14 J. lust. Brew.. January-February, 1976, Vol. 82, pp.

14-26

THE STEEPING OF BARLEY. A REVIEW OF THE METABOLIC CONSEQUENCES

By P. A. Brookes, D. A. Lovett
{Allied Breweries (.Production) Lid., Burton on Trent)

AND

I. C. MacWilliam
(Brewing Industry Research Foundation, Nutfield, Surrey)

Received 30 September 1975

The physiological and biochemical consequences of water uptake by the barley grain are reviewed,
and their significance in malting assessed.

Key words: barley, steeping, survey. uptake, water sensitivity docs."0 The malting quality of barleys
has been correlated with their rate of water uptake under fixed
Introduction conditions.4'-123-1" When developing techniques, stimulatory
The maltster aims to produce malt by modifying barley as substances, e.g. gibbercllic acid,*" kinetin," and other com
efficiently as possible. This modification is satisfactory only if pounds which influence the metabolic processes accompanying
the barley grain has a moisture content of 43-46%. During water uptake must be considered. Metabolic activity has
germination in the soil this moisture level is easily attainable received considerable attention but most of the work has been
but during commercial malting the moisture must be added directed at the germination process and is generally beyond
more rapidly by steeping the grain. Traditionally steeping was the scope of this review. It is, however, very difficult to draw
carried out by soaking grain in a tank for 2-3 days with two or a line between steeping and germination and such work that
three changes of steep liquor. This was the best method of has practical implications for the steeping process is discussed
increasing moisture content to the desired level but there was here.
no special provision for supplying oxygen to the grain. The
small amount dissolved in the steep liquor allowed respiration Properties of the Corn Influencing Water Uptake
to proceed slowly and prevented damage to the germinative
Initial moisture content.—It is generally accepted that barleys
power of the embryo. Increasingly research has been devoted
should have a moisture content between 12 and 14% during
to studying the effects of aeration in the steep and this has
prolonged storage. Higher moisture levels may have harmful
resulted in the use of fittings which allow aeration during
effects while the extra time and cost required to reach lower
steeping. Alternatively, the grain is exposed to the atmosphere
levels are not justified by the slight improvements in gcrmina-
or 'air-rested' before it has reached its optimum moisture
tive capacity and dormancy breaking which occur only at
content for germination."1-184-'08'"0
<S% moisture. Hence little work on the influence of initial
Some workers have advocated the use of flush steeping in
moisture content on water uptake has been carried out. Such
which the grain is alternately immersed in water and exposed
reports as do exist are contradictory. Thus, Baker & Dick8
to the air a number of times. If this process is not properly
observed that the rate of absorption and the total amount of
controlled it can lead to excessive rises in temperature and
water absorbed was greater when the initial water content of
excessive rootlet growth during germination and consequently
the grain was low, whereas Joyce147 found that drier barleys
high malting losses. However, if properly controlled it is a
(8% moisture) need longer steeping than normal ones (14%
much more rapid process which thus has economic advant
moisture) to yield a similar malt. It is 'agreed, however, that
ages.
old, dry grains absorb water much more slowly than fresh
Continuous processes have found little commercial applica
moist ones. In other seeds,308 it has been found that the
tion because elaborations in turning and aerating beds of
velocity of water uptake is inversely proportional to the
grain have made batch processes easier to control and yet more
amount of water previously absorbed and this relationship
flexible than they used to be. A particularly important
may also hold for barley grains with the moisture content at
development is the technique of malting abraded barley29 in
harvesting exerting influence on water uptake at steeping.
conjunction with acidulated steeping."1-2" In view of this
interest in new processes, it appeared opportune to survey and Corn size.—The uptake of water is influenced by corn size
assess present knowledge on the steeping of barley as exhaus in most cereals.94 Large corns absorb water more rapidly than
tively as possible. In some cases, relevant work on wheat and small ones initially,163"" although after 24 h the percentage
other seeds has been included if the line of research has not increase is similar. After prolonged steeping (88 h) smaller
been carried out on barley, but such references have been kept corns have a greater percentage of moisture than larger ones3"1
to a minimum. as the results given in Table I show. Levinge"1 and Maier*01
Many factors influence the uptake of water including the have also found that small corns reach their appropriate
size, nitrogenous content and initial moisture content of the moisture content for germination faster than larger ones. It is
corns. While dormancy"0 does not appear to affect water not usual to malt corns smaller than about 2-2 mm.

TABLE I. Uptake of Water by Corns of Different Size after 88 h. Steeping

Corn size (mm) 2-9 2-8 2-7 2-6 2-5 2-4 2-3 2-2 21 20 Original
Moisture content (%) 43-7 43-4 43-6 43-7 43-7 44-7 45-6 47-8 48-9 490 44-6
1,000 corn wt (g) 46-3 42-6 38-8 350 33-7 31-5 27-4 23-7 21-3 220 330
Wt of water absorbed (g) per
1,000 corns 360 32-6 29-7 27-3 26-7 25-5 230 21-8 200 19-2 26.1
Vol. 82, 1976] BROOKES, LOVETT AND MACWILLIAM: STEEPING OF BARLEY 15

Because of this difference in water uptake by corns of In practice, barley which is water sensitive, but does not
different sizes it might be advantageous to steep batches of have low germinativc energy, germinates successfully if the
grain of different size separately, as has recently been des moisture content is raised to 31-35 %, the steep liquor drained
cribed.03 and the grain left exposed to the air until it chits (18-24 h)
Nitrogen content.—Little systematic work has been devoted after which the grain can be immersed again without impairing
to uptake or water by samples containing different percentages germination. After reaching the usual moisture content (43-
of nitrogen. Slight311 states that the duration of steeping 46%) the water sensitive condition has been overcome and no
should bear a direct relationship to the nitrogen content of the further problems with germination are usually found.
barley. In a more systematic study Ehrich & Kneip73 separated Water sensitivity is frequently found in freshly harvested
a sample of barley of 1-89% nitrogen content into three frac barleys gathered in damp conditions and is more prevalent in
tions with respective nitrogen contents of 1-68, 1-89 and 2-11 %. maritime countries such as Britain than in warmer, drier
The sample with the lowest nitrogen absorbed water most Central European countries.259-265
rapidly and malted faster than the other two. Sallans & It is well known that dormancy as assessed by an increase
Anderson*01 noted that the starch content of barley was in germinative energy can be overcome by using high drying
positively correlated with the time required to reach a pre and storage temperatures,106 but some work has suggested
determined moisture content. Since the samples with higher that recovery from water sensitivity is influenced differently by
starch contents had lower nitrogen values and took up water storage conditions. High storage temperature was reported to
faster, their results agree with those mentioned above. In have no effect on water sensitivity but storage at low moisture
wheat, however, it appears that the rate of water uptake is content destroyed the condition.8*-83
independent of the nitrogen content.04 Recently, attempts have been made to find why water sensi
Dormancy and Water Sensitivity.—Barley which is dormant tive grains fail to germinate if steeped to normal moisture
is not usually malted unless it is treated with additives to contents. Crabb & Kirsop'9 found no significant difference in
overcome this condition. Early work suggested that dormancy the rate of oxygen consumption in water sensitive and insensi
was due to failure of water to pass through the outer layers of tive barleys, but at low oxygen tensions the germination of
the barley.128 These findings were based on the ability of barley sensitive barleys was less. It thus appears that water sensitivity
which had been dried at high temperatures to absorb water is a reflection of a higher oxygen requirement for germination
more rapidly than untreated grain.1"-3"0 Kirsop159 suggested by the embryo. In further work" Crabb & Kirsop showed
that dormancy comprises the two phenomena, low gcrmina- that extracts from the barley embryo induced in barley grains
tive energy and water sensitivity. The phenomenon of water a condition that was indistinguishable from water sensitivity.
sensitivity itself was first detected by Essery, Kirsop & Sucrose and raflinosc were partly responsible and other
Pollock*0 when it was found that certain barleys would not unidentified compounds were also present.
germinate under normal steeping conditions. Such barley does Gaber & Roberts investigated08 the effect of respiratory
not show low germinative energy under the conditions of the uncoupling agents, auxins, and auxin antagonists on the
accepted test"-202 but if the amount of water in the test is germination of barley. The results did not support the theory
increased from 4 ml to 8 ml germination is reduced and a that water sensitivity was the result of the accumulation of a
proportion of the corns do not grow. It was concluded that the large concentration of indole acetic acid which uncoupled
failure of water sensitive barley to grow was due to the inabi respiration. In other work99 they showed that the inhibitory
lity of the embryo to respond to the low internal oxygen effect of excess water could be overcome by adding mixtures
tension resulting from the presence of an unduly thick water of antibiotics. About 80% of the seeds that failed to germinate
film. Low gcrminative energy may also reflect a deficiency of contained large mixed populations of microorganisms. Both
oxygen in the vicinity of the embryo.189 Both aspects of the microflora and the excess water acted to reduce the level
dormancy can be overcome by increasing the availability of of oxygen reaching the embryo.
oxygen."0 Water sensitivity is influenced by the conditions prevailing
throughout ripening and is dependent on variety and storage
Nevertheless, low germinative energy and water sensitivity
temperature.285 Zastrow3*8 found that with water-sensitive
have been studied separately by many workers and the former
grain a steeping procedure using air rests resulted in a quick
has often been equated directly with dormancy and has been
start to germination and in general gave higher malting loss
shown to be controlled by metabolic factors whereas water
but better modification.
sensitivity is influenced by external conditions.
It thus appears that although dormancy and water sensi
It has been shown203 that during the initial 6 h of water tivity seem superficially to be related and that water sensitivity
uptake dormant seeds of barley and rice take up more oxygen can be conveniently considered as an aspect of dormancy, low
and give off more carbon dioxide than non-dormant seeds gcrminative energy and water sensitivity are the result of
although no differences in activity or distribution of respira different conditions in the grain. Low gcrminativc energy is a
tory enzymes were found. reflection of the metabolic state of the embryo and water
Dormancy can be broken by chemical treatment. It has been sensitivity is a manifestation of external effects on the
shown13-182 that, on addition of 500 mg/litre gibberellic acid or behaviour of the grain.
gibbcrcllic acid +0-1 % potassium nitrate, dormancy is broken.
An inhibitor of gibberellin activity, isolated from dormant
barley seeds was found to be progressively inactivated during Water Uptake and Malt Quality
storage.387 Germination was accompanied also by an increase Several attempts have been made to correlate these factors.
in the RNA and DNA contents of the seed and a negative Hartong & Kretschmer123-124 measured the water-uptake of
correlation was found between RNA synthesis and inhibitor different barleys during steeping and then analysed the
concentration in dormant seeds. It has been suggested resultant malts by the Hartong 4-mash procedure.121-1" They
further140 that soaking the seeds of barley activates a factor found that samples of grain that absorbed water faster gave
that stimulates DNA, RNA and protein synthesis. better malts than those absorbing water more slowly. Narziss
The control of seed dormancy was recently reviewed by & Kieninger234 could not demonstrate a mathematical
Wareing.347 From work on seeds other than cereals it has been correlation between rate of water uptake and modification of
concluded that the role of hormones in regulating dormancy malt, but there was some evidence that barleys which absorbed
may involve repression and derepression of the metabolic role water more rapidly gave malts of better proteolytic and
of DNA. Further work is required to establish whether this cytolytic modification. The effect of initial steeping time on
hypothesis applies to cereal germination. malt quality was studied by Meredith & Bettner,211 who found
 16 BROOKES, LOVETT AND MACWILL1AMJ STEEPING OF BARLEY [J. Inst. Brew.

that the effects of differences in time persisted throughout the Pericarp and
remainder of the steeping and germination process. An initial testa
Endosperm
IS h wet period produced the best modified malt but no
explanation was offered for the differences. Chapon41 found
\ Husk

that barleys which took up water most easily gave the more
friable and better modified malts. In this work Chapon
devised a technique for measuring the case of wetting giving Embryo
results as a 'steeping index'. Awn
In more recent work231 Narziss & Fried rich found that the
most favourable analytical properties for malt were produced Micropyle
by higher moisture levels (i.e. 46%). Vesclov has reported*11 Aleurone
Scutellar
that repeated moistening of the barley (keeping the moisture layer
epithelium
content at 48-50%) allowed malting to be completed in 5-5-6
days. It can be concluded that high out-of-steep moisture levels
Fig. 2. Longitudinal section of a barley corn (diagrammatic).
result in a better modified malt.

Physiological Aspects of the Uptake of Water Gaps exist in both the pericarp and testa through which
moisture can enter. After examination of the movement of
Water uptake by barley grain8 and other seeds85-212-322-349 at
acids, salts and dyes through the various layers of the corn,
normal steeping temperatures of I2-I5°C, is generally
several workers, including Collins,*1 Gruss110 and Krauss164
considered to occur in three phases as shown in Fig. I. Phase I,
have concluded that the pericarp has an outer layer of cork or
cuticle which is missing around the embryo and at the upper
tip of the corn. At these points water can be absorbed. Some
varieties320 have a heavier layer of cuticle in their pericarp and
50 thus have a lower permeability. At the brown stretch along
the ventral furrow of the grain and at the micropyle region the
structure of the testa is modified to allow passage of water,
most of which probably passes through the micropyle region,208
40 although Bleisch & Will" and Chapon" believe that more
passes through the ventral furrow as is the case in wheat.11'
IS It was early noted103 that during steeping the embryo half
of the grain absorbed water faster than the distal half. Later
3 30
workers38-37-43-51-89-298-370 found that the embryo itself was
3
responsible for this preferential uptake. The distribution of
water in germinating barley has been studied by Kirsop et alK0
.2
O 20 who found that, after steeping, the embryo contains 65-70%
of moisture and the endosperm approximately 41 %. The
individual grains are also surrounded by a thin film of mois
ture which accounts for 2-3 % of the whole grain. In the early
10 stages of germination the embryo absorbs water initially from
this film but later it abstracts water from the endosperm.
It has been shown05'"9 that if the surface water is removed,
germination occurs more rapidly but the final extract is less
10 20 30 40 50 than with the normally drained sample. Thus the amount of
Steeping time (h) water held by the fully saturated grain is not enough to
produce a full yield of extract and extra must be supplied.
Fig. 1. Water uptake during traditional steeping of barley (55°F). Blaim23 showed that coumarin inhibited the uptake of water
by the embryo but not the endosperm of wheat. It is the active
part of the uptake of water by the embryo which could be
inhibited by similar substances and also by factors responsible
of 6-10 h, of rapid water uptake is considered183 to be the time for water sensitivity.
when 60% of the total water is absorbed. Phase 2, of 10-20 h, The endosperm absorbs water more slowly and continues
is also referred to as the Bakcr-Oick break.8 In this phase the process until the grain is fully hydrated. Brown3' found
water uptake is very slow or ceases completely and some that both tissues compete for water and that as the water
workers1'07-78 have questioned the significance of this phase. content of the grain increases towards a maximum the rate of
Phase 3, of over 20 h, involves more rapid uptake of water absorption by the embryo increases relative to that of the
interrupted by a plateau, the 'steep-ripe plateau', steeping endosperm where water-absorbing capacity falls more rapidly
beyond which is said to cause a breakdown of the semi- than in the embryo. It is possible that the scutellum is the
permeable membrane of the grain. The three phases arc now first organ in barley to become metabolically active71 during
discussed in more detail. steeping as it is in maize.79
Phase 1.—During this short period of rapid water uptake Phase 2.—Here water uptake is very slow or ceases alto
the seed colloids, primarily proteins and carbohydrates,209 gether.8 During this period the conversion of starch to
imbibe water previously lost during ripening.118-119 This sugar212'519 takes place. Such hydrolytic processes probably
absorption is a purely physical process. The absorbed water produce increasing osmotic pressure which results in the
penetrates the outer layers of the grain more slowly. The uptake of more water by the tissues of the embryo. If the
pericarp (Fig. 2) is the principal barrier to the entry of water embryo of wheat is deprived of sugar, uptake of water
while the testa restricts entry only during the first few hours continues for 120 h and then ceases. If however, it is then
of steeping although it remains impermeable to many sub supplied with nutrients new meristcmalic tissue is formed and
stances. Brown" was the first to demonstrate that the testa more water is taken up.'00 It seems probable this happens also
acted as a semipcrmeable membrane. in barley in which the embryo, having reached a sufficiently
Vol. 82, 1976] BROOKES, LOVETT AND MACW1LLIAM: STEEPING OF BARLEY 17

moist state in about 6 h, has become metabolically active and traditional unaerated steeping the grain was not damaged to
requires nutrients which may be supplied by the endosperm. the extent that it did not germinate.
In certain other seeds,319 a two-step reaction during Phase 2 Both oxygen consumption and carbon dioxide emission rise
is considered to take place, in which substrate materials are during steeping if the steep liquor is continuously aerated.74
converted to biologically active intermediate compounds The former is closely controlled by the oxygen supply but the
whose formation then initiates the onset of growth. Some of latter is much less influenced by oxygen tension. The respira
these active compounds may well be gibberellins. tory quotient rises to a maximum (1-5) during the first six
Phase 3.—Here water is taken up at a steady linear rate which hours after moistening3 and falls after 24-48 hours of steeping
is correlated with metabolism67 particularly with initiation of to 0-95.338
activity of the cytochrome system.213 This stage is associated The intensity of respiration is related to the speed of growth
with the visible onset of germination when the root and shoot of the plumule,167 growth and respiration being most rapid in
elongate as cells in their meristematic regions take up water the zone of 3/8-5/8 the length of the grain. This 'index of
and become vacuolated.1-350 It is said that mitosis does not maximum growth' changes from one variety of barley to
take place during the first 46 hours after wetting and that all another and is independent of steeping and is not caused by
growth is due to cell enlargement.62 As in Phase 2 this uptake asphyxiation of the grain. It is claimed167 that the determina
of water only continues if there is a concurrent supply of tion of this index is useful in controlling malting and micro-
nutrients from the endosperm. malting conditions.
The third phase takes place only if the oxygen supply is Few attempts have been made to relate respiration directly
adequate and does not occur at very low or very high tempera with enzyme activity in barley. Respiration and a-amylase
tures or in dead grain. It is therefore described as a period of activities have, however, been studied by Tahara.324 The endo
active uptake rather than imbibition, although it seems that sperm was found to be the major area of amylase production
the two may go on side by side for a short while. Linko172 but was a minor area of respiration (in the preparations
considers that in wheat, anaerobic uptake of water continues studied aleurone layer tissue was probably included with the
until a moisture content of 40% is reached and that metabol embryo.) Rootlets and embryo showed profound respiration
ites also accumulate in this early period. But Dahlstrom, but they formed little enzyme.
Morton & Sfat64 consider that this first phase is not anaerobic Carbohydrate metabolism.—As early as 1895 Petit2*4 noted
in barley. that conversion of starch to sucrose took place during steeping
The three phases cannot be considered as separate from one and Bode26 and Leibu169 found that reducing sugars were
another as the metabolic processes taking place in them are formed. On the other hand, Loibl180 observed that glucose,
interrelated. fructose and sucrose decreased during steeping. Using
Swelling of the grain occurs during the first 24 h and then the chromatographic methods, Macleod, Travis & Wreay1"8
rate of swelling decreases. The change in thickness that results found that, in Ymer barley, sucrose, raffinose, gluco-di-fructose
varies similarly and eventually there is an increase in the ratio and low molecular weight fructosans decreased during steep
of length and thickness.163 ing, whilst maltose and maltotriose increased. However,
The importance of achieving the right moisture distribution MacWilliam1"9 observed that in Plumage-Archer barley both
within the corn has been realized but this is difficult to control glucose and fructose increased during steeping but that sucrose
with much of the steeping equipment available. In some and rafnnose were depleted. The picture was clarified in a
recent work with maize166 an attempt was made to influence subsequent study using Carlsberg barley by Hall, Harris &
moisture distribution to yield a product of greater commercial MacWilliam118 who found that glucose and fructose fell in
value. concentration during the early part of steeping (24 h) and then
rose towards the later phase (48-72 h). Similarly, in barley
Metabolic Changes during Steeping embryos kept moist under anaerobic conditions or treated
Thus steeping is a period when many physical and chemical with cyanide the raffinose concentration remained constant,
activities take place which influence further development of but in similar embryos with ample access to air the sugar
the grain.43-44-45-46-78-302-330-331-332 Refinement of analytical disappeared rapidly. In contrast, sucrose was consumed in
techniques in recent years has helped to provide much more each case.101
detailed information which is dealt with below. The disappearance of raffinose, also noted in other types of
Respiration.—The very low level of respiration in the resting seed,27-307 has resulted in a considerable amount of
corn rises rapidly when the grain is wetted.238-331 This increase study.118-1"-190 Recently Palmer847 found that during germina
is the first easily observed metabolic change in barley and in tion the rafnnose content of the embryo declined slowly and
most other seeds.01-136-*09-260-329 Oxygen is taken up exponent- it is suggested that raffinose may undergo degradation to
ially242-320 and in 1 h most of the oxygen has been removed galactose and sucrose, the galactose being enzymically
from the steep liquor by the grain.64 The increased respiratory epimerized to glucose before being metabolized.
activity is shared between embryo and aleurone layer.04 Early work118 revealed that the starch content diminished
Simultaneous with the increase in respiratory activity is the slightly during steeping as a result of diastatic activity.
influx of sugars from the endosperm337 although it is not clear Leonard & Martin170 observed that starch appears in the root,
whether or not the respiratory activity is due to the oxidation coleorhiza and plumule soon after wetting but break-down of
of sugars to provide energy. reserve starch in the endosperm cells adjacent to the scutellum
In some cases, within the first few minutes of wetting of the exceeds this synthesis.
grain, absorbed air or carbon dioxide is released by the seed Jaeger has reported133 that the moisture content significantly
tissues. Some workers118-213 consider that this release is merely affected the activities of a-and /3-amylase when preparing
a physical effect whilst others178 think that the carbon dioxide distilling malts. The best results were obtained with 3 days
is produced as the result of enzymic activity. wet/dry steeping (36 h wet 36 h dry) at 15°C to achieve a
In the absence of further supplies of oxygen, the embryo moisture content of 46%. Bettner & Meredith reported17 that
respires anaerobically to produce carbon dioxide43-163-330 and free 0-amylase activity at the end of the final steep decreased
alcohol,67-331 which may inhibit germination.45-330 Oxygen with increasing length of initial steep period. Linko & Enari
access is restricted by the envelopes of the grain831 and does have reported174 that a low moisture content (39%), gives high
not become adequate until chitting occurs.4' Similarly, a-amylase activities and an increase in malt yield. Similar
alcohol is said to accumulate under the coverings until they results were reported by Yomo & Iinuma.367 However,
are ruptured.43 It seems, however, that this inhibitory effect diastatic power remains lower than when using a normal
of alcohol and carbon dioxide is over-emphasised since in degree of steeping.
18 BROOKES, LOVETT AND MACWILUAM: STEEPING OF BARLEY [J. Inst. Brew.

Much fundamental work on the synthesis of a-amylase and changes in phosphorylation of the mitochondria were observed
the involvement of gibberellin in this synthesis has been in the presence of gibberellic acid.
reported.31-"'48-102-"4'1'5-196-19'-216-3*9-340 In a series of experi The relationship between water uptake and enzyme
ments Massart & Verbeek-Wyndaele206 reported that the activity has also been studied288 at a more practical level.
induction of a-amylase by mevalonic acid and gibberellic acid Initially during steeping, ribonuclease and phosphatase
was the greater the more the barley was soaked during activities rise following the water uptake. After the first 6 h
steeping. activities fall only to rise again if the steep liquor is changed
Mandl204 studied changes occurring in the enzymes of the or if the grain is exposed to the atmosphere.
glycolytic pathway during steeping and germination. The If orthophosphate solution containing radioactive phos
amylases, hexokinase and phosphofructokinase decreased in phorus is used for steeping, radioactivity rapidly appears in
activity during steeping but cellobiase increased. the phospholipid fraction, particularly phosphoryl choline.117
During the malting of wheat and barley212 the bulk of the A dehydrogenase, present in the embryo of resting grain18
starch solubilized is the amylopectin fraction. However, is one of the first systems to increase in activity,177 becoming
amylose is increasingly utilized as germination is prolonged. active when the moisture content of the grain exceeds 25 %.
It has been suggested140-141 that inhibition of growth in
Barwald10 has reported that some breakdown of /3-glucan
water-sensitive barley is caused by uncoupling of phosphoryl
can occur in steeping and this is taken to mean that barley
ation due to the production of abnormally large amounts of
endo p glucanase is active at this early stage of the process.
indole-3-acetic acid (IAA) during the early part of steeping.
Nitrogen metabolism.—It has been suggested250 that during
This excess is said to result from the presence of a lag factor
malting, proteases play a more significant role than poly-
in water-sensitive grains which causes a delay in the enzymic
saccharidases in the conversion of hard barley into friable
oxidation of IAA. In view of previous suggestions138-266 that
malt. Transaminase and protease244 activities and amino acid
sulphydryl groups are involved in water-sensitivity and that a
content258 increase in barley and other cereals as the water
correlation exists between bound disulphide groups and
content increases and aeration during steeping229 causes a rise
dormancy in wheat,14 it is interesting to note that cysteine,
in activity of peptidases. Recently it has been shown278 that
reduced glutathione and mercapto acetic acid create a lag in
l-leucyl-jS-naphthyl amide amidase decreases slightly in steep the oxidation of IAA140 although they do not in fact induce
ing and then increases in germination. The significance of
water-sensitivity in normal barley grains.
these changes in enzyme activity on final malt quality has not
been established.
Variation of Conditions
In wheat, when the moisture content was raised to 15%,
Duration of Steeping.—There is no easy answer to the
glutamate decarboxylase, protease and transaminase became
question of how long should barley be steeped. As stated
active,1"-179 and the level of a-amino butyric acid rose as the
earlier, it is generally accepted that the moisture content should
grain was moistened. When the moisture content of the wheat
be 43-46% at the beginning of the germination period, and
reached 40% a-oxoglutamate, oxaloacetate, glutamate and
that understeeping is less dangerous to growth than over-
aspartatc all increased in concentration indicating activity of
steeping. With understeeping, abnormal rootlet growth may
the tricarboxylic acid cycle and associated transaminases.179
occur131 and modification may not be optimal although this
The amino acid metabolism of germinating barley was can be remedied by sprinkling during the first day of germina
studied extensively by Jones & Pierce.143"146 Restriction of
tion. As early as 1896 Day85 noted that metabolism of the
oxygen during flooring causes the free assimilable amino acids
endosperm constituents is prevented by inadequate moisture
to increase throughout germination, which may be a result of
supply. Sims3™ found that germination only occurred when
a different mechanism from that holding in the steep where the moisture content of the grain lay between 30 and 45%
aeration increases peptidase activity.
while for optimum germination the barley he studied required
Belderok15 has studied the metabolism of sulphur-containing 37-5% moisture. Oversteeping cannot be so easily overcome
amino acids, in dormancy and after-ripening, while Kauppi- and the grain respires much more quickly once germination
nen154 and co-workers examined the variations of protein commences. Zila, Trkan & Skvor389 have noted that sound,
sulphydryl groups during malting and the activities of gluta- mature barley can be kept in the steep for as long as 15 days
thione reductase and cysteine reductase. with only short periods exposed to the air for weighing. Many
Attention is increasingly being paid to nucleotides and their workers22-25-134-296 have pointed out that excessive steeping
association with gibberellic acid in the control of barley without aeration damages the gcrminativc capacity of the
germination.42-70-"4'237 Few studies have, however, con grain leading to irregular growth.
centrated solely on the steeping process and most of the work It is not an easy matter to determine the moisture content
has been on the role of nucleotides in protein synthesis. of a piece of barley accurately during steeping but methods
There have been reports305-808'351 that in maize and some other have been described by Benton16 and De Clerck.66
cereals, guanosine triphosphate, uridine triphosphate and The most important factor governing the uptake of water
adenosine triphosphate accumulate after steeping, but little is temperature, and Macey188 has given the following relation
is known of the formation of these compounds in barley. ship as a practical guide: approximate time of steeping in
Microsomes isolated from steeped wheat embryos were hours = 110 — temperature of water in °F. Slight311 has listed
active in the incorporation of amino acids into proteins criteria by which the steep-iipeness can be judged by the
although a similar preparation from dry material was not maltster.
active.206 It has also been shown130 that there is a turnover of In most maltings using well water at a temperature of about
ribosomes in embryoless half seeds of barley when they are 52°F, steeping times are generally 55-80 hours. Since the
soaked in gibberellic acid. The aleurone cells contain ribo discovery of water sensitivity, maltsters have given air-rests
somes in the dry state but metabolism is not active until the during steeping much more frequently and present practice
seeds are wetted. both in Britain311 and on the Continent165-258-*99-300-301 employs
Metabolism of Other Compounds.—In seeds other than air-rests whether the grain shows water sensitivity or not.
barley125-236 phosphatase activity has been found to increase Temperature of steep water.—There has been considerable
as the grain is wetted. Srivastava316-317 has shown that after discussion as to the desirability of carrying out part of the
soaking barley seeds in water and gibberellic acid there was an steeping process at high temperature. It was early recognized323
increase in ribonuclease and phytase activity. The effect of that when colder water than normal was used a longer
gibberellin on the phosphorus metabolism of barley in the steeping time was necessary. On the other hand, water uptake
early stages of germination has been further studied148 and is more rapid at higher temperatures as shown in Table II.
Vol. 82, 1976] BROOKES, LOVETT AND MACWJLLIAM: STEEPING OF BARLEY 19

TABLE II. Moisture Contents of Barley Steeped at Different detailed study by Lubert & Pool181 who found that at higher
Temperatures. temperatues (<30°C) certain barleys became water sensitive
and failed to germinate evenly. But in most cases good malts
Temperature
were produced with low malting loss.
of water (°C) 10° 15-6° 21-3° It has not passed unnoticed that warm water steeping
preferentially inactivates certain parts of the grain. Thus
Initial moisture content 13-1 131 131 Moufang & Vetter222 in 1909 noted that by heating barley to
after 16 hours 29-5 32-8 34-2
55°C for 20 min and then malting in the normal manner, no
40 „ 36-4 39-3 42-1
63 „ 39-2 42-5 44-9 roots formed, malting loss was lower and extract higher.
87 „ 41-4 44-0 46-7 Systems of malting have been developed in which the embryo
112 „ 43-3 46-2 48-2 is inactivated, root growth is eliminated and malting loss is
decreased either by using warm water6-2"-2'1 or by freezing to
—20°C. Modification proceeds in the normal way. The use
Baker & Dick8 measured the uptake of water by barley at of root killing steeps is now accepted commercially parti
different temperatures and found that the total amount of cularly in the production of distilling malts. Initial practical
water absorbed was greater at higher temperatures and trials with root-killing steeps have been described by Griffin
although the temperature of the steep liquor had little effect & Pinner.110
on final growth, warm steeping transiently retarded germina It seems that important practical developments in the field
tion. The velocity of water uptake is an exponential function of warm water steeping may still arise; especially since warm
of temperature as is the case with other seeds.8-12'38-8*-*0 water can overcome dormancy4*-60 and thus lead to an earlier
Reports on the effects of warm water steeping on sub start to malting the season's barley after harvesting.
sequent germination have often been contradictory, and Steeping in very cold water has a retarding effect on germin
opinion is sharply divided as to whether this treatment is of ation but leads to very even growth.4 Improved enzymic
commercial value. Somlo313 claimed that as a result of steeping activity has also been found344 in barley steeped at 5°-10°C.
in warm water, both steeping and germination periods probably because the low temperatures minimize the influence
were shortened and 'difficult' barleys were easier to malt. of excess carbon dioxide and oxygen deficiency during
Moufang218-219-221 found that after a short steep at 40-50°C, steeping. More recent work has been carried out on the
the absorption of oxygen was greater, while growth was production of malt with little embryo growth'8 by treating
optimum after steeping at 45°C. But Goldacker101 found that abraded barley at low temperatures or with acid. The malts
although warm water was taken up initially at a greater rate, produced were highly modified and could be used with a much
no advantage accrued, whilst Bletsch24 found that after steep greater proportion of carbohydrate adjunct than normal
ing at higher temperatures growth was set back although it malts.
recovered later. Wegtnann348 observed that the diastatic power Influence of oxygen and carbon dioxide.—As already stated,
of malts steeped in warm water developed slowly during many authors22'"-134-296-331'36* have noted that excessive
flooring and remained low, while Schmid29' and Wust3*4 steeping without aeration damages the germinative capacity
reported definite loss of extract. Furnrohr" emphasized that of the barley and leads to uneven growth.84 In many modern
the time and temperature of warm water steeping was very mailings gentle aeration is applied through appropriate
critical as was aeration. The optimum temperature was 40°C. fittings in the steep tank81-301 Barley is sometimes aerated
Windisch361 agreed that time, temperature and aeration are vigorously, in some cases in special tanks,89-301 after water is
critical but fixed the temperature maximum at 30°C. added, but this is mainly useful as a cleansing and mixing
Emslander" commenced his steeping at the normal tempera operation.293
ture but after a period of about 24 h, added water at a temp In a series of pilot-scale tests, Macey189 found that aeration
erature of 36-5-45°C to bring the final temperature to 19°C. of the first steep had little effect on the subsequent growth of
This treatment accelerated the onset of germination and the barley. Luff187 found that aeration did not increase water
shortened the flooring period by 1 day. Piratzky2" claimed that uptake but did note hastened germination. The latter observa
warm steeping caused only minor stimulation but that heating tion is not confirmed by later work.2'0 With sluggish barleys
over 36°C inhibited growth. aeration is useful and although it has been reported that with
More recent work has tended to increase this confusion. normal barley chitting in the steep can lead to high malting
Sommer has extensively reviewed the correlation of steep loss1'9 this can be controlled and it is now standard mailing
temperature and malt quality,314 and he found that steeping at practice to try to get all barley to chit in the steep. Scriban303
temperatures over the range 12-20°C has no significant effect has noted that certain barleys which he described as 'sensitive
on malt quality though germination time was reduced at the to air' do not germinate properly if they are aerated during
higher temperature. On the other hand Navarro & Brandao236 steeping.
observed that high temperature steeping (25°C) made it Although Chambers & Lambie41 agreed that the presence
possible to improve productivity (significantly) while main of free oxygen in the steep liquor greatly accelerated the
taining malt quality. Pool has concluded272 that steep water changes which occur in barley, they considered that this led to
temperature played no part in determining the extract 'bolting'. They suggested that more even germination was
obtainable, but steeping time could be reduced if the tempera obtained if the concentration of carbon dioxide in the steep
ture was raised. In contrast, Narziss224-225 has reported that liquor was maintained at levels between 1400 and 1800 ppm
the temperature of steeping affects the speed and extent of by bubbling gas through the steep liquor.
enzyme formation in germination and thus presumably affects Eyben & van Droogenbroek have recently attempted to
malt quality. Steep water temperatures above 13°C resulted in study the effects of gaseous pressures of between 2 and 4 kg/
a lower degree of modification. Ward & Briggs346 steeped at cm2 on the steeping process.85-88-87 They found the pressure
25°C and sprayed with hydrogen peroxide and gibberellic acid undergone by the grain in steeping diminished germinative
to give a malt with a hot water extract of 103 Ib/qr with a energy and could delay the onset of modification. It was
malting loss of 7-5%. further shown86 that the oxygen pressure inside barley cells
Briggs30 has derived a relationship to calculate steeping and the accumulation of carbon dioxide and ethanol control
time to achieve a desired moisture level at different tempera the yield and quality of the finished malt. In similar studies by
tures which could be used to regulate the rate of water uptake Gomez,105 the effects of evacuation or imposition of pressure
so that steeping is completed in a convenient time. in the presence of air, nitrogen, carbon dioxide and oxygen
The use of warm water for steeping has been subjected to were examined. Application of vacuum causes an increase
20 BROOKES, LOVETT AND MACWILLIAMI STEEPING OF BARLEY [J. Inst. Brew.

in protein degradation. When pressure is applied the increase the grain by leaching in the steep liquor these losses were not
in proteolysis is more marked and is proportional to the significant2" and it was concluded that pneumatic steeping
increase in pressure, but there was no comparable increase in yielded malt of the highest quality.
amylolysis. In addition to multiple and flush steeping, Narziss &
The concept of ventilated steeping using a conical-bottom Friedrich have examined"0-"1'233 the re-steeping process in
vessel with a false perforated inner core to allow even air flow which grain is steeped during germination. Malts prepared by
through the grain has recently been described."6 this process gave beers with lower tannin and higher high-
Multiple and flush sleeping.—If the demands of the grain molecular-weight nitrogenous and viscous compounds than
for oxygen in the later stages of steeping arc greater than in did those produced by conventional malting. The possible
the earlier ones, germination will be more rapid if the grain is commercial advantages of this process have not been widely
removed from the steep and exposed to air. In the traditional exploited in the UK. Pool has reviewed the subject273-274'2™
process, the grain would have to be removed from the tank and it seems that the greatest advantages lie in increased
and in such cases it was more economical of effort to change yield of malt and shorlcncd malting time. Effects on malt
the steep liquor more frequently. However, the possibility of quality are not so clearly described and it is not certain that
speeding up the germination process by draining the steep re-steeped malts are entirely suitable for conventional
liquor and allowing the grain to 'air-rest' for a short time soon brewing procedures.
came under study. Such steeping has been said to simulate British patents have been granted for producing malt by
conditions in the soil4101 and to obtain a more ready response re-steeping290 and for an almost continuous steeping process."
from the seed. It is considered to afford a natural environment Spray-steeping and Reduction of Effluent.—As early as 1900,
for the barley seedling and not to involve forcing in any way.108 Windisch355 suggested the elimination of steeping and instead
There are innumerable combinations of steeping and air- supplying all the water by sprinkling. However, the use of a
resting and much of the work carried out at the turn of the short 6-12 h steep before commencing spraying is more
century was largely of an empirical nature and frequently led widely preferred.168-207-312-322 It appears that until the barley is
to discouraging difficulties. Windisch3"-3*8-3"1'3" was one of reasonably moist, sprayed water may not be absorbed by the
the first to use a flush steeping technique of alternative 4 h grain but simply channels through the grain bed. The combin
soaking and 4 h exposed to air. Emslander,75 however, was ation of a short initial steep followed by spraying whilst the
unable to obtain good malts by Windisch's methods and found grain moves slowly on belts or trays has found limited
that the grain overheated and was difficult to control. application312-322 on the commercial scale.
Windisch countered this criticism, saying that unless the grain Spray steeping has many attractions, not least that it
was circulated while steeping, germination was uneven. In diminishes effluent, and it is now clear that the technique is
addition, to control the temperature of the grain cold air was being widely used on the commercial scale.188'230'240'24*
blown through it. Haslinger116 and Reischbock288 also used a MacWilliam200 reported that, after spray steeping, the grain
flush type of steeping for the later stage of their process but germinated normally and the malts obtained were of good
gave a long initial steep. Improvements in germination and quality. Voborsky34' found that the response of barley to
extract were obtained. spray steeping was dependent on variety. It has been further
It was thus concluded that cold air must be provided reported289 that spraying lo a moisture content of 46%
and the grain must be circulated to gain the advantages yielded higher hot water extracts in the malt than spraying to
of this method. Water sensitivity may have gone un 44%. It was, however, concluded that it would not be
recognized in these earlier results and with grain exhibiting economic to produce malts on the commercial scale solely
this condition, the uptake of water must not exceed 35% by spray steeping.
during the first steep and the steep liquor must be drained Efforts to reduce the volume of effluent from makings have
from the grain to avoid impairing germination. Numerous become increasingly necessary recently since Governments
workers191-1"-247-804'888-309'310-330 have examined various combin began to legislate to protect the environment. Apart from
ations of steeping and air-resting to overcome water sensiti spray-steeping, as already discussed, there has been much
vity, and also for use in speeding water uptake and germin interest in the re-use of effluent steep water, with or without
ation in 'difficult' barleys.101 prior purification, to steep a fresh sample of barley.
Other investigators have used flush steeping with normal Two papers147-214 have discussed the use of adsorbents, such
grain. Razga291 used this technique to accelerate germination. as montmorillonite and polyacrylamide (Flygtol process) to
Schuster,301 in a review of German steeping methods, reduce the BOD level of steep liquors to that suitable for re
emphasized the saving of time achieved by the flush technique. use. But in the Flygtol process the amount of adsorbent
Sirot310 drained the grain periodically during steeping until required for purification was so high that 35-4 mz the process
it had been exposed to the air for only one quarter of the total was uneconomic.
steeping time. Longer aeration periods produced inferior A pilot scale settling filter21 consisting of 2 units each of
malts. This conclusion was not confirmed by other workers filter area has been used20 to examine the removal of solid
(see above) who exposed the grain to the air for at least 50% matter from steep liquors and to measure the degree of
of the total steeping time. pollution when the filtered liquor was used to steep further
Pool88*'2" preferred a multiple steeping technique in which, lots of barley. After steeping, the barleys were malted and
after an initial period of 6 h under water, the grain was drained analyses showed that the malts from the re-used liquors were
for 20-24 h at the end of which it had begun to chit. Water of similar quality to control malts.
entered much more rapidly during the second steep and only Re-steeping by the Huppmann process,129 incorporating a
1-2 h soaking at 65°F was required to increase the moisture combination of germination box and steep, has been des
content from 33-35% at the end of the first steep to 43-45% cribed. This process uses considerably less water than tradi
at the end of the second. Besides the saving in steeping time, tional ones.
germination was accelerated. From all this work it can be concluded that a short initial
Narziss & Friedrich228 have shown that when barley was steep to bring the moisture content to between 30 and 35% is
alternately steeped and exposed to air the activity of amylases, most effective. With a shorter steep insufficient water may
peptidases and glucanases all increased markedly during the prevent development in the embryo while with a longer one
air-rest periods. These authors also describe230 a pneumatic excess moisture will lower the germination of water-sensitive
steeping procedure (short steeps and extended air rests) which barleys. For subsequent operations, as has been described,
also resulted in a significant increase in enzyme activity. there is a wide choice of methods of raising the moisture in the
Although small amounts of enzyme activity were lost from grain to 44-46%.
Vol. 82, 1976] BROOKES, LOVETT AND MACWILLIAM: STEEPING OF BARLEY 21

Effect of Additives in the Steep Liquor inhibits a-amylase activity in wheat and j3-amylasc in barley,
Salts, oxidizing agents, antiseptics.—There arc frequent germination is slowed but dormancy is not broken.
reports that certain salts when dissolved in the steep liquor A different method in which mineral acid is applied to
stimulate the germination of seeds. Alkaline liquors dissolve abraded barley has recently been utilized by Palmer"1 -2M to
our resinous material and tannin more readily than do acids
produce a full extract in only 72 h malting time.240 The tech
or neutral salts"8 and increase water uptake.302 Kauert161 has
nique is not successful with non-abraded barley. The whole
subject of abrasion has recently been reviewed by Palmer.243
suggested that the pH of steep liquors should be maintained
between 8 and 10 by the addtion of weak alkali or by continu If commercially acceptable, the technique would seem to have
ously changing the steep liquor1" to prevent harm to the considerable potential for producing malt in an extremely
grain by the products of respiration. It has been suggested by short time.
Moufang'20 that barley should be steeped in warm I % sodium Additives to the steep liquor which have a marked effect on
germination include certain recognized oxidizing agents,
or potassium hydroxide to dissolve out phenolic substances.
which often stimulate both dormant and non-dormant grain.
The malts so obtained were believed to yield finer beers and
Pallas"" and Tarrassov3" found that the addition of potassium
smoother fermentations, but the claims seem exaggerated
since Stevens"1 found that 01-n alkali solutions reduce
permanganate had a beneficial effect on growth, thereby
germination to 25% and I-n solutions kill the grain com
reducing malting time. Hydrogen peroxide as a 0-1 or 1%
pletely. More recently, Mostek217 showed that a first steep solution, besides being effective on dormant grain, also
containing 0-38% sodium hydroxide was generally satisfac
stimulated non-dormant barley.aS2 Endcrs et al."" also found
tory. Kieninger15* carried out a series of trials where barley
that solutions of 0-5% hydrogen peroxide or 0-1% solutions
of lactic acid in 0-1 % hydrogen peroxide stimulated germina
was steeped in solutions of from 005 to 0-5% sodium
hydroxide. He found that the best tasting beers were obtained tion and effected improved modification and greater proteo-
after using 005 and 0-1 % sodium hydroxide. Other processes lysis. These results were confirmed by other workers18-81'328'370
have been described316 where the alkali has been used essentially
using concentrations of hydrogen peroxide between 0-5 and
1 %. However, the high cost of hydrogen peroxide precludes
to pre-wash the grain prior to normal steeping.
At one time the practice of steeping barley in lime water
its use on the commercal scale at present. Other compounds,
(10-20 g lime/hccalitre, approx. 0-5 oz/200-250 gal) was
for example, pcracctic acid108 and sulphur dioxide (0-02 %)280
common. This treatment was considered127'284'383'"64'366'3" to
have been found to be less effective than hydrogen peroxide.
Pollock101 has pointed out that many compounds have been
promote the germination of barley which was dormant or had
been harvested under adverse weather conditions and had dissolved in steep liquors with the object of stimulating
probably suffered microbiological contamination. Pollock260
germination and reducing malting time. Amongst compounds
has found, however, that the effect of lime water on dormant known to raise the gcrminativc energy of dormant barleys arc
thiourea,"-'64 hydrogen sulphide and many mercaptans,
barleys is slight. Sterilizing agents such as hypochlorites40 have
been used but they taint the flavour of the resulting malt and
hydroquinone, phenol and a number of substituted phenols,2*4
nitrous acid (005%), sodium carbonate (0-1 %) or potassium
beer and their use is thus unsuitable on the commercial
scale.0-340 For a similar reason, antiseptics containing free bromate (0-002 %),333 nitrite,260 nitrate,71'815-301 tungstate and
chlorine cannot be used since they give a phenolic taste to
manganous ions.216 Many of these compounds show only a
beer. marginal effect, as in the example of sodium chloride quoted
Several workers188-333 have found that soaking in 0-2% by Katagiri et a/.160
solutions of formalin increased germination. The use Studies have been made on the ability of various compounds
to overcome water sensitivity or dormancy. The distinction
of formaldehyde in steeping has been studied by many
workers192'163'201-3*13 and found to reduce the anthocyanogen between the two types of inhibition was first emphasized by
content of wort from treated malt and so increase non- the different effects of treating the grain with oxygen and
biological stability of the resulting beers. Formaldehyde hydrogen peroxide.263 Both these agents, if present in low
concentrations during germination, markedly reduce water
residues in finished beers were less than 0-2 ppm—no greater
sensitivity but dormancy is not broken. But higher concentra
than those in corresponding untreated control beers. Withcy
& Briggs363 showed that re-steeping in formaldehyde was more
tion of hydrogen peroxide do break dormancy. A number of
effective than re-steeping in water for reducing malting loss.
sulphydryl compounds, e.g. thiourca and hydrogen sulphide,
Further, there has been no demonstration of any hazard to destroy dormancy but not water sensitivity.284 Many phenolic
health that might arise from the use of formaldehyde at these compounds and growth regulators, e.g. kinetin and gibberellic
low levels, and in view of its considerable potential it would acid200 stimulate germinative energy.
seem that further work on these lines is worth carrying out. On the other hand, water-sensitivity but not dormancy can
The addition of acids to the steep may prove beneficial. be relieved by the action of solutions containing ferrous
Vesclov342 obtained maximum germination and maximum sulphate"7-142 uranyl acetate139 and other metal ions, parti
diastatic power spraying the grain with 01 % sulphuric acid, cularly those which co-ordinate sulphur, sulphydryl-binding
whilst Gracanin107 observed that I % phosphoric acid stimul reagents such as N-ethylmaleimidc and oxidizing agents such
ated germination in old barleys but did not exert any notice as potassium bromate.141 •**• Recent work" has shown that
able effect on normal ones. Pool & O'Connor2" agreed that mixtures of antibiotics arc effective in relieving water sensi
the addition of sulphuric acid to a steep containing gibberellic tivity. Germination is shortened by 2-4 days in the presence of
acid reduced malting loss and increased extract but found that 500 mg soluble fungicidin (nystatin) per 10 tons barley.11
the resulting wort had poor attenuation. Barley retains its Popov & Vasileva277 showed that Cu2+ and Co2+ ions stimul
germinative properties even after treatment with 50% ated respiration, germination rate and amylase activity, and
sulphuric acid for 3 h,3"-81 owing to the protection afforded by Fukui06 in a study with 27 different metallic chlorides found
the testa. that As3+, Cr2+, Hg2+, Ti3+ and V3+ inhibited germination
Acetic acid and inorganic acetates at concentrations of 100 completely.
mg/litre in steep water stimulate the germination of dormant The hardness of steep liquor docs rot affect the quality of
but not non-dormant barley.113 Spillanc & Briggs316 have found the finished malt135'308 but if iron salts are present in higher
that spraying acetic acid or steeping in solutions of acetic than usual concentrations the malt is discoloured.00
acid reduced germination time and malting loss. Phenolic Schuster & Eppinger302 found that deleterious effects are
acids, such as fcrulic and p-hydroxybenzoic acid at con associated with some ions particularly at high concentrations.
centrations of 100 mg/kg delay the synthesis of enzymes by In particular n/100 nitrite markedly reduced malt quality.
gibbcrcllic acid.111 It has been shown210 that salicylic acid Plant Regulators Including Gibberellic Acid.—The work of
22 BROOKES, LOVETT AND MACWILLIAM: STEEPING OF BARLEY [J. Inst. Brew.

Yomo384 and Paleg846 showed that gibbcrcllic acid at low as a spray after steeping at a rate of 100-200 mg/kg. It appears
conccnlraions (0-1-0-2 ppm) stimulated the a leu rone layer of to be taken up very rapidly by the germinating grain and be
barley to release the enzymes which modify barley. This com present in the endosperm within 4 h of application.34
pound is without doubt the most important additive used in
the malting process. Its use as a malting aid and its mechanism Leaching of Barley Constituents by Steep Liquors
of action have recently been reviewed by Palmer"3 and
Even when no additive is present in steep liquor it very
Briggs32 and it is thus not proposed to include a detailed dis
quickly becomes yellow-brown in colour soon after barley is
cussion of its mode of action in this review.
added. Novtil'nov & Ezhov241 consider that the coloured
Studies7'161-"2 on the most appropriate point in the malting
materials are flavanoid whilst other workers18*'184'"8'188'"3-8*0'3"
process at which to add gibbcrcllic acid have concluded that
present evidence that they are similar to 'testinic acid'. The pH
it should be sprayed on soon after the grain is removed from
of steep liquors decreases rapidly after barley is added due to
the steep.
the presence of acids dissolved from the grain, but this has
Since the pericarp is impermeable to externally applied
no effect on water absorption or germination.282 Most of the
gibbcrellic acid348 it must be applied when the grain is chitted,
naturally-occurring amino acids63'120-188 have been detected in
i.e. preferably as a spray after steeping. This observation
steep liquor but despite this loss Robbins, Chestcrs & Dick-
formed the basis of the development of abraded barley"
son"2 observed that the concentration of most amino acids in
which provides further sites for entry of gibberellic acid to
barley rises during steeping, probably as a result of enzymic
the germinating grain.
activity. Simple carboxylic acids, e.g. acetic, succinic*4'*0 and
Gibberellic acid (gibberellin A,) is only one of a series of
phenolic acids5-" are also present. Anthocyanogcns and other
natural gibbercllins which exhibit stimulatory activity on
polyphenolic compounds, e.g. catechins, arc leached through
plants. Several of the other gibbcrcllins have been found to
the testa of decorticated barley steeped in water but not t hrough
exert activity on barley.112 Other compounds such as kinetin38
the pericarp of inact grain.287
compounds such as kinetin28 (6-furfurylaminopurine) and the
Phosphates representing approximately one-tenth of the
tricyclic terpenc, stcviol, also stimulate the formation of
total originally present in barley have been detected in steep
enzymes in barley.201
liquors.3" Since simple sugars are also removed*3 together
Gibbcrellic acid has proved extremely successful as a malting
with betaine,** steep liquors provide a good medium for the
aid but the use of other plant hormones has not so far been
growth of micro-organisms and changes in the liquor arc
developed commercially. Prentice2™ found that chlorocholinc
obviously required to minimize such growth.
chloride, 3-amino-l,2,4 triazolc and kinetin stimulated the
Detailed studies201 have shown that about 1 % of their dry
modification of barley but only at concentrations in excess of
100 mg/litre. Other workers have also examined the action
weight is leached from European barley varieties during con
ventional steeping. Only half of this amount is removed when
of 2,4-dichlorophenoxyacetic acid6"'1"'1"-209 2,3-benzoxazolen-
onct7s,i;e an(j o-phenyl phosphates" on germination and malt an interrupted steeping procedure requiring only a total of
quality.
8-10 h immersion is employed even though the same final
moisture content is attained.
Scriban304 has described the action of 2-hydroxymethyl and
2-formyl-4-chlorophcnoxyacetic acid on barley germination.
These compounds exert effects at very low levels (< 1 -0 mg/kg) Conclusions
but since they reduce cytolytic activity they do not seem suit Although at the end of most sections, relevant lines of
able for commercial use as malting aids. future research have been indicated, these are perhaps
MacLeod19* has studied the involvement of indole acetic principally for laboratory study. Frequently, promising
acid (IAA, auxin) with gibberellic acid in barley germination. results are obtained but the difficulties in applying these to
IAA stimulates a-amylase production, but since, when IAA industrial practice may be very great. One of the most import
and gibbcrellic acid are added together, a-amylase production ant problems appears to be the scaling up of operations from
decreases, the use of IAA as a commercial malting aid seems laboratory to industrial scale. This is not a problem facing the
precluded. The mechanism of action of 1AA has been studied malting industry alone and it is likely that improvements in
by Vcrbeek et a/.341 but again it seems that there arc too many plant used in the chemical and food industry will be of value
side effects for the successful application of IAA solely to in malting. Pilot-scale-equipment, handling approx. 1 cwt of
increase a-amylase producion. malt which can be adequately controlled, is obviously
Inhibitors of germination.—Apart from strong acids and required. This equipment is needed both for use in the steeping
alkalis, attention has mainly been devoted to the inhibiting process and for the later stages of malting and brewing, so
effect of compounds isolated from steep liquors themselves. that the influence of the first process on the later ones can be
Although it was early noted that the germination of seeds was fully determined.
inhibited by aqueous extracts derived from them (see refer Among the most novel alterations to the steeping process
ences cited by Cook & Pollock65), van Laer et al.33i-33t were described in recent years have been the attempts to produce
the first workers to measure the inhibiting effects of barley malt from barley without embryo growth*8 or even without
steep liquors quantitatively. Cook & Pollock5**53-*1'" made a germination.325 Tahara3" described the production of
detailed study of the inhibitory substances in steep liquor and ungerminatedmaltby steeping barley in water for 24h at 16°C
found that acetic acid was present in significant amounts. and then in 0-25% ammonia solution for 5 h. After washing,
The pure acid together with other simple carboxylic acids the grain was sprayed with 2 mg/kg gibbcrellic acid and
were all found to inhibit germination at concentrations above maintained at 16°C for 6 days with aeration, at a moisture
2 g/litre. Many compounds, including some which are content of 40%. Control malts contained more fermentable
stimulating a low concentrations (100 ppm), arc probably sugars and soluble nitrogen but beers made from ungerminated
inhibitory at higher ones. Gilbert, Blum & Frieden100 consider grain were reported to have a satisfactory flavour. It is
that bacteria or their products, namely acetic and other acids, difficult to imagine, however, that a process of this type would
are responsible for the inhibition. have any commercial significance, but this does illustrate the
Mention should be made of the use of bromate applied as ingenuity of approach of workers on barley and malting which
sodium or potassium bromate to control the proteolysis of itself increases the chances of results of the research being of
germinating grain. Following the original work of Maccy & direct benefit to producion.
Stowell, the use of bromatc is now widespread as a means of To explain some of the difference in malting quality between
controlling the effects of gibbcrellic acid on protein modifica barley varieties fundamental work on water uptake is still
tion of germinating barley.100'182 Bromate is normally applied required. Further work should try to assess the mode of
Vol. 82, 1976] BROOKES, LOVETT AND MACWILLIAM: STEEPING OF BARLEY 23

uptake of water which might be relevant in establishing 18. Bcvilacqua, L. R., & Pignato, L. C, Atti del Accademia
optimum moisture contents for satisfactory malting of new Ligure di Scienze Lettere (Genoa), 1961, 17, 227.
19. Bishop, L. R., Journal of the Institute of Brewing, 1945, 51,
barley varieties. 215.
Insufficient attention has been paid to the influence of steep 20. Bitter, H., Brauwelt, 1966, 106, 172.
water composition on the germination process and malt 21. Bitter, H., Brauereitechniker, 1967, 19, 1.
quality. It was stated many years ago51 that the barley testa 22. Blaber, S., Brewer's Journal, 1907, 43, 359.
23. Blaim, K., Journal of Experimental Botany, 1960, 11, 377.
did not show perfect semi-permeability to any solute, and it 24. Blcisch, C, Journal ofthe Institute of Brewing, 1910,16, 682.
has recently been shown that bromate applied to the grain 25. Blcisch, C, & Will, H., Zeitschrift fiir Gesamte Brauwesen,
enters the grain rapidly and is present in the endosperm within 1902, 25, 17.
4 h of application.94 It seems that some ions can penetrate to 26. Bode, G., Journal of the Institute of Brewing, 1904, 11,505.
27. Bond, A. B., & Glass, R. L., Cereal Chemistry, 1963, 40,
the grain through an intact testa but that defects can also exist 459.
in the testa which allow passage of salts originally thought to 27. Boothby, D., & Wright, S. T. C, Nature (London), 1962,
be excluded from the grain. 196, 389.
The influence of steep water on malt composition has been 29. Brewing Patents Ltd., British Patent 1,264,822. Granted
23.2.72.
studied302 but with new harvesting and barley handling 30. Briggs, D. E., Journal of the Institute of Brewing, 1967, 73,
techniques which may affect grain permeability, rc-asscssment 33.
of this work is due. Such work might explain differences in 31. Briggs, D. E., Journal of the Institute of Brewing, 1972, 78,
germination behaviour between barley varieties which arc not 365.
32. Briggs, D. E., in Biosynthesis and its Control in Plants, Ed.
easily understood in terms of other malting process data. Milborrow, B. V. Academic Press, 1973.
Further work is undoubtedly required on the effects of re-use 33. Briggs, D. E., Cluttcrbuck, V. J., & Murphy, G., Bio
of steep water without treatment and the effects of spray- chemical Journal, 1971, 124, 2p.
steeping on germination behaviour and malt quality. This work 34. Brookes, P. A., & Martin, P. A., Journal of the Institute of
Brewing, 1974, 80, 294.
will increase in importance as in modern malting design the 35. Brown, A. J., Journal of the Institute of Brewing, 1907, 13,
tendency is towards the use of extremely large barley batch 658.
sizes (100-200 tonnes) with correspondingly large steeping 36. Brown, R., Annals of Botany, 1943, N.S.7, 93.
cisterns and demand for water. It will probably always be good 37. Buchingcr, A., Proceedings of the International Seed Test
Association, 1932, 4, 46.
practice to get rid of the effluent steep water from the first 38. Bunch, T. A., & Dclouchc, J. C, Proceedings of the Associa
wetting but second or third steep waters could be held in tion of Official Seed Analysts, 1959, 49, 142.
effluent tanks and then pumped back mixed with fresh water 39. Caldwcll, F., Journal of the Science of Food and Agriculture,
to provide the first wetting of the next batch of barley. These 1963, 14, 765.
40. Cerny, F., Journal of the Institute of Brewing, 1900, 6, 128.
processes would no doubt be economically attractive but their 41. Chambers, A. R., & Lambic, A. D. B., Journal of the Insti
acceptance would depend on the demonstration that there was tute of Brewing, 1960, 66, 159.
no adverse effect on malt quality. Spray steeping may require 42. Chandra, G., Biochemistry and Physiology of Plant Growth
as little as one tenth of the water volume required for immer Substances, Proceedings of the International Conference on
Plant Growth Substances, 1967, 723.
sion steeping and such reports as are available200 suggest that 43. Chapon, L., Brasserie, 1959, 14, 222.
malt quality is not impaired. It would be expected, however, 44. Chapon, L., Brasserie, 1960, 15, 214.
that the mechanism of water uptake might be different and 45. Chapon, L., Brauwissenschaft, 1961,14, 457.
that the water distribution in the corn might be altered. This 46. Chapon, L., Bulletin des Anciens Glives de L'Ecole de
Brasserie de Nancy, 1963, 356.
would seem to be an extremely valuable future line of research 47. Chapon, L., Bulletin des Anciens Elives de L'Ecole de
work. Brasserie de Nancy, 1963, 327.
Gibbcrcllic acid remains the only plant growth hormone 48. Chrispeels, M. J., & Varncr, J. E., Plant Physiology, 1967,
that has been successfully exploited in malting. However, in 42, 398.
49. Christiansen, M. N., & Moore, R. P., Agronomy Journal,
view of the work of Verbeek339 on the importance of the 1959, 51, 582.
phytohormonal balance in controlling the germination of 50. Christiansen, M. N., Moore, R. P., & Rhyne, C. L.,
barley the possibility of influencing the balance by application Agronomy Journal, 1960, 52, 81.
of other hormones both during and after steeping should not 51. Collins, E. J., Annals of Botany, 1918, 32, 381.
52. Cook, A. H., & Pollock, J. R. A., Journal of the Institute of
be overlooked. Brewing, 1952, 58, 407.
53. Cook, A. H., & Pollock, J. R. A., Journal of the Institute of
Brewing, 1952, 58, 325.
References 54. Cook, A. H., & Pollock, J. R. A., Journal of the Institute of
Brewing, 1953,59,313.
1. Abe, M., Yamagata Daigaku Kigs Shizen Kagaku, I960, 5, 55. Cook, A. H., & Pollock, J. R. A., Journal of the Institute of
209. Brewing, 1954, 60, 300.
2. Allerup, S., Phystologia Plantarum, 1958, II, 99. 56. Cook, A. H., Hodgson, H. C, & Pollock, J. R. A., Journal
3. Allerup, S., Physiohgia Plantarum, 1959, 12, 118. of the Institute of Brewing, 1954, 60, 292.
4. Anon., Brewers Journal, 1896, 20, 149. 57. Cossins, E. A., & Turner, E. E., Annals of Botany, 1962, 26,
5. Asen, S., & Emsweller, S. L., Phylochemislry, 1962, 1, 169. 591.
6. Associated British Maltsters, Belgian Patent 619,005, 1962. 58. Crabb, D., Proceedings of the American Society of Brewing
7. Ault, R. G., Journal ofthe Institute ofBrewing, 1961,67,405. Chemists, 1972, 36.
8. Baker, J. L., & Dick, W. D., Journal of the Institute of 59. Crabb, D., & Kirsop, B. H., Journal of the Institute of
Brewing, 1905,11, 372. Brewing, 1969, 75, 254.
9. Barth, G., Zeitschrift fur das Gesamte Brauwesen, 1903, 25, 60. Crabb, D., & Kirsop, B. H., Journal of the Institute of
739. Brewing, 1970, 76. 158.
10. Barwald, G., Brauwelt, 1973, 113,1872. 61. Crocker, W., & Barton, L. V., Physiology of Seeds, 1953,
11. Basarova, G., Strosova, J., & Icha, F., Czechoslovakian p. 72.
Patent 128,112 Granted 16.6.68. 62. Czosnowski, J., Ada Societatis Botanicorum Poloniae, 1962,
12. Becker, H. A., Cereal Chemistry, 1960, 37, 309. 31,135.
13. Bekcndam, J., & Bruinsma, J., Proceedings of the Inter 63. Czosnowski, J., Ada Societatis Botanicorum Poloniae, 1962,
national Seed Testing Association, 1966, 31, 779.
31, 683.
14. Belderok, B., Bericht Getreidechemiker-Tagung, Detmold,
64. Dahlstrom, R. V., Morton, B. J., & Sfat, M. R., Proceedings
1962,21.
of the American Society of Brewing Chemists, 1963, 64.
15. Belderok, B., Journal of the Institute of Brewing, 1968, 74,
333.
65. Day, F. C, Transactions and Proceedings of the Botanical
16. Bcnton, W. A., Journal of the Institute of Brewing, 1932, 38, Society of Edinburgh, 1896,492.
245. 66. De Clerck, J., Textbook of Brewing, Chapman, & Hall
17. Beltncr, R. E., & Meredith, W. O. S., Proceedings of the London, 1957, 132.
American Society of Brewing Chemists, 1970, 118. 67. Dcwey, }., Plant Physiology, 1964, 39, 240.
24 BROOKES, LOVETT AND MACWILLIAM; STEEPING OF BARLEY [J. Inst. Brew.

68. Dickson, A. D., & Burkhart, B. A., Cereal Chemistry, 1942, 113. Griffiths, C. M.. MacWilliam, I. C, & Reynolds, T.,
19.251. Journal of the Institute of Brewing, 1965, 71, 316.
69. Dickson, A. D., Shands, H. L., & Burkhart, B. A., Cereal 114. Groat, J. I., & Briggs, D. E., Phytochemistry, 1969, 8, 1615.
Chemistry, 1949, 26, 13. 115. Grosh, G. M., & Milner, M., Cereal Chemistry, 1959, 36,
70. Earle, K. M., & Galsky, A. C, Plant Cell Physiology, 1971, 260.
12.727. 116. Gruss, J., Journal of the Institute of Brewing, 1930, 36, 394.
71. Edclman, J., Shibko, S. I., & Keys, A. J., Journal of Experi 117. Haber, A. H., & Tolbcrt, N. E., Plant Physiology, 1959, 34,
mental Botany, 1959, 10, 178. 376.
72. Eflront, M. J., Moniteur SclentIfique, 1906, 20, 5. 118. Hall, R. D., Harris, G., & MacWilliam, I. C, Journal of
73. Enrich, E., & Kneip, E., Zeitschrift fiir das Gesamte the Institute of Brewing, 1956, 62, 232.
Brauwesen, 1931, 54, I, 9, 98. 119. Harlan, H. V., & Pope, M. N., Journal of Agricultural
74. Ekstrttm, D., Ccderquist, B., & Sandegrcn, E., Proceedings Research, 1923. 23, 333.
of the European Brewery Convention Congress, Rome, 1959, 120. Harris, G., & Pollock, J. R. A., Journal of the Institute of
11. Brewing, 1953, 59, 28.
75. Emslandcr, F., Zeitschrift fiir das Gesamte Brauwesen, 1902, 121. Hartong, B. D., Wochenschrift fur Brauerei, 1936,53, 81.
25, 721. 122. Hartong, B. D., Wallerstein Laboratories Communications,
76. Emslandcr, F., Zeitschrift fiir das Gesamte Brauwesen, 1909, 1940, 3, 107.
32, 65. 123. Hartong, B. D., & Kretschmer, K. F., Brauerei Wissen-
77. Endcrs, C, Nowak, G., Schncebauer, F., & Pfahlcr, A., schaftliche Bcilage, 1958, 11, 238.
Wochcnschrift fiir Brauerci, 1940, 57, 81. 124. Hartong, B. D., & Kretschmer, K. F., Proceedings of the
78. Engel, O. S., & Prokof'cv, A. A.. Fiziologiya Rastenii, 1960, European Brewery Convention Congress, Vienna, 1961, 69.
1,32. 125. Haskins, F. A.. Plant Physiology, 1955, 30, 74.
79. Ermilov, G. B., Fiziologiya Rastenii, 1960,1, 36. 126. Haslinger, C. Wochenschrift fur Brauerei, 1902,19,167,196.
80. Esscry, R. E., Kirsop, B. H.. & Pollock, J. R. A., Journal 127. Hauter-Spcycr, C, Journal of the Institute of Brewing, 1897,
of the Institute of Brewing, 1954, 60, 473. 3, 156.
81. Essery, R. E., Kirsop, B. H., & Pollock, J. R. A., Journal 128. Hiltncr, J., Mitteilungen der Dcutschen Landwirtschafts-
of the Institute of Brewing, 1955, 61, 25. gesellschaft, 1901, 32.
82. Esscry, R. E., & Pollock, J. R. A., Journal of the Institute of 129. Holbein, G.. Brewers Journal, 1969, 105, (2), 47.
Brewing, 1956, 62, 327. 130. Holmes, P. L., & Spcakman, P. T., Nature New Biology,
83. Esscry, R. E., & Pollock, J. R. A., Journal of the Institute 1973, 242, 190.
of Brewing, 1957.63,221. 131. Hopkins, R. H., Brewers Guardian, 1960, 89, (1), 17.
84. Evans, R. E., Journal of the Institute of Brewing, 1901, 7, 132. Insik, H., Malzindustrie, 1905, 49.
264. 133. Jaeger, P., Branntweinwirtschaft, 1971, 111, 145,168.
85. Eyben, D., & van Droogenbroeck, L., Proceedings of the 134. Jacob, O., Zeitschrift fiir das Gesamte Brauwesen, 1902, 25,
European Brewery Convention Congress, Interlaken, 1969, 289.
107. 135. Jalowetz, E., Journal of the Institute of Brewing, 1931, 37,
86. Eybcn, D., & van Droogenbroeck, L., Chemical Abstracts, 241.
1971,74,75179. 136. James, W. O., Plant Respiration, Clarendon Press, Oxford,
87. Eybcn, D., & van Droogenbroeck, L., Bulletin de VAssocia 1953.
tion Royale des Anciens Etudiants en Brasserie de L'Univer- 137. Jansson, G., Arkiv fur Kemi, 1959, 14, 161.
sitd de Louvain, 1970, 66, (3), 105. 138. Jansson, G., Arkiv fiir Kemi, 1961, 17, 281.
88. Fairclough. D., & Williams, R. P., British Patent 1,122,807. 139. Jansson, G., Svensk Kemisk Tidskrift, 1962, 74, 181.
Granted 7.8.68. 140. Jansson, G., Arkiv fiir Kemi, 1962, 19, 141.
89. Fan, L., Chung, D. S., & Shellenberger, J. M., Cereal 141. Jansson, G., Arkiv fiir Kemi, 1962, 19, 149.
Chemistry. 1961,38,540. 142. Jansson, G., Kirsop, B. H., & Pollock, J. R. A., Journal of
90. Fan, L., Chu, P., & Shellcnbcrgcr, J. M., Cereal Chemistry, the Institute of Brewing, 1959, 65, 165.
1963, 40, 303. 143. Jones, M., Brewers Digest, 1969, 44, (3), 60, 74.
91. Fchrmann, K., & Sonntag, M., Mechanische Technologic 144. Jones, M., & Pierce, J. S., Proceedings of the American
der Brauerei, (2), Paul Parcy, Berlin and Hamburg, 1962. Society of Brewing Chemists, 1966, 10.
92. Flcckcnstein, J. G., United Stales Patent, 3,086,921. 145. Jones, M., & Pierce, J. S., Proceedings of the Irish Maltsters
Granted 23.4.63. Technical Meeting, 1966, 53.
93. Forschungsinstitut fur die Gaerungsindustric, Enzymologic 146. Jones, M., & Pierce, J. S., Journal ofthe Institute of Brewing,
und Technischc Mikrobiologic, British Patent, 1,316,226., 1967, 73, 577.
Granted 9.5.73. 147. Joyce, C. H., Journal of the Institute of Brewing, 1930, 36,
94. Frazer, C. W., & Haley, W. L., Cereal Chemistry, 1932, 9, 131.
45. 148. Kalinin, F. L., & Pel'Tek, C. N., Chemical Abstracts, 1969,
95. Fritz, A., Ulonska, E., & Lcnz, W., Brauwissenschaft, 1966, 70, 2626.
19, 144. 149. Karimov, K. K., & Kadyrova, D. K., Fiziologiya Rastenii,
96. Fukui. T., Bulletin of Brewing Science, 1971, 17, 17. 1966, 13, 1081.
97. Furnrohr, O.. Journal of the Institute of Brewing, 1912, 18, 150. Katagiri, H., Ikcmiya, M., & Anyoji, H., Chemical Abstracts,
55. 1953, 47, 10800f.
98. Gabcr, S. D., & Roberts, E. H., Journal of the Institute of 151. Kaucrt, G., Fcrmentatio, 1955, 229.
Brewing, 1969, 75, 299. 152. Kauert, G., Petit Journal du Brasseur, 1959, 67, 730.
99. Gabcr, S. D., & Roberts, E. H., Journal of the Institute of 153. Kauert, G., Brauwelt, 1961, 101, 981.
Brewing, 1969, 75, 303. 154. Kauppinen. V., Nummi, M., & Enari, T. M., Brauwwisen-
100. Gilbert, S. G., Blum, P. H., & Fricden, A., Proceedings of schaft, 1967, 20, I.
the American Society of Brewing Chemists, 1954, 51. 155. Kessler, B., & Kaplan, B., Physiologia Ptantarum, 1972, 27,
101. Gloetzl, J., Brauwelt, 1960,100, 876. 424.
102. Goodwin, P. B., & Carr, D. J., Journal of Experimental 156. Kieninger, H., Brauwelt, 1971, 111, 778.
Botany, 1972,23,8. 157. Kieninger, H.. & Graf., H., Brauwelt, 1973,113, 643, 706.
103. Goodwin, P. B., & Carr, D. J., Journal of Experimental 158. Kirsop, B. H., Proceedings Irish Maltsters Technical
Botany, 1972. 23. 1. Meeting, 1966, 37.
104. Goldacker, G., Journal of the Institute of Brewing, 1910, 16, 159. Kirsop, B. H., Journal of the Institute of Brewing, 1967, 73,
683. 214.
105. Gomez, R., Proceedings of the European Brewery Conven 160. Kirsop, B. H., Reynolds, T., & Griffiths, C. M., Journal of
tion, Congress Estoril, 1971, 29. the Institute of Brewing, 1967, 73, 182.
106. Gordon, A. G., Journal of the Institute of Brewing, 1968, 74, 161. Kleber, W., Lindemann, M., & Schmid, P., Brauwelt, 1959,
355. 99, 1781.
107. Gracanin, M., Biochemische Zeitschrift, 1928, 195, 457. 162. Klopper, W. J., & Vermeire, M. A., Technical Quarterly of
108. Gracsser, F. R., & Stoddart, W. E., Wallerstein Labora the Master Brewers Association of America, 1968, 5, 235.
tories Communications, 1963, 26, 47. 163. Kopecky, O., & Almendingcr, V., Journal of the Institute
109. Green, J. W., & Sangcr, M. J., Journal of the Institute of of Brewing, 1935.41,75.
Brewing, 1956, 62, 170. 164. Krauss, L., Wochenschrift fur Brauerei, 1933, 50, 209.
110. Griffin, O. T., & Pinner, B. C, Journal of the Institute of 165. Kretschmer, K. F., Brauwelt, 1957, 97, 1436.
Brewing, 1965, 71, 324. 166. Kumar, M., Journal of Food Technology, 1973, 8, 407.
111. Griffiths, C. M.. & MacWilliam, 1. C, Journal ofthe Institute 167. Lang, J. L., Bulletin de VAssociation Royale des Anciens
of Brewing, 1967,73, 172. Etudiants en Brasserie de UUniversite' de Louvain, 1965, 61,
112. Griffiths, C. M., MacWilliam. I. C, & Reynolds, T., 1.
Nature (London), 1964, 202, 1026. 168. Lampe, B., Journal of the Institute of Brewing, 1929, 35, 81.
Vol. 82, 1976] BROOKES, LOVETT AND MACWILLIAM: STEEPING OF BARLEY 25

169. Lcibu, J., Journal of the Institute of Brewing, 1920, 26, 527. 223. Miindlcr, K., Zeitschrift fur das Gesamte Brauwesen, 1934,
170. Leonard, W. H., & Martin, J. H., Cereal Crops, MacMillan, 57, 35.
New York, 1963, p. 478. 224. Narziss, L., Technical Quarterly of the Master Brewers
171. Levingc, R., Irish Maltsters Conference Selected Papers, Association of America, 1966,3, 237.
1953. 225. Narziss, L., Proceedings of the European Brewery Conven
172. Linko, M., Suomen Kemistilehti, 1960, B33, 31. tion Congress, Interlaken, 1969, 77.
173. Linko, M., & Enari, T. M., Journal of the Institute of Brew- 226. Narziss, L., Wallerstein Laboratories Communications, 1970,
ing, 1960, 66, 480. 33, 77.
174. Linko, M., & Enari, T. M., International Brewer and 227. Narziss, L., Brauwelt, 1972, 112, 1337.
Distiller, 1966, 1, (I), 46. 228. Narziss, L., & Friedrich, G., Brauwelt, 1970, 110, 229.
175. Linko, M., Linko, P., & Enari, T. M., Cereal Chemistry, 229. Narziss, L., & Friedrich, G., Brauwelt, 1970, 110, 652.
1961,38,60. 230. Narziss, L., & Friedrich, G., Brauwissenschaft, 1970,23,133.
176. Linko, P., Soiimen Kemistilehti, 1959, B32, 266. 231. Narziss, L., & Friedrich, G., Brauwissenschaft, 1970,23,167.
177. Linko, P., Annales Academiae Scientiarmn Fennicae, 1960, 232. Narziss, L., & Friedrich, G., Brauwissenschaft, 1970,23,229.
98, 7. 233. Narziss, L., & Friedrich, G., Brauwissenschaft, 1970,23,265.
178. Linko, P., & Milner, M., Cereal Chemistry, 1959, 36, 274. 234. Narziss, L., & Kicningcr, H., Brauwissenschaft, 1966,19, 343.
179. Linko, P., & Milncr, M., Cereal Chemistry, 1959, 36, 280. 235. Navarro, j., & Brandao, J. L. N., Proceedings of the
180. Loibl, H., Journal of lite Institute of Brewing, 1923, 29, 672. European Brewery Convention Congress, Estoril, 1971, 73.
181. Lubcrt, D. J., & Pool, A. A., Journal of the Institute of 236. Newark, M. Z., & Wengcr, B. S., Archives of Biochemistry
Brewing, 1964, 70, 145. and Biophysics, 1960,89. 110.
182. Ludwig, H., Proceedings of the International Seed Testing 237. Nickells, M. W., Schaefer, G. M., & Galsky, A. G., Plant
Association, 1971,36,289. Cell Physiology, 1971, 12, (5), 717.
183. Liters, H., Die wissenschaftliche Gntndlagen von Miilzerei 238. Nielsen, N., Compte Rendu des Travaux du Laboratoire de
und Brauerei, Nurnberg, Vcrlag Hans Carl, 1950. Carlsberg, 1937,22,49.
184. LUcrs, H., Wochcnschrift fur Brauerei, 1930, 47, 557. 239. Northam, P. C, Brewers Guild Journal, 1965, 51, 229.
185. Liicrs. H., & Slaubcr, J., Wochenschrift fur Brauerei, 1931. 240. Northam, P. C, Brewers Guardian, 1965, 94, (5), 97.
48,103,117. 241. Novtil'nov, N. V., & Ezhov, J. S., Botanichnii Zhurnal,
186. Liiers, H., & Collignon, E., Wochenschrift fiir Brauerei, 1957, 42, 255.
1939, 56, 305. 242. Oota, Y., Physiologia Plantarum, 1957,10, 910.
187. Luff, G., Journal of the Institute of Brewing, 1901, 7, 497. 243. Oota, Y., Physiologia Plantarum, 1958,11, 710.
188. Macey, A., Brewers Guardian, 1960, 89, (1), 49. 244. Otis, O. J., & Olson, W. J., Proceedings of the American
189. Maccy, A., Proceedings of the European Brewery Convention Society of Brewing Chemists, 1953, 22.
Congress, Stockholm, 1965, 41. 245. Paleg, L. G., Plant Physiology, I960, 35, 902.
190. Macey, A., & Stowell, K. C, Journal of the Institute of 246. Pallas, E., Journal of the Institute of Brewing, 1933, 40, 43.
Brewing, 1957, 63, 391. 247. Palmer, G. H., Journal of the Institute of Brewing, 1969, 75,
191. Macey, A., & Stowell, K. C, Proceedings of the European 505.
Brewery Convention Congress, Rome, 1959, 105. 248. Palmer, G. H., Journal of the Institute of Brewing, 1969, 75,
192. Maccy, A., & Stowell, K. C, Journal of the Institute of 536.
Brewing, 1961, 67, 396. 249. Palmer, G. H., Journal of the Institute of Brewing, 1970, 76,
193. Maccy, A., & Stowell, K. C, Journal of the Institute of 65.
Brewing, 1966, 72, 29. 250. Palmer, G. H., Proceedings of the European Brewery
194. MacLeod, A. M., New Phytologist, 1957, 56, 210. Convention, Estoril, 1971, 59.
195. MacLeod, A. M.. New Phytologist, 1969, 68, 295. 251. Palmer, G. H., Journal of the Institute of Brewing, 1972, 78,
196. MacLeod, A. M., DufFus. J. M., & Horsfall, D. J. L., 81.
Journal of the Institute of Brewing, 1966, 72, 36. 252. Palmer, G. H., Brewers Guardian Supplement, 1972, 101, 9.
197. MacLeod, A. M., & Palmer, G. H., Nature London, 1967, 253. Palmer, G. H., Journal of the Institute of Brewing, 1974, 80,
216, 1342. 13.
198. MacLeod, A. M., Travis, D. C, & Wrcay, D. G., Journal 254. Petit, B., Journal of the Institute of Brewing, 1895, 1, 528.
of the Institute of Brewing, 1953, 59, 154. 255. Petit, B., Journal of the Institute of Brewing, 1923, 29, 984.
199. MacWilliam, I. C , Brewers Journal, 1963, 99, 68. 256. Piendl, A., & Wagner, D., Process Biochemistry, 1972,7, (4),
200. MacWilliam, I. C, Journal of the Institute of Brewing, 1965, 26.
71, 197. 257. Piratzky, W., Journal of the Institute of Brewing, 1958, 64,
201. MacWilliam, I. C, Proceedings of the European Brewery 429.
Convention Congress, Stockholm, 1965, 81. 258. Pohlmann, R., Brauwelt, 194, 98, 1837.
202. Maicr, F., Saatgutwirtschaft, 1950, 2, 228, 251. 259. Pollock, J. R. A., Nature (London), 1956, 178, 1359.
203. Major, W., & Roberts, E. H., Journal of Experimental 260. Pollock, J. R. A., Journal of the Institute of Brewing, 1959,
Botany, 1968,19, 90. 65, 334.
204. Mandl, B., Brauwisscnsclmft, 1970, 23, 175. 261. Pollock, J. R. A., in Barley and Mall, Ed. A. H. Cook,
205. Marcus, A., Federation Proceedings, Federation of American Academic Press 1962, p. 306.
Society of Experimental Biology, 1964, 23, 268. 262. Pollock, J. R. A., Esscry, R. E., & Kirsop, B. H., Journal
206. Massart, L., & Vcrbeck-Wyndaclc, R., Echo de la Brasserie, of the Institute of Brewing, 1955, 61, 295.
1970, 26, 498. 263. Pollock, J. R. A., Kirsop, B. H., & Essery, R. E., Proceedings
207. Mauclairc, D., Brasserie, 1959, 14, 134. ofthe European Brewery Convention Congress, Baden-Baden,
208. Mayer, A. M., & Evcnari, M., Journal of Experimental 1955, 20.
Botany, 1953, 4, 207. 264. Pollock, J. R. A., & Kirsop, B. H., Journal of the Institute
209. Mayer, A. M., & PoljakolT-Mayber, A., The Germination of of Brewing, 1956, 62, 323.
Seeds, Pergammon Press, 1963, p. 134. 265. Pollock, J. R. A., Kirsop, B. H., & Essery, R. E., Journal
210. Meredith, P., New Zealand Journal ofScience, 1971,14,480. of the Institute of Brewing, 1955, 61, 301.
211. Meredith, W. O. S., & Bettncr, R. E., Proceedings of the 266. Pollock, J. R. A., & Pool, A. A., Journal of the Institute of
American Society of Brewing Chemists, 1969, 70. Brewing, 1962, 68, 427.
212. Merritt, N. R., Journal of the Institute of Brewing, 1969, 75, 267. Pollock, J. R. A., Pool, A. A., & Reynolds, T., Journal of
277. the Institute of Brewing, I960, 66, 389.
213. Milncr, M., & Geddcs, W. F., Cereal Chemistrv, 1946, 23, 268. Pool, A. A., Journal of the Institute of Brewing, 1961, 67,
225. 312.
214. MSbius, C. H., Braitwelt, 1973,113, 1862. 269. Pool, A. A., Journal ofthe Institute ofBrewing, 1961,67,389.
215. Moorman, B., Kiihn-Archiv, 1943, 56, 41. 270. Pool, A. A., Journal of the Institute of Brewing, 1962,68,21.
216. Morimoto, K., & Yoshida, T., Report of the Research 271. Pool, A. A., Journal ofthe Institute ofBrewing, 1964,70,221.
Laboratories of the Kirin Brewery Co. Ltd., 1965, 27. 272. Pool, A. A., Proceedings of the Irish Maltsters Technical
217. Mostck, J., Journal of the Science of Food and Agriculture, Meeting, 1966, 64.
1969,20, ii-201. 273. Pool, A. A., Technical Quarterly of the Master Brewers
218. Moufang, E., Journal of the Institute of Brewing, 1909, 15, Association of America, 1967,4, 217.
720. 274. Pool, A. A., Proceedings of the Irish Maltsters Technical
219. Moufang, E., Journal of the Institute of Brewing, 1910, 16, Meeting, 1967, 69.
684. 275. Pool, A. A.. & O'Connor, T. N., Journal of the Institute of
220. Moufang, E., Das Bier, Sclbstverlag Kirn/Nahe, 1918. Brewing, 1963,69,382.
221. Moufang, E , Journal of the Institute of Brewing, 1912, 18, 276. Pool, A. A., & Pollock, J. R. A., Proceedings of the European
Brewery Convention Congress, Madrid, 1967, 241.
222. Moufang, E., & Vctter, L., Journal of the Institute of Brew- 277. Popov, I. D., & Vasileva, T. S., Chemical Abstracts, 1969,
ing. 1910, 16, 320. 71, 19574.
26 BROOKES, LOVETT AND MACWILLIAM: STEEPING OF BARLEY [J. Inst. Brew.

278. Prentice, N.. Burger, W. C, & Moellcr, M., Cereal Client- 325. Tahara.S., Bulletin ofBrewing Science, 1973,18,1.
istry, 1971.48, 587. 326. Tanda, Y., Proceedings of the Crop Science Society of
279. Prentice, N., Dickson, A. D., Buckhart, B. A., & Standridgc, Japan, 1962,31, 167.
N. N., Cereal Chemistry, 1963, 40, 208. 327. Tarrassov, L., Spirtovodochnaya Prommyslennost, 1939,16,
280. Raux, J., Journal of the Institute of Brewing, 1928, 24, 574.
281. Razga, Z., Brauwissenschaft, 1962, 15, 54. 328. Thunaeus, H., Wochenschrift fur Brauerci, 1938, 55, 129.
282 Reboul-Salzc, A., Journal of the Institute of Brewing, 1925, 329. Toolc, E. H., Hcndricks, J. B., Borthwick, H. A., & Toolc,
31, 632. V. K., Annual Review of Plant Physiology, 1956, 7, 299.
283. Reichard, A., Chemische Zeitschrift, 1897, 21, 21. 330. Urion, E., Brasserie, 1955,10, 81.
284. Reichard, A., Zeitschrift flir das Gesamtc Brauwesen, 1909, 331. Urion, E., & Chapon, L., American Brewer, 1955, 88, (3),
32, 145. 41, 84.
285. Reiner, L., Brauwissenschaft, 1969, 22, 47. 332. Urion, E., & Chapon, L., Brasserie, 1957, 12, 3.
286. Rcischbock, X., Journal of the Institute of Brewing, 1902, 8, 333. (Jrquhart, W. B. M., Journal of the Institute of Brewing,
749. 1953, 59, 56.
287. Rcjowski, A., & Kulka, K., Ada Societatis Botanicorum 334. van Lacr, M. H., & Duvinage, R., Bulletin de la Societe
Poloniae, 1967,36,221. Chimique de Belgique, 1923, 32, 355.
288. Reynolds, T, & MacWilliam, I. C, Journal of the Institute 335. van Laer, M. H., & Froschel, P., Biires el Biossons, 1943, 3.
of Brewing, 1966, 72, 166. 336. van Laer, M. H., Froschel, P., & Vandeshueren, J., Revue
289. Reynolds, T., Button, A. H., & MacWilliam, I. C, Journal des Fermentations et des Industries Alimentaires, 1946, 177.
of the Institute of Brewing, 1966, 72, 282. 337. van Roey, G., & Hupe, J., Bulletin de VAssociation Anciens
290. Rheinstahl Wanheim GmbH., British Patent, 1,158,428. Royale des £tudiants en Brasserie de L'Universite de
Granted. 16.7.69. Louvain, 1954, 50, 75.
291. Ricmann, J., Brauwissenschaft, 1969, 22, 81. 338. van Somcrc.C. F., Petit Journal du Brasseur, 1959, 67,799.
292. Robbins, G. S., Chesters, J., & Dickson, A. D., Proceedings 339. Verbeck-Wyndaele, R., Proceedings of the European
of the American Society of Brewing Chemists, 1963, 141. Brewery Convention, Salzburg, 1973, 75.
293. Rohde, H. W., Cereal Chemistry, 1935, 12, 610. 340. Verbeck-Wyndaele, R., & Gasper, T., Compte Rendu
294. Ruddart, M., Lang, A., & Moscttig, E., Naturwissen- Hebdomadaire des Seances de LAcademic des Sciences,
schaften, 1963, 50, 23. 1973,276, 1561.
295. Sallans, H. R., & Anderson, J. A., Canadian Journal of 341. Veerbcck-Wyndale R., van Onckclcn, H. A., & Gasper, T.,
Research, 1940, 18C, 35. Physiologia Plantarum, 1973, 29, 208.
296. Schjerning, H., Journal of the Institute of Brewing, 1910,16, 342. Veselov, I. A., Chemical Abstracts, 1942, 36, 2078.
574. 343. Veselov, I. Y., Prikladnaja Biokhimija I. Mikrobiologija,
297. Schmid, J., Journal of the Institute of Brewing, 1911,17, 171. 1972, 8, 948.
298. Schulz, K. G., Wochenschrift fur Brauerci, 1934, 51, 201. 344. Vilain, B., & Vilain, H., French Patent 1,313,200. 28.12.62.
299. Schulz, K. G., Wochenschrift fur Brauerei, 1935, 52, 118. 345. Voborsky, J., Lebensmittel-lndustrie, 1971, IB, 377, 427.
300. Schuster, K.. in Barley and Malt, Ed. Cook, A. H., Acad- 346. Ward, P., & Briggs, D. E., Journal of the Science of Food
emic Press, 1962, p. 277. and Agriculture, 1971, 22, 581.
301. Schuster, K., Die Bier Braucrci, Band 1, Die Technologic dcr 347. Wareing, P. F., Biochemical Journal, 1971, 124, 1.
Malzbercitung. Ferdinand Enkc Verlag, Stuttgart, 1963. 348. Wegmann, D., Journal of the Institute of Brewing, 1911,17,
302. Schuster, K., & Eppingcr, H., Brauwissenschaft, 1959, 12, 63.
162, 191. 349. Wellington, P. S., Annals of Botany (London), 1964,
303. Scrjban, R., Brauwissenschaft, 1963, 16, 141. n.s. 28, 113.
304. Scriban, R., Proceedings of the European Brewery Convcn- 350. Wellington, P. S., & Durharo, V. M., Annals of Botany
lion Congress, Stockholm, 1965, 36. (London), 1961, N.s. 25, 185.
305. Sebcsta, K., & Sorm, F., Chemicke Listy, 1954, 48, 1585. 351. West, S. H., Plant Physiology, 1962, 37, 565.
306. Sebcsta, K., & Sorm, F., Chemicke Listy, 1956, 50, 632. 352. Winkler, C. Brasserie et Malterie, 1911, 288.
307. Shiroya, T., Phytochemistry, 1963, 2, 33. 353. Windisch, W., Wochenschrift fur Brauerei, 1897,14, 520.
308. Shull, C. A., Botanical Gazette, 1920, 69, 361. 354. Windisch, W., Wochenschrift fur Brauerei, 1898, 15, 529.
309. Sims, R. C, Journal of the Institute of Brewing, 1959,65,46. 355. Windisch, W., Journal of the Institute of Brewing, 1900, 6,
310. Sirot, W., Monatsschrift ftir Brauerei, 1961,14, 18. 129.
311. Slight, A. M., Brewers Guild Journal, 1954, 40, 113. 356. Windisch, W., Wochenschrift fur Brauerei, 1901, 18, 546.
312. Solek, L., Kvasny Prumysl, 1962, 8, 145. 357. Windisch, W., Wochenschrift fur Brauerci, 1901, 18, 573.
313. Somlo, K. J., Journal of the Institute of Brewing, 1907, 13, 358. Windisch, W., Wochenschrift fiir Brauerei, 1902,19,81,93,
531. 359. Windisch, W., Wochenschrift fur Brauerci, 1902, 19, 769.
314. Sommer, G., Monatsschrift fiir Brauerei, 1971, 24, 205. 360. Windisch, W., Wochenschrift fiir Brauerei, 1903, 20, 61.
315. Spillanc, M. H., & Briggs, D. E., Journal of the Institute of 361. Windisch, W., Wochenschrift fur Brauerei, 1905, 22, 165.
Brewing, 1966, 72, 398. 362. Windisch, W., Wochenschrift fur Brauerei, 1913, 30, 68.
316. Srivastava, B. I., Canadian Journal of Botany, 1964,42,1303. 363. Withey, J. S., & Briggs, D. E., Journal of the Institute of
317. Srivastava, B. I., Journal of the Institute of Brewing, 1965, Brewing, 1966, 72, 474.
71,21. 364. Wiist, W., Journalofthe Institute of Brewing, 1915, 21,606.
318. Stadlaucr Malzfabrik, A. G., Austrian Patent 282,509, 365. Yomo, H., Hakko Kyo Kaishi, 1958, 16, 444.
25.6.70. 366. Yomo, H., Hakko Kyo Kaishi, 1960,18, 494.
319. Stanley, R. G., Plant Physiology, 1957, 32, 409. 367. Yomo, H., & linuma, H., Proceedings of the European
320. Stanley, R. G., Physiologia Planiarum, 1958,11, 503. Brewery Convention, Brussels, 1963, 520.
321. Stevens, R., Journal ofthe Institute of Brewing, 1958,64,470. 368. Zastrow, K., Monatsschrift fur Brauerei, 1969, 22, 325.
322. Stoddart, W. E., Graesser, F. R., & Wcsscn, J. P., Proceed- 369. Zila, V. V., Trkan, M., & Skvor, F., Wochenschift ftir
Ings of the European Brewery Convention, Vienna, 1961, 105. Brauerei, 1942, 59, 63.
323. Slopes, H., Malt and Malting, Syon, London, 1885, p. 328. 370. Zwetkova, J. S., Journal of the Institute of Brewing, 1936,
324. Tahara, S., Bulletin of Brewing Science, 1968, 14, 1. 42, 300.

Corrigendum: Fig. 1 (p. 16). For inhibition read imbibition.