254 J. Inst. Drew.. July-August, 1984. Vol. 90, pp.

254-259

A RAPID AND SIMPLE METHOD FOR THE DETERMINATION OF STARCH AND P-GLUCAN IN
BARLEY AND MALT

Bv Bhavna Ahluwaua and Elizaim-th E. Ei.i.is

(Chemistry Department. Plant Breeding Institute, Trumpington. Cambridge)

Received II S'owmber I9K3

A simple and precise method suitable for the routine determination of starch and p-glucan in barley and malt
is described. Perchloric acid (50 mM) was used to effect rapid (3 min) and exhaustive extraction of both glucans
which were then measured directly from this single extract by specific enzymic hydrolysis of the individual
glucans to glucose. The glucose was also measured enzymically. Little or no acid hydrolysis of starch or p-glucan
was observed under the extraction conditions used; most or all of the free glucose could be attributed to hydrolysis
of sucrose. Complete solubilisation of the gum and hemicellulosic components of p-glucan was achieved. Pre-
incubation of the acid extracts with protease prior to amyloglucosidase digestion resulted in higher measure
ments (approximately 4% w/w) of starch. The method was used to measure the levels of starch and p-glucan
in five varieties of barley with contrasting malting quality, in micro-malts prepared from these samples and in
commercial lager and ale malts.

Key words: perchloric acid, starch, p-glucan. barley, malt.
Introduction
The a-glucan, starch and the mixed linkage p-glucan of
starchy endosperm comprise the bulk of the polysaccharides
present in the barley grain. In addition to starch being the
main source of carbohydrate for the germinating embryo, it
is also the major source of brewer's extract. The (l-*3),
(l-»4) p-n-glucan (hereafter referred to as P-glucan). when
present in elevated levels can not only influence the rate at
which a barley modifies, but can also produce highly viscous
solutions which can lead to problems in the brewing pro
cess.4" High concentrations of P-glucan also adversely affect
the feed quality of barley." Thus a simple and reliable
method for the estimation of these glucans would be of value
in predicting the suitability of barley for malting and for feed
purposes and in breeding programmes aimed at lowering the
p-glucan and/or increasing the starch content of barley.
The methods available for the determination of starch are
not entirely satisfactory. This is due to difficulties in extrac
tion or gelatinisation and defects in assaying the extracted
starch. Extraction with hot water26 or chloral30 is inadequate
because of low recoveries of starch. Dimethyl sulphoxide
will extract starch from cereals" without loss of the integrity
of starch granules, but several extractions arc necessary to
achieve complete extraction. Perchloric acid (52 or 72% w/v)
has been found to be the most efficient solvent for starch
from plant tissues and has been used for the extraction of
starch from barley14"29 and from wheat." However, even
with this very high concentration of perchloric acid, dissolu
tion of material was only effected by stirring for 50 minM or
by inclusion of sand to facilitate the extraction "
Both non-specific and specific methods have been pro
posed for estimating starch. The non-specific methods
include precipitation with iodine"27-21* and measurement of
the blue value.29 This procedure can be inaccurate because
the iodine binding capacity of amylose is reduced in the pres
ence of other molecules31 and also because the blue value
varies with amylose content in starch.26 Hydrolysis and
measurement of glucose by anthrone in hot, concentrated
sulphuric acid is not only inconvenient but can be inaccurate
due to colour reactions with other sugars.26 Specific measure
ments of starch involve enzymic hydrolysis to glucose with
amyloglucosidase (AMG),1012 but steps must be taken to
eliminate any p-glucanase activity. Methods in which the
difference between total carbohydrate and reducing sugars is
considered as starch" do not represent an accurate alterna
tive because other glucans, e.g. cellulose and p-glucan, are
not taken into consideration.
Similarly, extraction procedures for p-glucan have also
proved unsatisfactory, primarily due to inadequate cxtrac-
methods determined only the P-glucan soluble in water at
40 or 65°C thus not accounting for water insoluble and alkali
or urea soluble components.4 Moreover, methods involving
precipitation by ammonium sulphate (30% w/v) do not pre
cipitate all the p-glucan1 and extraction under alkaline con
ditions'' docs not necessarily increase the solubility of the
hemicellulosic component.
Since measurements of p-glucan now mainly rely on direct
estimation by enzymic hydrolysis"1M any extraction pro
cedure should effect maximum solubilisation of the P-
glucan. Despite the autoclavingstep introduced by Anderson
et aix some p-glucan may still remain inaccessible to the
enzyme due to interactions with protein16 or to covalent
cross linking.1 Complete release of p-glucan into solution
was apparently achieved by treatment of barley flour with
hydrazinc16 and more recently this solvent28 has been used
for the determination of total P-glucan. However, in addition
to a 40 h extraction, the removal of hydrazinc by dialysis
(20 h) involves the loss ofP-linked oligosaccharides.
Any improved method should thus exhaustively extract
starch and p-glucan in a form accessible to enzymic attack.
In the proposed method perchloric acid56" has been
exploited for the simultaneous extraction of starch and
P-glucan from barley and malt. This treatment also dena
tures endogenous hydrolytic enzymes which would modify
the size and solubility of the two glucans. The individual
glucan contents were then determined enzymically from
buffered aliquots of the acid extract by minor modifications
to established procedures.
EXPERIMENTAL
Materials.—Wheat starch (Sigma Chemical Ltd). This
contained 900 mg glucose/g dry weight. Barley P-glucan
(Biocon UK Ltd) contained 960 mg glucose/g dry weight.
Cellulase from Ponicillium funiculosum was obtained from
J. E. Sturge Ltd; AMG, hexokinase/glucose-6-phosphate
dchydrogenase and phosphoglucoisomerase from Boehringer
Corporation, London and protease (Type XIV essentially
starch free) and porcine pancreatic a-amylase from Sigma
Chemical Co. Ltd. Barley samples were obtained from trials
grown at Ickleton near Cambridge. The varieties Ark Royal
(% w/w nitrogen, %N= 1-49, NIAB Malting Grade, MG=8)
and Triumph (%N=l-52, MG=9) were selected to rep
resent good malting varieties and Georgie (%N=l-53,
MG=I) and Egmont (%N=l-62, MG = 1) were chosen as
examples of poor malting types. Varunda (%N = l-90,
MG= I), also a poor malting variety, was a gift from Mr
A. P. Rhodes, Miln Marsters, Norfolk. Micro-malts were
prepared and kilned from each of these barleys as described
by Gothard el at.1* The lager and ale malts were a gift from
DrC. W. Bamforth, Bass PLC. Grain and malt samples were
\inr\ in n I IPtV mill fitt/»H u/ith <t ft.^mm ci*»v<»
Vol. 90, 1984] AHLUWALIA AND ELLIS: A MI-TIIOD I-OR STARCH AND p-GLUCAN
Endosperm cell walls were isolated from pearled and milled
grain of barley varieties Ark Royal and Varunda, essentially
by the method of Palmer" using repeated maceration of the
flour in 70% (v/v) cthanol. Cell walls were dried in vacuo at
40°C after three exchanges in alcohol and ether. To deter
mine the monosaccharide concentration in the cell walls,
5 mg was hydrolyscd with I m H2SO4, neutralised with
barium hydroxide and the acctylated sugars were converted
to alditol acetate derivatives before analysis by gas-liquid
chromatography (GLC).3* Chromatography was conducted
on a Pye Unicam 204, fitted with a 2-8 mx2-2 mm column
containing 3% OV-225 on JJ's diatomite CQ (100-200
mesh).
Methods.—
Extraction of p-glucan and starch.—Barley and malt
samples (20 mg) were incubated in covered test tubes with
50 mM perchloric acid (40 ml) at 96°C for 3 min. After cool
ing, the extract was ccntrifuged, the pellet washed with cold
50 mM perchloric acid (I ml) and re-centrifuged. The supcr-
natants were bulked and made to a volume of 10 ml with
distilled water to give the extract for p-glucan and starch
determination. Wheat starch (6 mg) and barley P-glucan
(2 mg) were separately included in each batch to verify com
plete recovery. Malt extracts were dialysed against 0-02%
sodium azide at room temperature for 2 h 30 min.
P-Glucan determination.—A mixture of 250 ul of barley
or dialysed malt extract, 650 ul of sodium acetate buffer
(0-2 m, pH 5-4) and 100 ul of'treated1 cellulase from Peni-
cilliumfuniculosum'' was incubated at 40°C for 2 h. Appro
priate substrate and enzyme blanks were included to correct
for any free glucose not emanating from p-glucan. Check
samples of starch tested under identical conditions as p-
glucan showed the 'treated' cellulasc to be free of any AMG
activity. Glucose released by cellulasc action was measured
enzymically, in duplicate, using hexokinase/glucosc-6-
phosphate dehydrogenase.828 Results were expressed as
percentage P-glucan of the dry weight of the barley and malt.
Starch determination.—Protease (2-5 mg/ml) was dis
solved in 0-2 m Tris, 0-1 m NaCl (pH 7-2) and self digested
for 30 min at 30°C. Barley or dialysed malt extracts (250 ul)
were incubated with 200 ul of the protease and 200 ul of
0-2 m Tris (pH 7-2) at 30°C for 30 min. After this incubation
period the digests were boiled for 5 min to inactivate the pro
tease. Aliquots (100 ul) from the protease digest were made
to 10 ml with sodium acetate buffer (0-2 m, pH 4-8). AMG
(2-8 U)10 was added and the mixture incubated at 55°C for
40 min. The same substrate blanks used to correct P-glucan
measurements were also used to correct for any free glucose
not emanating from starch. The AMG preparation used
did not hydrolyse Biocon barley p-glucan under these con
ditions. The glucose released by hydrolysis of starch was
estimated as before, in duplicate, using hexokinase/glucosc-
6-phosphate dehydrogenase. The results were expressed as
percentage starch of dry weight.
Determination offree glucose andfructose in the perchloric
acid extracts.—Extracts of barley and malt (undialysed
100 ul) were each incubated with sodium acetate buffer, pH
4-8 (900 ul) at 55°C for 2 h (as for starch determination)
and pH 5-4 at 40°C for 2 h (as for P-glucan determination).
Fructose and glucose were measured using hcxokinasc/G-6-
phosphate dehydrogenase with and without the addition
of phosphoglucoisomerase8-9 respectively. Temperature
and pH had no influence on the glucose and fructose
concentrations measured.
Gel filtration chromatography.—Characterisation of the
neutralised perchloric acid extracts of endosperm cell walls
255
(5 mg), barley (30 mg) and malt (30 mg) (undialysed and
dialysed), all from variety Varunda, was carried out on a
12 x 1 -5 cm column of Sephacryl S-300. Neutralised extracts
(500 ul) were loaded onto the column and eluted with 001 m
NaCl, 002 M Tris, 002% (w/v) sodium azide pH 7-2. Void
and totally included volumes were determined using Blue
Dcxtran 2000 (0-5% w/v) and potassium dichromate (0-1%
w/v) respectively. P-Glucan and starch in the fractions were
determined from 500 ul and 250 ul aliquots respectively as
described above.
Rksults and discussion
Development ofthe method.—The extraction of starch and
P-glucan from barley and malt with 50 mM perchloric acid
at 96°C for 3 min yielded as much p-glucan and starch as
after extraction for 10 min (Fig. 1). Re-extraction of the pel
let with 50 mM perchloric acid for a further 3 min at 96CC
did not yield any additional glucans. Therefore the dilute
acid at the higher temperature effected extremely rapid and
maximum extraction of both starch and P-glucan from bar
ley and malt, without the lengthy extraction procedures
reported previously with perchloric acid.14-27 Indeed the
extraction of the glucans was markedly quicker than any
reported earlier using other solvents, e.g. extraction with
hydrazine (40 h),28 with sodium carbonate buffer (20 h)35 or
by autoclaving (I h).1 The use of sodium acetate buffer
enabled direct and rapid measurements of starch and P-
glucan from the acid extract without any further need to
neutralise the perchloric acid. Under these conditions re
covery of Biocon p-glucan was 99-4 + 3-2% and wheat
starch 98-1 ±2-4%.
Glucans are susceptible to proton catalysed hydrolysis, so
any extraction procedure using acid may result in degrada
tion of starch or p-glucan. Thus the increasing concentration
of free glucose with extraction time in barley and malt
extracts (Fig. 1) may have resulted from hydrolysis of starch
and/or p-glucan, but would be considered in the substrate
blanks as glucose of non-glucan origin. Indeed about 2-3%
(w/w) of wheat starch and <0-5% of barley P-glucan was
hydrolysed by perchloric acid after 10 min treatment (Fig. 2).
However, over the proposed extraction time of 3 min only
0-2% of starch was degraded and <0-l% of P-glucan was
detectable as free glucose. No free glucose was measured in
extracts of endosperm cell walls. Since cell walls arc the
major source of p-glucan, this indicated no acid hydrolysis
of the native polymer (Fig. 2).
However, a small amount of glucose, equivalent to about
1-5% (w/w) in barley and about 4% (w/w) in malt in the five
varieties, was released by the proposed perchloric acid
extraction (Table I). Only nominal levels of glucose exist in
ungerminated barley'1'-24 and very little glucose was released
by acid hydrolysis of the two polysaccharides. The levels of
free fructose, which were greater than those of free glucose
in the extracts, (Table I) suggested that sucrose (subject to
acid hydrolysis) was the main source of this free glucose. The
additional fructose was probably derived from raftinosc, the
next abundant sugar to sucrose;24 and from fructosans in
barley.25 The levels of free fructose which were similar to
the levels of free glucose in the undialysed acid extracts of
malt, once again suggested sucrose as the main source of the
free glucose (Table I). No additional free fructose was
obtained from raffinose since it completely disappears during
malting.25
The sucrose levels in the five varieties ranged between 2
and 3% (w/w) in barley and 6-5 and 9% (w/w) in malts, the
lowest concentration being that in the variety Varunda,
reflecting less development of the rootlets and plumule.19
That higher absolute levels than those reported previously
were measured here is probably due to the more sensitive
methods used and the different malting regime. Certainly,
Macleod el al.2i reported the final sucrose content in malt
256 AHLUWALIA AND ELLIS: A METHOD FOR STARCH AND P-GLUCAN
-*-*—*•
I 500-
1 and husked barley (Table II) determined by the proposed
O
a 200'
joo;
«-*-"?
2 1 6 8 10
Time (mini
Fig. 1. The extraction of starch and P-glucan and appearance of
glucose as a function of time. Barley (closed symbols) and malt
(open symbols), variety Ark Royal (10 mg) was treated with
perchloric acid (50 mM, 2 ml) at 96°C and starch (•, O),
p-glucan (■, D) and free glucose (A, A) were determined as
described in Methods. The results represent means ± standard
errors of 4 separate experiments.
25-
20-
lfl
Time (min)
Fig. 2. The perchloric acid hydrolysis of starch, P-glucan and iso
lated endosperm cell walls as a function of time. Wheat starch
(•, 8 mg), P-glucan (■, 5 mg) and cell wall (A, S mg) were
hydrolysed with perchloric acid (50 mM) at 96°C and free
glucose released determined as described in the Methods.
to be some five times that of barley. To avoid errors in
measurements arising from the higher concentrations of free
'non-glucan' glucose; the malt extracts were dialysed before
incubation with AMG or cellulase. This yielded levels of the
large molecular weight p-glucan (> 12-15 glucose residues),
sometimes referred to as 'troublesome P-glucan', remaining
in malt. Similarly additional information as to the level of
this P-glucan in barley can be obtained on dialysis of the
respective acid extracts.
[J. Inst. Brew.
Alternatively, apparently high values of p-glucan might
result from glucose released by cellulase from the husk cellu
lose. Husk, hand dissected from barley and extracted with
50 mM perchloric acid for 3 and 10 min, produced no glucose
with or without cellulase incubation. Close agreement of the
p-glucan contents of dehusked (in 50% (v/v) sulphuric acid)
method also confirmed that cellulose from the husk did not
contribute to the p-glucan measurements.
Since the most meaningful methods for estimation of total
p-glucan content arc those that measure both gum and hemi-
cellulose fractions, complete extraction of P-glucan from
isolated barley endosperm cell walls by the proposed
perchloric acid method, positively established solubilisation
of the hemicellulosic component as well. The concentration
of p-glucan in the cell walls of varieties, Ark Royal (71-2%
w/w) and Varunda (73-3% w/w) obtained by perchloric acid
extraction agreed extremely well with that determined
by GLC of alditol acetates of the total cell wall mono-
saccharides (Table HI) in the present investigation and that
reported by other workers.216 Of the extraction methods
reported to date only that using hydrazine completely dis
solves hemicellulose, but as indicated above this method
involves extensive dialysis and is therefore inconvenient for
routine determinations.
Exhaustive extraction of starch by the proposed method
was confirmed by negative iodine staining of the extracted
pellet. Complete enzymic hydrolysis of the extracted starch
is essential for accurate measurements. However, protein
associated with both the large and small starch granules
in barley has been shown to limit enzymic attack.39 The
granules isolated from malt also have some associated pro
tein although less than that found in barley.34 Pre-incubation
of the perchloric acid extracts with protease was found to
increase the recovery of starch from both barley and malt
when measured by AMG (Table IV). Pre-incubation of the
acid extracts with pancreatic a-amylase (0-5 tng/ml in 0-2 m
Tris buffer pH 7-2) in the presence of NaCI (0-1 M) and
calcium acetate (0-02 m) at 25°C for 30 min resulted in
no increase in the recovery of starch and is therefore not
necessary.
Gel filtration chromatography was used to investigate the
effect of perchloric acid treatment on size of the native p-
glucan and starch. Characterisation of the perchloric acid
extracts of the isolated endosperm cell walls on Scphacryl
S-300 (Fig. 3) showed that all of the cell wall p-glucan eluted
with the void volume, indicating little if any hydrolysis of
the p-glucan polymer. Reports of the size of native P-glucan
vary4 and are confounded by the presence of cross linking
between molecules, but exclusion from S-300 suggests the
extracted glucans have a degree of polymerisation of > 2000.
The p-glucan extracted from barley was resolved into two
size classes (Fig. 4), most of the P-glucan being of high
molecular weight, and the remainder probably representing
native p-linked oligosaccharides which eluted before the
total column volume. Similarly, P-glucan extracted from
malt resolved into two size classes (Fig. 4), but, in this case
a smaller proportion was in the high molecular weight frac
tion. Most of the low molecular weight P-linked oligosac
charides from malt were dialysable after 2 h 30 min (Fig. 4)
and were probably the hydrolytic products of p-glucanases
produced during malting.5 The undialysable oligosac
charides may be typical only of the poor malting barley
variety, Varunda, investigated here.
Almost all of the starch which was extracted from barley
by perchloric acid elutcd with the void volume (Fig. 5). The
'long shoulder' of high molecular weight dextrans may
suggest some hydrolysis, supporting the observation of
greater susceptibility of starch to acid hydrolysis than p-
glucans (Fig. 2). In contrast, starch extracted from the malt
separated into two fractions according to size (Fig. 5), a small
proportion being of low molecular weight and eluted with
Vol. 90, 1984] AIII.UWAI.IA AND I-I.I.IS: A METHOD FOR STARCH AND [1-GI.UCAN 257

TABLE I. Levels of Free Glucose and Fructose after Perchloric Acid

Extraction of Five Varieties of Barley and Malt

Variety Free glucose (mg) Free fructose (mg)

026'
Barley
Ark Royal 013 ±0008 0-21 ±0011
Triumph 014 ±0008 0-24 ±0008
Georgie 013 ±0007 018 ±0008 020

Egmont 0l7±00ll 0-21 ±0018

Varunda 010 ±0009 O-I8±OO12
s
Malt I 0 15- VI
Ark Royal 0-43 ±0008 0-45+0015 1
Triumph 0-42 ±0021 0-45+0021
Georgie 0-42 ±0015 0-44 ±0006
Egmont 0-47 ±0004 0-5O±0021
Varunda O-35±OOI6 0-3O±0015 0 10

Barley and malt samples (10 mg) were treated with perchloric acid
(50 mM, 2 ml) for 3 min at 96°C and free glucose and fructose were 005-

determined as described in the Methods. Results are means ± stan

dard error of 5 separate experiments.

10 IB 20
TABLE II. p-Glucan Content in Husked and Dehusked Barley Fraction number

Fig. 3. Molecular weight patterns of P-glucan in perchloric acid

% (w/w) p-Glucan content extract of barley, variety Varunda, endosperm cell wall as frac
tionated on Scphacryl S-300. The sample was prepared and
Variety Husked Dchusked loaded (200 ug glucose equivalent) as described in Methods and
eluted with 0-01 m NaCl, 002m Tris, 002% sodium azide
pH 7-2.
Ark Royal 4-3 + 0-21 40±015
Varunda 57 ±030 5-5 ±0-22

Barley was dehusked in 50% v/v sulphuric acid and B-glucan

extracted and estimated as described in Methods. 013

D11
TABLE III. Determination of p-Glucan Content of Isolated Barley
Endosperm Cell Walls
0 09'

P-Glucan as mg equivalents of
glucose/100 mg dry weight cell wall
0 07
Vt
1
Perchloric acid
Variety extraction method GLC

Ark Royal 7l-2±2-47 69-8 0O3

Varunda 73-3 ±1-51 75-3

P-Glucan was estimated by (i) perchloric acid extraction (3 separate 001

experiments) and (ii) GLC 6f alditol acetates in two barley varieties

as described in Methods and Materials. 10 IS 20
Fraction number

TABLE IV. Effect of Protease Incubation prior to AMG Digestion Fig. 4. Molecular weight patterns of p-glucan in perchloric acid
on the Starch Content in Five Varieties of Barley and Malt extracts of barley and malt as fractionated on Sephacryl S-300.
Barley (•) and malt, undialyscd (A) and dialyscd (A) samples,
variety Varunda, were prepared and loaded (90 and 60 ug glu
% (w/w) Starch cose equivalents for barley and malt respectively) as described.
in Methods and eluted with 001 M NaCl, 002m Tris, 002%
sodium azide pH 7-2. The absorbancc of free glucose cluting at
Variety AMG Protease+AMG Vt, representing the substrate blank was subtracted from the
rest.

Barley
Ark Royal 63-1 ±2 23 67-5 ±2-10
Triumph 621 ±1-84 66-4 ±204 the dialysable (i-linked oligosacchandcs. These represent
Georgie 59-3 ±1 64 63-8±l-5l some of the products of the starch-degrading enzymes during
Egmont 570 ±201 61-1 ±2-6 malting.34 Thus perchloric acid treatment almost certainly
Varunda 56-8±lll 60-2 ±2-10
Mai x
did not significantly alter the size of the polymers, by proton
1V1AI.I

Ark Royal 52-2± 118 550±l-28

catalysed hydrolysis, the malt oligosaccharides representing
Triumph 49-5 ±2-17 54-5 ±0-69 the hydrolytic products of enzymes during modification.
Georgie 53-6± 118 560 ±2-13
Egmont 52-3 ±209 550 ±2-96
Varunda 47-8 ±2-41 52-5 ±2-30 Application of the method.—Of the methods reported to
date, no single extraction procedure has been used for the
Barley and malt extracts (250 til) were pre-incubatcd with protease
determination of both barley a and P-glucans. The proposed
prior to AMG digestion and the starch content compared with the
respective extracts (100 ul) digested with AMG (2-8 U) only as method with perchloric acid, coupled with minor modifica
described in Methods. Results represent means ± standard error of tions to the already published enzymic hydrolysis of {$-
3 separate experiments. glucan6 and starch,10 was both reproducible and precise. The
258 AHLUWALIA AND ELLIS: A MI-TltOI) FOR STARCH AND p-GLUCAN [J. Inst. Brew.
Vo
mean standard error of the five barley varieties was ±019
1
for p-glucan and ±2-75 for starch. Equivalent standard
errors for the malt samples were ±0-24 and ±2-25. These
values, together with the measured concentrations (Table V)
show the precision of the method.
Since neither extensive ethanol extraction1 -M nor dialysis28
of low molecular weight carbohydrates was necessary for
barley at any stage, the total P-glucan measurements
reported here (Table V) represent the total including the
P-linked oligosaccharides (Fig. 4). The total P-glucan con
tents of the barleys ranged between 4-3% (w/w) in Ark Royal
Vt and 6-0% (w/w) in Varunda (Table V). The varieties con
e 06' 1 sidered to be best suited for malting tended to have the lower
P-glucan contents. Although genotypic variations in barley
p-glucan have been reported previously,"728 comparisons
with these values are not strictly valid as different methods
were used and glucan concentration might also be influenced
by environment.30
The p-glucan contents of the micro-malts ranged between
1 -7% (w/w) in Triumph and 3-9% (w/w) in Varunda; with
the content of lager and ale malts being 1-6 and 1-5% (w/w)
respectively. The amount of P-glucan remaining in malt also
5 10 15 20 depended on variety3 and barleys rich in p-glucan and of
Fraction number
low malting potential, e.g. Varunda, gave malts with high
Fig. 5. Molecular weight patterns of starch in perchloric acid P-glucan levels (Table V). Thus the extent to which p-glucan
extracts of barley and malt as fractionated on Scphacryl S-300. was degraded by various enzymes during malting in the five
Barley (•) and malt, undialyscd (A) and dialyscd (A) samples, varieties could be monitored.
variety Varunda, were prepared and loaded (900 and 750 ug
The total starch content of the barleys ranged between
glucose equivalents for barley and malt respectively) as
described in Methods. The absorbancc of free glucose eluting 57% (w/w) in Varunda and 68% (w/w) in Triumph (Table
at Vt, representing the substrate blank was subtracted from the VI). Although these values agree in general with those pub
rest. lished previously21-40 they represent more accurate and
specific measurements since no additional glucose was
measured from the enzymic hydrolysis of p-glucan. Hence,
the varieties with a high malting potential, Ark Royal and
Triumph, tended to have higher starch contents and thus
TABLE V. B-Glucan Content of Barley and Malt as Determined by higher potential for a brewer's extract. A varietal influence
Perchloric Acid Extraction Method
on starch content in barleys has been reported previously.21 -40
During malting the decrease in starch content varied
(i-Glucan content % (w/w)
between 5% in Varunda and 15% in Triumph (Table VI),
with the maximum degradation occurring in good malting
Variety Barley Malt types. In comparison the starch content consumed during a
9 day flooring period of malt varied between 15 and 18% for
Ark Royal 4-310-11 1-910-21 several varieties.1* In the present investigation barley was
Triumph 4-4 ±0-30 1-7+0-21 micro-malted for 5 days.
Georgie 5-6+0-15 2-710-25 The extraction method developed is well suited for routine
Egmonl 5-4 ±009 3-3 ±0-23 analysis since it provides a rapid means of specifically deter
Varunda 60 ±0-30 3-9 ±0-31
Lager l-6±0-25
mining both starch and p-glucan in barley and malt directly
Ale 1-5 ±008 from a single extract. No significant acid hydrolysis of the
glucans occurred and solubilisation of both gum and hemi-
B-Glucan in barley and malt was extracted and estimated as cellulosic components of P-glucan and complete extraction
described in Methods. Results represent a mean ± standard error of of starch was achieved.
10 separate experiments.

Acknowledgements.—The authors wish to acknowledge

D. B. Smith and T. J. Riggs for critical evaluation of the
TABLE VI. Starch Content of Barley and Malt as Determined by manuscript. D. E. Quain, C. W. Bamforth, M. P. Cochrane
Perchloric Acid Extraction Method and F. Renwick arc thanked for valuable discussions. We are
grateful to the HGCA for funding the project.
Starch content % (w/w)

Variety Barley Malt

References
1. Anderson, M. A., Cook, J. A. & Stone, B. A., Journal of the
Institute of Brewing. 1978, 84, 233.
Ark Royal 66-4 ±3-II 53-911-34 2. Ballance, G. M. & Manners, D. J.. Carbohydrate Research.
Triumph 68-2 ±305 53012-81 1978,61,107.
Georgie 63-1 ±2-81 54111-81 3. Bamforth, C. W. & Martin, H. L., Journal of the Institute of
Egmont 62-4±2-19 53-412-30 Brewing. 1981,87,365.
Varunda 57-3 ±2-61 52-013-01 4. Bamforth, C. W., Brewers Digest. 1982, 57, 22.
Lager 55-812-11 5. Bamforth, C. W. & Martin, H. L., Journal of the Institute of
Ale 51110-50 Brewing. 1983.89,303.
6. Bamforth, C. W., Journal of the Institute of Brewing. 1983. 89,
Starch in barley and malt was extracted and estimated as described 391.
in Methods. Results represent means 1 standard error of a minimum 7. Bass, E. J. & Meredith, W. O. S.. Cereal Chemistry. 1955. 32,
of 10 separate experiments. 374.
Vol. 90. 1984] AHI.UWAI.IA AND I-I.I.IS: A MI-THOI) I OR STARCH AND (!-GI.UCAN
8. Bergmcycr, H. U.. Bernt. E., Schmidl. F. & Stork, H.. Methods
of Enzvmatic Analysis, cd: H. U. Bergmeyer. Academic Press:
London. 1974. 3. 1196-1201.
9. Bcrnl, R. & Bergmeyer. H. U.. Methods of Enzymatic Analysis.
cd: H. U. Bergmeycr. Academic Press: London. 1974. 3.
1304-1307.
10. Beullcr. H. O.. StiirkeStarch. 1978. 30. 309.
11. Burnctl. G. S.. British Poulirv Science. 1966. 7. 55.
12. Chiang. B Y. & Johnson. J. A.. Cereal Chemistry. 1977. 54,
429.
13. Chu, L-J. C. & Shannon. J. C, Crop Science . 1975. 15, 814.
14. Clegg, K. M.. Journal of the Science of Food and Agriculture,
1956.7,40.
15. Finchcr. G. B., Journal of the Institute of Brewing. 1975. 81,
!I6.
16. Forrest. I. S. & Wainwright. T.. Proceedings of the huropean
Brewinz Convention Congress. Amsterdam. 1977. 401.
17. Gill. A. A., Morgan. A. G. & Smith. D. B.. Journal of the
Institute of Brewing. 1982.88, 317.
18. Gothard, P. G.. Morgan. A. G. & Smith. D. B.. Journal of the
Institute of Brewing. 1980, 86, 69.
19. Hall. R. D., Harris. G. & MacWilliam. I. C. Journal of the
Institute of Brewing. 1956.62. 232.
20. Harris. G. & MacWilliam. I. C, Journal of the Institute of
Brewing. 1954.60,387.
21. Harris. G.. Barlev and Mall, ed: A. H. Cook, Academic Press:
London, 1962.435.
22. Jenkins. L D.. Loncy. D. P. & Meredith. P.. Cereal Chemistry.
1974.51,718. ' 39. Slack. P. T. Baxter, E. D. & Wainwrighl, T. J.. Journal of the
23. Jcnner, C. F., Australian Journal of Biological Science. 1968.
21,597. ' 40. Torp. J., Journal of the Science of Food and Agriculture. 1980,
24. MacLeod, A. M., Journal of the Institute of Brewing. 1952, 58.
363.
259
25. MacLeod. A. M.. Travis. D. C. & Wreay. D. G.. Journal of the
Institute of Brewing. 1953.59, 154.
26. Macrae. J. C. & Armstrong. D. G., Journal of the Science of
Food and Agriculture. 1968. 19, 578.
27. MacWilliarri. I. C, Hall. R. D. & Harris. C... Journal of the
Institute of Brewing. 1956, 62, 226.
28. Martin. H. L. & Bamforth. C. W.. Journal of the Institute of
Brewing. 1981.87.88.
29. Mcrrit. N. R. & Walker. J. T.. Journal of the Institute of
Brewing. 1969.75. 156.
30. Morgan, A. G. & Riggs, T. J., Journal of the Institute of
Brewing. 1981.87,88.
31. Morrison, N. R. & Laignclct, B.. Journal oj Cereal Science.
1983,1,9.
32. Narziss. L., Proceedings of the European Brewing Convention
Barlev and Malt Symposium. Helsinki. 1981.99.
33. Palmer, G. H.,' Proceedings American Society Brewing
Chemists. 1975.33, 174.
34. Palmer. G. H. & Bathgatc, G. N.. Advances in Cereal Science
and Technology, ed: Y. Pomeranz, American Association of
Cereal Chemistry Inc: Minnesota, 1976, Volume 1,237-324.
35. Prentice. N.. Babler. S. & Faber. S.. Cereal Chemistry. 1980.57.
198.
36. Pucher. G. W.. Leavenworth. C. S. & Vickcry. H. B.. Analytical
Chemistry. 1948,20,850.
37. Quain, D. E., Journal ofthe Institute ofBrewing, 1981,87,289.
38. Selvendran, R. R.. March. J. F. & Ring, S. G., Analytical
Biochemistry. 1979, 96, 282.
Institute of Brewing. 1979.85, 112.
31, 1354.