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Journal of Food Quality ISSN 1745-4557

COMBINED EFFECT OF XYLANASE, ASCORBIC AND CITRIC

ACID IN REGULATING THE QUALITY OF BREAD MADE FROM
ORGANICALLY GROWN SPELT CULTIVARS
BOJANA FILIPČEV1, OLIVERA ŠIMURINA and MARIJA BODROŽA-SOLAROV
Institute of Food Technology, University of Novi Sad, Novi Sad, Serbia

1
Corresponding author. ABSTRACT
TEL: +381 21 485 3778;
FAX: +381 21 450 725; The present study was aimed at improving bread-making performance of two
EMAIL: Bojana.filipcev@fins.uns.ac.rs spelt cultivars from organic production differing in dough rheological properties.
For this purpose, a small composite design was employed to study the combined
Received for Publication March 8, 2013
effect of xylanase (XYL), ascorbic (ASC) and citric (CIT) acid on bread character-
Accepted for Publication January 13, 2014
istics and, finally, to optimize their doses. In both cultivars, XYL exerted improv-
10.1111/jfq.12081 ing effects on specific volume and crumb firmness. In stronger spelt cultivar, there
was a significant ASC/XYL synergistic effect on improving loaf volume and crumb
properties. CIT showed crumb softening effect in both cultivars and increased
crumb resilience (weaker spelt being more sensitive). Optimized formulation con-
tained (in mg/kg): 50 ASC/90 CIT/60 XYL for stronger and 20 ASC/10 CIT/120
XYL for weaker spelt cultivar. The optimized formulations allowed substantial
increases in specific volume (by 45%) and crumb resilience (30%) for stronger
and crumb softening (36%) for weaker spelt cultivar.

PRACTICAL APPLICATIONS
Spelt wheat is an important cereal in organic cultivation and spelt products are
highly demanded by health-conscious customers. Most commonly, spelt is pro-
cessed into flour and further into bread. Technologically, spelt is impaired in com-
parison to common bread wheat, although high diversity exists in bread-making
potential within cultivars. In practice, it is not rare that spelt bread-making results
in production of unappealing, small bread loaves with dense, crumbly structure.
Considering the much higher price of spelt bread, low quality of loaves may
diminish its valorization in spite of its unique nutritional and health properties.
This work presents that by combining improvers based on enzyme (xylanase),
ascorbic and citric acid, which are permitted in organic foods, it is possible to
substantially improve spelt bread volume and crumb properties.

spelt cultivation because it is readily consumed as an alter-

INTRODUCTION
Although cultivars of conventional winter wheat specially
developed for low-input systems occupy majority of organic
cereal acreage in well-developed organic farming systems
such as those established in some European countries,
U.S.A. and Canada, the importance of spelt wheat (Triticum
aestivum spp. spelta) as an organic crop has been continu-
ously increasing (Wolfe et al. 2008). Spelt wheat possesses
agronomic characteristics which make it suitable for organic
cultivation. Many certified organic producers decide on
native cereal by those suffering from various allergies. More-
over, it is becoming popular among organic producers in
developing countries, having been usually the only wheat
alternative in the absence of conventional wheat varieties
adapted for organic production.
Spelt wheat is suitable for processing into different kinds
of food. In the cost–benefit analysis of Rozman et al. (2006),
it was found that the most beneficial spelt processing solu-
tion for an organic farmer is husk production, followed by
cookie, flour and bread production. Even though the spelt

Journal of Food Quality 37 (2014) 185–195 © 2014 Wiley Periodicals, Inc. 185
REGULATING THE QUALITY OF ORGANIC SPELT BREAD
product palette has been broadening, spelt is mostly mar-
keted in the form of flour and bread. Spelt emerges as a
suitable ingredient for organic bread production. Organic
bread is promoted for its organic ingredients, for its relative
lack of harmful residues and synthetic chemical preserva-
tives and as a good alternative for health-conscious consum-
ers. A recent study reported that organic breads were less
variable in ingredients but contained more nutritionally
dense ingredients (more whole grains and whole meal
ingredients), and were consistently lower in preservatives/
additives and pesticide residues in comparison to conven-
tional counterparts (Smith et al. 2012). Bread is such an
important part of our everyday nutrition that it is natural to
expect its healthiness and wholesomeness. Organic breads
meet these expectations, in contrast to conventional breads
whose nutritional quality has been frequently questioned.
For use in bread-making, organically produced spelt
should fulfill the technological requirements of bakers.
Conversely to agricultural and nutritional advantages, the
bread-making ability of spelt is rather diminished in com-
parison to conventional wheat. Doughs prepared from
spelt wheat exhibit poor rheological properties, which are
reflected in lower stability, increased stickiness and softness,
difficult handling properties, and low oven rise (Schober
et al. 2006; Pruska-Kedzior et al. 2008). The produced
breads are low in volume and have underdeveloped crumb
with rough pore structure (Ranhotra et al. 1995). However,
some studies showed similar or better bread-making perfor-
mance of spelt than common wheat (Bojňanská and
Frančáková 2002; Pasqualone et al. 2011). Lacko-Bartošová
and Rédlová (2007) reported that genetically pure spelt
varieties are of lower processing quality, whereas spelt vari-
eties crossed with T. aestivum L. genome are characterized
with suitable bread-making ability. Schober et al. (2006)
suggested a classification of spelt varieties into three groups:
group 1 are those most similar in characteristics to modern
wheat; group 2 are typical spelt varieties; and group 3 are
weak spelt cultivars, not suitable for most food applications.
The loaf volume, crumb hardness and elasticity of spelt
breads were correlated to dough strength and tenacity, but
not to protein content (Filipčev et al. 2013). Actually, bread
quality depends on the ability of dough to retain gas during
fermentation and baking, while this depends on the quality
of gluten proteins. Schober et al. (2002) concluded that the
baking quality of spelt cultivars is largely determined by
gluten properties. In practice, bread-making with spelt
wheat requires certain adaptations, which usually include
application of higher doses of ascorbic acid (ASC), shorter
mixing times and longer dough resting periods at lower
temperature with frequent rounding (Reiter et al. 2000).
Dough conditioners can improve dough handling prop-
erties and bread quality. Oxidants are well-known strength-
eners of gluten network. L-ASC, in spite of being a reducing
186
B. FILIPČEV, O. ŠIMURINA and M. BODROŽA-SOLAROV
agent itself, is one of the most widely used improver with
strong oxidizing effect on dough. In addition, it is the most
acceptable additive in bread, as being a vitamin (Wieser
2003). ASC improves bread volume and crumb structure. It
is typically used in doses from 50 to 75 mg/kg flour (Wieser
2003), but wider ranges of 10–200 mg/kg are also proposed
(Selomulyo and Zhou 2007). However, as for most dough
conditioners, its effect is not only dose-dependent but
dependent on the initial quality of flour and the type of
bread-making method (Pečivová et al. 2011). Horvat et al.
(2009) reported different actions of combined dough-
strengthening improver (L-ASC + DATEM E472e) on wheat
cultivars differing in gluten strength. It was found that the
additive improved loaf volume, loaf shape and crumb struc-
ture in cultivars with medium-strength gluten, whereas it
decreased the quality of bread made from cultivars having
strong gluten. However, when studying the ASC response of
various baking quality flours, Aamodt et al. (2003) did not
find significant differences between strong and weak baking
quality cultivars. When testing several dough-strengthening
improvers on spelt cultivars differing in bread-making
performance, Filipčev et al. (2013) found that the cultivars
showed the highest response to the action of ASC, with
more pronounced effects on bread quality in stronger spelts.
Use of citric acid (CIT) in bread-making has been
rarely considered. However, the occurrence of wheat flour of
extremely low quality (extensigraph area [flour energy]
around or below 10%; Šimurina et al. 2012) has led to its
use as a compound in combined dough conditioners. In an
earlier work, Kefalas et al. (2009) indicated that CIT makes
gluten more inclined to strengthening. This effect may be
closely associated with dough acidification during which
hydrogen anions from acid bind to negative parts of amino
acids, changing the total charge of protein macromolecule
positive hence making it prone to strengthening (Galal et al.
1978; Damodaran and Kinsella 1982). According to Liu
(1970), depending on dose and substrate type, CIT may
exert oxidizing character.
Xylanases (XYLs) are pentosan-modifying enzymes
which reduce the water-binding capacity of pentosans, thus
increasing the availability of water for gluten development.
In addition, XYLs break down the pentosan network, which
limits the aggregation of glutenin polymers. Therefore,
the action of XYL strengthens the gluten (Si and Drost-
Lustenberger 2002). XYLs having larger action on insoluble
pentosans by turning them into soluble ones positively
affect dough rheology by stabilizing gas cells and improving
their expansion capacity during proofing and baking (Gan
et al. 1995; Steffolani et al. 2010). Although different types
of XYL preparations may exert different effects on pento-
sans, good baking XYL gives dry dough, improves gluten
extensibility and elasticity, improves oven spring and loaf
volume and softens crumb (Butt et al. 2008).
Journal of Food Quality 37 (2014) 185–195 © 2014 Wiley Periodicals, Inc.
B. FILIPČEV, O. ŠIMURINA and M. BODROŽA-SOLAROV
The aforementioned additives are suitable for production
of natural and organic products. ASC and CIT are generally
recognized as safe (GRAS)-listed and permitted in organic
food within the EU, whereas enzyme additives replace
potentially harmful chemical food additives such as emulsi-
fiers and bromates.
Application of various dough improvers and their combi-
nations in conventional wheat has been extensively studied
in the past decades; however, little data can be found with
regard to spelt wheat. Therefore, the present study was
focused on spelt material from organic production charac-
terized with different baking qualities, with the objective to
elucidate the effect of XYL, ASC and CIT combinations on
bread quality. Another goal was to develop an optimized
improver formulation using response surface methodology
(RSM).
MATERIALS AND METHODS
Spelt Samples and Baking Ingredients
The material consisted of two spelt cultivars grown in an
environment where organic cultivation practices have been
employed (organic farm “Jevtić” in Bačko Gradište, Serbia).
One cultivar was Eko-10 originating from Hungary, and
the other was not identified originating from Austria
or Germany, here designated as western spelt. Spelt was
dehulled and milled on a farm-scale dehuller (Heger
type DS I 400S, Herrenberg D-7033, Heger GmbH&Co. KG,
Germany) and roll mill (A 500 MSM KOMBI Getreide
Mühle, Osttiroler Getreidemühlen, Stribach-Dölsach,
Austria), to flour with moisture and ash contents of 10.23
and 0.95% dry basis (d.b.; western spelt) and 10.39 and
1.30% d.b. (Eko-10), respectively.
Commercial brand of XYL preparation BXP 5000BG
(Bakezyme DSMProduct, DSM Food Specialities B.V., Delft,
Herleen, The Netherlands) was used. L-ASC and anhydrous
CIT were from BASF (Ludwigshafen, Germany). Fresh
bakery yeast was from Alltech-Fermin (Serbia).
Bread-Making Procedure
A modified bread-making procedure adjusted to spelt wheat
was used (Filipčev et al. 2013). The basic dough formulation
was 700 g flour, 17.5 g yeast, 14.5 g salt and water. Dough
improvers doses were varied according to the experimental
design displayed in Table 1. The water used in formula-
tions was according to farinograph absorption (Table 2).
The dough was mixed in a laboratory-scale Diosna mixer
(Dierks&Söhne GmbH, Osnabrück, Germany) at low speed
(85 rev/min) for 1 min and at high speed (120 rev/min) for
4 min. The final dough temperature was 25C. Dough was
left to rest in bulk for 1 h at 25C, with manual rounding
Journal of Food Quality 37 (2014) 185–195 © 2014 Wiley Periodicals, Inc.
REGULATING THE QUALITY OF ORGANIC SPELT BREAD
TABLE 1. EXPERIMENTAL DESIGNa
Actual levels (mg/kg)
Coded Ascorbic acid Citric acid Xylanase
levels ASC CIT XYL
−1.41 20 0 50
−1 50 10 60
0 125 50 90
+1 200 90 120
+1.41 230 100 130
a
Design comprised 15 runs in random order. The centre point was
replicated five times; factorial and star points were not replicated.
every 20 min. Dough was then divided into 350-g portions,
manually rounded, rolled and put into baking pans
(24.5 × 9 × 6.5 cm) to final proofing for 50 min (western
spelt cultivar) and 40 min (cv. Eko-10). Baking was carried
out for 20 min at 220C. Bread quality characteristics were
assessed 24 h after baking. Per trial entry, three loaves
were analyzed. Cooled loaves were packed in polyethylene
pouches and stored at ambient temperature.
Bread Evaluation
Loaves were weighted and loaf volume was measured by
millet seed displacement method (AACC Standard 10-05,
2000). Loaf specific volume was calculated as loaf volume
(mL)/loaf weight (g).
Crumb texture features were determined instrumentally
on a texture analyzer TA.XTplus (Stable Micro Systems,
England, UK) according to AACC method 74-10A modified
to record crumb resilience in addition to crumb hardness.
A 36-mm aluminum cylindrical probe and a 5-kg load
cell were used. Pre-test speed, test speed and post-test speed
were 1.0 mm/s, 1.7 mm/s and 1.7 mm/s, respectively. Breads
were evaluated at 40% compression strain. Crumb hardness
was recorded as the peak force during compression. Crumb
resilience was calculated as the area during the withdrawal
of the first compression divided by the area of the first com-
pression, and is an indicator of the speed of crumb recovery.
Flour Analyses
Standard tests were used for the determination of proteins
(ICC Standard No. 105/2, 1994), Zeleny’s sedimentation
value (ICC Standard No. 116/1, 1994), wet gluten content
and gluten index (ICC Standard No. 155, 1994) and falling
number (ICC Standard No. 107/1, 1995). Gluten index after
60 min resting (GI 60) was as described by Aja et al. (2004)
with a slight modification related to the incubation of a
dough piece instead of wet gluten. Empirical rheological
properties were analyzed by farinograph (ICC Standard No.
187
REGULATING THE QUALITY OF ORGANIC SPELT BREAD B. FILIPČEV, O. ŠIMURINA and M. BODROŽA-SOLAROV

TABLE 2. SPELT FLOUR CHARACTERISTICS

Quality parameters Western spelt cultivar Spelt cv. Eko-10
HMW-GS 1* 6.1 + 22.1 5 + 10 1* 6 + 8 2 + 12
Ash (% d.b.) 0.95a 1.30b
Protein (% d.b.) 12.77a 13.87b
Wet gluten (%) 37.05a 46.3b
Gluten index (%) 86.50b 36.50a
Gluten index after 1 h dough resting (%) 52.0b 0.15a
Zeleny sedimentation (mL) 34.4a 50.2b
Water absorption (%) 58.05a 61.95b
Dough stability (min) 0.75b 0.00a
Dough weakening (BU) 77.5a 130.0b
Extensigraph area (cm2) 47.5 –
Resistance, R (BU) 132.5 –
Extensibility, E (cm) 218.5 –
R/E 0.61 –
W (10−4 J) 46.0b 34.0a
P (mm H2O) 26.0a 27.5a
L (mm) 144.0b 66.0a
P/L 0.18a 0.42b
G 26.7b 18.1a

Different letters in a row denote significant difference at P < 0.05 (Tukey’s honest significant dif-
ference [HSD] test).
Standard bakery requirements for bread wheat (Ðaković 1997; Pasqualone et al. 2011; Šimurina
2013):
Gluten index: 60–90%; Farinograph stability: 0.5–10 min; Dough weakening: 75–125 BU
medium quality; Extensigraph area: >50 cm2; Resistance: 200–400 BU; R/E: 1.80; P/L: <0.6;
W: 130–200 × 10−4 J; G: 20–23.
HMW-GS, high molecular weight glutenin subunit.

115/1, 1992), extensograph (ICC Standard No. 114/1, 1992)
and alveograph (ICC Standard No. 121, 1996).
Statistics
To compare the flour characteristics as well as the control
breads with the two spelt cultivars, one-way analysis of vari-
ance (ANOVA) was used. Tukey’s test was applied at a 5%
significance level to compare means. Statistical analysis
software package STATISTICA 10 (StatSoft Inc., Tulsa, OK;
released in 2010) was used.
To study the simultaneous effects of XYL, ASC and CIT,
response surface methodology (RSM) was employed. A
small central composite rotatable design was prepared
(Table 1). The doses of dough improvers were selected
according to preliminary trials and previous publica-
tions (Hilhorst et al. 1999; Selomulyo and Zhou 2007;
Pescador-Piedra et al. 2009; Ghorbel et al. 2010; Šimurina
et al. 2012). Responses were bread attributes described
under bread evaluation: loaf specific volume, crumb hard-
ness and resilience. Model selection (linear, two-factor inter-
action [2FI] or quadratic) for each response was made on
the basis of sequential model sum of squares, lack-of-fit
tests and the multiple correlation coefficients (R2). Sug-
gested model was the highest degree model for which the
F tests showed significant effects (P < 0.05, 0.01 or 0.001),
with insignificant lack-of-fit test and R2 close to 1. The sta-
tistical significance of coefficients for all terms in the model
was tested.
The optimization of dough improvers’ doses was based
on the technique of numerical optimization of multiple
responses. The desirability function approach was used
(Myers and Montgomery 2002). All three responses were
used for optimization. The goal for volume was maximum,
with the relative importance of five; the goal for crumb
hardness was minimum, with the relative importance
of four, and the goal for resilience was maximum, with
the relative importance of four. All calculations were per-
formed with Design-Expert 5 statistical package (Stat-Ease
Corporation, Minneapolis, MN; released in 1996).
RESULTS
Characterization of the Spelt Flours and
Control Breads (Without Improvers)
Table 2 shows the flour characteristics of the two spelt
cultivars used in the study. The two spelt cultivars had dif-
ferent high molecular weight glutenin subunit (HMW-GS)
protein compositions. In comparison to standard bakery

188 Journal of Food Quality 37 (2014) 185–195 © 2014 Wiley Periodicals, Inc.
B. FILIPČEV, O. ŠIMURINA and M. BODROŽA-SOLAROV REGULATING THE QUALITY OF ORGANIC SPELT BREAD

TABLE 3. QUALITY ATTRIBUTES FOR CONTROL SPELT BREADS showed significant difference and it was better in stronger,
(WITHOUT IMPROVERS) western spelt cultivar, which had significantly softer crumb.
Quality parameter Western spelt cultivar Spelt cv. Eko-10
Loaf specific volume (mL/g) 2.54 ± 0.02a 2.57 ± 0.02a
Effect of Improver Combination on
Hardness (g) 1,411.47 ± 36.17a 2,454.8 ± 56.97b
Resilience (%) 22.90 ± 1.08a 23.96 ± 0.48a
Bread Characteristics

Different letters in a row denote significant difference at P < 0.05
(Tukey’s honest significant difference [HSD] test).
requirements, the used cultivars formed weaker doughs
with low stabilities, which is not unexpected for spelt
doughs. However, there were differences between the two
cultivars. The western spelt cultivar had lower protein
content but better gluten quality characteristics (higher
gluten indices), and consequently formed stronger dough
(lower dough softening, higher deformation energy). The
Eko-10 cultivar had poorer dough characteristics (higher
dough softening, lower deformation energy) and it was
even not possible to record extensigrams. This performance
was in agreement with their genetic potential reflected by
HMW-GS composition (Table 2). Presence of HMW-GS
5 + 10 subunit is associated with higher baking perfor-
mance, whereas HMW-GS 2 + 12 with poorer technological
characteristics. However, the breads produced from the
two spelt cultivars did not significantly differ in specific loaf
volume and crumb resilience (Table 3). But crumb hardness
Table 4 displays the fit summaries for the best fitting
models and the significance of model coefficients for the
selected bread characteristics. Figure 1 displays the three-
dimensional response surface plots and perturbation plots
illustrating the effects of factors on loaf specific volume,
crumb hardness and resilience for both spelt cultivars.
Loaf volume is one of the most important quality criteria
for consumers. Loaf specific volume was fitted with linear
models for both spelt cultivars (Table 4). Positive effect
of XYL was evident from the perturbation plot (Fig. 2) for
Eko-10 cultivar. However, for the western spelt cultivar,
no significant linear effects on specific loaf volume were
observed, but significant interaction effects existed (Table 4,
Fig. 2). Within the selected range of factor levels, the inter-
action of ASC and XYL positively affected on loaf volume,
whereas interactions with CIT had a negative effect on
loaf volume. For Eko-10 cultivar, higher loaf volumes were
obtained with higher ASC doses.
Crumb texture is an important bread attribute because
it is directly linked to consumers’ acceptance. In both culti-
vars, the added improvers showed improving linear effects

TABLE 4. ANALYSIS OF VARIANCE AND REGRESSION COEFFICIENTS FOR THE RESPONSES

Responses

Loaf specific volume (mL/g) Crumb hardness (g) Crumb resilience (%)
ANOVA Western cultivar cv. Eko-10 Western cultivar cv. Eko-10 Western cultivar cv. Eko-10
Suggested model 2FI Linear Quadratic Quadratic Quadratic Quadratic
Model F value 5.66** 4.26** 32.26** 17.87*** 4.17* 2.24 n.s.
R2 0.81 0.54 0.98 0.97 0.88 0.80
R2 0.67 0.41 0.95 0.91 0.67 0.44
Lack-of-fit n.s. n.s. n.s. n.s. n.s. n.s.
C.V. (%) 2.73 2.12 3.63 3.38 2.22 2.29
Predictors Coded coefficient estimates
Intercept 3.31 2.84 1,656.60 2,049.37 27.49 25.00
ASC −0.0033 0.032 −15.83 −20.67 −0.89** −0.11
CIT 0.074 −0.0002 −120.68** −53.99 0.053 0.29
XYL 0.011 0.069*** −114.07** −187.07*** −0.56 −0.58
ASC × CIT −0.16** – 236.63*** 5.26 −0.87* −0.85
ASC × XYL 0.21** – 351.65*** 80.70 1.04* −0.08
CIT × XYL −0.13* – 177.53** 149.91** −1.82*** −0.11
ASC2 – – 46.38 20.47 0.02 0.01
CIT2 – – 71.16** −167.98** −0.10 0.72**
XYL2 – – 182.15*** 8.10 −0.43 −0.12

* P < 0.1; ** P < 0.05, *** P < 0.01.

R2 coefficient of determination.
R 2 adjusted coefficient of determination.
2FI, two-factor interaction model; ASC, ascorbic acid; CIT, citric acid; C.V., coefficient of variation; n.s., not significant; XYL, xylanase .

Journal of Food Quality 37 (2014) 185–195 © 2014 Wiley Periodicals, Inc. 189
REGULATING THE QUALITY OF ORGANIC SPELT BREAD B. FILIPČEV, O. ŠIMURINA and M. BODROŽA-SOLAROV

Western
spelt
cultivar

Spelt cv.
Eko-10

FIG. 1. THREE-DIMENSIONAL RESPONSE SURFACE PLOTS ILLUSTRATING THE EFFECTS OF XYLANASE, ASCORBIC AND CITRIC ACID ON LOAF SPE-
CIFIC VOLUME (ml/g), CRUMB HARDNESS AND RESILIENCE FOR SPELT CULTIVARS
On each plot, the effects of a combination of two factors (ascorbic and citric acid) are given whereas the third factor (xylanase) is kept constant on
the medium level (0 in coded terms).

on crumb hardness of which the effect of XYL was signifi-
cant. All synergistic effects between the added components
were significant in the case of western spelt wheat, whereas
in the case of Eko-10 synergistic effect between CIT and
XYL existed. Synergistic effect between ASC and XYL had
a decreasing effect on crumb hardness in western spelt,
whereas other interactions showed a crumb hardening
trend. CIT and XYL produced a significant quadratic effect
on crumb hardness increase in the case of western spelt
cultivar. In the case of weaker cultivar Eko-10, CIT also
exhibited a significant but opposite quadratic effect on
crumb hardness; the maximum of crumb hardness was
reached at medium levels of CIT. ASC showed a slight trend
on improving crumb hardness in both cultivars. XYL clearly
decreases hardness in the case of weaker spelt cultivar,
whereas in stronger spelt cultivar it exhibits crumb soften-
ing up to an optimum point, in agreement with significant
quadratic effect (Fig. 3). CIT decreases crumb hardness in

Western spelt cultivar Spelt cultivar Eko-10

FIG. 2. PERTURBATION DIAGRAMS ON THE EFFECT OF COMPOUNDS ON LOAF SPECIFIC VOLUME (ml/g) IN TESTED SPELT CULTIVARS
(A) Ascorbic acid. (B) Citric acid. (C) Xylanase.

190 Journal of Food Quality 37 (2014) 185–195 © 2014 Wiley Periodicals, Inc.
B. FILIPČEV, O. ŠIMURINA and M. BODROŽA-SOLAROV
Western spelt cultivar
FIG. 3. PERTURBATION DIAGRAMS ON THE EFFECT OF COMPOUNDS ON CRUMB HARDNESS (g) IN TESTED SPELT CULTIVARS
(A) Ascorbic acid. (B) Citric acid. (C) Xylanase.
stronger spelt but in weaker spelt, higher doses of CIT
would be required for substantial crumb softening effect
(significant quadratic effect) (Fig. 3).
Crumb resilience indicates the rate of crumb recovery
after the removal of loading. In stronger spelt, four signifi-
cant effects on crumb resilience were recorded at various
levels of significance: negative effects of ASC and interac-
tions CIT–ASC and CIT–XYL, and a positive effect of inter-
action between ASC and XYL. In the case of weaker spelt
cultivar, a low R 2 value and a non-significant F value indi-
cated that the model was not significant and only a major
Western spelt cultivar
FIG. 4. PERTURBATION DIAGRAMS ON THE EFFECT OF COMPOUNDS ON CRUMB RESILIENCE (%) IN TESTED SPELT CULTIVARS
(A) Ascorbic acid. (B) Citric acid. (C) Xylanase.
Journal of Food Quality 37 (2014) 185–195 © 2014 Wiley Periodicals, Inc.
REGULATING THE QUALITY OF ORGANIC SPELT BREAD
Spelt cultivar Eko-10
quadratic effect of CIT was found significant with improv-
ing trend. Perturbation diagrams (Fig. 4) showing the effect
of various levels of independent factors on crumb resilience
reveal the higher sensitivity of weaker spelt to the action of
CIT regarding crumb resilience.
Optimization
Numerical optimization by the RSM procedure resulted in
minimal ASC and CIT dose with higher XYL dose for weak
spelt cultivar (Eko-10), i.e., medium-low ASC dose, high CIT
Spelt cultivar Eko-10
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REGULATING THE QUALITY OF ORGANIC SPELT BREAD B. FILIPČEV, O. ŠIMURINA and M. BODROŽA-SOLAROV

TABLE 5. COMPARISON OF PREDICTED VERSUS MEASURED VALUES FOR OPTIMIZED FORMULATION OF BREAD MADE FROM TWO SPELT
CULTIVARS

Western spelt cultivar Spelt cv. Eko-10

% improvement % improvement
Predicted compared with Predicted compared with
value Measured value control value Measured value control
Loaf specific volume (mL/g) 3.87 3.69 ± 0.07 45.3 2.94 2.91 ± 0.020 13.23
Hardness (g) 1,199.94 1,216.65 ± 20.68 16.01 1,681.75 1,805.75 ± 16.69 35.94
Resilience (%) 32.22 29.58 ± 2.591 29.2 25.4686 24.866 ± 0.483 3.80

and medium-low XYL dose for strong, western spelt culti-
var. Therefore, the optimized improver formulations were:
50 mg/kg ASC, 90 mg/kg CIT, 60 mg/kg XYL for western spelt
cultivar and 20 mg/kg ASC, 10 mg/kg CIT, 120 mg/kg XYL for
Eko-10. Optimized formulations yielded better quality breads
for both spelt cultivars (Table 5, Fig. 5). Although the predicted
values were not completely reached, generally the measured
responses corresponded favorably. In comparison to corre-
sponding control breads, improved formulations contri-
buted to substantial increases in specific volume (by 45%) and
crumb resilience (by almost 30%) for western spelt cultivar,
and crumb softening (almost 36%) for Eko-10 spelt cultivar.
Steffolani et al. (2010) reported a 37% volume increase in
bread with optimized formulation containing the combination
of glucose-oxidase, XYL and amylase.
DISCUSSION
XYL had improving effect on loaf volume and crumb hard-
ness in both cultivars. High specific volume increase in the
optimized bread formulation for strong spelt cultivar
obtained in this study might be due to high initial dough
extensibility enhanced by the action of XYL. It was also
observed here a significant ASC/XYL synergistic effect on
improving loaf volume and crumb resilience, as well as sig-
nificant crumb hardening effect in stronger spelt cultivar.
Dagdelen and Gocmen (2007) reported that the best effect
on bread loaf volume was exerted by glucose-oxidase/
hemicellulase combinations. This shows that XYL has
synergistic effect with dough oxidizing agents. In practice,
it is known from earlier report that combinations of
hemicellulase or XYL with amylase have synergistic effect
(Si and Drost-Lustenberger 2002). XYL alone as well as
in combinations with amylase showed positive effects on
the overall quality of chappati produced from both good
and poor quality wheat flour (Hemalatha et al. 2010).
Grausgruber et al. (2008) reported significant improv-
ing individual effect of XYL and synergistic effects with
amylase, transglutaminase, and/or emulsifiers on loaf
volume and crumb firmness of einkorn bread.

Spelt cultivar Control sample Optimized formulation

Western
spelt

Eko-10

FIG. 5. CROSS-SECTIONS OF CONTROL SPELT

BREADS AND OPTIMIZED FORMULATIONS

192 Journal of Food Quality 37 (2014) 185–195 © 2014 Wiley Periodicals, Inc.
B. FILIPČEV, O. ŠIMURINA and M. BODROŽA-SOLAROV
Within the selected range of factor levels, CIT decreased
crumb hardness in both cultivars but with significant linear
effect in stronger spelt and significant quadratic effect in
weaker spelt cultivar. Also, CIT had a tendency to increase
crumb resilience in both cultivars but with significant qua-
dratic effect only with weak spelt cultivar. However, none of
the interactions of CIT with other factors showed improv-
ing effects on volume, hardness or resilience in both culti-
vars. These interactions were significant for stronger spelt
cultivar but not for weaker one. CIT was shown to linearly
increase extensibility in doughs made from extremely low-
quality flour (extensigraph energy 4 cm2) from conventional
wheat but with negative quadratic effect (Šimurina et al.
2012). Moreover, Šimurina et al. (2012) reported that the
addition of CIT significantly decreases the content of free
thiol groups in dough. The same authors found a significant
synergistic effect between CIT and ASC in improving dough
rheology of substandard quality wheat. The optimum doses
to increase dough energy and extensibility in previously
mentioned substandard flour were 97 mg/kg ASC and
100 mg/kg CIT. As mentioned earlier, the addition of CIT in
combination with ASC may enhance dough oxidation by
making it more prone to oxidation.
ASC had a tendency to reduce crumb resilience in both
cultivars but significant effects were registered only with
stronger spelt cultivar. This is probably due to the gluten-
strengthening effect of ASC. But, the synergistic effect of
XYL with ASC was positive and significant for this cultivar
regarding crumb resilience.
The above results suggest that treating both strong and
weak spelt cultivars with XYL, ASC and CIT in combina-
tion produced a significant improvement on the quality
of breads. Better results were obtained with stronger spelt
cultivar (western spelt). This supports the observation of
Dagdelen and Gocmen (2007) that the effects of dough
improvers and their combinations are highly dependent on
dose and the initial flour quality.
CONCLUSIONS
The results showed that the tested spelt cultivars differed
in gluten quality, which mainly manifested as significant
difference in dough softening, extensibility and alveograph
deformation energy. However, their bread-making perfor-
mance did not differ significantly except in crumb hardness,
which was significantly lower in stronger spelt (western
spelt cultivar).
XYL had improving effect on loaf volume and crumb
hardness in both cultivars. In stronger spelt cultivar, there
was a significant ASC/XYL synergistic effect on improving
loaf volume and crumb properties. CIT decreased crumb
hardness in both cultivars (with higher doses more efficient
in weaker spelt) and increased crumb resilience in weak
Journal of Food Quality 37 (2014) 185–195 © 2014 Wiley Periodicals, Inc.
REGULATING THE QUALITY OF ORGANIC SPELT BREAD
spelt cultivar (at higher doses). Optimization showed that
combination of medium-low ASC dose, high CIT and
medium-low XYL dose was optimal for western spelt
(stronger cultivar), whereas minimal ASC and CIT dose
with higher XYL dose was most suitable for cultivar Eko-10
(weaker spelt). In comparison to control formulations,
optimal formulations yielded substantial increases in spe-
cific volume (by 45%) and crumb resilience (by almost
30%) for western spelt cultivar, and significant crumb soft-
ening (almost 36%) for Eko-10 spelt cultivar. Evidently,
combinations of XYL, ASC and CIT can be used to improve
the quality of spelt breads, which are important commodi-
ties in the natural and health product markets.
ACKNOWLEDGMENTS
This paper resulted as a part of research work supported
financially by the Ministry of Education, Science and Tech-
nological Development (Republic of Serbia) under contract
No. III 46005. The authors also thank Srd̄an Šolaja (Barentz
d.o.o., Belgrade, Serbia) for providing the enzyme sample.
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