Research Article

Evaluation of nutritional characteristics and consumers’ acceptance

of gluten‑free sweet biscuits made from rice‑based pregelatinized
composite flours containing orange pomace and soy protein isolate
Caroline Alves Cayres1 · José Luis Ramírez Ascheri2 · Maria Antonieta Peixoto Gimenes Couto3

Received: 31 August 2020 / Accepted: 12 January 2021 / Published online: 23 January 2021
© The Author(s) 2021  OPEN

Abstract
Gluten-free bakery products are usually deficient in dietary fiber and inferior sources of protein. Studies that have assessed
sensory characterization of products prepared with orange pomace are scarce. This study evaluated the employment of
30% of rice-based pregelatinized composite flours (containing 15, 25 or 35% orange pomace blended with 5% soy protein
isolate) on gluten-free biscuits. The target was the investigation of the nutritional profile and consumer’s acceptance of
three short dough sweet biscuit samples. Thermoplastic extrusion was an efficient unit operation for the elaboration of a
new gluten-free ingredient in which a by-product rich in total dietary fiber and a component with a high level of protein
were employed. All pregelatinized composite flours in this study can be labeled as protein sources and can be claimed as
high in total dietary fiber content, as well as two biscuit samples that can be labeled as total dietary fiber source. Also, all
pregelatinized composite flours and biscuits presented low moisture and water activity, being microbiologically stable
to room temperature (25 ± 2 °C). Complementarily, the results of the microbiological evaluations [(coagulase-positive
Staphylococcus, thermotolerant coliform (45 °C) and Salmonella sp. (absence in 25 g)] of the biscuits and their comparison
with the Brazilian microbiological standards legislation attested their safety. Concerning the sensory analysis, adults’
results showed that the flavor attribute might be responsible for increasing the overall impression score of a sample.
In contrast, children’s results showed that they did not distinguish (p ≥ 0.05) two of three samples of biscuits analyzed.

Keywords Dietary fiber · Extrusion cooking · Industrial by-product · Physicochemical characterization · Protein
ingredient · Sensory evaluation

1 Introduction gives them a long shelf-life. They serve as a source of

energy for many people [1]. Attempts are being made to
Biscuits are one of the most popular bakery items con- improve biscuits’ nutritive value by modifying their nutri-
sumed by nearly everyone, appealing to children, teen- tive composition. This objective is very often achieved by
agers, adults, and elderly ones. These products are con- increasing the ratios of different types of dietary fiber and
venient, ready-to-eat, inexpensive and affordable snacks, protein in an elementary formulation. The goal is to serve
being able to present a great variety of flavors, prepared healthier and more functional food market demand [2].
through baking into a low moisture content (< 5%), which

* Caroline Alves Cayres, caroline.a.cayres@gmail.com; José Luis Ramírez Ascheri, jose.ascheri@embrapa.br; Maria Antonieta Peixoto
Gimenes Couto, gimenes@eq.ufrj.br | 1Graduate Program in Engineering of Chemical and Biochemical Processes at School of Chemistry,
Technology Center, Federal University of Rio de Janeiro, Avenida Athos da Silveira Ramos 149, Ilha do Fundão, Rio de Janeiro, RJ 21941‑909,
Brazil. 2Embrapa Agroindústria de Alimentos, Avenida das Américas 29501, Guaratiba, Rio de Janeiro, RJ 23020‑470, Brazil. 3Technology
Center, School of Chemistry, Federal University of Rio de Janeiro, Avenida Athos da Silveira Ramos, 149, Ilha do Fundão, Rio de Janeiro,
RJ 21941‑909, Brazil.

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Gluten and related proteins found in grains contrib- [1]. Hence, there are two strong reasons to rescue orange
ute to a cluster of diseases, not limited to celiac disease pomace into the food industry: improve the nutritional
[3]. Underneath the umbrella of gluten-related disorders, quality of food products to which it can be added and
there are also people with wheat allergy, non-celiac gluten reduce the disposal of waste in the environment con-
sensitivity, dermatitis herpetiformis and gluten ataxia [4]. comitantly. Also, the protein quality improvement of
For them, gluten could also be considered a “toxin” based gluten-free bakery products with sources of protein-rich
upon its mechanism of action in the small bowel and per- ingredients seems to be unavoidable. Soy protein isolate
haps even in the central nervous system [5]. Besides the is one of soy protein products with high protein content,
people needing to follow a gluten-free diet for health rea- without anti-nutritional factors such as trypsin inhibitor,
sons, a new segment of consumers has arisen who con- lipoxygenase, and urease [2]. Moreover, soy protein pre-
sume gluten-free products as a lifestyle choice [4]. How- sents a health effect on calcium metabolism and lowering
ever, gluten-free bakery products are generally deficient cholesterol [16].
in dietary fiber due to the use of pure refined flour and/or Thermoplastic extrusion is a method in which starch
starch as primary raw materials for their preparation [6]. and/or protein are transformed into plastic polymers by
Gluten-free products are also usually inferior sources of adding little water, and they are cooked with a high degree
protein [7]. The importance to simultaneously improve of mechanical energy. Functional ingredients which have
both the quality and the nutritional profile of gluten-free an unpleasant taste alone can be mixed with other com-
products has been stimulating researchers to investigate ponents into a unique food item through thermoplastic
new ingredients and technologies to be applied in gluten- extrusion. Also, extrusion can reduce anti-nutritional
free bakery products [8]. Rice flour is the most suitable compounds, promoting safer and more nutritious foods.
source of starch for gluten-free bakery applications due Extrusion improves protein digestibility via denaturation,
to its bland taste, white color, digestibility, and hypoaller- while total dietary fiber can be affected in different ways
genic properties [6]. Furthermore, it is entirely feasible to by this process [8]. Regarding dietary fiber, Huang and Ma
employ rice flour in short dough biscuit formulation. The [17] investigated the effect of extrusion processing (barrel
addition of soy proteins to rice flour improves the amino temperatures: 115–135 °C; feed moistures 10–18%) on the
acid balance of the new ingredient since legume and physicochemical properties of extruded orange pomace.
cereal proteins are complemented in the essential amino They found a redistribution of insoluble dietary fiber to
acids lysine and methionine. The rice flour obtained by soluble dietary fiber in the extrusion cooking, in which
thermoplastic extrusion presents increased cold viscosity, the soluble dietary fiber fraction of the extruded orange
and reduced peak viscosity and tendency to retrograde pomace was significantly (p < 0.05) higher than that of
as the amount of gelatinized starch is higher than the rice unextruded pomace. Hence, the extruded orange pomace
flour without any thermal treatment [9]. flour may contribute to heightening the insoluble/soluble
As the nutritional value has also become an essential dietary fiber ratio in food formulations. The thermoplastic
aspect of gluten-free foodstuffs [10], the addition of fruit extrusion can be an efficient unit operation for processing
pomace to biscuits is one of the possible approaches that a by-product rich in dietary fibers and an ingredient with
could be adopted to increase their dietary fiber and miner- a high level of protein, through the incorporation of these
als contents. For instance, orange pomace, an abundant two components into a starch-rich matrix.
agricultural by-product, is a fibrous residue of orange left Consumer acceptance is an indicator for prospective
over after the extraction of the juice. Brazil, the world’s purchase intentions and intake as well [18]. Nevertheless,
largest producer of oranges, produced more than seven current research in extruded ingredients is focused mainly
thousands of tonnes of orange pomace in 2019 [11], which on reformulation strategies rather than sensory aspects,
represents 50% of the mass of oranges employed in Brazil- and this gap needs to be addressed [14]. Studies that have
ian orange juice industry. Orange pomace is an excellent assessed sensory characterization of products prepared
source of dietary fiber, considering that 83% of orange with orange pomace flour are scarce. From the literature
pomace’s total dietary fiber is the insoluble fraction and its review, there is scanty information about any attempt to
soluble fraction is mostly composed of pectin [12], besides produce gluten-free biscuits fortified with dietary fiber
being rich in minerals [13]. So, orange pomace could be and protein by the employment of orange pomace flour
used as an excellent source of nutritionally enhancing and soy protein isolate to the biscuit dough. Therefore, the
and eco-friendly compounds [14]. Using orange waste present study aimed at the physicochemical characteriza-
by-products as a raw material in the food industry will tion of pregelatinized composite flours (PCFs), formulated
bring lots of health promotion like preventing obesity, with white rice flour, orange pomace flour, and soy pro-
overweight and diabetes due to its fiber content [15], tein isolate. The goal was to develop nutritionally enriched
as this component impacts all aspects of gut physiology gluten-free biscuits in terms of dietary fiber, minerals, and

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protein by the incorporation of PCFs as a source of the and 40.14% carbohydrates) and soy protein isolate (com-
abovementioned compounds. The biscuits also went for position: 5.02% moisture, 85.15% protein, 0.72% lipid,
microbiological analysis (coagulase-positive Staphylococ- 4.30% ash and 4.81% carbohydrates). These ingredients
cus, thermotolerant coliform (45 °C) and Salmonella sp.) were mixed on a powder homogenizer Chopin (Tripette &
to ensure their safety for consumption. Finally, adults’ and Renaud; Villeneuve la Garenne, France) and subsequently
children’s acceptance of these biscuits was assessed. conditioned. The proportion of raw materials in the com-
posite flour, the moisture conditioning, and the tempera-
ture in the third zone of the extruder were described in
2 Material and methods Table 1. For each variable, the levels were established
according to preliminary experiments (unpublished
2.1 Material results).
Conditioning moisture was adjusted by the condition-
White rice was purchased in the local market in the city ing of 2 kg composite flour. The determination of moisture
of Rio de Janeiro (Brazil). The by-product (consisted of was carried out, in duplicate, using AACC Method 44-16.01
flavedo, albedo, and seeds) remained after orange (Cit- [19]. Distilled water was slowly added to the sample in the
rus sinensis variety Osbeck) juicing was collected from an process of homogenization in blender HMT (Guangzhou
industrial extractor Otto 1800 (Organização Industrial Cen- Panyu Jianye; Guangzhou, China). After the addition of
tenário Ltda; Limeira, Brazil) at the food processing indus- water, the samples were stored into sealed polyethylene
try Bora Bora Comércio e Indústria Ltda (Rio de Janeiro, bags, in refrigeration, for 17 h.
Brazil). Soy protein isolate utilized originates from Marsul The extrusion was conducted using a single-screw
Proteínas LTDA (Montenegro, Brazil). extruder DO-CORDER Brabender (Brabender; Duisburg,
The ingredients used in the preparation of the biscuits Germany). The extrusion profile was: feed section speed:
were: pregelatinized composite flour (PCF) (prepared as 40 rpm; shear rate: 5:1; screw speed: 180 rpm; feed sec-
described in Sect. 2.1.2) and whey powder (Laticínios Porto tion temperature: 80 °C; compression section temperature
Alegre, Ponte Nova, Brazil). Corn starch, vegetal shorten- 100 °C; circular die diameter: 3 mm.
ing, refined sugar, brown sugar, egg, soy lecithin, ammo- The extrudate was dehydrated in a fan oven Macan-
nium bicarbonate, sodium bicarbonate, and salt were pur- uda Hauber DMS-G (Macanuda Hauber; Joinville, Brazil)
chased in local markets in the city of Rio de Janeiro (Brazil). at 60 °C for 4 h and subsequently milled on a disc mill
LM3600 (Perten Instruments AB; Huddinge, Sweden), set
2.1.1 Preprocessing of raw material to aperture 0, and, afterward, on a hammer mill LM3100
(Perten Instruments AB; Huddinge, Sweden), with 0.8 mm
White rice flour was produced through milling this cereal sieves. The obtained product will henceforth be called as
on the knife and hammer granulator mill (TREU S.A.; Rio pregelatinized composite flour (PCF).
de Janeiro, Brazil), with 1 mm sieves.
The by-product of orange juicing was ground on a 2.2 Assessment of physical characteristics
multiprocessor Cadence Mix for You MPR853 right after of pregelatinized composite flours
collecting (JCS Brasil Eletrodomésticos SA; Balneário Piçar-
ras, Brazil), dispersed on trays, and dehydrated in a turbo- Two hundred g of PCF were sieved for 10 min in the sieve
electric oven FTT 390G (Tedesco; Caxias do Sul, Brazil) at shaker ROTAP RX-29-10 (WS Tyler; Ohio, USA), utilizing
60 °C for 5 h. After dehydration, the by-product was milled steel mesh sieves, which frame is 8″ × 2″, with 212 μm and
in a disc mill LM3600 (Perten Instruments AB; Huddinge, 106 μm screen sieve sizes. The material of particle size infe-
Sweden), set to aperture 6. This product will henceforth be rior to 212 μm [20] was used to standardize particle sizes.
called orange pomace flour. The samples were packed into polyethylene bags, sealed,
labeled, and stored in a cool (25 ± 2 °C), dry place until the
2.1.2 Formulation, conditioning, and extrusion analyses were carried out.
of composite flours
2.2.1 Hydration properties
Three composite flours were elaborated by mixing the
raw materials: white rice flour (composition: 10.83% Water solubility index (WSI) and water absorption index
moisture, 6.37% protein and 82.80% carbohydrates; not (WAI) of PCFs were determined according to the method-
detectable contents of lipid, ash, and total dietary fiber), ology described by Anderson et al. [21], in quadruplicate.
orange pomace flour (composition: 2.35% moisture, 4.76% WSI indicates the amount of sample that is solubilized in
protein, 1.82% lipid, 3.77% ash, 47.16% total dietary fiber water at 30 °C relative to the initial sample after shaking

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and centrifuging. In comparison, WAI reports the amount

composite flour with 15% of orange pomace flour in its composition; PCF-25, pregelatinized composite flour with 25% of orange pomace flour in its composition; PCF-35, pregelatinized
463 ± 38a 466 ± 12a 71 ± 11a 395 ± 1a 227 ± 1a 158 ± 8a
388 ± 3a,b 411 ± 12a,b 79 ± 4a 332 ± 8b 207 ± 3a 132 ± 3b
301 ± 4b 348 ± 11b 69 ± 3a 279 ± 8c 211 ± 5a 144 ± 3a,b

mean ± standard deviation, n = 2; means followed by different superscript letters within each column are statistically different according to Tukey’s test (p < 0.05); PCF-15, pregelatinized

composite flour with 35% of orange pomace flour in its composition; WSI, water solubility index; WAI, water absorption index; PT, pasting temperature; MV25, maximum viscosity at 25 °C;
of water absorbed at 30 °C related to the sample that did

(cP)
SBB
not solubilize under shaking and centrifugation.

(cP) 2.2.2 Pasting properties

FVB

The viscosity of PCFs was determined, in an aqueous

BDB
(cP)

medium, under heating and stirring, using the equipment

Rapid Visco Analyser (RVA) (Newport Scientific; Warriewood,
Australia), according to AACC Method 76-21.01 [22], with
(cP)
TRB

modifications. The flour was initially stirred during 10 s, at

the initial temperature of 25 °C and 960 rpm. The remain-
der of the procedure, which lasted 20 min, was conducted
(cP)
PVB

at 160 rpm. The initial temperature was held for 2 min and
gradually raised, heating to 95 °C, at a constant rate of
Table 1  Formulation of composite flours and results from the assessment of physical characteristics of pregelatinized composite flours

14 °C min−1. Then, the temperature was held constant for

MV25B

3 min and cooled to 25 °C, at a constant rate of 14 °C min−1,

(cP)

which was held during the last 5 min of the analysis.

13.07 ± 0.39a 6.48 ± 0.17a 56.8 ± 3.1a
13.10 ± 1.21a 6.07 ± 0.12a 54.0 ± 3.1a
13.34 ± 1.56a 5.45 ± 0.22b 55.8 ± 3.1a

The readings from the paste curve generated were:

pasting temperature (PT), maximum viscosity at 25 °C
(°C)
PTB

(MV25), peak viscosity (PV), trough (TR), breakdown (BD),

final viscosity (FV), and setback (SB), obtained through the
register carried out by the software Thermocline (Newport
(g ­g−1)
WAIA

Scientific; Warriewood, Australia). The measurements were

conducted in duplicate.

2.3 Short dough sweet biscuit production

Temperature WSIA
tein isolate ing moisture in the third (%)

Preliminary baking tests were made to adjust the biscuit

extruder (°C)
zone of the

formula. Gluten-free flour was elaborated by mixing corn

starch and PCF in a fixed proportion of 7:3. This proportion
120
140
160

PV, peak viscosity; TR, trough; BD, breakdown; FV, final viscosity; SB, setback

was chosen according to the study of Ostermann-Porcel

et al. [23], in which the gluten-free cookies prepared with
Condition-

flour containing 30% of okara flour, in substitution to man-

ioc flour, were preferred by the consumers. The biscuit for-
(%)

mula, on a gluten-free flour weight basis, was: 35% water,

16
18
20

30% vegetal shortening, 20% refined sugar, 20% brown

sugar, 5% egg, 2% soy lecithin, 2% ammonium bicarbo-
Soy pro-

nate, 2% whey powder, 1% sodium bicarbonate, and 1%

(%)

salt. To produce the biscuit, gluten-free flour, refined sugar,

5
5
5

brown sugar, soy lecithin, ammonium bicarbonate, whey

powder, sodium bicarbonate, and salt were mixed during
flour (%)
pomace
Orange

2 min at medium speed in a planetary mixer (HMT, China).

mean ± standard deviation, n = 4
15
25
35

Vegetal shortening and egg were added to this mixture

and mixed for 5 min. Water was added little by little to the
White rice

above cream and mixed for 3 min at medium speed to get

flour (%)

the biscuit dough. After mixing, biscuit dough was sheeted

80
70
60

manually to a thickness of 4 mm, cut using a knife, to form

rectangles of 7 cm × 1 cm. Cut dough was transferred to
composite

stainless trays and then baked in an electric oven (Suggar,

latinized

PCF-15
PCF-25
PCF-35
Prege-

China) at 200 °C for 15 min. Finally, biscuits were kept at

flours

room temperature (25 ± 2 °C) for 1 h before analyses.

A

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2.4 Assessment of physicochemical characteristics following the rules of the Ethics Committee. There was
of pregelatinized composite flours and biscuits no incentive for attending the test.
The three biscuit samples (approximately 7 g each) were
2.4.1 Proximate composition served to the consumers on monadic presentation, in ran-
dom and balanced order [27]. Samples were coded with
The proximate composition was carried out using Asso- three-digit numbers and served to consumers in white
ciation of Official Analytical Chemists methods [24], in plastic plates. Water and crackers were provided for pal-
duplicate measurements: moisture content (Method ate cleansing.
925.09), total nitrogen (Method 2001.11, a conversion
factor of 5.75 was used to convert total nitrogen to pro- 2.6.1 Adults
tein content), lipid content (Method 945.38), ash content
(Method 923.03), and total dietary fiber (Method 985.29). 2.6.1.1 Participants A total of 151 consumers [28, 29]
Carbohydrates were determined by difference. participated in the test. Participants (86 women and 65
men, in which 66% were aged between 18 and 24 years
2.4.2 Mineral composition old, 28% between 25 and 35 years old, 3% between 36
and 50 years old, and 3% more than 51 years old) were
The quantification of minerals sodium (Na), potassium recruited from the staff and the students present in the
(K), magnesium (Mg), calcium (Ca), phosphorus (P), university environment during the test.
manganese (Mn), iron (Fe), zinc (Zn) and copper (Cu)
was determined by Inductively Coupled Plasma Optical 2.6.1.2 Affective test An acceptance test was carried
Emission Spectrometry (ICP-OES) (Optima 2100DV, Per- out to evaluate appearance, aroma, flavor, crunchiness,
kin Elmer, USA) after complete digestion of the sample in and overall impression, using a 9-cm hedonic unstruc-
nitric and perchloric acids, following the Methods 975.03 tured scale anchored at 0.5 cm on both ends with “dis-
and 990.08 of Association of Official Analytical Chemists like extremely” at the left and “like extremely” at the right
methods [24], in duplicate measurements. (Fig. 1) [30]. The position on the line scale indicated by
consumers was converted to a number between 0 (left-
most position) and 9 (rightmost position).
2.4.3 Water activity
2.6.1.3 Purchase intention Just after the acceptance test,
Water activity (25 °C) was measured in AquaLab LITE participants were asked about how much they were likely
(Decagon; Pullman, USA), in duplicate measurements. to purchase the biscuit that gave rise to the coded sample.
Purchase intention was assessed using a 5-point struc-
2.5 Microbiological analysis of biscuits tured scale ranging from “would certainly buy” to “would
certainly not buy” [30].
The microbiological evaluations (coagulase-positive
Staphylococcus, thermotolerant coliform (45 °C) and Sal-
monella sp. (absence in 25 g) were performed according
to the American Public Health Association [25]. These
species were selected following the Brazilian legislation
for biscuits [26].

2.6 Sensory evaluation

Tests were carried out at the School of Chemistry of the

Federal University of Rio de Janeiro (UFRJ) and Colégio de
Aplicação (UFRJ) and were approved by Ethics Commit-
tee (CAAE 0027.0.313.000-11). All adult participants and
those responsible for the children received written infor-
mation about the study before giving their informed
consent. This procedure was adopted to avoid risks
(mainly food allergy or intolerance) to the participants,
Fig. 1  Scales used in affective tests. a adults: 9-cm hedonic unstruc-
tured scale, b children: 5-point facial hedonic scale

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2.6.2 Children This combination of these parameters of the thermoplastic

extrusion process was responsible for the WSI values of all
2.6.2.1 Participants A total of 111 consumers [28, 29] par- PCFs, which did not differ statistically (p ≥ 0.05) (Table 1). It
ticipated in the test. Participants (54 girls and 57 boys, in is possible to conclude that although the concentrations
which 20% were aged 8 years old, 31% 9 years old, 36% of orange pomace flour are different in each of the PCFs,
10 years old and 14% 11 years old) were recruited from the degree of degradation suffered by this ingredient’s
the students present in the elementary school environ- TDF, as well as the relative proportion of damaged starch
ment during the test, with the prior permission of those granules was the same in all PCFs produced. Soy protein
responsible. isolate and TDF, along with conditioning moisture, may
have caused a damping effect on shear rate.
2.6.2.2 Affective test An acceptance test was carried out The damaged starch performs a higher capacity for
to evaluate the overall impression, using a 5-point facial holding water at room temperature and swelling in con-
hedonic scale (Fig. 1). Values from 1 to 5 are assigned to trast to the native starch, increasing WAI [33]. Consider-
each facial expression of the hedonic scale presented to ing that the native starch does not absorb water since the
children [30]. extruded starch absorbs water quickly and form a gel at 30
°C or lower temperature [31], WAI estimates the amount of
2.7 Statistical analyses water absorbed by the starch. It can be used as an index
for the gelatinization degree of the sample [21]. The lowest
Differences between the mean values for the analyses WAI value presented for PCF-35 (Table 1) is probably due
made on PCFs and biscuits were assessed by the analysis of to its most moderate content of starch. Similar results were
variance (ANOVA; p < 0.05) and followed by the Tukey test reported by Altan et al. [34], on barley extrudates contain-
(p < 0.05, only when significant differences were observed ing different concentrations of tomato and grape pom-
in ANOVA). Correlations between hydration properties and ace, and by Cortez et al. [35], on two types of blue corn
pasting properties of pregelatinized composite flours and extrudates combined with orange pomace. The result of
physicochemical characteristics of pregelatinized compos- the present study could indicate that in composite flours
ite flours and biscuits were done by Pearson correlation whose proportions of rice flour are less than 70%, TDF and
coefficients. A p-value < 0.05 (2-sided) was considered as proteins present in the formulation protect the starch from
statistically significant. For the aforementioned statistical gelatinization.
analyses, it was employed Statistica software 11.0 for Win-
dows (StatSoft Inc.; Tulsa, USA). 3.1.2 Pasting properties

Pasting properties are considered as one of the most criti-

3 Results and discussion cal indices in the evaluation of starch-rich ingredients and
raw materials. The viscosity of a paste mainly relies on the
3.1 Physical characteristics of pregelatinized degree of gelatinization of the starch granules and the rate
composite flours of molecular breakdown [36].
Higher values of MV25 indicates a more significant
3.1.1 Hydration properties starch breakdown, which forms a three-dimensional net-
work at 25 °C as a result of more hydrophilic sites of the
WSI is an adequate index to measure the intensity of the sheared starch molecules exposed to water; therefore,
extrusion conditions; likewise, the transformation that indicating high shearing during extrusion [36]. It is pos-
occurs in starch molecules [31]. This index still discloses sible to infer that MV25 parallels to WAI (R = 0.997, p < 0.05).
the degradation of total dietary fiber (TDF) components Hence, the differences (p < 0.05) observed between the
submitted to severe processing conditions, which entails MV25 values presented for PCF-15 and PCF-35 (Table 1)
the production of compounds having a low molecular can be explained for the same reasons abovementioned
weight [17]. Furthermore, lower shear rates occur during for WAI (Sect. 3.1.1).
the thermoplastic extrusion in which higher conditioning In the course of the heating cycle of the pasting vis-
moistures are employed [32]. Commonly, the increase in cosity profile, PV is the maximum viscosity presented by
the barrel temperature manages to the rise in WSI values, the sample. This property coordinates to the quality of
which result in more water-soluble compounds due to the the final product because PV indicates the maximum
increment in damaged starch granules [31, 32]. As can be swelling of the starch granules due to the sample water-
seen in Table 1, the higher the conditioning moisture, the binding capacity [37]. The highest PV value presented
higher the temperature in the third zone of the extruder. for PCF-15 (Table 1) could be assigned to its most top

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content of starch, and also more starch granules with a caused a protective effect against starch retrogradation
high swelling capacity. and syneresis.
Amylose leaching out and alignment are the results of
further starch granules disruption due to high tempera- 3.2 Physicochemical characteristics
ture and mechanical shear stress to which the sample of pregelatinized composite flours and biscuits
is submitted during the holding period of the viscosity
test. Generally, this period is followed by a breakdown 3.2.1 Proximate composition
in viscosity [37]. As BD is the difference between PV and
MV25, it is expected that high values of BD are associ- The nutrient content (Table 2) of the pregelatinized
ated with high PV. This implication can be observed in white rice flour fortified with different proportions of
Table 1. Moreover, it is conceivable to find another phe- orange pomace flour and soy protein isolate significantly
nomenon occurring in parallel. As suggested by Kim and increased in comparison to the white rice flour. There was
Walker [38], lipids may inhibit water transfer into the an increase of at least 56% of protein content in PCFs
granules due to a film cover, which increases the hydro- when compared to that of white rice flour, in addition to
phobicity of starch. The observed highest BD (Table 1) the enrichment in ash and TDF contents (Table 2), which
related to the lowest lipids content in PCF-15 (Table 2) were not even detected in white rice flour. Therefore, for-
supports this explanation. tification of orange pomace flour, as well as soy protein
The difference between FV and TR represents SB. The isolate, could be used in the production of PCFs enhanced
viscosity of the sample presents a variation during its reas- in protein, ash, and TDF contents.
sociation at the stage of RVA cooling, which reveals the According to the CODEX Alimentarius [40], the results
degree of starch retrogradation. Accordingly, SB ties into for the protein content (Table 2) show that PCF-25 can
the texture properties of the final product [39]. Lower SB be labeled as a protein source ingredient/food because it
values (p < 0.05) (Table 1) are related to higher protein and contains at least 5 g of protein per 100 g. Similarly, PCF-
lipid contents (p < 0.05) (Table 2). These two components 15 and PCF-35 can be labeled as high in protein content
could have surrounded the starch granules and may have considering that they contain more than 10 g of protein

Table 2  Composition, energy value and water activity of pregelatinized composite flours and biscuits
Components Pregelatinized composite flours Biscuits
PCF-15 PCF-25 PCF-35 B-15 B-25 B-35

Moisture (g/100 g) 7.86 ± 0.04a 7.47 ± 0.16a 7.61 ± 0.01a 3.99 ± 0.01B 4.53 ± 0.04A 3.43 ± 0.06C
Protein (g/100 g) 10.18 ± 0.04a 9.95 ± 0.00b 10.00 ± 0.04a,b
2.47 ± 0.00A
2.36 ± 0.08A
2.70 ± 0.08A
Lipid (g/100 g) 0.19 ± 0.01c 0.64 ± 0.04a 0.36 ± 0.01b
18.71 ± 0.15A
18.35 ± 0.10B
18.28 ± 0.14B
Ash (g/100 g) 1.20 ± 0.01c 1.53 ± 0.01b 1.69 ± 0.03a
1.44 ± 0.03A
1.43 ± 0.01A
1.51 ± 0.03A
Sodium (Na) (mg/100 g) 67.88 ± 0.96a 68.09 ± 0.15a 66.16 ± 0.19a
435.60 ± 4.78 A
401.29 ± 6.65 A
408.97 ± 4.56A
Potassium (K) (mg/100 g) 258.84 ± 2.18c 359.76 ± 2.42b 414.91 ± 0.28 a
98.32 ± 1.10B
115.50 ± 2.15 A
123.03 ± 2.81A
Magnesium (Mg) (mg/100 g) 56.07 ± 0.60b 60.08 ± 0.04a 60.04 ± 0.27a
16.49 ± 0.14A
16.92 ± 0.53A
17.10 ± 0.36A
Calcium (Ca) (mg/100 g) 111.35 ± 2.96c 180.11 ± 4.54b 223.43 ± 4.23a 48.90 ± 1.41B 57.99 ± 0.69B 68.84 ± 1.71A
Phosphorus (P) (mg/100 g) 186.60 ± 2.50a 183.97 ± 1.12a 176.36 ± 0.43 a
77.49 ± 0.83A
75.81 ± 1.60A
73.42 ± 0.72A
Manganese (Mn) (mg/100 g) 0.98 ± 0.01a 0.95 ± 0.02b 0.91 ± 0.02c
0.28 ± 0.01A
0.27 ± 0.01A
0.27 ± 0.01A
Iron (Fe) (mg/100 g) 1.42 ± 0.01b 1.48 ± 0.01b 1.64 ± 0.01a
1.81 ± 0.05A
1.62 ± 0.11A
1.94 ± 0.01A
Zinc (Zn) (mg/100 g) 1.64 ± 0.07a 1.43 ± 0.02b 1.33 ± 0.03b
0.39 ± 0.01B
0.44 ± 0.01A
0.38 ± 0.01B
Cooper (Cu) (mg/100 g) 0.29 ± 0.01b 0.29 ± 0.01b 0.31 ± 0.01a
0.10 ± 0.01A
0.09 ± 0.01A
0.07 ± 0.01A
Total dietary fiber (g/100 g) 7.51 ± 0.02c 11.81 ± 0.01b 13.84 ± 0.04a
1.97 ± 0.02C
3.59 ± 0.01B
5.47 ± 0.03A
Carbohydrates (g/100 g) 73.06 ± 0.02a 68.60 ± 0.02b 66.50 ± 0.04b
71.42 ± 0.01A
69.74 ± 0.02A
68.61 ± 0.02A
Energy value (kcal/100 g) 334.76 ± 0.05a 319.96 ± 0.83b 309.27 ± 0.21 c
464.00 ± 0.88 A
453.57 ± 0.41 B
449.76 ± 0.37C
Water activity (25 °C) 0.370 ± 0.004 a 0.341 ± 0.007 a,b 0.330 ± 0.001b 0.410 ± 0.000 A 0.272 ± 0.016 B 0.223 ± 0.004 C

mean ± standard deviation, n = 2; means followed by different superscript lower case within each row are statistically different according to
Tukey’s test (p < 0.05); means followed by different superscript capital letter within each row are statistically different according to Tukey’s
test (p < 0.05); PCF-15 = pregelatinized composite flour with 15% of orange pomace flour in its composition; PCF-25 = pregelatinized com-
posite flour with 25% of orange pomace flour in its composition; PCF-35 = pregelatinized composite flour with 35% of orange pomace flour
in its composition; B-15 = biscuit formulated with PCF-15; B-25 = biscuit formulated with PCF-25; B-35 = biscuit formulated with PCF-35

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per 100 g. A key ingredient to achieve, on average, 10% 3.2.2 Mineral composition

of protein content in PCFs was the use of soy protein
isolate, even if in a smaller amount (5%) in the formula- White rice flour, which is part of the composition of PCFs,
tion of composite flours. and corn starch have no detectable contents of ash/min-
Furthermore, all PCFs can be claimed as high in TDF erals. Both of them are typically employed in gluten-free
content (Table 2) because of containing more than 6 g bakery product formulation. Hence, orange pomace flour
of TDF per 100 g, while B-25 and B-35 can be claimed as and soy protein isolate contributed to the enrichment in
a TDF source because they contain more than 3 g of TDF macro- and microminerals to PCFs and, consequently, to
per 100 g, according to the CODEX Alimentarius [40]. The biscuits (Table 2).
enrichment in TDF content is due to the use of orange Potassium was the most abundant mineral in all PCFs,
pomace flour, which is mostly comprised of celluloses, followed by phosphorus and calcium (Table 2). The addi-
hemicelluloses, and pectin [41]. In their study, Huang tion of PCFs in food products would then increase the
et al. [42] showed that thermoplastic extrusion is a novel amount of the minerals mentioned above. PFC-25 and
strategy for developing orange pomace extrudate with PCF-35 can be claimed as calcium sources (Table 2),
enhanced hypoglycemic, hypocholesterolemic, and fer- according to the CODEX Alimentarius [40], because they
mentative capacities. The reason is due to this functional contain more than 150 mg of calcium per 100 g.
ingredient exhibited stronger effects than did orange It should be noted that one serving (30 g) of biscuits
pomace flour in retarding the rate of in vitro glucose dif- (Table 2) provides, on average, approximately 6% of
fusion, delaying the rate of glucose absorption, inhibit- the Nutrient Reference Values (NRVs) of iron and cop-
ing the α-amylase activity, binding cholesterol or bile per, each, for children aged between 7 to 10-year-old
acid capacities, and increasing the total short-chain fatty [45] (representing the children consumers’ age group in
acids production. the present study). All PCFs can be claimed as a copper
Since the energy value (Table 2) is calculated by source (Table 2), according to the CODEX Alimentarius
the sum of each energy source (protein—4 kcal/g, [40], because they contain more than 0.135 mg of copper
lipid—9 kcal/g, and carbohydrates—4 kcal/g), it is per 100 g.
expected that the amount of calories provided by each It is essential to highlight that one serving (30 g) of
biscuit formulation will be different if these components biscuits (Table 2) provides, on average, 3.84%, 3.62% and
are presented in higher or lower concentrations. Also, 3.30% of the NRVs for adults of iron, manganese, and
it is observed in Table 2 that the energy value of each phosphorus, respectively [45]. All PCFs can be claimed
biscuit is inversely proportional to its TDF content. This as high in manganese content (Table 2), according to the
result comes from the formulation of the PCFs (Table 1), CODEX Alimentarius [40], because they contain more than
where the higher the amount of orange pomace, the 0.90 mg of manganese per 100 g. Also, all PCFs can be
lower the amount of white rice flour, and consequently, claimed as a phosphorus source (Table 2), according to
the less is the available carbohydrates for energy the CODEX Alimentarius [40], because they contain more
conversion. than 105 mg of phosphorus per 100 g.
Finally, one way to assure that the broad population Additionally, all PFCs can be claimed as a magnesium
receives adequate amounts of protein and TDF in the source (Table 2) according to the CODEX Alimentarius [40]
diet is to fortify foodstuffs that would not commonly because they contain more than 46.5 mg of magnesium
be correlated with these fortifications. Some examples per 100 g.
are foods often eaten as snacks or at breakfast, such as Considering the intake of 30 g (1 serving) a day of bis-
biscuits, consumed by adults and children. For instance, cuits, sodium intake would be around 125 mg (Table 2),
Arribas et al. [43] presented a gluten-free snack-like corresponding to 6.25% of the intake level not to exceed
extruded product with increased levels of protein and (2,000 mg/day) [46].
dietary fiber, employing pea and carob as enriching
ingredients. Also, Morales et al. [44] showed that extru- 3.2.3 Water activity
sion promoted an increase in soluble fiber in some glu-
ten-free lentil-based flour formulations, fiber-enriched All PCFs and biscuits presented water acitivity (­ Aw) lower
with corn or apple fibers. Developing and commercial- than 0.6 (Table 2); hence, they are enclosed in the category
izing tempting protein- and TDF-enriched ingredients/ of microbiologically stable foods [47]. Foods that present
foods is a way by which the food scientists and indus- ­Aw varying from 0.40 and 0.70 must be stored in packs
try can support consumers in meeting their nutritional that prevent moisture uptake or loss [48], and do not need
needs. refrigerated storage.

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It is observed, both in PCFs and biscuits, that ­Aw is Table 4  Acceptance means for affective tests carried out with
inversely proportional to the content of TDF (Table 2) adults and children
(R = 0.997, p < 0.05 for PCFs and R = 0.952, p < 0.05 for bis- Acceptance Biscuits
cuits). TDF possesses excellent properties of water absorp-
B-15 B-25 B-35
tion and swelling power, which cause high water retention
[49], and a subsequent decrease in ­Aw. Appearance 4.7a 4.5a 4.6a
Aroma 5.0a 4.8a 5.0a
3.3 Microbiological analysis of biscuits Flavor 6.2a 4.9b 5.0b
Crunchiness 7.2a 6.1b 6.9a
Microbial spoilage is known as the most critical factor Overall impression (adults) 6.5a 5.0b 5.3b
affecting the safety and shelf-life of food products pre- Overall impression (children) 4.0A 3.9A 3.6B
pared with fruits [50], and, consequently, to those that are Means followed by different superscript lower case within each row
produced with their by-products. are statistically different according to Tukey’s test (p < 0.05), using
The results of the microbiological analysis of the bis- a 9-cm hedonic unstructured scale from “dislike extremely” to “like
cuits and their comparison with the microbiological stand- extremely”. Means followed by different superscript capital letter
within each row are statistically different according to Tukey’s test
ards (Table 3) allow the characterization of the products as (p < 0.05), using a 5-point facial hedonic scale; B-15 = biscuit for-
microbiologically adequate since the values for all micro- mulated with pregelatinized composite flour with 15% of orange
organism counts are under Brazilian legislation [26]. There- pomace flour in its composition; B-25 = biscuit formulated with
fore, the biscuits can be considered safe for consumption, pregelatinized composite flour with 25% of orange pomace flour in
its composition; B-35 = biscuit formulated with pregelatinized com-
which indicates that good manufacturing practices have posite flour with 35% of orange pomace flour in its composition
been adopted throughout the entire production chain
(manufacture of orange pomace flour, PCFs, and biscuits).
with sour cassava starch and orange albedo flour, the
3.4 Sensory evaluation means for all the sensory attributes evaluated (appear-
ance, aroma, flavor, texture and overall impression) were
3.4.1 Adults between 5.0 and 7.3, on a 9-point structured scale. These
results correspond to the hedonic terms “neither like nor
The sensory profile presented in Table 4 demonstrates that dislike” and “like moderately”. Except for the appearance
appearance and aroma did not show any significant differ- attribute, which obtained means of less than 5.0, the pre-
ence (p ≥ 0.05) between the three samples of biscuits ana- sent work got similar scores of the studies mentioned
lyzed (Fig. 2). The flavor of sample B-15 is reported as the above.
preferred one (p < 0.05), while the crunchiness of samples Probably because it reached the best acceptance
B-15 and B-35 achieved the best score. mean in the flavor attribute, sample B-15 obtained the
Santos et al. [51], in a study where gluten-free choc- highest overall impression score, being indirectly pre-
olate biscuits were prepared with cassava starch and ferred among the three samples analyzed. The flavor is
orange albedo flour, the biscuits obtained acceptance more critical than crunchiness for these products. Fla-
means ranging from 5 (neither like nor dislike) to 8 (like vor means may have been lower in samples B-25 and
very much), using a 9-point structured scale. In a study B-35 due to the higher amount of orange pomace flour
reported by Santos et al. [52], in which chocolate biscuits in their formulations, compared to B-15 (p < 0.05), as it is
were prepared with partial replacement of wheat flour an ingredient that consumers are not used to.

Table 3  Microbiological analysis of biscuits

Micro-organisms Biscuits Brazilian ­legislationa
B-15 B-25 B-35

Thermotolerant coliforms (MPN/g) <3 <3 <3 10 MPN/g

Coagulase-positive Staphylococcus (CFU/g) < 1 × 101 < 1 × 101 < 1 × 101 5 × 102 CFU/g
Salmonella sp./25 g Absent in 25 g Absent in 25 g Absent in 25 g Absence in 25 g

MPN most probable number, CFU colony-forming unit

a
Brazil (2001); B-15 = biscuit formulated with pregelatinized composite flour with 15% of orange pomace flour in its composition; B-25 = bis-
cuit formulated with pregelatinized composite flour with 25% of orange pomace flour in its composition; B-35 = biscuit formulated with
pregelatinized composite flour with 35% of orange pomace flour in its composition

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Fig. 2  Biscuit samples. B-15 = biscuit formulated with prege- posite flour with 25% of orange pomace flour in its composition;
latinized composite flour with 15% of orange pomace flour in its B-35 = biscuit formulated with pregelatinized composite flour with
composition; B-25 = biscuit formulated with pregelatinized com- 35% of orange pomace flour in its composition

Due to the reading of the consent term, where it was the lowest mean (p < 0.05) presented by sample B-25 in
exposed that biscuits were formulated with orange the crunchiness attribute (Table 4).
pomace flour, an expectation effect may have occurred,
because too much information about the sample can 3.4.2 Children
influence the consumers’ response.
The purchase intention test, which aims to comple- Means presented in Table 4 reveal that there was no signifi-
ment sensory analysis, was also applied to adult con- cant difference in acceptance (p ≥ 0.05) between samples
sumers. It can be seen in Fig. 3 that the sample B-15 B-15 and B-25 and also that these samples were indirectly
received 66.2% of the evaluations between “would cer- preferred in this assessment. From this outcome, it is sug-
tainly buy” and “would probably buy”, comprising most gested that B-25 should be the biscuit offered to children,
of the positive attitudes among the three biscuit samples given that its nutrient content is better than the B-15 one,
analyzed. These results corroborate with the result of the as B-25 presented less lipids and more potassium, zinc and
affective test, where the sample B-15 was indicated as TDF (Table 2).
the preferred one, according to the overall impression In the study presented by Seo et al. [53], in which five
evaluation. The B-25 sample received most of the nega- different types of foods containing wild blueberry pow-
tive positions, which correspond to the answers “would der were prepared for sensory testing, children liked and
probably not buy” and “would certainly not buy”, with wanted to consume the cookies more when compared to
31.1% of the judgments. This result may be related to beverages, ice pops, and/or gummies. Their results cor-
roborate the idea that biscuits can be appropriate food

Fig. 3  Purchase intention 70%

of biscuits. B-15 = biscuit
formulated with pregelatinized 60%
composite flour with 15% of
orange pomace flour in its 50%
Consumers (%)

composition; B-25 = biscuit

formulated with pregelatinized 40%
composite flour with 25% of
orange pomace flour in its 30%
composition; B-35 = biscuit
formulated with pregelatinized 20%
composite flour with 35% of
orange pomace flour in its 10%
composition
0%
Would certainly buy Unsure Would probably not buy
+ +
Would probably buy Would certainly not buy
B-15 B-25 B-35

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products to be enriched with fruit pomace powders, indicated otherwise in a credit line to the material. If material is not
to achieve better means of sensory acceptance within included in the article’s Creative Commons licence and your intended
use is not permitted by statutory regulation or exceeds the permitted
children. use, you will need to obtain permission directly from the copyright
holder. To view a copy of this licence, visit http://creat​iveco​mmons​
.org/licen​ses/by/4.0/.

4 Conclusions

The present study delineated a strategy of the utilization References

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