ARTICLE IN PRESS

LWT 40 (2007) 860–870

www.elsevier.com/locate/lwt

Optimization of ingredients and baking process for

improved wholemeal oat bread quality
L. Flander, M. Salmenkallio-Marttila, T. Suortti, K. Autio
VTT Biotechnology, P.O. Box 1500, 02044 VTT, Finland
Received 24 October 2005; received in revised form 28 April 2006; accepted 2 May 2006

Abstract

Baking technology for tasty bread with high wholemeal oat content and good texture was developed. Bread was baked with a straight
baking process using whole grain oat (51/100 g flour) and white wheat (49/100 g four). The effects of gluten and water content, dough
mixing time, proofing temperature and time, and baking conditions on bread quality were investigated using response surface
methodology with a central composite design. Response variables measured were specific volume, instrumental crumb hardness, and
sensory texture, mouthfeel, and flavour. The concentration and molecular weight distribution of b-glucan were analysed both from the
flours and the bread. Light microscopy was used to locate b-glucan in the bread. Proofing conditions, gluten, and water content had a
major effect on specific volume and hardness of the oat bread. The sensory crumb properties were mainly affected by ingredients,
whereas processing conditions exhibited their main effects on crust properties and richness of the crumb flavour. b-glucan content of oat
bread was 1.3/100 g bread. The proportion of the highest molecular weight fraction of b-glucan was decreased as compared with the
original b-glucan content of oat/wheat flour. A great part of b-glucan in bread was located in the large bran pieces.
r 2006 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.

Keywords: Oat bread; Baking; Optimization; b-glucan

1. Introduction heat treatment, do not possess the unique visco-elastic

Bread is an ideal functional food product, since it is an
important part of our daily diet. The taste of oat bread is
nutty, mild, and pleasant, and it could compete successfully
as a healthy alternative to consumers, who are used to
eating white wheat bread. Oat has excellent moisture
retention properties that keep breads fresher for longer
periods of time (McKechnie, 1983). Addition of oat, oat
starch, or oat lecithin to wheat bread might retard also the
staling rate of the bread (Forssell, Shamek, Härkönen, &
Poutanen, 1998; Prentice, Cuendet, & Geddes, 1954;
Zhang, Moore, & Doehlert, 1998).
The main problem in the use of oat in higher quantities is
the inferior baking quality (Brümmer, Morgenstern, &
Neumann, 1988; Gormley & Morrissey, 1993; Oomah,
1983), because oat proteins, normally denaturated by a
Corresponding author. Tel.: +358 20 722 5842; fax: +358 20 722 7071.
E-mail address: laura.flander@vtt.fi (L. Flander).
properties of wheat gluten. An addition of 20 g oat/100 g
wheat flour allows the bread to be labelled as ‘‘oat bread’’
in Germany (Brümmer et al., 1988). Exceeding this amount
of oat leads easily to tight, moist, and gummy breads. The
effect of oat on dough properties and bread quality have
been studied so far with addition of oat bran, flakes or
flour from 10 to 25/100 g wheat flour (Degutyte-Fomins,
Sontag-Strohm, & Salovaara, 2002; Krishnan, Chang, &
Brown, 1987; Oomah, 1983; Zhang et al., 1998). In all these
cases, breads with 10 g oat/100 g wheat flour had better loaf
volume, crumb grain, and texture than breads with 15–25 g
oat/100 g wheat flour. Brümmer et al. (1988) attained good
quality of oat bread by adding 20 g oat flakes/100 g wheat
flour, although the bread volume was 10% smaller than
volume of the control white wheat bread.
The poor baking performance of oat can be compen-
sated for by adding dry gluten into the dough (Gormley &
Morrissey, 1993). If the amount of added fibre is higher
than 10 g oat flour/100 g wheat flour, a flour protein

0023-6438/$30.00 r 2006 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.lwt.2006.05.004
 ARTICLE IN PRESS
L. Flander et al. / LWT 40 (2007) 860–870 861

content of about 16/100 g flour is recommended and the 2. Materials and methods
amount of water used should be sufficient to properly
hydrate both the added gluten and the fibre components 2.1. Flour properties
(Stear, 1990). Gluten strengthens the protein matrix and
enhances the structure of oat bread. According to Oomah Oat flour (Helsingin Mylly Ltd., Järvenpää) and white
and Lefkovitch (1988), loaf volume of wheat breads wheat flour (Raisio Group plc, Raisio) were obtained from
containing up to 15 g oat flour/100 g flour can be optimized Finnish mills. White wheat flour contained ascorbic acid.
by a sufficiently high water level (68–72/100 g flour) in the All chemical analyses of the flours were made in duplicate
dough. (Table 1). Moisture content was determined according
Whole grain oat contains high amounts of valuable to standard method 44/15A, and wet gluten with method
nutrients such as soluble fibres, proteins, unsaturated fatty 38/12A (AACC, 2000). Protein content was determined by
acids, vitamins, minerals, and phytochemicals. The dietary the Kjeldahl method (AACC standard 46/11A), and ash
fibre complex with its antioxidants and other phytochem- content was analysed according to standard method
icals may protect us from cardiovascular disease and 08-01 (AACC, 2000). The sieve analysis of oat flour was
some types of cancer (Jacobs, Marquart, Slavin, & Kushi, carried out by sieving two 100 g portions of the flour for
1998a, b; Slavin, Marquart, & Jacobs, 2000; Thompson, 10 min.
1994). Whole grain oat contains significant amounts
of dietary fibre and especially water soluble (1-3), 2.2. Experimental design
(1-4)-b-D-glucan. The b-glucan content in oat (Avena
sativa) varies between 2.3 and 8.5/100 g (Welch, Brown, & A fractional factorial design was used to screen the most
Leggett, 2000). On the basis of numerous clinical studies, important factors (ingredients and processing conditions)
the US Food and Drug Administration (FDA) permitted affecting the specific volume, instrumental crumb hardness
the use of a claim that oat-soluble fibre has the ability to and sensory properties of the breads and to choose the
reduce the risk of coronary hearth disease (FDA, 1996). most significant ones and their appropriate range for
The required dose of b-glucan for a single food is optimization tests (data not shown). The specific volumes
0.75 g/serving. The highly viscous b-glucan fraction of oat of these breads varied from 1.9 to 3.1 cm3/g and their
has been related to the ability to lower blood cholesterol instrumental crumb hardness after 2 and 72 h varied
and the intestinal absorption of glucose (Mälkki, 2001; between 0.2 and 0.4 kg, and between 0.2 and 0.7 kg,
Wood, 1993). In order to be physiologically active and respectively.
form viscous solutions in the gut, b-glucan must be soluble,
and the concentration and molecular weight must be
2.2.1. Optimization of ingredients
sufficiently high (Åman, Rimsten, & Andersson, 2004;
A central composite face-centered design (CCF) was
Autio, Myllymäki, & Mälkki, 1987; Doublier & Wood,
used with two variables and four replicates at the centre
1995). The molecular weight of b-glucan in oat/barley
point, for a total of 12 experiments (Table 2). The two
products has been reported to be smaller than the
recipe variables optimized were the gluten (G, g/100 g flour)
molecular weight of b-glucan in the raw material (Åman
and water content (W, g/100 g flour). Experimental
et al., 2004; Beer, Wood, Weisz, & Fillion, 1997;
conditions at the centre point were G ¼ 11.8 g/100 g and
Kerckhoffs, Hornstra, & Mensink, 2003; Sundberg et al.,
W ¼ 88/100 g. The scaled values were x1 ¼ (G–11.8)/3.45,
1996). The molecular weight of b-glucan in oat bread was
reduced as compared with the molecular weight of
b-glucan in oat bran, and no significant changes in LDL-
Table 1
cholesterol levels of hypercholesterolemic subjects were
Chemical analyses of flours
detected (Kerckhoffs et al., 2003; Törrönen et al., 1992).
Raw material, endogenous b-glucanase activity, proces- Attribute Oat flour, Helsingin Wheat flour,
sing, and storage conditions exhibit an effect on the Mylly Ltd. Raisio Group
plc
amount, solubility, molecular weight, and structure of
b-glucan in the products (Beer et al., 1997; Degutyte- Moisture (g/100 g flour) 10.6 12.1
Fomins et al., 2002; Zhang, Moore, & Doehlert, 1998). Protein (g/100 g, db)a 19.0 12.9
The aim of this study was to develop wholemeal oat Ash (g/100 g, db) 2.8 0.7
bread baking technology by using experimental design Wet gluten (g/100 g) — 28.1
Sieves, mm (%)
and response surface model (RSM) to attain a bread 630 21 —
with good quality, high wholemeal oat content (51 g whole 475 12 —
grain oat flour/100 g wheat flour), and high b-glucan 355 8 —
content (0.75 g/portion). The possible depolymerization 250 7 —
of oat b-glucan in bread was also studied and light 132 50 —
o95 0.5 —
microscopy was used to locate oat b-glucan in the bread
matrix. a
Nx5.7 for wheat and Nx6.26 for oat.
 ARTICLE IN PRESS
862 L. Flander et al. / LWT 40 (2007) 860–870

where G ranged from 8.3 to 15.2/100 g flour; x2 ¼ (W–88)/ are given in Table 3. The scaled values (1, 0 and +1) were
10, where W ranged from 78 to 98/100 g flour. x1 ¼ (tm–6)/2, where tm ranged from 4 to 8 min; x2 ¼
(ti–12)/7.5, where ti ranged from 5 to 20 min; x3 ¼ (tf–65)/
2.2.2. Optimization of baking process 10, where tf ranged from 55 to 75 min; x4 ¼ (Tf–35)/5,
A CCF was used with five process variables and four where Tf ranged from 30 to 40 1C; and x5 ¼ (Tb–195)/15,
replicates at the centre point, for a total of 30 experiments where Tb ranged from 180 to 210 1C. A verification
(Table 3). The five process variables studied were the experiment was performed in the experimental domain
mixing time (tm, min), the intermediate proofing time (floor (experiment 31).
time) (ti, min), the final proofing time (tf, min), the final
proofing temperature of the cabinet (Tf), and the baking 2.3. Baking
temperature (Tb). Experimental conditions at the centre
point were tm ¼ 6 min, ti ¼ 12 min, tf ¼ 65 min, Tf ¼ 35 1C, 2.3.1. Optimization of ingredients
and Tb ¼ 195 1C. The experimental number and real values The amounts of tap water and gluten (Raisio Group plc,
Raisio, Finland) were according to the experimental design
(Table 2). The formula consisted of oat flour (51 g), wheat
Table 2 flour (49 g), sugar (3.2 g), salt (2.3 g), shortening (3.2 g)
Optimization of the ingredientsa (Sunnuntai, Raisio Group plc, Raisio, Finland), and
pressed baking yeast (3.2 g) (Finnish Yeast Ltd., Lahti,
Run Scaled value Real value
Finland). Amounts of the ingredients were based on the
X1 X2 Gluten (g/100 g Water (g/100 g weight of oat and wheat flour mixture (100 g). Total weight
flour) flour) of the dough batch was 2500 g. Wheat flour was blended
thoroughly with the sugar and salt and placed in a spiral
1 0.0 0.0 11.8 88.0
2 0.0 1.0 11.8 78.0 mixer (Diosna SP 12 F, Dierks and Söhne GmbH,
3 1.0 1.0 8.3 78.0 Osnabrück, Germany). The yeast was suspended in water
4 0.0 +1.0 11.8 98.0 (24 1C) and added to the mixture with the tempered
5 0.0 0.0 11.8 88.0 shortening. The optimal mixing time of the dough was
6 1.0 0.0 8.3 88.0 based on the results of screening tests. The dough was
7 +1.0 +1.0 15.2 98.0
8 0.0 0.0 11.8 88.0 mixed with fast speed (200 rpm) for 6 min and the oat flour
9 1.0 +1.0 8.3 98.0 with the rest of the water was added to the dough and
10 0.0 0.0 11.8 88.0 mixed at low speed (100 rpm) for 6 min. By this method it
11 +1.0 0.0 15.2 88.0 was possible to minimize the detrimental effect of oat on
12 +1.0 1.0 15.2 78.0 the formation of gluten network. The temperature of
Amount of water and gluten in oat breads in scaled and real values. dough at the end of mixing was kept at 30 1C by controlling
a
x1 ¼ (G–11.8)/3.4, where G ranged from 8.3 to 15.2/100 g flour; the temperature of the added water. After intermediate
x2 ¼ (W–88.0)/10, where W ranged from 78.0 to 98.0/100 g flour. proofing at 28 1C, 80% rh for 12 min, the dough was

Table 3
Optimization of processing conditions

Run Mixing Int. proofing Proofing Proofing Baking Run Mixing Int. proofing Proofing Proofing Baking
time (min) (min) time (min) temp. (1C) temp. (1C) time (min) (min) time (min) temp. (1C) temp. (1C)

1 8 5 55 30 180 16 8 12 65 35 195
2 6 12 75 35 195 17 6 12 65 35 195
3 4 5 55 30 210 18 6 12 65 40 195
4 6 12 65 35 210 19 8 5 75 30 210
5 4 20 55 40 210 20 4 20 55 30 180
6 8 20 75 40 210 21 6 5 65 35 195
7 4 20 75 40 180 22 4 20 75 30 210
8 4 5 55 40 180 23 8 20 75 30 180
9 4 5 75 40 210 24 8 20 55 40 180
10 6 12 65 35 195 25 8 5 55 40 210
11 6 12 65 30 195 26 4 12 65 35 195
12 8 20 55 30 210 27 4 5 75 30 180
13 6 12 65 35 180 28 6 12 55 35 195
14 6 12 65 35 195 29 8 5 75 40 180
15 6 20 65 35 195 30 6 12 65 35 195
31 6 12 65 39 210

Processing conditions of oat breads in real values.

 ARTICLE IN PRESS
L. Flander et al. / LWT 40 (2007) 860–870
divided into six 400 g pieces, rounded, and moulded by
hand to tempered pans which were sprayed with pan grease
and proofed at 80% rh, 35 1C for 65 min. The breads were
baked at 195 1C for 30 min (Rack Oven 9000, Sveba
Dahlen Ab, Sweden) and with 5 s of steam in the beginning.
2.3.2. Optimization of baking process
Water and gluten content of the dough were fixed
according to the results attained by optimization of the
recipe to 91.5 and 15.2/100 g flour, respectively. The
experimental design for the processing parameters are
presented in Table 3.
2.4. Analyses of bread characteristics
After baking, the loaves were cooled for 2 h before being
weighed. Loaf volume was determined by the rapeseed
displacement method (Siebert, 1971). Crumb hardness was
measured at 2 and 72 h after baking by the TA-XT2
Texture Analyser (Stable Micro Systems, Godalming, UK)
using the Texture Profile Analysis test. Six 25 mm thick
slices were used for the analysis, two slices were taken from
the middle of each of the three different breads. The crust
of the slices was removed so that only textural parameters
from the crumb were measured. The slices were compressed
by 10 mm (40%) with a speed of 1.7 mm/s. A trained
descriptive sensory panel (n ¼ 5) evaluated the character-
istics of the breads. Attribute intensities were rated on
5-unit, verbally anchored intensity scales (Table 4).
Altogether, nine attributes were selected to describe
texture, mouthfeel, and flavour of the breads.
2.5. b-glucan content and molecular weight distribution
b-glucan was determined by the enzymatic method of
McCleary and Codd (1991) using an assay kit obtained
from Megazyme International Ltd. (Bray, Ireland). The
average molecular weights (Mw) of b-glucan of flours and
Table 4
Sensory attributes and verbal anchors of the rating scales
Attribute Anchors
Bread crust
Evenness Not at all even–very even
Intensity of the colour Very light–very dark
Thickness Very thin (1–2 mm)–very thick (45 mm)
Intensity of the flavour No flavour–very intense flavour
Crispness Not at all crispy–very crispy (a crispy sound
during biting)
Bread crumb
Softness Not at all soft–very soft
Elasticity Not at all elastic (slice breaks immediately,
when bended)–very elastic
Moistness Not at all moist (in the mouth)–very moist
Richness of the flavour Very lean flavour–very rich flavour
863
bread were analysed after stirring 1 g of the sample
overnight with magnetic stirrer in 1 l of 0.1 N NaOH
containing 0.1% NaBH4. The samples were analysed by
HPLC-SEC with calcofluor staining by using right-angle
laser light scattering for detection (Suortti, 1993).
2.6. Microscopy
Pieces of bread crumb (0.5 cm) were taken from the
middle of the loaf, embedded in 1% agarose, fixed in 1%
glutaraldehyde in 0.1 mol/l phosphate buffer, pH 7.0,
dehydrated with ethanol, embedded in Historesin (Leica,
Heidelberg), and sectioned (4 mm) with a Leica microtome
(Heidelberg, Germany). For the fluorescence microscopic
examination, the bread sections were stained with specific
fluorochromes (Fulcher & Wong, 1980; Parkkonen,
Härkönen, & Autio, 1994; Wood, Fulcher, & Stone,
1983). Protein was stained with aqueous 0.1% (w/v)
Fuchsin acid for 1 min (Gurr, BDH Ltd., Poole, England),
and b-glucan was stained with aqueous 0.01% (w/v)
calcofluor White for 1 min (Fluorescent brightener 28,
Aldrich, Germany). Calcofluor stains intact cell walls blue.
Fuchsin acid stains proteins red. Starch remains unstained
and appears black.
2.7. Data analyses
Maximum and minimum ingredient and process variable
levels for the optimization tests were chosen by carrying
out preliminary screening tests (data not shown). A central
composite design made it possible to approximate the
measured data (yobs) using an RSM (Eq. (1)) expressed in
unscaled variables:
Y obs ¼ b0 þ b1 x1 þ b2 x2 þ b3 x3 þ b11 x1 x1
þ b22 x2 x2 þ b33 x3 x3 þ b12 x1 x2
þ b13 x1 x3 þ b23 x2 x3 þ e, ð1Þ
e ¼ yobs  ycalc , (2)
where b0 is a constant; b1, b2, and b3 express the main effect
of each process variable b12, b13, and b23 show the
interaction effects between the variables and the square
coefficients; and b11, b22, and b33 reveal whether any of the
variables give a maximum or minimum within the experi-
mental domain. The difference between the experimental
data (yobs) and the model (ycalc) gives the residual (e) in
Eq. (2) (Lindgren, Sjöström, Hellsten, & Lif, 1995). The
results were examined by the computer program MODDE
version 4.0 (Umetrics Ab, Umeå Sweden). The RSM were
estimated by partial least squares (PLS) for the 12 and 30
experiments in the CCF design. The centre point made it
possible to estimate the pure error of the analyses, which
was used to predict whether the models gave significant lack
of fit (Carlsson, 1992). The reliability of the models was
evaluated by calculating the R2 and Q2 values for each
model, where R2 is the variation of the response explained
 ARTICLE IN PRESS
864 L. Flander et al. / LWT 40 (2007) 860–870

by the model and Q2 is the fraction of the variation of the
response that can be predicted by the model (MODDE
version 4.0). Q2 should be 40.5 if conclusions are to be
drawn from the model (Lindgren, 1995). Generally, a model
is considered excellent if R2 and Q2 exceed 0.9 (Lindgren
et al., 1995). A verification experiment was performed to
estimate the predictive capacity of the models.
Tables 2 and 3 show the combination of predictor
variable levels used in the central composite design. To
study five factors (predictor variables) at three levels
would require 35, i.e., 243 experiments, whereas use of
the central composite design required 30 runs of experi-
ments. The centre point in the design was repeated four
times to calculate the reproducibility of the method.
The response variables measuring baking performance
were specific volume, hardness after 2 and 72 h and
sensory attributes of the bread crust (evenness, colour,
thickness, intensity of the flavour, and crispness) and
crumb (softness, elasticity, moistness, richness of the
flavour) (Table 4).
For each of the response variables, model summaries
and lack of fit tests were analysed for linear, quadratic,
and cubic models. From this information, the most
accurate model was chosen, which in all cases was
quadratic. Two-dimensional response surface plots were
generated for each quality parameter. Calculation of
optimal processing conditions for baking performance of
oat bread was performed using a multiple response
method called desirability. This optimization method
incorporates desires and priorities for each of the variables.
Specific volume, evenness, and crispness of the crust, and
softness, elasticity, and richness of the bread crumb were
specified as maximum level desirable. Hardness was
specified as minimum desirable. The desirable levels of
intensity of the colour and flavour as well as thickness of
the crust and moistness of the crumb was fixed to
intermediate level (score 3 on the sensory scale) (Table 4).
Optimization was carried out at recipe first and the
optimized recipe was used to carry out the optimization
of the process.
3. Results and discussion
3.1. Baking quality
3.1.1. Optimization of ingredients
The results of the 12 experiments were evaluated
statistically by PLS method. Complete RSM were esti-
mated. The achieved mathematical models, expressed in
unscaled variables are presented in Table 5. Using these
tables, the specific volume of the bread can be predicted by
the equation for the experimental area under consideration,
e.g.,
Specific volume ¼  83:2 þ 2:8G þ 3:6W
 0:044W 2  0:06GW .
The gluten and water significantly influenced specific
volume and hardness of the oat bread crumb. The evenness
of the bread crust as well as sensory softness, elasticity, and
moistness of the bread crumb were also affected signifi-
cantly by gluten and water contents. Gluten and water
contents did not affect significantly to the intensity of the
crust colour, thickness or crispness of the crust, or intensity
of the crust flavour.
Response surface plots of specific volume and hardness
of the breads are shown in Fig. 1A–C. As can be seen in
Fig. 1A–C, water had a slightly more pronounced effect on
these quality attributes than gluten due to a wider range.
According to the response surface plots, a water content of
90–92.5 g and a gluten content of 14.2–15.2/100 g flour
were required for maximal specific volume and minimal
instrumental hardness measured after 2 and 72 h, respec-
tively. The maximal specific volume (3.6 cm3/g) and
minimal hardness (0.1 kg after 2 h, and 0.2 kg after 72 h)
were attained by adding gluten 15.2 g and water 91.5/100 g
of flour weight to the dough.
From the sensory attributes tested, the gluten and water
contents mainly affected crumb properties, whereas pro-
cessing conditions exhibited their main effects on crust
properties. Optimal evenness of the crust and optimal

Table 5
Effects of ingredient factors, expressed as their corresponding coefficients obtained in the models for texture and sensory attributesa

Factorb Specific Hardness, Hardness, Evenness Softness Elasticity Moistness

volume 2h 72 h

Constant 83.2 23.1 59.4 31.1 84.9 17.1 92.3

G 2.8 0.6 1.7 2.6 3.2 1.1 4.5
W 3.6 1.0 2.5 1.2 3.6 0.4 ns
GW 0.06 0.02 0.04 0.06 0.04 0.05 ns
G2 ns ns ns ns 0.11 0.09 0.05
W2 0.04 0.01 0.03 0.01 0.04 ns ns
R2 0.93 0.97 0.96 0.85 0.87 0.89 0.80
Q2 0.66 0.73 0.68 0.62 0.57 0.60 0.74
a
Only values of significant coefficients are presented (95% confidence level); ns, no significant effect at the 5% level.
b
G, gluten; W, water; G  W, interaction between gluten and water; G2, quadratic effect of gluten; W2, quadratic effect of water; R2, measure of fit of the
model: Q2, predictive power of the model.
 ARTICLE IN PRESS
L. Flander et al. / LWT 40 (2007) 860–870 865

98 98

94 94

Water (g/100g flour).

Water (g/100 g flour.)

3.6 90
90
0.15
3.4
86 86 0.20
3.20 0.25
3.0 0.3
82 82
2.8
2.6 0.4 0.35
78 78
8.3 9.4 10.6 11.8 12.9 14.0 15.2 8.3 9.4 10.6 11.8 12.9 14.0 15.2
(A) Gluten (g/100 g flour) (B) Gluten (g/100 g flour)

98
0.24

94
Water (g/100 g flour)

0.24

90
0.37

86 0.50
0.64
82 0.78
0.92
1.04
78
8.3 9.4 10.6 11.8 12.9 14.0 15.2
(C) Gluten (g/100 g flour)

Fig. 1. Response surface plots of recipe optimization. Effects of gluten and water on (A) specific volume (cm3/g) of the bread, (B) instrumental hardness
(kg) of the crumb after storage of 2 h, (C) instrumental hardness (kg) of the crumb after storage of 72 h.

moistness of the crumb were achieved with the same bread (64.9/100 and 87/100 g flour, respectively). All of the
concentrations of water and gluten as the optimal volume ingredients in barley bread were blended with a slow-speed
and hardness of the oat bread. The softest bread was mixer with shorter fermentation time and at a lower
obtained when the gluten content of the bread was 12/100 g temperature than our breads.
flour and the water content was 90/100 g flour. Most elastic
bread was attained with the smallest gluten and water 3.1.2. Optimization of baking process
contents (8 and 84/100 g flour, respectively). The processing conditions were optimized by using
According to Oomah and Lefkovitch (1988), loaf 15.2 g gluten and 91.5 g water/100 g flour to the dough.
volumes of wheat–oat breads can be optimized by a The results from the measurements were evaluated
sufficiently high amount of water. Also, Gormley and statistically by PLS. Complete RSMs, including the 30
Morrissey (1993) obtained the largest bread volumes by the experiments in the design, were estimated. The achieved
addition of gluten and extra water to bread containing mathematical models for texture and sensory attributes are
5–20 g oat flakes/100 g flour. The suppression of loaf presented in Table 6. The final proofing temperature and
volume by soluble oat dietary fibre is probably related to time, as well as baking temperature significantly influenced
the inhibition of gluten strength. Increasing its content the specific volume of the oat bread. The final proofing
without a concomitant increase in vital gluten content temperature and time as well as intermediate proofing time
results in a reduction in loaf volume and even in together with final proofing temperature had significant
unsatisfactory loaves (Rudel, 1990). Barley is another effects on the hardness of the oat bread. Also, mixing time
cereal with a high content of soluble b-glucan. Cavallero, affected the crumb hardness significantly. The significant
Empilli, Brighenti, and Stanca (2002) evaluated the bread- effect of proofing time on specific volume and hardness of
making quality of mixed barley breads containing 50 g wheat sourdough bread has also been reported by Clarke,
whole grain barley flour/100 g wheat flour by evaluations Schober, Angst, and Arendt (2003).
of the bread volume and sensory acceptance in order to get The intensity of the crust colour and richness of the
test meals for glycemic index analysis. The volume of the crumb flavour was affected most by baking temperature.
bread was low (specific volume was about 2.5 ml/g) and the Thickest crust with most intensive flavour was obtained
bread received quite low flavour/aroma scores by the panel. with highest baking temperature. The most significant
The barley bread had lower water contents than our oat variables affecting the crispness of the crust, were final
 ARTICLE IN PRESS
866 L. Flander et al. / LWT 40 (2007) 860–870

Table 6
Effects of processing factors, expressed as their corresponding coefficients obtained in the models for texture and sensory attributesa

Factorb Texture attributes Sensory attributes

Specific volume Hardness, 2 h Hardness, 72 h Crust Crumb

Colour Thickness Flavour Crispness Richness

Constant 5.457 496.164 1142.089 3.746 2.963 6.058 52.554 5.364

M 0.020 89.093 130.410 0.981 1.111 0.715 0.245 0.771
PI 0.004 31.769 39.924 0.000 0.294 0.060 0.197 0.193
PTi 0.085 10.960 25.486 0.159 0.050 0.067 0.095 0.105
PTe 0.044 11.066 25.801 0.272 0.681 0.005 0.132 0.141
B 0.026 6.974 15.338 0.032 0.094 0.031 0.504 0.074
M  PI ns ns ns 0.009 0.004 0.009 0.018 ns
M  PTi ns ns ns ns 0.003 0.013 ns ns
M  PTe ns ns ns 0.009 0.004 ns ns ns
MB ns 0.426 0.587 0.002 0.003 ns ns 0.003
PI  M ns ns ns ns 0.009 ns ns ns
PI  PTi ns 0.131 0.053 0.001 0.001 ns 0.002 0.002
PI  PTe ns 0.464 0.839 0.003 0.004 ns ns 0.001
PI  B 0.001
PTi  PTe ns ns ns ns 0.005 ns 0.003 0.001
PTi  B 0.001 0.076 0.170 0.001 ns ns ns 0.001
PTe  B ns ns ns 0.001 0.001 ns ns 0.001
M2 ns ns ns 0.009 0.015 ns ns 0.010
PI2 ns 0.267 0.237 0.001 0.001 ns ns 0.001
PTi2 ns ns ns 1.814 0.001 ns ns 0.002
PTe2 ns ns ns 0.001 0.002 ns ns 0.001
B2 ns ns ns 0.000 0.000 ns 0.001 0.000
R2 0.85 0.83 0.86 0.85 0.93 0.84 0.84 0.84
Q2 0.66 0.52 0.50 0.58 0.72 0.66 0.57 0.61
a
Only values of significant coefficients are presented (95% confidence level).
b
M, mixing time; PI, intermediate proofing time; PTi, proofing time; PTe, proofing temperature; B, baking temperature; M2, quadratic effect of mixing
time; M  PI, interaction between mixing time and intermediate proofing time; R2, measure of fit of the model: Q2, predictive power of the model.

proofing time, temperature, and baking temperature.
Processing did not significantly affect the evenness of the
crust, softness, elasticity and moistness of the breads.
Of the five processing factors, proofing temperature and
time had the greatest effects on bread quality. Response
surface plots for specific volume and hardness after 2 and
72 h storage are shown in Fig. 2A–C. Proofing time had a
slightly more pronounced effect than proofing temperature
on the specific volume and hardness of the breads.
According to the response surface plot, the maximal
specific volume 3.7 (cm3/g) and minimal hardness (0.1 kg
after 2 h, and 0.3 kg after 72 h) could be achieved by longest
proofing times of 71–75 min and highest temperatures of
39–40 1C, respectively.
Of the sensory attributes evaluated, the processing
conditions significantly affected the crust properties and
richness of the crumb flavour. The thickness, intensity of
flavour and crispness of the crust were attained in
conditions which were the same as the optimal conditions
for bread volume and hardness. The optimal colour of the
crust and richness of the crumb flavour were attained in a
slightly shorter proofing time (64 min) than other sensory
attributes.
3.1.3. Optimized oat bread
The water content and the gluten content for the
optimized bread were 91.5 and 15.2/100 g flour, respec-
tively. The dough was mixed for 6 min and the intermediate
proofing time was 12 min. The final proofing time was
slightly reduced from 70 to 65 min in order to obtain
optimum richness of the crumb flavour and crust colour to
the bread. The final proofing temperature was 39 1C and
the baking temperature was 210 1C. The specific volume of
the bread baked for the verification was somewhat higher
than the predicted value, but fell within the confidence
interval (Table 7). Because of the higher specific volume,
the hardness of the stored bread in the verification
experiment was lower than the predicted value. Fig. 3
shows the difference between the oat breads before and
after optimization of the bread quality. Tasty breads with
high quality are most important factors to increase the
popularity of healthier breads among consumers.
3.2. b-glucan content and molecular weight distribution
The b-glucan content of the bread was 2.4/100 g bread
(db) (Table 8), so the amount of b-glucan in fresh bread
 ARTICLE IN PRESS
L. Flander et al. / LWT 40 (2007) 860–870 867

40 40
3.7 0.16

Proof temperature ( C)

Proof temperature ( C)
38 3.6 38
0.18
3.4
36 36
3.3 0.20

34 3.2 34 0.21
3.1 0.23
32 32 0.24
2.9 0.26
2.8
30 30
55 60 65 70 75 55 60 65 70 75
(A) Proof time(min) (B) Proof time (min)

40 0.34
Proof temperature( C)
38
0.38

36
0.43
34 0.47
0.52
32 0.56
0.6
30
55 60 65 70 75
(C) Proof time (min)

Fig. 2. Optimization of the process conditions. Response surface plots at 14.4 g gluten/100 g flour and 87 g water/100 g flour, 4 min mixing time, 20 min
intermediate proofing time, and 210 1C baking temperature. Effects of final proofing time and temperature on (A) specific volume (cm3/g) of the bread, (B)
instrumental hardness (kg) of the crumb after storage of 2 h, (C) instrumental hardness (kg) of the crumb after storage of 72 h.

Table 7 Ellis et al., 1997; Henry, Martin, & Stewart, 1989) and
Measured and predicted valuesa for specific volume, hardness and sensory average molecular weight of barley b-glucan (Rimsten,
attributes for the verification experimentb 2003). Åman et al. (2004) found that large particle size of
Measured value Predicted value the oat bran and short fermentation time limited the
b-glucan degradation during baking. Molecular weights of
Specific volume (cm3/g) 3.6070.07 3.4070.1 b-glucan in oat breads and muffins have shown to be
Hardness, 2 h 0.1470.02 0.1970.02
Hardness, 72 h 0.2970.04 0.4470.04
smaller than in oat flour or bran (Beer et al., 1997;
Kerckhoffs et al., 2003; Sundberg et al., 1996). Kerckhoffs
a
The values were calculated by response surface modelling using the et al. (2003) analysed the molecular weight distribution by
computer program MODDE 4.0 (Umetri AB, Umeå, Sweden). HPLC-SEC. The proportion of the highest molecular
b
Experiment 31: mixing time 6 min, intermediate proof 12 min, proofing
weight (over 1 000 000) of b-glucan was only 30% and 20%
time 65 min at 39 1C, baking temperature 210 1C.
in the oat bran concentrate and oat bran. About 50% of
the b-glucan exceeded this size in oat flours used in this
study. Possible differences in variety, crop year, and/or
was 1.3/100 g. This means that a portion (two slices á 30 g) processing of the oat may have caused these differences in
of the bread contains 0.78 g b-glucan. FDA allows the molecular weight distribution. The amount of b-glucan in
health claim for products containing a minimum of 0.75 g the oat bran–wheat bread of Kerckhoffs et al. (2003) was
of b-glucan per portion. The lowest suggested daily intake 3.3/100 g and in our oat flour-wheat bread 1.3/100 g. In
of b-glucan for achieving the health effects is 3 g per day, both breads, only about half of the original proportion of
which requires four portions with 0.75 g of b-glucan. b-glucan with the highest molecular weight remained after
The molecular weight distribution of b-glucan indicated baking; 10–15% in oat bran–wheat breads and 30% in oat
that the proportion of very high molecular weight b-glucan flour-wheat breads, respectively. This is in accordance with
(Mw41  106) had decreased and the proportion of the other studies on barley and oat bran breads (Andersson
lower molecular weight b-glucan had increased during the et al., 2004; Åman et al., 2004). The amount of high
baking process (Table 8). The reason for the partial molecular weight b-glucan (41 000 000) was about the
degradation of b-glucan in oat bread may be the same; 0.3–0.5/100 g in oat bran–wheat bread and 0.4/100 g
b-glucanase activity of the wheat flours. Several studies oat flour-wheat bread. The amount of middle-size mole-
have shown that endogenous b-glucanase activity will cular weight b-glucan (200 000–1 000 000) was 30% in both
decrease the content (Degutyte-Fomins et al., 2002; breads. The middle-size b-glucans were higher in oat
 ARTICLE IN PRESS
868 L. Flander et al. / LWT 40 (2007) 860–870

Fig. 3. Oat bread from preliminary screening test (A), after optimization of the recipe (B) and after optimization of the process (C).

Table 8
b-glucan analyses of oat flour and bread

Whole grain White wheat Oat bread

oat flour flour

b-glucan content (g/ 5.9870.04 0.2670.01 2.4070.18

100 g dry weight)
b-glucan content (g/ 5.35 0.23 1.34
100 g fresh weight)
Molecular weight distribution of b-glucan
Mw41 000 000 60 25 30
Mw, 200 000–1 000 000 30 30 30
Mwo200 000 10 45 45
 Moisture contents: oat flour 10.6/100 g, white wheat flour 12.1/100 g,
bread 44.3/100 g.

bran–wheat bread 1.0/100 g than in our bread 0.4/100 g.

Törrönen et al. (1992) prepared oat breads without wheat
for clinical studies. They did not detect any changes in
molecular weight of b-glucan during baking or storage of
those oat breads. This indicates that b-glucanase from
wheat could be the main reason for degradation of high
molecular weight b-glucan during baking (Åman et al.,
2004; Andersson et al., 2004).
Microstructural study of the oat breads showed that the
b-glucan was located mainly in insoluble form in the cell
walls of large bran particles (Fig. 4). b-glucan is the major
endospermic cell wall polysaccharide in oat, constituting
approximately 85% of the cell wall (Miller, Fulcher, Sen, &
Arnason, 1995). The bran fraction of the grain is rich in
b-glucan, as the cell walls of the subaleurone layer are very
thick (Fig. 4). It appears that the use of coarse flour is
advantageous for protection of b-glucan from enzymatic
degradation. Hydration of large particles is slower than
hydration of small particles, and the complex structure and
thickness of the cell walls slow down the solubilization of
b-glucan and other cell wall polymers. The mixing method
probably affects the distribution of water between wheat
and oat flour components and especially the added gluten
in the dough. This may have had protective effect against
degradation of high molecular level b-glucans, because the
Fig. 4. Microstructure of oat bread. The b-glucan in the cell walls has
been stained with calcofluor and appears blue.
oat flour was incorporated into the dough with slow mixing
speed after formation of a proper gluten network with fast
mixing speed. The degrading effect of b-glucanase and
mixing might have been stronger, if all of the ingredients
had been mixed at the same time with fast speed. Long
fermentation of the dough at high temperature could have
activated the b-glucanase of the wheat flour and degraded
part of the b-glucan. The incorporation of oat groat or
grain instead of oat flour to the bread dough, or
inactivation of b-glucanase of wheat flour could be the
solutions to reduce the degradation of b-glucan during
processing of the oat bread.
4. Conclusions
The concentrations of gluten and water in oat breads
were optimized first and these concentrations were used in
 ARTICLE IN PRESS
L. Flander et al. / LWT 40 (2007) 860–870
the optimization test for process conditions. Maximal
specific volume 3.7 (cm3/g) and minimal hardness (0.1 kg
after 2 h, and 0.2 kg after 72 h) were attained by adding
gluten (15.2/100 g flour) and water (91.5/100 g flour) to the
dough. The optimal evenness of the crust and moisture of
the crumb were attained with the same concentrations of
gluten and water as those used for maximal volume and
minimal hardness. Of the five processing conditions, bak-
ing temperature, proofing time and temperature exhibited
the greatest effects on bread quality. The maximal specific
volume 3.7 (cm3/g) and minimal hardness (0.1 kg after 2 h,
and 0.3 kg after 72 h) were attained by proofing the bread
at 40 1C for 75 min and baking it at 210 1C. Of the sensory
attributes evaluated, the processing conditions significantly
affected the crust properties and richness of the crumb
flavour. The optimal thickness, flavour, and crispness of
the crust were attained in the same conditions as maximal
volume and minimal hardness of the bread.
The b-glucan content of oat bread was 2.4/100 g bread
(db) corresponding to the amount 1.3 g b-glucan/100 g of
fresh bread. This means that a portion (two slices á 30 g) of
the bread contains 0.78 g of b-glucan, which meets the
required amount of 0.75 g b-glucan per portion for a health
claim (FDA, 1996). A slight decrease in molecular weight
of b-glucan was observed. It is probable that the b-glucan
hydrolysis was caused mainly by endogenous enzymes
present in wheat flour.
From the data presented, it is evident that bread with
51/100 g flour could be baked and that good taste and
structure as well as long shelf-life could be obtained by
optimizing recipe and processing parameters. These breads
with elevated levels of fibres, nutrients and b-glucan can be
produced on an industrial scale.
Acknowledgments
We thank the Ministry of Agriculture and Forestry of
Finland for financial support and Arja Viljamaa for her
skilful technical assistance.
References
Åman, P., Rimsten, L., & Andersson, R. (2004). Molecular weight
distribution of b-glucan in oat-based foods. Cereal Chemistry, 81,
356–360.
American Association of Cereal Chemists (ACCC). (2000). Approved
methods of the AACC, 10th ed. Methods 08-01, 38-12, 44-15A, and
46-11A. St. Paul, MN: The Association.
Andersson, A. A., Armö, E., Grangeon, E., Fredriksson, H., Andersson,
R., & Åman, P. (2004). Molecular weight and structure units of (1-3,
1-4)-b-glucans in dough and bread made from hull-less barley milling
fractions. Journal of Cereal Science, 40, 195–204.
Autio, K., Myllymäki, O., & Mälkki, Y. (1987). Flow properties of
solutions of oat b-glucans. Journal of Food Science, 52, 1354–1366.
Beer, M., Wood, P., Weisz, J., & Fillion, N. (1997). Effect of cooking and
storage on the amount and molecular weight of (1-3)(1-4)-b-
D-glucan extracted from oat products by an in vitro digestion system.
Cereal Chemistry, 74, 705–709.
869
Brümmer, J.-M., Morgenstern, G., & Neumann, H. (1988). Herstellung
von Hafer-, Gerste-, Mais-, Reis-, Hirse- und Buchweizenbrot.
Getreide, Mehl und Brot, 5, 153–158.
Carlsson, R. (1992). Response surface methods. In R. Carlsson (Ed.),
Design and optimization in organic synthesis (pp. 249–324). Amsterdam:
Elsevier Science.
Cavallero, A., Empilli, S., Brighenti, F., & Stanca, A. M. (2002). High
(1-3, 1-4)-b-glucan barley fractions in bread making and their
effects on human glycemic response. Journal of Cereal Science, 36,
59–66.
Clarke, C. I., Schober, T. J., Angst, E., & Arendt, E. (2003). Use of
response surface methodology to investigate the effects of processing
conditions on sourdough wheat bread quality. European Journal of
Food Research and Technology, 217, 23–33.
Degutyte-Fomins, L., Sontag-Strohm, T., & Salovaara, H. (2002). Oat
bran fermentation by rye sourdough. Cereal Chemistry, 79, 345–348.
Doublier, J.-L., & Wood, P. J. (1995). Rheological properties of aqueous
solutions of (1-3) (1-4)-b-D-glucan from oat (Avena sativa L.).
Cereal Chemistry, 72, 335–340.
Ellis, R. P., Swanston, J. S., Rubio, A., Perez-Vendrell, A. M., Romagosa,
I., & Molina-Cano, J. L. (1997). The development of b-glucanase and
degradation of b-glucan in barley grown in Scotland and Spain.
Journal of Cereal Science, 26, 5–82.
FDA. (1996). Food labeling: Health claims; Oat and coronary heart
disease. Federal Register, 61(3) January 4.
Forssell, P., Shamek, S., Härkönen, H., & Poutanen, K. (1998). Effects of
native and enzymatically hydrolysed soya and oat lecithins in starch
phase transitions and bread baking. Journal of the Science of Food and
Agriculture, 76, 31–38.
Fulcher, RG., & Wong, S. J. (1980). Inside cereals—A fluorescence
microchemical view. In E. Inglett, & L. G. Munck (Eds.), Cereals for
food and beverages (pp. 1–25). New York: Academic Press.
Gormley, T. R., & Morrissey, A. (1993). A note on the evaluation of
wheaten breads containing oat flour or oat flakes. Irish Journal of
Agricultural and Food Research, 32, 205–209.
Henry, R. J., Martin, D. J., & Stewart, B. G. (1989). Cel-wall
polysaccharides of rye-derived wheats: Investigations of the biochem-
ical causes of dough stickiness. Food Chemistry, 34, 309–316.
Jacobs, D. R., Jr., Marquart, L., Slavin, J., & Kushi, L. H. (1998a).
Whole-grain intake and cancer: An expanded review and meta-
analysis. Nutrition and Cancer, 30, 85–96.
Jacobs, D. R., Marquart, L., Slavin, J., & Kushi, L. H. (1998b). Whole-
grain intake may reduce the risk of ischemic heart disease death in
postmenopausal women: The Iowa Women’s Health Study. American
Journal of Clinical Nutrition, 68, 248–257.
Kerckhoffs, D., Hornstra, G., & Mensink, R. (2003). Cholesterol-l
owering effect of b-glucan from oat bran in mildly hyper-
cholesterolemic subjects may decrease when b-glucan is incorporated
into bread and cookies. American Journal of Clinical Nutrition, 78,
221–227.
Krishnan, P., Chang, K., & Brown, G. (1987). Effect of commercial oat
bran on the characteristics and composition of bread. Cereal
Chemistry, 64, 55–58.
Lindgren, Å. (1995). Multivariate structure–property-relationships of
non-ionic surfactants. PhD thesis, Umeå University, Umeå, Sweden.
Lindgren, Å., Sjöström., M., Hellsten, M., & Lif, A. (1995). Modelling of
detergency performance for some technical nonionic surfactants.
Tensile Surfactants Detergents, 32, 300–309.
Mälkki, Y. (2001). Physical properties of dietary fiber as keys to
physiological functions. Cereal Foods World, 46, 196–199.
McCleary, B., & Codd, R. (1991). Measurement of (1-3)(1-4)-b-
D-glucan in barley and oat: A streamlined enzymic procedure. Journal
of the Science of Food and Agriculture, 55, 303–312.
McKechnie, R. (1983). Oat products in bakery foods. Cereal Foods World,
28, 635–637.
Miller, S. S., Fulcher, R. G., Sen, A., & Arnason, J. T. (1995). Oat
endosperm cell walls: I. Isolation, composition, and comparison with
other tissues. Cereal Chemistry, 72, 421–427.
 ARTICLE IN PRESS
870 L. Flander et al. / LWT 40 (2007) 860–870
Oomah, B. D. (1983). Baking and related properties of wheat–oat
composite flours. Cereal Chemistry, 60, 220–225.
Oomah, B. D., & Lefkovitch, L. P. (1988). Optimal oxidant requirement
of wheat–oat composite flours. Die Nahrung, 32(6), 527–538.
Parkkonen, T., Härkönen, H., & Autio, K. (1994). The effect of baking on
microstructure of rye cell walls and proteins. Cereal Chemistry, 71,
58–63.
Prentice, N., Cuendet, L. S., & Geddes, W. F. (1954). Studies on bread
staling. V. Effect of flour fractions and various starches on the firming
of bread crumb. Cereal Chemistry, 31, 188–206.
Rimsten, L. (2003). Extractable cell-wall polysaccharides in cereals, with
emphasis on b-glucan in steeped and germinated barley. Doctoral thesis,
Swedish University of Agricultural Sciences, Uppsala.
Rudel, H. (1990). A composition of flours containing vital gluten and soluble
oat dietary fiber and a baked product produced therefrom. US Patent
4961937.
Siebert, G. (1971). Determination of volume in bakery products. Bäcker
und Konditor, 19, 103–104.
Slavin, J., Marquart, L., & Jacobs, D., Jr. (2000). Consumption of whole-
grain foods and decreased risk of cancer: Proposed mechanisms.
Cereal Foods World, 45, 54–58.
Stear, C. A. (1990). High-fibre breads. In C. A. Stear (Ed.), Handbook of
breadmaking technology (pp. 524–525). London: Elsevier Science
Publishers Ltd.
Sundberg, B., Wood, P., Lia, Å., Andersson, H., Sandberg, A.-S.,
Hallmans, G., et al. (1996). Mixed-linked b-glucan from breads of
different cereals is partly degraded in the human ileostomy model 1–3.
American Journal of Clinical Nutrition, 64, 878–885.
Suortti, T. (1993). Size-exclusion chromatographic determination of beta-
glucan with postcolumn reaction detection. Journal of Chromatogra-
phy, 632, 105–110.
Thompson, L. U. (1994). Antioxidants and hormone-mediated health
benefits of whole grains. Critical Reviews in Food Science and Nutrition,
34, 473–497.
Törrönen, R., Kansanen, L., Uusitupa, M., Hänninen, O., Myllymäki, O.,
Härkönen, H., et al. (1992). Effects of oat bran concentrate on serum
lipids in free-living men with mild to moderate hypercholesterolaemia.
European Journal of Clinical Nutrition, 46, 621–627.
Welch, R., Brown, J., & Leggett, J. (2000). Interspecific and intraspecific
variation in grain and groat characteristics of wild oat (Avena) species:
Very high groat (1-3),(1-4)-b-D-glucan in an Avena atlantica
genotype. Journal of Cereal Science, 31, 273–279.
Wood, P. J. (1993). Physicochemical characteristics and physiological
properties of oat (1-3) (1-4)-b-D-glucan. In P. J. Wood (Ed.),
Oat bran (pp. 49–82). St. Paul, MN: AACC Inc.
Wood, P. J., Fulcher, R. G., & Stone, B. A. (1983). Studies on the
specificity of interaction of cereal cell wall components with Congo
Red and calcofluor. Specific detection and histochemistry of
(1-3),(1-4)-b-D-glucan. Journal of Cereal Science, 1, 95–110.
Zhang, D. C., Moore, W. R., & Doehlert, D. C. (1998). Effects of oat
grain hydrothermal treatments on wheat–oat flour dough properties
and breadbaking quality. Cereal Chemistry, 75, 602–605.