applied

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Article
Isolation and Characterization of Starch from Different Potato
Cultivars Grown in Croatia
Mario Kovač 1 , Boris Ravnjak 2, * , Drago Šubarić 3 , Tomislav Vinković 2 , Jurislav Babić 3 , Ðurd̄ica Ačkar 3 ,
Ante Lončarić 3 , Antonija Šarić 3 , Vesna Ocelić Bulatović 4 and Antun Jozinović 3

1 Faculty of Agriculture and Food Technology, University of Mostar, Biskupa Čule bb,
88000 Mostar, Bosnia and Herzegovina; [email protected]
2 Faculty of Agrobiotechnical Sciences Osijek, Josip Juraj Strossmayer University of Osijek, Vladimira Preloga 1,
31000 Osijek, Croatia; [email protected]
3 Faculty of Food Technology Osijek, Josip Juraj Strossmayer University of Osijek, Franje Kuhača 18,
31000 Osijek, Croatia; [email protected] (D.Š.); [email protected] (J.B.); [email protected] (Ð.A.);
[email protected] (A.L.); [email protected] (A.Š.); [email protected] (A.J.)
4 Faculty of Chemical Engineering and Technology, University of Zagreb, Marulićev trg 19,
10000 Zagreb, Croatia; [email protected]
* Correspondence: [email protected]

Abstract: Starch is a polysaccharide that is widely used in food and other industries; therefore, due
to its great potential, it is attempted to be maximally isolated from various foods rich in starch.
Commonly, potatoes are used for starch production due to the relatively high starch content in tubers,
and the process itself is complex and includes several steps. The aim of this study was to isolate and
characterize starch from eight potato varieties. First, the basic chemical composition of the potato
samples was determined, and then the isolation was carried out under laboratory conditions. The
isolated starch was air dried, then ground and sieved through a 400 µm sieve. The basic chemical
composition, amylose content, starch color, swelling capacity and solubility index, clarity of starch
pastes, texture of starch gels and thermo-physical properties (gelatinization and retrogradation) were
determined in the obtained starch samples. The results showed that the SL 13–25 potato variety had
Citation: Kovač, M.; Ravnjak, B.; the lowest starch content, while the Stilleto variety had the highest starch content. The content of
Šubarić, D.; Vinković, T.; Babić, J.; protein, fat, ash and crude fiber was relatively low in all of the isolated starches, indicating their
Ačkar, Ð.; Lončarić, A.; Šarić, A.; high purity. Also, the difference in the color of the isolated starches was difficult to see. The highest
Bulatović, V.O.; Jozinović, A. Isolation amylose content had starch from Saprodi, while the lowest was observed in starch from SL 13–25.
and Characterization of Starch from Starch from the cv. Dartiest had the highest, while starch from the cv. Sereno had the lowest gel
Different Potato Cultivars Grown in
strength. The starch of the cv. Dartiest also had the highest clarity value. The retrogradation transition
Croatia. Appl. Sci. 2024, 14, 909.
temperatures and enthalpies were lower compared to the gelatinization temperatures and enthalpies.
https://doi.org/10.3390/
With the increasing temperature, both the swelling capacity and the solubility index of all the samples
app14020909
increased. According to the obtained differences, this study confirmed the significant influence and
Academic Editor: Alessio Adamiano role of different cultivars on starch characteristics.
Received: 20 November 2023
Revised: 16 January 2024
Keywords: potato; starch; isolation; characterization
Accepted: 19 January 2024
Published: 21 January 2024

1. Introduction
Potatoes are the fourth most important food crop in the world after wheat, rice and
Copyright: © 2024 by the authors.
maize, with a global annual production of approximately 300 million tons. The chemical
Licensee MDPI, Basel, Switzerland.
composition and ratio of nutrients differ depending on the potato cultivar, but also on a
This article is an open access article
whole series of ecological and production factors (soil, fertilization, climatic conditions,
distributed under the terms and
etc.) and the method of preparation [1–3].
conditions of the Creative Commons
Potato is a raw material rich in starch, which is found in the potato tuber and is formed
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
by dividing the ends of the stolons, after which the cell swells and the tuber develops [4].
4.0/).
Potato starch is an essential product obtained through the process of the separation of the

Appl. Sci. 2024, 14, 909. https://doi.org/10.3390/app14020909 https://www.mdpi.com/journal/applsci

Appl. Sci. 2024, 14, 909 2 of 12

solid components—starch and fiber—from the liquid potato juice. The goal of the starch
production industry is to obtain starch with as few impurities as possible—i.e., to obtain
the highest yield of starch—and this is a complex and demanding process that includes a
series of steps [5,6].
The process of producing starch from potatoes can be conducted in three ways; namely,
by separating the three components during production: the starch itself, fibers and potato
juice. The economy over a long period of processing should also be taken into account, as
well as parameters such as the dry matter, starch and protein content [7].
This research aimed to examine the influence of different potato cultivars on the
starch properties: starch color, swelling capacity and solubility index, clarity of starch
pastes, texture of starch gels and thermophysical properties of starch (gelatinization and
retrogradation). For this purpose, starch was isolated from eight potato cultivars grown in
the area of Osijek-Baranja County.

2. Materials and Methods

2.1. Materials
Eight different starchy potato cultivars (Saprodi, Sofista, Stilleto, Dartiest, Sereno,
Senata, Scala and future cultivar (breeding clone) with code name SL 13–25) were grown
in 2021 at the Experimental Station of the Faculty of Agrobiotechnical Sciences in Osijek,
Croatia, located near the suburban settlement Tenja (45.518770, 18.774791), Osijek-Baranja
County. Starchy potato cultivars were obtained from starch potato seeds producer Royal
ZAP-Semagri Holland BV (Wieringerwerf, The Netherlands). Potato seeds were delivered
to the faculty to test the quality and possibility of their production in the area of Eastern
Europe (temperate to dry climate). Planting was performed mechanically on 3 May 2021
(plant spacing 32 × 70 cm; plant density 4.46 plants per square meter). The trial was
set in a randomized block design, where each cultivar was planted in four repetitions,
each containing 100 plants. For sampling, tuber harvesting was performed manually on
29 September 2021. After the harvest, potato tubers were stored for three months at 4 ◦ C,
and this was the time of tuber maturation. Afterwards, laboratory analysis and starch
isolation were performed.

2.2. Method of Isolation of Starch from Potatoes

The process of isolating starch from potatoes began with a thorough washing of the
potato tubers in tap water, after which the potato tubers were washed in distilled water. The
potatoes were then cut into small pieces and ground using a laboratory blender (Kenwood)
with distilled water (0.5 kg potatoes/250 mL water) for about 25 s (at speed level 5). The
next step was to manually wash the starch from the potato pulp. Potato pulp was poured
onto starch gauze with the symbol 11xxx [132 µm] and the starch was rinsed with distilled
water. The process of washing the starch was repeated 4–5 times until the starch was no
longer slippery and the water became clear. The obtained starch milk was left overnight
(12 h at room temperature) for starch sedimentation, and then the water was poured off
over the sediment (decantation). Then, the starch was mixed with a new portion of distilled
water (the process took place in a bucket, and the amount of distilled water was 5 L). This
operation was performed two more times (starch was rinsed 3 times in total). In the fourth
mixing of starch with distilled water, refining was performed—i.e., a suspension of starch
and distilled water was poured through 13xxx [104 µm] starch gauze—and the starch was
allowed to sediment again (12 h). After the final pouring of water over the precipitate, the
wet starch was placed in plastic trays to dry. The samples were dried in a laboratory dryer,
Memmert UFE 500, Schwabach, Germany, at a temperature of 30 ◦ C. The dried starch was
ground in an IKA M20 laboratory grinder, Staufen, Germany, with a grinding time of 6 s,
and then sieved through a sieve with an opening size of 400 µm. The starches were packed
in polyethylene bags and then their properties were determined.
Appl. Sci. 2024, 14, 909 3 of 12

2.3. Determination of Basic Chemical Composition and Amylose Content

The chemical composition was determined according to the following ISO standards:
determination of dry matter content through drying to constant mass at 105 ◦ C (ISO 6540),
determination of ash content through burning the sample in a muffle furnace at 550 ◦ C (ISO
5984), determination of fat content using the Soxhlet method (ISO 6492), determination
of protein content using the Kjeldahl method (ISO 5983-2), determination of crude fiber
content using the method with intermediate filtration (ISO 6865). Amylose content was
determined according to the Megazyme method using a commercial amylose/amylopectin
enzymatic kit K-AMYL 06/18 (Megazyme Ireland International, Ltd., Bray, Ireland).

2.4. Determination of the Color of Starch

Chroma Meter CR-300, Konica Minolta, Tokyo, Japanwas used to determine the
color of the starch. Before measuring the color in the CIELab and CIELCh systems
(L*—black/white; a*—red/green; b*—yellow/blue; C—chroma, h—hue), the device was
calibrated using a calibration plate. Five measurements were performed for each sample.

2.5. Determination of Paste Properties by Brabender’s Micro Visco-Amylograph

Determination of paste properties was carried out using Brabender’s micro visco-
amylograph (Brabender, Duisburg, Germany). The device is connected to a computer that
manages the operation of the device and processes the obtained results. When measuring
the rheological properties, the samples were subjected to a temperature program:
1. Heating from 30 to 92 ◦ C, heating rate 7.5 ◦ C/min.
2. Isothermal at 92 ◦ C, 15 min.
3. Cooling from 92 to 50 ◦ C, cooling rate 7.5 ◦ C/min.
4. Isothermal at 50 ◦ C, 15 min.

2.6. Determination of Swelling Power and Solubility Index of Starches

The isolated starches were weighed in 50 mL centrifuge cuvettes with a conical bottom
to prepare a 1.0% suspension (25 g suspension). The cuvettes were covered with a cap
and mixed well, and the suspension was then heated at temperatures of 55, 65, 75 and
85 ◦ C for 30 min in a water bath with a shaker (150 rpm), with occasional manual mixing
to homogenize the sample. Gelatinized starch was then cooled as soon as possible in cold
water to room temperature and centrifuged at 4000 rpm for 15 min. After centrifugation,
the supernatant was decanted into pre-dried and weighed aluminium drying containers
and dried at 105 ◦ C to a constant mass.

2.7. Determination of Clarity of Starch Pastes

Paste clarity of the starches was determined based on the method of Kerr and Cleve-
land (1959). The calculated mass of starch and distilled water up to a mass of 20 g was
added to previously weighed empty cuvettes, and the samples were then homogenized on
a Vortex shaker. A 1% suspension of each sample was heated in a boiling water bath for
30 min with constant stirring. The cuvettes with the samples were then removed from the
bath and allowed to cool at room temperature for 1 hour. If there is a loss in the mass of the
suspension, it is compensated by the addition of water that has evaporated, and the mixing
on the Vortex is repeated. Finally, the samples were measured for transmittance at 650 nm
using a laboratory LLG Uni Spec 2 spectrophotometer.

2.8. Determination of the Texture of Starch Gels

The TA-XT Plus device, Stable microsystem, UK, was used to determine the textural
properties of the samples. The sample was analyzed after 24 h to remove air bubbles
incorporated during the preparation of the gels. The gel structure analysis procedure is
carried out using a cylindrical probe with the following dimensions: diameter = 25 mm;
height = 35 mm.
Appl. Sci. 2024, 14, 909 4 of 12

Measuring method:
1. Speed before measuring—3 mm/s;
2. Speed of measuring (during penetration)—2 mm/s;
3. Speed after measuring—10 mm/s.
The penetration depth of the measuring body during measurement is 20 mm, and the
measuring force is 1 g.

2.9. Determination of Thermophysical Properties

To determine the thermophysical properties of the samples, a Mettler-Toledo DSC
model 822e differential scanning calorimeter (DSC) (Mettler Toledo, Giessen, Germany)
was used, and the measurements were carried out in a nitrogen atmosphere of purity 5.0
(Linde). The results were displayed and processed using the STARe Evaluation V6_V12
Conversation software. The control or verification (so-called check) of the reliability of
the instrument (module), which determines the difference between the measured and
determined reference value of the temperature or heat flow of the tested reference material,
was carried out using indium. Cooling was carried out using liquid nitrogen (100 L
container, Messer, Frankfurt, Germany).

2.10. Statistical Analysis

Data presented in this work are expressed as the mean value ± SD (standard deviation)
from three parallel repetitions for all analysis. One-way analysis of variance (ANOVA) and
Duncan’s least significant difference (LSD) test were used (Statistica 13.5, TIBCO Software
Inc., Palo Alto, CA, USA) to assess the significance of the differences between the mean
values at a significant level of p < 0.05.

3. Results and Discussion

3.1. Basic Chemical Composition of Tested Potato Cultivars
The chemical composition of potatoes is very important for the potato starch industry.
The most important parameter is the amount of starch present in the potato so that at
the end of the process, the economy and utilization are as high as possible. The type
and cultivar of the potato itself, as well as the growing conditions (soil, climate, fertilizer
addition, etc.), should also be considered; for instance, potatoes grown on silty soil have
the highest, while those grown on sandy soil have the lowest proportion of dry matter [8].
For this research, starches were isolated from eight potato cultivars, whose properties are
shown in Table 1. As Table 1 shows, the Stilleto cultivar had the highest proportion of dry
matter, and thus the highest proportion of starch, which could have been assumed as most
of the dry matter consists of starch. The Senata cultivar had the lowest proportion of dry
matter. However, the lowest proportion of starch was found in the SL 13–25 cultivar. The
content of dry matter in potatoes is a very important factor, with an average value of 22.3%,
and can vary significantly depending on the cultivar, conditions during growth and the
degree of maturity [7]. Changes also occur during storage itself. The share of water in all of
the cultivars was above 78.34%. Grommers and van der Krogt [7] state that, on average,
water makes up more than 70% of the total mass of potatoes, which was confirmed by the
above results. Potatoes used for processing into starch must be healthy, must not sprout
and must contain at least 14% starch [9]. In this research, the proportion of starch ranged
between 17.26 ± 0.37 and 22.49 ± 0.71%, which confirms the suitability of these cultivars
for starch production. The proportion of protein ranged between 1.79 ± 0.09 to 3.47 and
0.16%. The Stilleto cultivar had the highest protein content, while Senata had the lowest.
The highest proportion of fat was found in the SL 13–25 cultivar, and the lowest proportion
in the Senata cultivar. The Sereno cultivar had the highest, while the Sofista cultivar had
the lowest share of crude fiber.
Appl. Sci. 2024, 14, 909 5 of 12

Table 1. Chemical composition of the investigated potato cultivars.

Dry Matter Protein Fat Ash Starch Crude Fiber

Sample
[%] [%] [%] [%] [%] [%]
Saprodi 25.05 ± 0.29 c 2.53 ± 0.11 c 0.09 ± 0.01 a,b 1.24 ± 0.01 d,e 19.78 ± 0.17 b 1.08 ± 0.01 d
Sofista 24.43 ± 1.99 b,c 2.15 ± 0.13 b 0.09 ± 0.00 a,b 1.24 ± 0.01 d,e 19.91 ± 0.12 b 0.88 ± 0.04 a
Stilleto 28.54 ± 0.69 d 3.47 ± 0.16 d 0.10 ± 0.00 b 1.02 ± 0.01 b 22.49 ± 0.71 c 0.96 ± 0.00 b,c
Dartiest 21.95 ± 1.14 a,b 2.37 ± 0.06 b,c 0.10 ± 0.00 b 0.96 ± 0.01 a 17.42 ± 0.00 a 1.08 ± 0.01 d
Sereno 24.48 ± 0.76 b,c 3.37 ± 0.03 d 0.09 ± 0.00 a 1.20 ± 0.01 d 17.63 ± 0.43 a 1.18 ± 0.04 e
Senata 21.66 ± 0.97 a 1.79 ± 0.09 a 0.08 ± 0.00 a 1.11 ± 0.01 c 17.36 ± 0.32 a 1.07 ± 0.05 d
Scala 23.57 ± 0.84 a,b,c 2.15 ± 0.08 b 0.12 ± 0.00 c 1.14 ± 0.02 c 19.06 ± 0.70 b 0.90 ± 0.03 a,b
SL 13–25 23.48 ± 0.72 a,b,c 2.33 ± 0.09 b,c 0.16 ± 0.00 d 1.25 ± 0.03 e 17.26 ± 0.37 a 1.03 ± 0.04 c,d
Values with different letters in the same column are significantly different at p < 0.05.

3.2. Basic Chemical Composition of Isolated Potato Starches

Potato cultivars differ in the proportion of starch they possess, and the proportion of
starch itself defines their applicability in nutrition [10]. Starches were isolated from eight
potato cultivars, listed in Table 1, and their chemical composition is shown in Table 2. As
stated by Martínez et al. [11], the chemical composition of starches depends on the potato
cultivar, as well as on the starch isolation process.

Table 2. Chemical composition of potato starch obtained from the tested probes.

Dry Matter Protein Fat Ash Starch Crude Fiber Amylose

Sample
[%] [%] [%] [%] [%] [%] [%]
Saprodi 84.66 ± 0.02 c 0.09 ± 0.00 a 0.01 ± 0.00 a 0.22 ± 0.00 a,b 82.67 ± 0.27 b 1.01 ± 0.03 d 22.42 ± 0.67 f
Sofista 85.33 ± 0.02 e 0.10 ± 0.01 a 0.01 ± 0.00 a 0.26 ± 0.00 c 83.25 ± 0.41 b,c 0.77 ± 0.01 b 20.67 ± 0.61 c,d
Stilleto 84.67 ± 0.01 c 0.09 ± 0.01 a 0.01 ± 0.00 a 0.22 ± 0.01 a,b 82.96 ± 0.06 b,c 0.95 ± 0.04 c,d 21.16 ± 0.29 d,e
Dartiest 85.69 ± 0.02 f 0.10 ± 0.00 a 0.01 ± 0.00 a 0.21 ± 0.00 a,b 83.75 ± 0.13 c 0.70 ± 0.01 a 20.28 ± 0.37 b,c
Sereno 84.33 ± 0.08 b 0.15 ± 0.03 b 0.01 ± 0.00 a 0.23 ± 0.01 b 82.51 ± 0.40 b 0.70 ± 0.02 a 21.40 ± 0.48 e
Senata 84.92 ± 0.03 d 0.10 ± 0.01 a 0.01 ± 0.00 a 0.26 ± 0.00 c 83.32 ± 0.58 b,c 0.90 ± 0.01 c 19.93 ± 0.34 b
Scala 84.95 ± 0.01 d 0.09 ± 0.00 a 0.01 ± 0.00 a 0.25 ± 0.00 c 82.95 ± 0.42 b,c 0.90 ± 0.01 c 16.97 ± 0.37 a
SL 13–25 83.51 ± 0.13 a 0.09 ± 0.00 a 0.01 ± 0.00 a 0.21 ± 0.01 a 81.23 ± 0.10 a 1.02 ± 0.05 d 16.82 ± 0.34 a
Values with different letters in the same column are significantly different at p < 0.05.

The protein content of the isolated potato starches in this study varied between
0.09 and 0.15%, and these values are very low compared to the protein values in the
starches isolated from three autochthonous potatoes of the Andean region [11], which
indicates high purity [12]. The proportion of fat is the same in all of the starches and
amounts to 0.01%, and this proportion is lower than that in the Mexican potato starch [13]
and potato starch from Venezuela [14]. The ash content ranged between 0.21 and 0.26%,
which is similar to the values for the starches from autochthonous potatoes of the Andean
region [11] and potato starch from Venezuela [14]. The raw fiber content ranged between
0.70 and 1.02%, while the amylose content ranged from 16.82% in SL 13–25 cultivar to
22.42% in Saprodi cultivar.

3.3. The Color of Potato Starches

Starch color is an important parameter of starch quality. Table 3 shows the color
parameters of the isolated potato starches. In the CIELab system, the intensity of white
color—i.e., the brightness of starch—is expressed through the L* value. The values of the
L* parameter range from 0 (black) to 100 (white), and the values obtained for all of the
starches were close to 100, which indicates a white color of high intensity. The parameter
a* describes the color domain; if the value is positive, it is in the domain of red, and if the
value is negative, it is in the domain of green. As the table shows, all of the obtained values
are negative, which means that the samples are in the domain of green.
Appl. Sci. 2024, 14, 909 6 of 12

Table 3. Color properties of the isolated potato starches.

Sample L* a* b* C h◦
Saprodi 94.85 ± 0.31 c −1.45 ± 0.19 a 2.29 ± 0.12 a 2.72 ± 0.05 b,c,d 122.36 ± 4.59 e
Sofista 94.08 ± 0.45 a,b −1.32 ± 0.02 a,b 2.33 ± 0.07 a 2.67 ± 0.06 b,c 119.46 ± 0.88 d,e
Stilleto 93.71 ± 0.17 a −1.21 ± 0.02 b 2.49 ± 0.04 b 2.77 ± 0.04 c,d 115.94 ± 0.59 b,c,d
Dartiest 94.49 ± 0.30 b,c −1.15 ± 0.29 b 2.52 ± 0.14 b 2.79 ± 0.14 d 114.40 ± 6.06 b,c
Sereno 93.81 ± 0.56 a,b −1.31 ± 0.04 a,b 2.65 ± 0.01 c 2.96 ± 0.02 e 116.35 ± 0.75 c,d
Senata 93.67 ± 1.19 a −1.18 ± 0.26 b 2.93 ± 0.05 d 3.17 ± 0.11 f 111.86 ± 4.25 a,b
Scala 94.18 ± 0.25 a,b,c −1.32 ± 0.02 a,b 2.31 ± 0.07 a 2.65 ± 0.05 b 119.77 ± 0.96 d,e
SL 13–25 94.85 ± 0.04 c −0.79 ± 0.03 c 2.27 ± 0.04 a 2.41 ± 0.04 a 109.16 ± 0.51 a
Values with different letters in the same column are significantly different at p < 0.05.

From the obtained results, it is evident that the values of the parameter b* of all the
tested samples are positive, which means that all of the samples are in the yellow domain.
Parameter C expresses color saturation, and the highest value, as well as saturation, was in
the starch obtained from the potatoes of the Senata cultivar. The h◦ parameter represents
the color tone and is expressed in ◦ , where 0◦ indicates red, values over 90◦ indicate yellow,
over 180◦ indicate green and values up to 270◦ indicate a blue tone.
According to the obtained results, shown in the table, it can be seen that all of the
samples are in the yellow domain. Compared to the results of Sit et al. [15], the results for
the L* value were slightly lower. Pérez Sira and Amaiz [16] state that L* values above 90 are
satisfactory and confirm the purity of the starch. In general, high L* values and low values
of the a* and b* parameters confirm that starch can be used in products that require a clean
and uniform color.

3.4. Paste Properties by Brabender’s Micro Visco-Amylograph

Among all commercial starches, potato starch has the highest swelling power and
provides the highest viscosity of starch pasting properties [17]. Matvejev et al. [18] con-
cluded that the thermodynamic melting properties of starches are directly correlated with
the amylose content. The pasting properties of starch are usually determined based on
changes in the starch viscosity. The results of the properties of the pastes determined using
Brabender’s micro visco-amylograph are shown in Table 4. The highest peak viscosity value
was found in the starch sample obtained from the Scala cultivar 1673.5 ± 20.5 BU, while the
lowest value was in the starch sample obtained from the Stilleto cultivar 1410.0 ± 2.8 BU.
After shearing at 92 ◦ C, the viscosity of all eight potato cultivars decreased because the
structure of the paste was damaged due to shearing. Cooling to 50 ◦ C showed an increase
in the viscosity of the potato starch paste.

Table 4. Pasting properties of the isolated potato starches.

Viscosity after Viscosity at Viscosity after

Peak Viscosity Viscosity at Breakdown Setback
Sample Shearing at 50 ◦ C Shearing at 50 ◦ C
[BU] 92 ◦ C [BU] [BU] [BU]
92 ◦ C [BU] [BU] [BU]
Saprodi 1505.5 ± 33.2 b 1130.5 ± 6.4 f 600.0 ± 2.8 f 1046.0 ± 18.4 e 854.5 ± 13.4 e 905.5 ± 30.4 a 446.0 ± 15.6 d
Sofista 1499.5 ± 19.1 b 1016.0 ± 7.1 d 539.0 ± 8.5 d 924.5 ± 3.5 c 758.5 ± 0.7 c 960.5 ± 27.6 b,c 385.5 ± 4.9 b,c
Stilleto 1410.0 ± 2.8 a 1006.0 ± 18.4 d 514.5 ± 12.0 c 898.0 ± 18.4 c 745.5 ± 33.2 c 895.5 ± 9.2 a 383.5 ± 6.4 b,c
Dartiest 1446.5 ± 9.2 a 904.5 ± 16.3 b 468.0 ± 7.1 a 834.5 ± 19.1 b 693.0 ± 11.3 b 978.5 ± 2.1 b,c 366.5 ± 12.0 b
Sereno 1518.5 ± 21.9 b 1048.0 ± 11.3 e 562.5 ± 6.4 e 962.5 ± 19.1 d 799.0 ± 15.6 d 956.0 ± 28.3 b 400.0 ± 12.7 c
Senata 1490.0 ± 1.4 b 888.5 ± 9.2 a,b 487.0 ± 4.2 b 806.5 ± 6.4 a,b 641.5 ± 10.6 a 1003.0 ± 5.7 c,d 319.5 ± 2.1 a
Scala 1673.5 ± 20.5 c 974.0 ± 9.9 c 521.0 ± 4.2 c 915.0 ± 4.2 c 738.0 ± 4.2 c 1152.5 ± 16.3 e 394.0 ± 0.0 c
SL 13–25 1519.0 ± 9.9 b 870.0 ± 9.9 a 474.0 ± 5.7 a,b 796.0 ± 0.0 a 663.0 ± 7.1 a,b 1045.0 ± 4.2 d 322.0 ± 5.7 a
Values with different letters in the same column are significantly different at p < 0.05.

According to Chung et al. [19], the increase in viscosity during cooling occurs due to
retrogradation—that is, the connection of dissolved starch molecules. Breakdown, which
indicates the stability of the starch paste during shearing at high temperatures, is calculated
 Appl. Sci. 2024, 14, 909 7 of 12

from the difference in the viscosity value after 15 min of mixing at 92 ◦ C, and the peak
viscosity value. The Scala cultivar had the highest breakdown value, which indicates that it
is less stable during shearing at high temperatures. The cultivars Senata and SL 13–25 had
similar breakdown values. From the setback results, which indicate the tendency of starch
paste to retrograde, it is noticeable that the starch obtained from the Saprodi cultivar is
slightly more susceptible to retrogradation compared to the other starches.

3.5. Swelling Power and Solubility Index

The swelling power and the solubility index show the interconnection of starch chains
within the crystalline and amorphous regions of the starch granule, which is determined by
the ratio of amylopectin and amylose, their conformation and the degree of branching [20].
Figures 1 and 2 show the swelling power and solubility index of the starches isolated
from different potato cultivars at temperatures of between 55 ◦ C and 85 ◦ C. As suggested
by the results in the figures, there is a distinct tendency to increase the swelling power, as
well as the solubility index, with an increase in the treatment temperature. At the highest
temperature, the starch of the Sereno cultivar showed the highest, and the starch of the
SL 13–25 cultivar showed the lowest value of swelling power, while the highest value of
the solubility index was found in the starch of the Stilleto cultivar, and the lowest in the
Senata cultivar. The swelling power and solubility index depend on the type of starch
and the ratio of amylose and amylopectin. The values of the swelling power decrease
with an increase in the amylose content, and as the amylopectin content increases, the
swelling power increases too, and such starch is more stable to freeze-thaw processes,
which significantly accelerates the processes of retrogradation and syneresis. Potato starch
x FOR PEER REVIEW consists of relatively large granules that have relatively high swelling power values [21].8 of 13
The morphological characteristics of the granules can also influence the swelling power
and solubility index [22].

Figure 1. Swelling power

Figure of thepower
1. Swelling isolated
of thepotato starches
isolated potato at different
starches at differenttemperatures. Different
temperatures. Different letters letters
above the bars indicate that the values are significantly different at p < 0.05.
above the bars indicate that the values are significantly different at p < 0.05.
 Figure
Appl. Sci. 2024, 14, 909 1.
Swelling power of the isolated potato starches at different temperatures. Different
8 of 12letters
above the bars indicate that the values are significantly different at p < 0.05.

Figure 2. Solubility index

Figure of the isolated
2. Solubility potato
index of the isolatedstarches at different
potato starches at differenttemperatures. Different
temperatures. Different lettersletters
above the bars indicate that the values are significantly different at p < 0.05.
above the bars indicate that the values are significantly different at p < 0.05.

3.6. Clarity of Starch Paste

3.6. Clarity of Starch Starch
Pastepaste clarity is affected by the amylose content, molecular weight and starch
granule structure, which directly affects starch granule swelling [23]. The transparency
Starch paste clarity is affected by the amylose content, molecular weight and starch
data, expressed as % of transmission at 650 nm, are shown in Figure 3. The starch of the
granule structure, which
Dartiest directly
cultivar had theaffects starchvalue,
highest clarity granule swelling
followed [23].cultivar,
by the Senata The transparency
while the
data, expressed as %
other
Appl. Sci. 2024, 14, x FOR PEER REVIEW
of transmission
cultivars had lower and at 650
similarnm, are
clarity shown
values. in Figure
The Dartiest 3.
cultivarThe starch
had the lowest of the
9 of 13
proportion of mineral substances and the highest proportion of starch, which is a possible
Dartiest cultivar had the highest clarity value, followed by the Senata cultivar, while the
reason for the high clarity.
other cultivars had lower and similar clarity values. The Dartiest cultivar had the lowest
proportion of mineral substances and the highest proportion of starch, which is a possible
reason for the high clarity.
With gelatinization, starch granules swell and dissociate and let more light through—
i.e., their clarity increases [24]. The differences in the starch clarity values may be due to
several factors such as the amylose content, lipid and protein content, granule particle
size, amylose and amylopectin chain length [22] and phosphate monoester content [25,26].

Figure3.3.Paste
Figure Pasteclarity
clarity(%
(%T)
T)ofofthe
theisolated
isolatedpotato
potatostarches.
starches.Different
Differentletters
lettersabove
abovethe
thebars
barsindicate
indicate
that the values are significantly different at p < 0.05.
that the values are significantly different at p < 0.05.
Appl. Sci. 2024, 14, 909 9 of 12

With gelatinization, starch granules swell and dissociate and let more light through—
i.e., their clarity increases [24]. The differences in the starch clarity values may be due to
several factors such as the amylose content, lipid and protein content, granule particle size,
amylose and amylopectin chain length [22] and phosphate monoester content [25,26].

3.7. Texture of Starch Gels

The results of the texture analysis of the starch gels are shown in Table 5. It is known
that the strength of the starch gel depends on the size of the “net” formed by the dissolved
amylose molecules and on the deformation property of the dissolved starch granules [27].
The Dartiest cultivar had the highest, while the Sereno cultivar had the lowest gel strength.
Higher breaking force values were observed for the starch gels of the Sereno and Stilleto
cultivars compared to the Dartiest and Senata cultivars. Adhesion refers to the stickiness of
the gel with the contact surface. From the results shown in Table 5, it is evident that the
Dartiest cultivar had significantly lower adhesion values than the other starch samples.
The values for the gel strength are lower than in the study by Sandhu et al. [28], and higher
than in the study by Gałkowska et al. [29].

Table 5. Texture properties of starch gels of the isolated potato starches.

Gel Strength Rupture Strength Adhesiveness

Sample
[g] [g] [g sec]
Saprodi 3.14 ± 0.08 b,c 928.89 ± 18.84 d −219.78 ± 6.00 b,c
Sofista 2.98 ± 0.01 b 840.18 ± 17.90 b −202.52 ± 4.58 d
Stilleto 3.25 ± 0.08 b,c 959.33 ± 6.77 e −224.69 ± 4.04 b
Dartiest 3.30 ± 0.16 c 776.63 ± 9.16 a −73.02 ± 9.50 e
Sereno 2.48 ± 0.23 a 998.99 ± 10.61 f −348.49 ± 2.23 a
Senata 3.08 ± 0.00 b,c 758.10 ± 4.46 a −219.27 ± 8.41 b,c
Scala 3.03 ± 0.08 b,c 912.11 ± 17.51 d −192.56 ± 5.68 d
SL 13–25 3.08 ± 0.00 b,c 873.06 ± 1.98 c −205.13 ± 8.11 c,d
Values with different letters in the same column are significantly different at p < 0.05.

3.8. Determination of Thermophysical Properties

3.8.1. Properties of Gelatinization
Gelatinization is an important functional property of potato starch. This process is
required to produce the desired functionality, such as the thickening and swelling of potato
starch in food products.
Table 6 shows the gelatinization parameters, determined through DSC. The onset tem-
perature of gelatinization ranged from 63.73 ◦ C for the starch obtained from the Dartiest
cultivar to 69.18 ◦ C for the starch obtained from the SL 13–25 cultivar. The gelatinization
peak temperatures ranged from 66.21 ◦ C for the starch obtained from the Dartiest cultivar to
71.89 ◦ C for the starch obtained from the Senata cultivar, and the final gelatinization temper-
ature ranged from 70.24 ◦ C for the starch obtained from the Dartiest cultivar up to 77.05 ◦ C
for the starch obtained from the Scala cultivar. The starch isolated from the Dartiest cultivar
had the lowest gelatinization temperature, while the highest gelatinization temperatures
were observed in the starches from the SL 13–25, Senata and Scala cultivars. The gelatiniza-
tion temperatures of the isolated starches are higher than the gelatinization temperatures
of the starches obtained from autochthonous potatoes of the Andean region [11], as well as
from different potato starches from India [30]. The differences between the transition tem-
peratures of the gelatinization of the starches obtained from different potato cultivars can be
attributed to differences in the degree of crystallinity. A high degree of crystallinity enables
a stable structure of the granule and makes it more resistant to gelatinization, which is why
the gelatinization temperatures are higher [31]. Higher gelatinization temperatures can also
be caused by a high double helical arrangement between amylose and amylopectin chains
and strong interactions between amylose–amylose chains and amylopectin–amylopectin
chains [11]. The gelatinization enthalpies of the potato starches are significantly lower
Appl. Sci. 2024, 14, 909 10 of 12

compared to the enthalpies of the starches obtained from indigenous potato cultivars of the
Andean region, and are also different from potato starches from India [11,30].

Table 6. Gelatinization parameters of suspensions of the isolated potato starches.

Sample To (◦ C) Tp (◦ C) Te (◦ C) ∆H (J/g)
Saprodi 64.80 ± 0.11 b 70.32 ± 0.36 d 74.24 ± 0.02 c,d 2.16 ± 0.04 c
Sofista 63.80 ± 0.18 a 67.56 ± 0.28 b 73.80 ± 0.13 b 1.22 ± 0.06 a
Stilleto 64.22 ± 0.42 a,b 68.27 ± 0.33 c 76.73 ± 0.21 f 1.15 ± 0.04 a
Dartiest 63.73 ± 0.16 a 66.21 ± 0.13 a 70.24 ± 0.08 a 2.27 ± 0.08 c,d
Sereno 67.08 ± 0.06 c 70.11 ± 0.06 d 74.62 ± 0.25 d 2.32 ± 0.05 d
Senata 68.45 ± 0.28 d 71.89 ± 0.12 e 75.94 ± 0.08 e 2.01 ± 0.08 b
Scala 67.52 ± 0.59 c 71.41 ± 0.06 e 77.05 ± 0.17 f 2.30 ± 0.04 c,d
SL 13–25 69.18 ± 0.15 e 71.84 ± 0.17 e 75.81 ± 0.49 e 2.21 ± 0.02 c,d
Values with different letters in the same column are significantly different at p < 0.05. Gelatinization parameters:
T0 —onset temperature; Tp —peak temperature; Te —endset temperature; ∆H—gelatinization enthalpy.

3.8.2. Properties of Retrogradation

Retrogradation is a second important functional property of starch, and it is related
to the stability of the starch paste during storage. Table 7 shows the values of the initial
temperature (T0 ), peak temperature (Tp ), end temperature (Te ) and retrogradation enthalpy
(∆H) of the isolated potato starch gels after 14 days of storage at 4 ◦ C. The starch gel of the
SL 13–25 cultivar had the highest initial temperature, and the Senata cultivar starch gel
had the lowest. The starch gel of the Stilleto cultivar had the highest peak temperature,
and the starch gel of the Senata cultivar had the lowest. The Dartiest starch gel had the
highest final temperature, and the Saprodi starch gel had the lowest. The retrogradation
enthalpy values ranged from 0.11 J/g for the Sereno cultivar starch gel to 0.42 J/g for
the SL 13–25 cultivar starch gel. The transition temperatures, as well as the enthalpies of
retrogradation at the end of the storage period, were significantly reduced compared to
the transition temperatures and enthalpies during gelatinization. The results are consistent
with the results of Karim et al. [25], where the transition temperatures and retrogradation
enthalpies are lower compared to the gelatinization temperatures. After gelatinization,
there is a reconnection of starch molecules and their recrystallization, whereby they form
less ordered structures, and therefore less energy is needed to melt the restructured crystal
in the retrograded starch [25].

Table 7. Retrogradation parameters of gels of the isolated potato starches after 14 days of storage at
4 ◦ C.

Sample To (◦ C) Tp (◦ C) Te (◦ C) ∆H (J/g)
b d a
Saprodi 44.57 ± 0.43 53.11 ± 0.49 58.89 ± 0.12 0.38 ± 0.00 c,d
Sofista 44.80 ± 0.27 b 51.23 ± 0.24 a,b 62.06 ± 0.07 b,c 0.35 ± 0.02 c
Stilleto 44.76 ± 0.26 b 55.56 ± 0.59 e 63.19 ± 0.17 d 0.28 ± 0.01 b
Dartiest 44.39 ± 0.70 b 55.11 ± 0.04 e 63.27 ± 0.29 d 0.29 ± 0.01 b
Sereno 44.70 ± 0.13 b 53.39 ± 0.24 d 62.23 ± 0.28 c 0.11 ± 0.04 a
Senata 42.40 ± 0.39 a 50.72 ± 0.35 a 61.53 ± 0.56 b 0.36 ± 0.01 c
Scala 45.27 ± 0.42 b 51.99 ± 0.13 b,c 63.09 ± 0.21 d 0.15 ± 0.01 a
SL 13–25 49.19 ± 0.08 c 52.64 ± 0.47 c,d 62.25 ± 0.08 c 0.42 ± 0.03 d
Values with different letters in the same column are significantly different at p < 0.05. Retrogradation parameters:
T0 —onset temperature; Tp —peak temperature; Te —endset temperature; ∆H—retrogradation enthalpy.

4. Conclusions
The highest dry matter content was found in the Stiletto, while the lowest dry matter
content was in the Senata cultivar, and the lowest starch content was found in the SL
13–25 cultivar. The isolated starches from all the cultivars had a high level of purity, which
is evident from the analysis of the basic chemical composition of the obtained starches,
Appl. Sci. 2024, 14, 909 11 of 12

where the content of protein, fat, ash and crude fiber is relatively low. The starch of the
Saprodi cultivar had the highest and the starch of the Senata cultivar had the lowest value
of the L* parameter, although the differences between the values of the L* parameter were
very small, so the difference in the color of the starches isolated from different potato
cultivars is difficult to see. The starch of the Senata cultivar had the highest color saturation,
and the starch of the SL 13–25 cultivar had the lowest. Starch from the Saprodi cultivar had
the highest, while starch from the SL 13–25 cultivar had the lowest amylose content. The
Dartiest cultivar had the highest, while the Sereno cultivar had the lowest gel strength. The
starch gels of the Sereno and Stilleto cultivars had higher breaking force values compared
to the Dartiest and Senata cultivars. With an increase in temperature, an increase in the
swelling capacity was found in all samples, and a proportional increase in the solubility
index with an increase in temperature was also recorded. The starch of the Dartiest cultivar
had the highest clarity value, followed by the Senata cultivar, while the other cultivars
had lower and similar clarity values. The Dartiest starch had the lowest transitional
gelatinization temperatures, and the starches obtained from the SL 13–25, Senata and Scala
cultivars showed the highest gelatinization temperatures. The transition temperatures
and enthalpies of retrogradation were lower compared to gelatinization. The highest
transitional retrogradation temperatures were obtained by the starch gels of the SL 13–25,
Stilleto and Dartiest cultivars, while the lowest values were found in the starch gels of the
Senata and Saprodi cultivars.

Author Contributions: Conceptualization, A.J., D.Š., T.V. and M.K.; methodology, A.J., M.K., T.V. and
B.R.; formal analysis, M.K., A.J. and A.L.; investigation, Ð.A., J.B., D.Š. and V.O.B.; writing—original
draft preparation, M.K., A.J. and T.V.; writing—review and editing, A.Š., Ð.A., J.B., A.L., B.R. and
V.O.B.; supervision, D.Š., T.V. and A.J. All authors have read and agreed to the published version of
the manuscript.
Funding: This research was funded by Osijek-Baranja County under the project “Isolation and
characterization of potato starch produced in the Osijek-Baranja County”.
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: The data presented in this study are available on request from the
corresponding author.
Acknowledgments: We want to acknowledge to Royal ZAP/Semagri Holland BV for the donation of
potato cultivars.
Conflicts of Interest: The authors declare no conflicts of interest.

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