pale ale .. 330 + 5-0 9-5 B -6-3 13
-2-5 2 B -6-4 17
- 12-0 70 B -6-5 14-5
A -6-8 18
Brown ale 328 + 5-0 8 A - 7-1 10
-2-6 4
- 12-0 02» Mean IB- 1
Standard deviation ±2- 8
* Side scam opened.
B - 18-8 1
B - 190 4
method, finely-minced ice, obtained from B - 19-0 8*
water saturated with carbon dioxide, was B -20-5 5-5

allowed to equilibrate with carbon dioxide in a

constant-temperature room and, when satura
tion was thought to be complete, the carbon
dioxide was removed with air as above.
appeared that the first method might give
high results owing to occlusion of carbon
dioxide as bubbles in the ice, whilst the
second might give low results owing to in
complete saturation of the ice.
As might be expected, the figures obtained
are somewhat variable, but it seems clear
that, in the region from — 3° to — 7° C, the
solubility falls to about 1/20 of the value
obtained with water at 0° C; in the region of
— 20° C, it falls to about 1/100 of the water
value.
By far the greater part of the increase in
pressure on freezing is therefore due to
expulsion of carbon dioxide from solution,
although the particular manner in which the
Mean (omitting*) .. 3-5
Standard deviation ±2-3
* CO, apparently trapped under pressure in the ice.
It
Whilst these phenomena are of little or no
importance in this country, they might well
be significant when beer, in cask, bottle, or
can, is exported to regions where winter
temperatures of — 20° C. are attained.
Acknowledgements.—Thanks are due to
White, Tomkins & Courage, Ltd., for gifts of
hop-resin concentrate, complete hop-concen
trate, and essential oil of hops. Miss J.
Perret and Miss J. Trevena, and Messrs. H. J.
Shephard, K. B. Parry and L. E. Harrison,
assisted with the experimental work. For
one of us (R. G.), this work formed part of a
programme devised by the Food Investiga
tion Organization of the Department of
Scientific and Industrial Research.