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ORIGINAL ARTICLE
Development of gluten-free muffins utilizing squash seed dietaryfiber
Marıa I. Palacio1,2 • Analıa I. Etcheverrıa2 • Guillermo D. Manrique1
Revised: 21 April 2018 / Accepted: 9 May 2018
� Association of Food Scientists & Technologists (India) 2018
Abstract Gluten-free muffins with squash seed flour (SSF)
were developed for contribute to reduce nutritional defi-
ciencies and improving the health of the celiac population.
Physicochemical and sensory properties of muffins were
evaluated. SSF was incorporated at two levels (10 and 20%
w/w) in commercial gluten-free premix (control). Incor-
poration of SSF increased total dietary fiber content, pro-
tein and unsaturated fatty acids. The addition of SSF at
10% resulted in a muffin that did not differ significantly
from the control muffin; and also that formulations with
SSF at 20% caused an increase in the browning index.
Browning was favored by the increase of SSF with higher
levels of fiber content. Incorporation of SSF at 20% had a
significant effect on the textural parameters (firmness and
chewiness) of the muffin. Also, both formulations con-
taining SSF showed a higher overall acceptability, partic-
ularly muffins with 20% of SSF that rendered the highest
scores for sponginess, texture, taste and colour.
Keywords Gluten-free � Muffins � Squash seed flour �Dietary fiber � Bake
Introduction
In recent years there has been an increasing demand of
gluten-free foods as about 1% of the world population is
suffering from celiac disease (CD) (Shevkani and Singh
2014; Singh et al. 2016). CD is an inflammatory disease of
the small intestine triggered by gluten proteins from cere-
als, particularly wheat, barley and rye. Cereal proteins
comprise up to 80% of gluten proteins which play an
important role in the baking industry (Drabinska et al.
2016). Gluten is composed of two main protein fractions:
gliadins, which contribute essentially to the viscosity of the
batter, and glutenins, which are responsible for batter
elasticity (Wieser 2007). The only known treatment for CD
patients is strict lifelong gluten-free diet (Farrell and Kelly
2002; Fasano and Catassi 2001). Most of the gluten-free
products are generally made from flours that are deficient
in dietary fiber (DF) (Thompson et al. 2005; Majzoobi et al.
2016). Fiber supplements do not have the same health
benefits as naturally occurring high-fiber foods. Several
studies suggested that people should consume more natural
fiber which is found in whole grains, seeds, fruits and
vegetables (Parisi et al. 2002; Flint et al. 2009; Brownlee
2011). Reportedly, DF has been known to improve and
maintain human health. According to Meghwal and
Kadeppagari (2016) there is fair evidence that DF from
whole foods lowers blood pressure, improves serum lipids
and reduces inflammation. Another study reported a range
of cardiovascular benefits associated with DF (Flint et al.
2009). Nevertheless, DF intake is commonly lower than
recommended (Rubel et al. 2014).
& Marıa I. Palacio
1 Departamento de Ingenierıa Quımica, Facultad de Ingenierıa,
Universidad Nacional del Centro de la Provincia de Buenos
Aires (UNCPBA), Av. Del Valle 5737, B7400JWI Olavarrıa,
Argentina
2 Consejo Nacional de Investigaciones Cientıficas y Tecnicas
(CONICET), Facultad de Ciencias Veterinarias, Centro de
Investigacion Veterinaria de Tandil (CIVETAN), Laboratorio
de Inmunoquımica y Biotecnologıa, UNCPBA,
B7000 Tandil, Argentina
123
J Food Sci Technol
https://doi.org/10.1007/s13197-018-3213-z
Muffins are widely-consumed sweet, spongy breakfast
or evening snacks prepared traditionally from wheat flour,
sugar, oil/fat, milk and eggs, which means that people
having CD cannot consume them (Ciesarova et al. 2016).
However, gluten-free muffins are of interest in present time
not only for people who have CD, also for people who are
interested in gluten-free products (Singh et al. 2015).
Squash (Cucurbita maxima) seeds are gluten-free and can
be used by those with CD. Many of the health benefits of
squash seeds have been attributed to the following factors:
large amounts of lipids of the type of polyunsaturated and
monounsaturated fatty acids, a-galactosides with prebiotic
activity, proteins with well-balanced amino acids and high
biological value, phenolic compounds, and high dietary
fiber content (Stevenson et al. 2007; Habib et al. 2015;
Palacio et al. 2014; Srbinoska et al. 2012; Alfawaz 2004).
Squash seed flour (SSF) seems to be a potential source of
natural and functional ingredients related to health-pro-
moting and/or CD prevention and can be used for enriching
fiber gluten-free muffins. There is no available published
data on muffin formulation containing SSF blended with
commercial gluten-free flour. So, the objective of the pre-
sent study was to develop a gluten-free muffin containing
various levels of SSF (0, 10 and 20%). Because the addi-
tion of fibers may cause a significant effect on the final
muffin quality, the physicochemical and sensory properties
were also evaluated and compared with those obtained
using commercial gluten-free flours.
Materials and methods
Materials
Squash seeds (Cucurbita maxima) were purchased from
‘‘For Good’’ in a local gluten-free market (Tandil, Argen-
tina), sealed and stored in polyethylene bags until used.
The SSF was obtained from the milling of squash seeds to a
fine powder in a cereal mill (Mod. TDMC, TecnoDalvo,
Argentina) and sieved through a 32 mesh sieve (500 lm).
The resulting flour showed the following characteristics:
fats 40.16%; proteins 28.72%; total dietary fiber (TDF)
25.69%; ashes 3.63%; and carbohydrates 1.80%. A ready-
to-bake dry chocolate gluten-free premix (GFP) (Kapac,
Argentina) was used as control. The ready-to-bake dry GFP
ingredients were: powdered sugar, cornstarch, rice flour,
whole milk powder, cocoa powder, whole egg powder,
whipping agent (INS 471, INS 477, milk proteins, glucose
solids), and salt. Leavening agents: INS 500ii INS 541i.
Thickener: INS 412. Flavoring: vanillin. Other ingredients
used were vanilla essence (Castillo, Argentina) and water,
both at the same volume for the three muffin formulations
(1 and 50 ml, respectively).
Batter formulation and baking procedure
Three muffin formulations were prepared replacing GFP
with 0, 10 and 20% of SSF, respectively, 50 mL of water
and 1 mL of vanilla essence gluten-free. The sample
without SSF was called control (C). Two samples with 10
and 20% of SSF, respectively, were called SSF-10 and
SSF-20. The choice of these levels was based on the
proximate composition of SSF and on the most common
values used in other studies of fiber enriched baked prod-
ucts (Costantini et al. 2014; Bialek et al. 2015). Ingredients
were individually weighed using electronic scale (Model
V-350, ACCULAB, USA).
For the development of the muffins, the dry ingredients
and the specified amount of water (50 mL) were placed
into the mixer of a bread maker machine (HP4030E,
ATMA, China) using the program ‘‘batter’’ during 10 min
in order to standardize the mixing procedure for all for-
mulations. Then, approximately 12 g of the resulting
preparation were placed into individual paper cups. Muf-
fins were baked at 190 �C for 10 min in a convection
electric oven (DELI-4, Moretti, Argentina). Finally, muf-
fins were cooled at room temperature for 1 h and packed
individually in heat-sealed food grade plastic bags until
used. Muffin parameters were assessed at 20 �C 3-h after
baking.
Proximate composition
Moisture, protein, total fat, and ash content of muffin
samples were done according to AACC (Methods: 44-19,
Kjeldahl, 30-25.01, 08-01, 2000). Muffins were defatted
with hexane and dried for analysis of and TDF. Soluble,
insoluble and TDF was determined by commercial kit
following AOAC method 985.29 (2000). Digestible car-
bohydrate content was obtained by difference method.
Results were expressed as g/100 g of product (fresh basis).
Calorie contents were also calculated using the following
specific energy factors: 9 for fats, 2 for TDF, and 4 for
carbohydrates and proteins, respectively (AAC 2004) and
changed to kJ.
Muffin quality evaluation
The quality of the muffins obtained for each formulation
was evaluated by determining parameters related to vol-
ume, moisture, crumb structure, alveoli, texture and colour:
Specific volume Specific volume (Vs) was determined as
a ratio of volume and weight (AACC 2000). Four muffins
of each formula were analyzed.
Moisture content Moisture content of the muffin crumb
was determined by oven drying for 2 h at 135 �C. For eachmeasurement, approximately 3 ± 0.001 g of crumb were
J Food Sci Technol
123
taken from a central slice of the piece. Four muffins of each
formula were analyzed.
Crumb structure Two central and vertical slices (12 mm
thick) were cut from each muffin and placed in a light-box.
Colour images of slices were acquired using a Nikon
D3100 digital camera (35 mm, 1/8, f/5, ISO 100), with a
resolution of 300 pixels per inch. A square field of view of
40 9 40 mm2 of the slice was considered for the structure
analysis. The image was converted to 8-bits in grey scale
(ranging from 0 to 255) and then was binarized (converted
from gray-level to black and white) using the algorithm
Isodata (ImageJ 1.47 v software, National Institute of
Health, Bethesda, MD, USA). Black dots represent alveoli.
Image imperfections were considered with an alveolar
threshold of 0.005 cm2 (Salinas et al. 2015). The numbers
of cell per area (N) (cells/mm2), mean cell area (AM)
(mm2) and the void fraction or total area occupied by
alveoli (AT) (%) were calculated.
Crumb texture Texture profile analysis (TPA) of muffin
slices was performed using a texture analyzer INSTRON
(Model 3343, USA) equipped with a 50 N load cell. From
the middle part of each muffin two slices 2 cm high were
obtained. Slices were subjected to a double compression
cycle (deformation: 40%, crosshead speed: 0.5 mm/s) with
a cylindrical probe (diameter: 25 mm). The parameters
determined were firmness, cohesiveness, springiness and
chewiness. Eight measurements were performed for each
formulation sample (4 muffins 9 2 slices).
Crumb colour Colour measurements were performed on
the crumb using a tri-stimulus colour analyzer (Chroma
Meter CR-400, Konica Minolta, Japan). Ten measurements
per formula were analyzed. Reflected colour was measured
at 108-observer angle with D65 illuminant. Values of L*,
a* and b* were measured and used to calculate the
browning index (BI) parameter (Buera et al. 1985) with the
following equations:
BI =100ðX � 0:31Þ
0:172ð1Þ
X =a�þ1:75L�
5:645L� + a��0:12b�ð2Þ
Sensory evaluation
Finally, muffins prepared from C, SSF-10 and SSF-20
formulations were subjected to sensory evaluation by an
untrained panel of twenty celiac persons (12 females and 8
males with ages ranging from 16 to 74 years). Three
muffins (one of each formulation, coded with three digits
and presented in random order) were served to each pan-
elist accompanied with water to cleanse their palates
between sample tasting. Panelists were asked to evaluate
attributes as crumb colour, taste, sponginess (defined as
porosity), crumb texture and general acceptability. Also,
they were asked about the possibility of buying the product
or not. They were asked to score each attribute on a 9-point
hedonic scale where one is Dislike Extremely and nine is
Like Extremely (Watts et al. 1989). Friedman Test (paired
data) was used to analyze sensory results (InfoStat
Software).
Statistical analysis
Results were expressed as the mean ± standard deviation
(SD) for each parameter. Differences were tested by one-
way analysis of variance (ANOVA) followed by Tukey test
to determine significant differences (p\ 0.05) using
Infostat version 2011. Different letters were used to label
values with statistically significant differences among
them.
Results and discussion
Proximate composition of muffins
The proximate composition of C, SSF-10 and SSF-20 is
shown in Table 1. The addition of SSF to an all-purpose
ready-to-bake dry premix in muffin formulation resulted in
nutritional differences of the final products obtained. The
ash contents of all the samples did not differ significantly
(p\ 0.05). Also the results showed that both SSF-10 and
SSF-20 formulations presented significantly higher fat,
protein and TDF contents as compared to C sample
(p\ 0.05). These increased values in muffins formulated
with SSF occurred concomitantly with a decrease in
assimilable carbohydrates compared to C (p\ 0.05), giv-
ing rise to products with a higher protein content and a lipid
fraction enriched with polyunsaturated and monounsatu-
rated fatty acids (Stevenson et al. 2007; Habib et al. 2015).
In addition, the observed increase in TDF in both formu-
lations containing SSF compared to C, allows these foods
to be included in the label the attributed ‘‘High in dietary
fiber,’’ according to the current national legislation (for this
claim the Argentinean Alimentarius Codex establishes that
a solid food must contain a minimal of 6 g of dietary fiber
per 100 g) (AAC 2004). The higher content of TDF in
muffins formulated with SSF is mainly due to the higher
proportion of insoluble fiber. On the other hand, in a pre-
vious study we have demonstrated that a-galactosidesextracted from Cucurbita maxima seeds exhibited a posi-
tive in vitro prebiotic activity using L. paracasei as a
probiotic microorganism (Palacio et al. 2014). From the
above, it is clear that muffins made with SSF, in addition to
providing compounds of a-galactoside type (soluble fiber)
J Food Sci Technol
123
with prebiotic activity, have improved the nutritional pro-
file with respect to the product obtained with the ready-to-
bake premix (control), while maintaining the characteristic
of being free from gluten-forming proteins.
In agreement with our results, Majzoobi et al. (2016)
found that the replacement of gluten-free flour with carrot
pomace poder at various levels (10, 20 and 30%) and two
particle size (210 and 500 lm) increased fiber, ash and
protein contents when compared to control samples. Other
authors (Gularte et al. 2012) reported the same results in
their studies, in which rice flour was substituted with dif-
ferent legumes (chickpea, pea, lentil and bean) on the
development of gluten-free cake products.
Quality evaluation of muffins
Technological quality of muffins was evaluated by deter-
mining the Vs, crumb colour, crumb structure, and crumb
properties such as moisture, alveoli and texture.
The Vs value represents the relationship between the
volume and weight of the product, parameter generally
recognized as useful for the evaluation of bread and bakery
products (Capriles et al. 2016). Higher volumes exert
positive economic effects for bakery product because
products with higher volume often attract consumers. Final
volume depends on batter expansion during baking pro-
cedure and on the ability of the matrix structure to retain
gas. In this study, according to the Tukey test no statistical
differences of Vs values for the three-sample muffin were
observed (p\ 0.05) (Fig. 1a). It can be thus concluded that
the substitution of the premix by SSF up to a 20% does not
alter the rheological properties of the muffins since it
allows a level of expansion and gas retention comparable to
that of the control formulation.
The final crumb moisture content of the muffins is
related to the water absorption during batter preparation
and the water loss during baking procedure. In fact, it has
been suggested that crumb morphology strongly affects the
Table 1 Proximate
composition (g/100 g dry
weight) and energetic value of
gluten-free muffins
Parameter (g/100 g dry weight) C SSF-10 SSF-20
Ash 3.44 ± 0.28 a 3.60 ± 0.02 a 3.70 ± 0.04 a
Fat 3.70 ± 0.07 a 8.62 ± 0.26 b 13.68 ± 0.16 c
Total dietary fibre 0 a 9.04 ± 0.45 b 11.11 ± 0.55 c
Soluble 0 a 2.92 ± 0.14 b 2.89 ± 0.14 b
Insoluble 0 a 6.12 ± 0.37 b 8.22 ± 0.41 c
Protein 5.45 ± 0.14 a 7.94 ± 0.20 b 10.76 ± 0.04 c
Carbohydratesa 87.41 ± 0.07 a 70.80 ± 0.08 b 60.75 ± 0.15 c
K caloriesb 161 164 172
kJb 674 686 720
Mean ± SD of triplicate analysis. Different letters in the same file indicate significant differences between
means (p\ 0.05)
SSF squash seed flour, C control, SSF-10 (10 g SSF/100 g gluten-free premix (GFP)), SSF-20 (20 g SSF/
100 g GFP)aCalculated by difference methodbValues of 40 g of edible portion
Fig. 1 Parameter of muffin quality: a specific volume, b crumb moisture, c crumb browning index. Different letters in the same graphics indicate
significant differences (p\ 0.05)
J Food Sci Technol
123
rate of water transport (Roca et al. 2006). As shown in
Table 2, the control sample showed the highest cell/total
area ratio. This probably sped up moisture migration and
increased loss during baking. Smaller pore size is expected
to be involved in slowing down the moisture migration on
SSF-20 muffin with respect to C. The addition of SSF to
muffin formulations increased the moisture content of
samples. The differences in moisture content between C
and the samples containing SSF, might also be due to the
high fiber content (Uthumporn et al. 2014). Therefore, the
C muffin crumb was the driest one and the addition of SSF
caused a significant increase in crumb moisture (p\ 0.05)
(Fig. 1b).
Colour of the muffin crumb is the most important
parameter taken into account by the consumers when
purchasing the product. The crumb colour depends mainly
on the type of flour, the quality and quantity of the ingre-
dients used, and the temperature and time of baking. The
BI indicates the level of brown colour and was reported as
an important parameter of quality related to Maillard
reactions (Saricoban and Yilmaz 2010). The extent of
browning also determines the flavor of the final product
(Ureta et al. 2014). Figure 1c shows the BI obtained for all
the formulations baked for the same time and at the same
temperature. Since these variables were the same for all
formulations, the results allow to conclude that the addition
of SSF at 10% resulted in a muffin that did not differ
significantly from the control muffin; and also that for-
mulations with SSF at 20% caused an increase in the BI
compared to the control formulation (p\ 0.05). Browning
was favored by the increase of SSF with higher levels of
fiber content. This increase may be due to the pigments
from the SSF that influenced the colour of the crumb.
According to Salinas et al. (2015), the BI is favored by the
presence of prebiotics on bakery products. Similarly, Fru-
tos et al. (2008) also reported that the increase of over 12%
of artichoke fiber caused darker bread colour.
Images of the structure of the muffins and respective
crumb centers of all formulations can be observed in
Fig. 2. Values of the structure parameters (number of
alveoli per area, mean alveolus area and percentage of total
air trapped) of muffin crumb for C, SSF-10 and SSF-20
formulations are shown on Table 1. Anova results showed
that muffin enrichment with SSF had a significant effect on
the structure of the crumbs. The addition of 20% of SSF per
100 g of mix resulted in the crumb with the smallest mean
cell area and cell density with significant differences
compared to C (p\ 0.05). Considering the sample with
10 g of SSF per 100 g of total mix, the crumb obtained had
a significantly smaller mean cell area accompanied by non-
significant differences in cell density, compared to the C
muffin (p\ 0.05). Finally, the addition of 20 g of SSF
caused a significant decrease on the cell/total area ratio
(void fraction) compared to the C sample.
TPA consists of the double compression of a food
sample imitating the action of the jaw. Figure 3 represents
the TPA of crumb of the three experimental samples.
Firmness was defined as the highest force required to
compress the muffin crumb under a constant deformation.
Another textural parameter considered was cohesiveness
that quantifies the internal resistance of the muffin structure
under some compression. A product with high cohesion is
more tolerant to handling and packaging, and thus it will be
presented to the consumers in its expected state (Singh
et al. 2016). Springiness is also important in baked prod-
ucts, a measure that is defined as the elasticity of the
muffins, calculated by measuring the extent of recovery
between the first and second compression. This textural
parameter is a desirable muffin property associated with an
elastic and fresh aerated product (Shevkani and Singh
2014). Anova results showed that the addition of SSF at
20% had a significant effect on the firmness and chewiness
textural parameters of the muffin crumb (p\ 0.05).
Beginning with firmness, we observed an increase in
comparison to C sample. Secondly, the C muffin crumb
had no significant differences with either SSF muffin for-
mulation on values corresponding to cohesiveness and
springiness, respectively. Finally, a significant increase on
chewiness was observed for SSF-20 muffin (compared to
C). These results showed the same trend as firm-
ness:chewiness values increased at increasing fiber con-
centrations, with SSF-20% having a stronger effect than
SSF-10%. It is worth to remind that chewiness is directly
proportional to firmness. Possibly, the type and concen-
tration of fiber used to produce the gluten-free product as
well as its effects on moisture retention in the muffin might
influence the specific effects observed on the texture of the
products, a fact which requires further investigation.
The effect of the addition of different flours to gluten-
free products on crumb textural properties has been dealt
with in several studies. In agreement with our results, Shih
et al. (2006) reported that the firmness and chewiness of the
Table 2 Effect of squash seed flour addition (0, 10 and 20 g/100 g of
ready-to-bake dry gluten-free premix) on the muffin crumb
Samples N (cells/cm2) AM (mm2) AT (%)
C 26.4 b 149 c 66 b
SSF-10 24.2 ab 136 b 60 ab
SSF-20 22.8 a 127 a 57 a
GFP gluten-free premix, SSF squash seed flour, C Control, SSF-10
(10 g SSF/100 g GFP), SSF-20 (20 g SSF/100 g GFP), N numbers of
cells per area, AM mean cell area, AT void fraction
Different letters in the same column indicate significant differences
between means (p\ 0.05)
J Food Sci Technol
123
Fig. 2 Muffin crumb structure images: a control, b SSF-10, c SSF-20
Fig. 3 Textural parameter of muffin crumb: a firmness, b cohesiveness, c springiness, d chewiness. Different letters in the same graphics
indicate significant differences (p\ 0.05)
J Food Sci Technol
123
gluten-free cakes improved with the addition of up to 40%
of sweet potato flour. However, Singh et al. (2016) develop
a gluten-free rice muffin with black carrot pomace dietary
fiber concentrate (three levels) and xanthan gum (0.5%)
and observed a reduction in muffin firmness with the
addition of 3, 6 and 9% of black carrot pomace fiber. Wang
et al. (2002) also found an increase in breadcrumb
chewiness with inulin addition and no significant effect on
cohesiveness and springiness (in agreement with our
results).
Sensory evaluation of muffins
Sensory evaluation results of muffins are shown in Table 3.
The colour, taste and texture are important parameters for
the acceptability of food products by consumers (Kaur
et al. 2012). SSF-addition significantly increased the col-
our, sponginess, taste, texture and overall acceptability of
gluten-free muffins. Colour scores of muffins containing 0,
10 and 20% of SSF ranged from 7 to 9; while sponginess,
taste, texture and overall acceptability scores ranged from 5
to 9. Results analyzed by Friedman Test showed that celiac
consumers indicated that the muffins prepared with 20% of
SSF had the higher value scores with a higher overall
acceptability when compared to C muffin. Panelists scores
agree with the instrumental analysis that showed SSF-ad-
dition improved muffin quality by yielding better moisture
content, browning index, firmness and chewiness; all of
which enhance the sensory characteristics of gluten-free
muffins containing SSF.
Conclusion
In this study, the development of a gluten-free muffin with
compounds of a-galactoside type (soluble fiber) with a
functional and acceptable profile was obtained. Muffins
enriched with SSF exhibited higher TDF, protein and
unsaturated fat contents when compared to muffins pre-
pared with GFP. Also, both formulations containing SSF
showed higher overall acceptability, particularly muffins
with 20% of SSF that rendered the highest scores for
sponginess, texture, taste and colour. The enrichment with
SSF can be used as a strategy to develop not only for celiac
population, but also for other consumers.
Acknowledgements Authors would like to thanks CONICET for the
graduate fellowship granted to Marıa I. Palacio, Celibrand (healthy
food store) for the collaboration on the elaboration of gluten-free
muffins for the sensory evaluation and to the Department of Food
Technology-FCV (UNCPBA) for the collaboration on the use of the
texturometer.
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Texture Friedman
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Overall
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C 9
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SF-10 9
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SF-20 9
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(8–9)
2.43 b 9 (8–9) 2.58 b 9 (9–9) 2.40 b
p value 0.0182 \ 0.0001 \ 0.0001 \ 0.0001 \ 0.0001
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