Journal of Ecological Engineering

Journal of Ecological Engineering 2023, 24(12), 130–142 Received: 2023.09.13

https://doi.org/10.12911/22998993/173009 Accepted: 2023.10.21
ISSN 2299–8993, License CC-BY 4.0 Published: 2023.11.04

Possibility of Water Saving in Processing of Snack Pellets

by the Application of Fresh Lucerne Sprouts: Selected Aspects
and Nutritional Characteristics

Maciej Combrzyński1, Agnieszka Wójtowicz1, Beata Biernacka1, Tomasz Oniszczuk1*,

Marcin Mitrus1, Jakub Soja1, Renata Różyło2, Karolina Wojtunik-Kulesza3,
Kamila Kasprzak-Drozd3, Anna Oniszczuk3
1
Department of Thermal Technology and Food Process Engineering, University of Life Sciences in Lublin,
Głęboka 31, 20-612 Lublin, Poland
2
Department of Food Engineering and Machines, University of Life Sciences in Lublin, Głęboka 28,
20-612 Lublin, Poland
3
Department of Inorganic Chemistry, Medical University of Lublin, Chodźki 4a, 20-093 Lublin, Poland
* Corresponding author’s e-mail: [email protected]

ABSTRACT
New types of extruded snack pellets of wheat-corn blend base and fresh lucerne sprouts were developed. The aim
of the study was to examine the effect of fresh lucerne sprouts addition on the water consumption, processing ef-
ficiency and the specific mechanical energy during production of wheat-maize snack pellets. Additionally, the total
phenolic content and antiradical activity, as well as the water absorption and water solubility indices in samples
processed under variable processing conditions were tested. The extrusion-cooking of blends consisted of 10,
20 and 30% of lucerne sprouts was carried out using a single screw extruder at screw speeds of 60 and 100 rpm,
and at moisture contents of 32, 34 and 36%. Replacement of wheat-corn flour blends by fresh lucerne sprouts at
various levels (10, 20 and 30%) enabled to sufficiently reduce technological water which is needed in extrusion-
cooking process of snacks pellets. The limitation of water was from 89 to 100% if fresh lucerne sprouts were used,
depending on the recipe and dough moisture level tested. Total phenolic content and antioxidant activity increased
significantly due to lucerne sprouts addition. Furthermore, higher water absorption and water solubility index were
noted if increased initial moisture content was applied during the processing of snack pellets. It can be concluded
that fresh lucerne sprouts can be valuable additives, enabling to save the technological water in production process
and to obtain nutritionally valuable supplemented wheat-corn-based snack pellets.

Keywords: technological water; lucerne; Medicago sativa L.; extrusion; snack pellets; antioxidants; functional food

INTRODUCTION on water and energy. One of the solutions to this

problem could be hydroponic plant cultivation,
One of today’s greatest challenges is to pro- which saves up to 95% water compared to tradi-
vide valuable food for the growing number of tional methods, as well as the direct use of fresh
people on our globe with no negative environ- fruits, vegetables and especially plant sprouts in
mental impact. Due to the climate change, among the agri-food industry to produce nutritionally
others, the limited access to fresh water is ob- valuable and environmentally friendly food and
served in many countries. Scientists around the feed products [Elmulthum et al., 2023; Stachows-
world are faced with the enormous problem of en- ki et al., 2023]. Highly efficient agriculture gener-
suring food security and the availability of fresh- ates huge amounts of biomass every year, which
water resources. Animal husbandry and cultiva- can be used e.g. for methane fermentation. Agri-
tion of traditional crops involve huge demands cultural biogas plants can utilize surplus biomass

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 Journal of Ecological Engineering 2023, 24(12), 130–142

for energy purposes [Kalinichenko and Havrysh, diabetes, and cancer. Antioxidants, both synthetic
2019]. Shah et al. [2022]. The energy generated and natural, are given to the diet to prevent the
in this way can be used in the processing and pro- aforementioned conditions [Labhar et al., 2023].
duction of food with specific quality parameters, Apart from the seeds or extracts, lucerne
where production technology is very energy and sprouts are well-known products with high nu-
water consuming. Sustainability policies require tritional value. Previous research has discovered
control of water and energy flows in the food in- that sprouting of lucerne seeds enhanced the con-
dustry. By using innovative food processing and tent of vitamins, minerals, and phytoestrogens,
production technologies, a significant reduction boosting their nutritional and health value when
in water and energy demands can be achieved compared to fresh plant material. Lucerne sprouts
[Lisiecka and Wójtowicz, 2020]. include a wide range of flavonoids, phenolic ac-
One of the valuable plants with high produc- ids and terpenoids, with calycosin, methyl gallate
tion efficiency and low cultivation requirements and epicatechin 3-gallat being the top three ele-
is lucerne. Lucerne is used as a functional food vated. Lucerne sprouts are characterized by high
in the form of protein concentrates, extracts or antioxidant activity [Zincă et al., 2013] and are
dried leaf powders. For consumption purposes, also an excellent source of estrogenic bioactive
lucerne seeds undergo various processing tech- chemicals [Hong et al., 2011]. Despite the proven
niques (wet heating, microwave treatment, dry nutritional value of lucerne and derived products,
heating, soaking or extrusion) in order to reduce they are rarely used as a functional food additive
the content of anti-nutritional substances and for human consumption [Aloo et al., 2021]. At-
improve the technological properties and bioac- tempts have been made to add lucerne leaf pow-
tive potential [Lai et al., 2020]. der to vegan chocolate; however, lucerne pro-
Lucerne has been utilized in traditional medi- tein isolate has been included within cereal bars
cine and phytotherapy. Studies have indicated [Sahni et al., 2022]. Fresh sprouts are nutritious
that lucerne concentrate can be used as a dietary foods that are low in fat and high in phytochemi-
supplement to help humans counter malnutrition, cals, but they have a short shelf life, necessitating
ischemia and numerous diseases. Research indi- the development of processing procedures that
cates that the constituents of lucerne concentrate retain their nutritional value [Hong et al., 2011].
strengthen human resistance to infections and To prevent the short shelf-life of sprouts, dry-
many diseases, and that the iron concentration ing into powder by various methods can be ap-
improves the hemoglobin content in blood, hence plied, but this generates additional energy con-
aiding in the control of anemia caused by hemo- sumption. A new way to circumvent this loss
globin shortage. In various studies, the saponins could be an incorporation of fresh sprouts into
and flavonoids in lucerne concentrate have been products processed via HTST (high temperature
found to have a positive effect on human health short time) treatment such as through the extru-
and help strengthen the immune system [Gaweł sion-cooking [Shirazi et al., 2020]. This technolo-
et al., 2017]. In addition to proteins, lucerne ex- gy allows an intensive treatment with a very short
tracts and concentrates contain a variety of vita- processing time, so it has low energy require-
mins (A, B, C, D, E, K), minerals (Ca, Cu, Fe, ments and moreover a limited destructive effect
Mg, Mn, P, Zn, Si), phytochemical substances on the biologically active substances present in
(carotene, chlorophyll, coumarins, isoflavones, the extruded materials. Additionally, this treat-
alkaloids, saponins) and antinutritional compo- ment technology may increase the extractability
nents (phytates, L-canavanine, saponins).What is of bioactive compounds. This is very profitable
more, this plant is high in dietary fiber and other when fruit, vegetable or herbs are added as func-
active components [Shah et al., 2020]. Caunii tional components, because of limited changes in
et al. [2012] discovered that lucerne extract in- bioavailability of the extruded components, espe-
cluded free hydroxyl groups (hydrogen donors), cially when low temperature extrusion is applied
which conferred the product with substantial an- as in the processing of snack pellets [Soja et al.,
tioxidizing activity in the human body. It should 2023]. Expanded puffs can be directly created as
be noted that the presence of oxidizing substances a result of low-moisture (12–18%) high-shear ex-
in the human body causes cell damage (e.g., DNA trusion-cooking at high temperature, but also they
mutation) and the development of numerous can be produced as indirectly expanded crisps
chronic diseases such as arteriosclerosis, arthritis, from snack pellets created through the use of

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low-temperature semi-moisture (25–36%) extru- to desired moisture level of 32, 34 and 36% of
sion technology [Prabha et al., 2021]. The snack water content by the addition of a proper amount
pellets may be then expanded by many ways, the of tap water [Lisiecka and Wójtowicz 2020].
most popular being hot oil frying or low-fat ex- Blends were then manually transferred into the
pansion by toasting or microwaving [Kraus et al., extruder hopper. Snack pellets were extruded
2014; Lisiecka et al., 2021]. with a single screw prototype modular extruder
As stated by Lisiecka and Wójtowicz [2020], EXP-45–32 [built by Zamak Mercator, Skawina,
the use of fresh vegetables can significantly re- Poland] with barrel configuration of L/D=20.
duce the water volume needed in the technologi- The samples without additive (as control) and the
cal process and the energy needed for the drying samples supplemented with fresh lucerne sprouts
process. Thus, the incorporation of fresh lucerne pulp were processed at temperatures in individual
sprouts as an additive to snack pellets processed sequential extruder sections: dosing section –
via extrusion-cooking may have many advantag- 80 to 85°C, plasticizing section – 90 to 105°C,
es, both processing and nutritional. What is more, cooling section – 50 to 65°C, forming die – 60
after drying, the pellets have moisture content be- to 75°C. Two variable screw speeds were ap-
low 11%, making the products stable and safe for plied: 60 and 100 rpm. The extruded material was
long storage. shaped with a flat forming die of 0.6 mm x 30 mm
The aim of the study was to develop new type as a strip, followed by an external cutting device
of wheat-maize-based snack pellets fortified with to create tetragon samples of 30 x 30 mm. The cut
fresh lucerne sprouts addition and to evaluate the snack pellets were dried at room temperature for
effects of lucerne sprouts level as well as process- 24 h (final moisture content below 11%) and then
ing conditions on water consumption, processing stored in hermetically sealed plastic bags.
aspects and selected physiochemical characteris-
tics of the obtained snack pellets. There are no Calculation of water consumption
current studies on the use of fresh lucerne sprouts
as an additive to extruded snack pellets; thus, the The moisture content of ingredients was test-
present study is a first. ed utilizing an air-oven by following the AACC
44–15.02 standard procedure [Kasprzycka et al.,
2018]. Calculation of the water amount needed
MATERIALS AND METHODS to crerate the desired initial moisture content of
blends was based on the following formula (1)
Raw materials and snack pellets processing [Lisiecka and Wójtowicz, 2020]:
The main components of snack pellets were
wheat flour type 450 and corn flour (50:50), sugar 𝑀𝑀𝑓𝑓 − 𝑀𝑀𝑖𝑖
(2%) and salt (1%), purchased at the local market. 𝑊𝑊 = 𝑚𝑚𝑠𝑠 ( ) (1)
100 − 𝑀𝑀𝑓𝑓
Basic composition of the used wheat flour was:
moisture 11.40%, protein 12.26%, fat 0.98%, ash where: W – demand of water (kg/kg),
0.45%, fiber 1.30%. That of corn flour was: mois- ms – mass of sample (kg),
ture 9.88%, protein 5.13%, fat 1.41%, ash 0.45%,
Mf – final moisture content of blend (%)
fiber 2.00% (producers’ data in dry mass). Fresh
Mi – initial moisture content of blend (%).
lucerne sprouts purchased from Uniflora Sp. z o.
o. (Częstochowa, Poland) were used in amounts The difference between the water amount
of 10, 20 and 30% of the total blend mass as re- needed to moisten the dry components and that
placement of flours in the recipe. Fresh lucerne needed for blends with fresh vegetable addition
sprouts (basic composition: moisture 92.10%, was calculated.
protein 3.99%, fat 0.69%, ash 0.40%, fiber 2.50%,
producers’ data in edible sprouts) were ground to Processing efficiency and energy
a particle size lower than 1 mm by using a kitchen consumption
blender to create a pulp. Dry components were
mixed and fresh lucerne sprouts pulp was added The processing efficiency (Q) was determined
w/w and mixed with dry blend to create a homog- as a ratio of the obtained pellets mass at the defined
enous mass. The mixed blends were tested for time. The process efficiency was calculated using
moisture content, and samples were moistened up the formula proposed by Matysiak et al. [2018]:

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measured with the use of UV-VIS spectropho-

𝑚𝑚
𝑄𝑄 = (kg/h) (2) tometer (Genesys 20 UV-VIS, Thermo Scientific,
𝑡𝑡
Waltham, MA, US) at 760 nm. The level of phe-
where: Q – process efficiency (kg/h), nolic compounds was expressed as μg gallic acid
m – mass of fed extrudate (kg), equivalents (GAE) per g of dry mass (d.m.). The
t – measurement time (h). free radicals scavenging activity of the obtained
extracts was determined spectrophotometrically
The measurement was repeated three times by using the 2,2-diphenyl-1-picrylhydrazyl free
for each trial, the arithmetic mean of the measure- radical (DPPH) method based on a method of
ments was adopted as the result. Burda and Oleszek [2001]. The following param-
The measurement of the extruder active power eters were applied: 517 nm wavelength, measure-
was recorded by a measuring system installed in ments every 5 min. for 30 min. The results are ex-
the machine. The value of energy was converted pressed as Trolox equivalent antioxidant activity
into the specific mechanical energy (SME) using (TEAC) calculated at 15 minutes from reaction
the formula provided by Matysiak et al. [2018]: initiation for all samples, and the TEAC values
were calculated on the basis of a standard curve:
𝑛𝑛 𝑂𝑂 𝑃𝑃 y = -0.0214x + 0.471.
𝑆𝑆𝑆𝑆𝑆𝑆 = 𝑛𝑛𝑚𝑚
× 100 × 𝑄𝑄 (kWh/kg) (3)

Water absorption and water solubility index

where: SME – specific mechanical energy
(kWh/kg), Water absorption index (WAI) and water sol-
n – extruder rotational speed measured ubility index (WSI) were assessed according to
with a tachometer (s), the methods described by Lisiecka et al. [2021]
nm – extruder nominal speed measured in triplicate. The ratio of the gel weight to the dry
with a tachometer (s), sample weight was defined as WAI. The ratio of
O – extruder motor load versus maximum the weight of dry solids in the supernatant to the
load (%), weight of the dry sample was derived as WSI.
P – nominal power indicated from control
panel (kW), Statistical analysis
Q – process efficiency described above
(kg/h). The obtained results were subjected to statis-
tical analysis using the Excel software to calcu-
Phenolics content and antioxidant activity late means of the results and standard deviations
of multiple measurements.
The extracts from ground supplemented
snack pellets were prepared using ultrasound-
assisted extraction (UAE) by means of an ultra- RESULTS AND DISCUSSION
sonic bath [Bandelin Electronic GmbH & Co.
KG, Berlin, Germany] at 60°C. In the first step
Calculation of water consumption
4 g of ground snack pellet were mixed with 80%
ethanol (80 mL) and extracted (40 min). The ex- Dry components as wheat flour and corn flour
traction was carried out in two identical cycles. applied in the control recipe (50/50) were charac-
Both parts of extracts were combined, evaporated terized by the moisture content of 10.64%, where-
and dissolved in methanol (10 mL). The mea- as fresh lucerne sprouts showed 92.10% of mois-
surement of total content of polyphenolic com- ture content. The initial moisture content of raw
pounds (TPC) was performed by applying the materials for snack pellets processing should vary
Folin-Ciocâlteu (FC) method (modified) [Burda from 25 to 36%, required to proper gelatinization
and Oleszek, 2001]. In the initial step, 200 μL of of starchy components under the extrusion-cook-
extract was mixed with 1.8 mL of distilled water. ing conditions used in the experiment [Matysiak
Afterwards, 200 μL of FC reagent was added. Af- et al., 2018; Lisiecka and Wójtowicz, 2020].
ter mixing the mixture was left for 5 min. Subse- In the experiment, three levels of initial mois-
quently, 2 mL of 7% Na2CO3 was added and then ture were applied at levels of 32, 34 and 36%.
incubated (for 60 min at 40°C). Absorbance was Each blend, both control one and supplemented

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with fresh lucerne sprouts have to be moistened very important both for producers, make the prod-
before treatment, so proper amount of techno- uct easier to prepare, and for environment by lim-
logical water added must be calculated based on iting or completely avoiding of supply the tech-
the equation (1). The results of the technological nological water into processing of supplemented
water added to each tested mixture are presented snack pellets. With the production capacity of
in Figure 1. Replacement of wheat-corn flour 1000 kg/h, the water needed for the control recipe
blends by fresh lucerne sprouts at various levels should be 396 kg/h if 36% of initial moisture is
(10, 20 and 30%) allowed sufficient reduction of set. With daily work of commercial processing
technological water which is needed in the extru- line from 10 to 12 hours, the water amount is
sion-cooking process of snacks pellets. If control 3960–4752 L daily. Dry starchy components used
sample was moistened, at least 314 to 396 mL of in the recipe were characterized the moisture con-
water per kg of dry components need to be added tent of 11.40% for wheat flour and 9.88% for corn
to reach 32 and 36% of raw material blends mois- flour, so the control mixture was 10.64% of initial
ture before the extrusion, respectively. Applica- moisture content in blend (50/50 wheat and corn
tion of 10% of fresh lucerne sprouts enabled to flour). To obtain snack pellets with proper qual-
limit water added to 201–258 mL per kg of dry ity, the technological water must be supplied to
components and for 20% of fresh additive the wa- reach the desired moisture levels from 25 to 36%
ter amount was limited to 101–170 mL per kg, [Panak Balentić et al., 2018].
respectively to obtain 32 and 36% of initial mois- Replacement of technological water in snack
ture of components before the extrusion process. pellets composition by the natural water coming
The most significant water saving was from fresh lucerne sprouts (92.10% of moisture)
reached when 30% of fresh lucerne sprouts were made it possible to obtain well-processed snack
applied and when initial moisture was set at 32%, pellets without any addition of water, which was
because in this case any additional technologi- confirmed by the proper structure and stability
cal water was needed. Similarly, only 10 mL per of products, so daily limitation is possible from
kg and 42 mL per kg was needed in the samples 4 to almost 5 m3 of technological water, saving
supplemented with 30% of fresh lucerne sprouts the environment and the money. This developed
if the initial moisture of blends was set to 34 solution to replace the starchy components by
and 36%, respectively. Therefore, the limitation fresh lucerne sprouts seems to be also benefi-
of water was from 89 to 100% if fresh lucerne cial due to the increasing amount of nutritionally
sprouts were used, depending on the recipe and valuable component from lucerne sprouts in final
dough moisture level tested. pellets, which would be confirmed by the analy-
Looking for these numbers obtained in the sis of nutritional characteristics of the obtained
laboratory scale, it seems not much but on the in- snack pellets.
dustrial scale, when the processing line efficiency Similar tendencies were reported by Lisiecka
is even up to 1000 kg per h, these savings may be and Wójtowicz [2020], they found the possibility

Figure 1. Water amount needed to reach the initial moisture of snacks pellets
supplemented with 10, 20 and 30% of fresh lucerne sprouts

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to save the water and energy by the addition of production of snack pellets using the addition of
fresh vegetables into potato-based snack pellets. fresh lucerne sprouts allowed the development of
They reported the possibility to limit the tech- technological guidelines for a new range of snack
nological water addition coming from fresh products. The addition of fresh lucerne sprouts to
vegetables as the moisture content of fresh veg- the raw material blends showed a differential ef-
etable pulps was as follows: carrot–87.10%, fect on process efficiency (Table 1). The efficien-
kale–85.89%, leek–88.62% and onion–87.35%. cy of the extrusion process mostly depended on
Therefore, the water amount needed to moisten the screw speed of the extruder, the sprout con-
the dough for pellets to 33% of initial moisture tent in the mixture and initial moisture content of
content allowed limiting at least 0.334 kg wa- components. The highest extrusion process effi-
ter per kg of raw materials if 30% of additives ciency was observed for the blends with an initial
were used. Thus, the use of vegetable additives moisture content of 32% extruded at the screw
in the amount of 30% reduced the amount of wa- speed of 100 rpm. Matysiak et al. [2018] showed
ter consumed in the range from 97.92% (kale) to similar trends. The applied functional additive in
99.5% (leek). the form of fresh lucerne sprouts had a significant
According to the European Commission (EC) effect on the extrusion process efficiency. The ad-
Council Directive 96/61/EC on Integrated Pollu- dition of fresh vegetables influences the physico-
tion Prevention and Control (IPPC), the strategy chemical composition of the raw material mix-
to save the water is strongly advisable, espe- tures, which directly translates into the extrusion
cially as part of the Food Industry Sustainability process efficiency. The highest process efficiency
Strategy (FISS) prepared by the Department of (32.72 kg/h) was observed for the raw mate-
Environment, Food and Rural Affairs [DEFRA, rial blends containing 10% added fresh lucerne
London, UK]. They recommended as mandatory sprouts. Lisicka and Wójtowicz (2019) conducted
a 10–15% reduction of water in food industry by an extrusion study of corn snacks and obtained an
2020, but there is no information if these recom- extrusion process efficiency of 32–36 kg/h. This
mendations have been fixed. The application of may be due to the lower amount of water in the
water efficiency management systems or waste- raw material mixture, which directly influences
water re-use after treatment make it possible to the flow speed of the mass inside the extruder.
save 30–50% of fresh water in industry [Nikma- The lowest extrusion efficiency (9.12 kg/h)
ram and Rosentrater, 2019]. was observed for the blends produced at 60 rpm
This water saving is important to producers, of the extruder screw speed with the additive
so many activities were done to limit the tech- level of 30% and blend moisture content of 36%.
nological water needs in food industry, such as The study also found that the extrusion efficiency
dry peeling instead of wet peeling of fruits and is significantly influenced by the moisture content
vegetables, high pressure-low volume cleanup of the raw material mixture. This was confirmed
systems, dry pre-cleaning prior to washing, or op- by the results of the extrusion efficiency of the
timization of water flow by flow restrictors [Öl- control sample without the addition of fresh lu-
mez, 2013]. Advisable reduction of fresh water in cerne sprouts. As the moisture content of the raw
food industry should be up to 90%, so the finding materials increased, the efficiency of the extru-
presented in this paper completely fulfill these sion process decreased.
requirement and can be recommended directly The results of SME was low as calculated
to the producers as an environmentally friendly per kg of product. The range of specific en-
technological solution ready to implement on an ergy consumption varied from 0.035 to 0.179
industrial scale. kWh/kg. Higher SME values were found if
greater initail moisture was applied during
Processing efficiency and energy processing of snack pellets but the highest re-
consumption sults were found for control wheat-corn snack
pellets without additives. Increasing amount
The efficiency of the extrusion-cooking of fresh lucerne sprouts in the recipe lowered
process depends on many factors, including energy consumption which could be the effect
the raw materials used, their moisture content, of lower starch content and thus lower viscos-
granulation, functional additives, extrusion pro- ity of the treated mass undergoing the gelati-
cess temperature and extruder screw speed. The nization process [Prabha et al., 2012]. Also,

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Table 1. Efficiency and energy consumption values obtained for new generation snack pellets fortified with fresh
lucerne sprouts (data expressed as mean value of n = 3 ± SD standard deviation)
Fresh lucerne sprouts Efficiency SME
Screw speed (rpm) Moisture content (%)
content (%) (kg/h) (kWh/kg)
32 15.28 ± 0.54 0.086 ± 0.002
60 34 27.81 ± 0.25 0.137 ± 0.002
36 13.04 ± 0.32 0.142 ± 0.004
0
32 31.44 ± 1.26 0.090 ± 0.003
100 34 14.48 ± 1.46 0.137 ± 0.002
36 25.36 ± 0.63 0.179 ± 0.005
32 20.80 ± 0.72 0.088 ± 0.003
60 34 17.92 ± 2.01 0.090 ± 0.003
36 16.48 ± 0.24 0.104 ± 0.002
10
32 32.72 ± 0.98 0.045 ± 0.002
100 34 20.72 ± 0.85 0.081 ± 0.004
36 22.76 ± 0.56 0.061 ± 0.001
32 20.08 ± 0.72 0.035 ± 0.003
60 34 17.84 ± 0.28 0.094 ± 0.003
36 9.28 ± 0.24 0.181 ± 0.002
20
32 27.84 ± 0.98 0.035 ± 0.002
100 34 27.36 ± 2.01 0.061 ± 0.002
36 22.76 ± 0.56 0.061 ± 0.001
32 15.36 ± 0.26 0.096 ± 0.002
60 34 10.24 ± 0.28 0.153 ± 0.003
36 9.12 ± 0.24 0.163 ± 0.004
30
32 26.24 ± 0.84 0.068 ± 0.002
100 34 22.01 ± 0.08 0.090 ± 0.004
36 14.96 ± 0.60 0.118 ± 0.004

as compared to other snacks processess, the Total content of polyphenolic compounds

extrusion-cooking is less energy consuming, and free radical scavenging activity
especially when semi or high-moisture condi-
tions are applied. Meng et al. [2010] reported An excellent method of improving the health
and nutritional properties of plants is to germinate
the SME increase along with screw speed and
their seeds. The sprouts from early plant’s growth
decreasing barrel temperature by applying
cycle, so nutrient content is extremely high.
twin screw extruder, but the feed moisture had
Sprouts are a very rich source of bioactive com-
no effect on SME (calculated SME varied from
pounds (e.g. polyphenols), minerals and vitamins.
87.9 to 115.8 Wh/kg).
Plants belonging to the genus Medicago,
Matysiak et al. (2018) reported higher en-
such as lucerne, were among the first cultivated
ergy consumption during the processing of by humans. This group of plants is characterized
wholegrain cereal raw materials than potato- by having high nutritional properties, as well
based recipies, because the higher SME usu- as enhanced vitamin (C, E, from group B, beta
ally is the result the greater degree of starch carotene) and mineral (potassium, calcium, mag-
gelatinization. Observations done by other au- nesium, silicon, iron) composition. Additionally,
thors suggesting that semi-moisture extrusion due to the high level of chlorophyll, plants of the
requires less energy, but as presented by da Medicago genus may have a detoxifying effect
Silva et al. [2014] the SME values (ranged be- for human body [Krakowska-Sieprawska et al.,
tween 32.20 and 58.88 kJ/kg) of the extruded 2021]. European Food Safety Authority accepted
products processed under experimental condi- plants from this group as safe dietary ingredients.
tions (moisture 11–21%, screw speed 333–378 For now, their consumption by humans is still
rpm) showed that none of the variables studied low, but there has been an increasing interest in
(screw speed, moisture and bean level) signifi- using lucerne as additives for salads, supplements
cantly influenced the SME values. and functional food.

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The contents of polyphenolic compounds and different quantities of lucerne sprouts (0, 10, 20,
antioxidant properties of snack pellets enriched 30%) produced at various screw speeds and mois-
with Medicago sativa L. (lucerne, alfalfa) sprouts ture contents by using extrusion-cooking showed
were tested in this paper to indicate the contained high content of polyphenolic compounds increas-
bioactive compounds and to establish their pro- ing significantly due to the higher addition of lu-
health potential. The first stage of the analysis cerne sprouts (Table 2).
was to isolate the polyphenolic compounds con- Results indicated the highest total content of
tained within the pellets. Recent directions indi- polyphenols for the pellets with 30% addition
cated by “green chemistry principles” in extrac- of the lucerne sprouts processed at screw speed
tion methods have focused on the improvement 60 rpm and at moisture content 36%, while the
of the efficiency of the process and reduction lowest was seen in a control sample without such
of solvent consumption [Krakowska-Siepraws- functional additive that was extruded at the screw
ka et al., 2021]. A recognized environmentally speed of 60 rpm and moisture content at 36%.
friendly technique is ultrasound-assisted extrac- Significant differences in the level of polyphe-
tion (UAE) with ethanol. UAE at elevated tem- nolic compounds in the produced snack pellets
peratures and with 80% ethanol was previously due to the effect of screw rotational speed and
used by Kasprzak-Drozd et al. [2022] or Soja et initial moisture content were observed. How-
al. [2023] to obtain polyphenolic extracts from ever, the discovered proportional increase in the
the snacks enriched with vegetables and herbs. level of polyphenolic compounds indicates the
In these papers, UAE was demonstrated to be an fact that extrusion technique did not degrade the
optimal technique for extracting phenolics from a active compounds present in pellets enhanced
wide group of functional foods, therefore, it was with M. sativa sprouts. This process may reduce
applied in this experiment. The tested pellets with the content of bioactive substances in the tested

Table 2. Total phenolic content and TEAC values obtained for new generation snack pellets fortified with fresh
lucerne sprouts (data expressed as mean value of n=3 ± SD standard deviation)
Fresh lucerne Moisture content TPC content TEAC value
Screw speed (rpm)
sprouts content (%) (%) (μg GAE /g d.w. product) (µg/g d.w. product)
32 62.10 ±1.20 111.45 ± 4.38
60 34 61.23 ± 1.45 112.21 ± 5.26
36 60.60 ±1.90 113.08 ± 5.12
0
32 78.10 ± 2.10 106.54 ± 6.11
100 34 71.25 ± 1.98 102.78 ± 5.14
36 60.80 ± 1.80 103.50 ± 4.98
32 102.10 ±7.62 117.76 ± 5.09
60 34 104.23 ± 6.14 112.58 ± 4.48
36 106.30 ± 5.12 116.12 ± 5.23
10
32 105.10 ± 5.16 113.32 ± 4.99
100 34 106.12 ± 4.69 118.45 ± 3.68
36 107.70 ± 4.87 125.12 ± 5.89
32 135.12 ± 6.24 114.25 ± 5.12
60 34 141.14 ± 7.12 116.85 ± 6.98
36 139.24 ± 5.47 128.45 ± 8.02
20
32 139.26 ± 4.89 121.21 ± 8.47
100 34 146.69 ± 6.98 129.31 ± 6.51
36 142.44 ± 6.47 126.98 ± 3.98
32 170.70 ± 7.56 126.92 ± 6.45
60 34 175.23 ± 9.11 129.71 ± 8.11
36 175.10 ± 7.12 126.75 ± 6.22
30
32 170.70 ± 6.98 117.29 ± 5.98
100 34 168.45 ± 7.12 121.65 ± 3.99
36 169.50 ± 8.02 126.17 ± 4.98
Note: TPC – total phenolics content; TEAC – Trolox equivalent antioxidant activity; a – d – data in columns with
similar letters did not differ significantly at α=0.05 by Tukey post-hoc test.

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Journal of Ecological Engineering 2023, 24(12), 130–142

samples, but simultaneously can increase their damage to cells. Therefore, the higher the poly-
bioavailability to the human body. Chiriac et al. phenol content in a snack pellet, the greater its
[2020], for example, identified a total of 59 poly- potential to act as an antioxidant. It is important
phenols, e.g. fenolic acids, isoflavones, flavones, to note that the specific types and concentrations
flavonones, and flavonols in alfalfa sprouts. After of polyphenols can vary among different snacks,
analyzing the presented data, sprouts of the third depending on their ingredients and processing
day of germination were used in the conducted methods. Considering the positive correlation
experiment. The biological actions of Medicago between polyphenol content and antioxidant ac-
may be due to the presence of polyphenols and the tivity, incorporation of ingredients rich in poly-
function as free radical scavengers and/or metal phenols, such as fruits, nuts, or whole grains, into
chelators [Krakowska-Sieprawska et al., 2021]. snacks can enhance their antioxidant properties
Therefore, the DPPH free radical scavenging po- [Francis et al., 2022; Lotfi Shirazi et al., 2020;
tential of the pellets was analyzed. The obtained Lourenço et al., 2016; Soja et al., 2023].
results presented as Trolox equivalent antioxidant
activity (TEAC, Table 2) confirmed that the anti- WAI and WSI of snack pellets
oxidant properties of products increased with the
addition of fresh lucerne sprouts. Water absorption index (WAI) indicates the
The highest antiradical properties were found ability of a treated component to absorb and
in the pellets supplemented with 30% of lucerne hold external water and it is a measurement of
sprouts produced using 60 rpm and with either the amount of gel formed by processed starch,
34 and 36% of moisture content. In general, the protein and fibers after swelling. During the
samples extruded at 60 rpm had higher antioxi- extrusion-cooking a starch gelatinization and
dant activity than those produced at 100 rpm. protein denaturation occur depending on pro-
It is possible that the shear forces prevailing at cessing conditions [da Silva et al., 2014; Meng
100 rpm may inactivate a part of the antioxidant et al., 2010]. Gelatinized starch absorbs more
compounds present in the pellets [Kasprzak-Dro- water trapped in the materials, thus increas-
zd et al., 2022]. However, upon noticing slight ing the WAI values. When material is treated
differences in TEAC for both rotational speeds, too intensively, the starch granules may be de-
this process must occur only to a small extent. stroyed and lose their water absorption ability
Moreover, the increasing moisture content had [Ruiz-Armenta et al., 2019]. During the extru-
a positive effect upon the antioxidant properties sion process, starch gelatinization is presented
of snack pellets processed at 100 rpm. Thus, the allowing a greater capacity of However; the
moisture content of snack pellets can have an im- presence of proteins or fibers in the extrudate
pact on their antioxidant properties; high mois- composition may have effect on WAI due to
ture can prevent the degradation of the antioxi- changes affecting the hydrophilic/hydrophobic
dants, reducing shear forces inside the extruder. balance of the processed components [Louren-
When analyzing the obtained study results, it was ço et al., 2016]. WSI is a particularly important
noted that both 34% and 36% moisture content parameter of extrusion-cooking intensity and
did not have negative influence on the health-pro- allows predicting raw material behavior during
moting activity if processed at 60 rpm. However, processing. In the tested samples, low water
increase in rpm when lower (32%) moisture con- absorption in control wheat-corn snack pel-
tent was applied during processing caused more lets was noted (3.15–4.43 1/g), but, at the same
intensive shearing and thus lower antioxidant ac- time, these samples showed the highest level of
tivity due to a higher shear stress. Hence, appro- WSI (10.34–14.72%). All results are presented
priate choice of selected production parameters in Table 3. An increasing amount of lucerne
may allow the protection of sensitive components sprouts significantly enhanced water absorp-
affecting pro-health values of the extruded food. tion of the tested snack pellets supplemented
In the conducted study, the antioxidant activity of with fresh sprouts, and more than twice inten-
the snack pellets was positively correlated with sive water absorption ability was evaluated if
the polyphenols content (r = 0.81). Polyphenols 30% of additive was used, as compared to the
exhibit antioxidant activity by scavenging and control sample.
neutralizing free radicals, which are highly reac- In the conducted study, high WSI values in-
tive molecules that can cause oxidative stress and dicated that components were not sufficiently

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 Journal of Ecological Engineering 2023, 24(12), 130–142

integrated in the pellet structure. This outcome products of both 60 and 100 rpm processing
was probably due to the high content of fiber screw speeds. As WSI increased with increas-
and the low amount of starch needed to stick ing moisture and with higher screw speeds, this
together all the components supplemented with outcome indicates that the components were
fresh lucerne sprouts. The fiber fractions which not sufficiently integrated in the pellet struc-
are non-processable at low temperature can, ture. This was probably due to the enhanced
however, be easily rinsed out from the snack content of protein, fiber and other healthy com-
pellet sample. Therefore, WSI was low in ponents present in fresh sprouts, and thus low-
snack pellets with 10% of sprouts, but the low- ered amount of starch in the recipes.
est value was obtained for sample with 30% of During the extrusion-cooking, due to the
lucerne processed at 60 rpm and 32% of initial presence of water, heat and shear-forces the
moisture. Indeed, in all samples processed with reactions between proteins, amylose, and amy-
these settings (32% m.c., 60 rpm) WSI was the lopectin generate several changes that lead to
lowest. Therefore, an improving effect of lu- reduction of WAI. In the research of Lisiecka
cerne sprout addition on the WAI of snack pel- et al. [2021] a negative correlation was demon-
lets was perceived. What is more, processing strated between WAI value and protein content,
conditions showed similar effects on the WAI soluble dietary fiber content and ash content of
and WSI of the tested samples. The higher WAI the snacks supplemented with fresh beetroot
and WSI were observed when increased initial pulp as a starch substitution. They found that
moisture content was applied during snack pel- extrusion-cooking induced an increase in the
let processing. Similarly, higher initial mois- insoluble dietary fiber content and WAI in the
ture influenced increased WSI values for the obtained potato-based pellets.

Table 3. Results of WAI and WSI in new generation snack pellets fortified with fresh lucerne sprouts (data
expressed as mean value of n=3 ± SD standard deviation)
Fresh lucerne sprouts content
Screw speed (rpm) Moisture content (%) WAI (1/g) WSI (%)
(%)
32 3.15 ± 0.03 10.34 ± 0.07
60 34 3.64 ± 0.04 12.17 ± 0.06
36 3.82 ± 0.05 14.14 ± 0.13
0
32 3.43 ± 0.01 12.47 ± 0.43
100 34 3.99 ± 0.02 13.47 ± 0.32
36 4.43 ± 0.14 14.72 ± 0.05
32 3.76 ± 0.03 8.96 ± 0.01
60 34 4.01 ± 0.02 9.12 ± 0.05
36 4.12 ± 0.01 9.87 ± 0.08
10
32 3.95 ± 0.03 9.03 ± 0.01
100 34 4.16 ± 0.05 10.11 ± 0.02
36 4.39 ± 0.12 10.84 ± 0.04
32 5.21 ± 0.02 9.28 ± 0.04
60 34 5.01 ± 0.04 10.11 ± 0.06
36 5.26 ± 0.02 11.14 ± 0.06
20
32 6.52 ± 0.12 10.13 ± 0.12
100 34 6.21 ± 0.19 9.24 ± 0.24
36 6.98 ± 0.14 10.31 ± 0.15
32 6.29 ± 0.04 7.78 ± 0.04
60 34 6.98 ± 0.03 8.58 ± 0.07
36 7.08 ± 0.13 10.46 ± 0.11
30
32 6.79 ± 0.11 11.93 ± 0.15
100 34 7.12 ± 0.04 12.23 ± 0.41
36 7.46 ± 0.05 13.07 ± 0.20
Note: WAI – water absorption index; WSI – water solubility index; a–j
– data in columns with similar letters did
not differ significantly at α=0.05 by Tukey post-hoc test.

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Journal of Ecological Engineering 2023, 24(12), 130–142

After processing the material in the pro- CONCLUSIONS

posed process conditions, higher WAI and WSI
index were recorded for the obtained beetroot- Replacement of wheat-corn flour blends
supplemented pellets in relation to the unpro- by fresh lucerne sprouts at various levels (10,
cessed control sample. The maximum WSI 20 and 30%) allowed sufficient reduction of
value of 8.63% was determined for the pel- technological water which is needed in the
lets with 10% of fresh beet pulp extruded at extrusion-cooking process of snacks pellets.
80 rpm, but the lowest value of the WSI index The limitation of water was from 89 to 100%
was 1.42% for the pellets from the control po- if fresh lucerne sprouts were used, depending
tato sample produced at similar screw speed. on the recipe and dough moisture level test-
It was concluded in the work that very low ed. Replacement of technological water in the
solubility may be the effect of lower starch ge- composition of snack pellets by the natural wa-
latinization temperature of potato components ter coming from fresh lucerne sprouts (92.1%
compared to wheat and corn raw materials. of moisture) made it possible to obtain well
Ruiz-Armenta et al. [2019] observed an in- processed snack pellets, which was confirmed
crease in WSI after extrusion in corn starch- by proper structure and stability of products,
based pellets supplemented with naranjita and without any technological water added if 30%
milk powder. They held the opinion that this of lucerne sprouts were applied. In an indus-
effect was linked to the loss of the semicrystal- trial practice with high-capacity processing
line form of native starch granules after pro- line, the daily saving from 4 to 5 m3 of tech-
cessing. In their study, the maximum WAI val- nological water may be done if this technol-
ue of pellets and snacks with carrot pulp addi- ogy will be adopted. The addition of fresh lu-
cerne sprouts had a significant impact on the
tion was 4.90 1/g when 5% of carrot pulp was
nutritional quality of developed wheat-corn
added during the extrusion at 80 rpm. How-
blend-based snack pellets. The almost triple
ever, significant differences in WAI were ob-
increase of total phenolics content at 30% of
served in the pellets with the maximum content
additive was observed as compared to the base
(30%) of fresh carrot pulp in the composition,
control sample. The addition of fresh lucerne
as compared to the control sample extruded at
sprouts to the basic recipe at the level of 10,
the same speed of the extruder screw.
20 and 30% also had significant effect on the
In the work, minimal WSI was found to be
snack pellets WAI and WSI. Moreover, WAI
1.42% in snack pellets with 5% of vegetable
increased if the initial moisture level was high-
additive produced at the minimum rotational
er and if higher screw speed was applied dur-
speed of the extruder screw. Ruiz-Armenta et ing snack pellets extrusion. On the basis of the
al. [2019] noted that the WAI of pellets was presented results, 36% initial moisture content
negatively correlated with soluble fiber and the and 60 rpm screw speed were recommend as
WSI of pellets was positively correlated with the appropriate processing conditions to pro-
the content of protein, ash, soluble and total duce acceptable lucerne sprout fortified snack
fiber, and negatively with fat content. Lisiecka pellets for the market. However, the developed
et al. [2021] tested snack pellets with the ad- snack pellets are only semi-finished products,
dition of fresh vegetables of Allium genus and so further research is needed in terms of final
they discovered that WAI ranged from 3.04 g/g expansion energy requirements depend on the
in pellets with 20% of added leek pulp, ex- expansion method used and sensory analysis
truded at 100 rpm, to 4.65 g/g, in the pellets and consumer acceptance before the produc-
with 2.5% of leek additive content produced at tion of a ready-to-eat expanded snack that con-
the highest screw speed, while the WSI values tains lucerne for the market.
were determined in the range from 0.20% (pel-
lets with the highest level of WAI) to 7.48%
Acknowledgements
(pellets with content of 2.5% of onion pulp
produced at 60 rpm). The statistical analysis This research was funded by The National Cen-
confirmed that the level of fresh vegetable tre for Research and Development (NCBiR), grant
pulp had a significant negative impact on the number LIDER/29/0158/L-10/18/NCBR/2019
WAI and WSI values of all pellet products. entitled “Development of a Comprehensive

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 Journal of Ecological Engineering 2023, 24(12), 130–142

Technology of Obtaining High-Quality Extruded 10, 8–16. DOI: 10.1016/j.hermed.2017.07.002

Snacks Based on Minimally Processed Vegetable 10. Hong Y.H., Wang S.C., Hsu C., Lin B.F., Kuo Y.H.,
and Animal Raw Materials”. Huang C.J. 2011. Phytoestrogenic compounds in
alfalfa sprout (Medicago sativa) beyond coumes-
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