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

mipalacio@vet.unicen.edu.ar

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

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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

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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)

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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)

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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)

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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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Table 3 Sensory characteristics (maximum score of 9) of gluten-free muffin

Samples Colour Friedman

test

Sponginess Friedman

test

Taste Friedman

test

Texture Friedman

test

Overall

acceptability

Friedman

test

C 9

(7–9)

1.80 a 7 (5–9) 1.28 a 7

(5–9)

1.25 a 7 (5–8) 1.10 a 7 (5–9) 1.18 a

SF-10 9

(7–9)

2.03 ab 8 (7–9) 2.18 b 9

(7–9)

2.33 b 8 (7–9) 2.33 b 9 (8–9) 2.43 b

SF-20 9

(7–9)

2.18 b 9 (8–9) 2.55 b 9

(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

C control, SF-10 (10 g squash flour/100 g gluten-free premix (GFP)), SF-20 (20 g squash flour/100 g GFP)

Different letters in the same column indicate significant differences between means (minimum–maximum value) analyzed by Friedman test

(p\ 0.05)

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