Краснообск, Новосибирская область, Россия
Краснообск, Новосибирская область, Россия
Краснообск, Новосибирская область, Россия
Краснообск, Новосибирская область, Россия
Введение. Морские звезды (Asteroidea) насчитывают более 160 видов, что делает их ценным сырьем для производства белков и жиров. Настоящее исследование позволило определить химический состав морских звезд и доказало целесообразность использования этого ресурса в качестве коммерческого источника жиров. Объекты и методы исследования. В ходе исследования были определены оптимальные параметры экстракции липидной фракции Lysastrosoma anthosticta сверхкритическим диоксидом углерода, а также описан качественный состав полученных экстрактов. Результаты и их обсуждение. Выход жирных кислот, полученных со сверхкритическим сорастворителем диоксида углерода, был в 1,8 раза выше, чем при стандартной экстракции по методу Фолча. Содержание примесей оказалось ниже, чем в образцах, где использовалась система хлороформ-метанол. Полиненасыщенные жирные кислоты, выделенные из L. anthosticta, принадлежали к ω-3 (18,0 %), ω-6 (11,7 %), ω-7 (21,2 %), ω-9 (10,1 %) и ω-11 (6,5 %). Остальное составляли насыщенные жирные кислоты: пальмитиновая (до 14 %) и миристиновая (до 6 %). Качественный состав липидной фракции не отличался от метода выделения. Однако сверхкритическая экстракция увеличила выход продукта, скорость экстракции и качество экстракционного остатка. Сверхкритический диоксид углерода оставил твердый осадок, который не имел характерного запаха и был достаточно хрупким для дальнейшего измельчения. В будущем такой остаток можно использовать для получения белкового концентрата. Выводы. Сверхкритическая экстракция хлороформом может быть рекомендована для выделения жирных кислот из морских организмов при 60°C и 400 бар.
Морская звезда, иглокожие, ненасыщенные жирные кислоты, липиды, море
Introduction
The population of the earth is growing every year,
which makes technologies for food obtaining and
processing very important for humanity. Efficient
processing technologies produce more useful products
while doing less harm to the environment. The oceans
are the least explored part of the earth. Every year,
dozens of new compounds are isolated from marine
aquatic organisms around the world. Many of them
possess various beneficial biological properties that can
be used, for instance, in pharmacology. Every year, new
secondary metabolites of great practical and fundamental
interest are extracted from echinoderms.
Starfish (Asteroidea) are widespread marine
echinoderms of more than 160 species. Starfish are
predators that damage shellfish plantations and coral
reefs. In the XX–XXI centuries, world fisheries have
been busy looking for new sources of nutrients, especially
for marine carriers of biologically active substances that
can be used to obtain highly effective medicines [1].
The present research featured Lysastrosoma anthosticta;
the research objective was to select a promising method
of supercritical extraction.
Supercritical fluid extraction and supercritical fluid
chromatography have been used since the late 1970s
in food analysis for determining lipids and toxicants.
Supercritical fluid extraction is an effective means of
natural product extraction. The supercritical extraction
process has potential advantages over conventional
extraction processes, such as shorter extraction time,
reduced organic solvent volume, and more selective
extraction [2].
Study objects and methods
Samples of Lysastrosoma anthosticta Fisher, 1922
were harvested in the Peter the Great Bay (Russia,
Sea of Japan) in 2020. Starfish with a total weight of
42 000 g were gutted, cut with scissors into small pieces
(about 1 cm long), and stored in a plastic bag at –70°C.
Frozen samples were lyophilized and crushed. The dry
weight was 3630 g. After that, 30 g of aliquots was used
for extraction. All experiments were done in triplicates.
Extraction with supercritical CO2 was performed using
THAR SFC 500 (USA). Figure 1 shows the technological
scheme of the supercritical extraction unit. Extraction
was carried out using supercritical CO2: pressure – 200,
400, and 600 bar per square inch; t – 30, 40, 50, 60,
and 70°C; flow rate – 20 g/min. At the second step,
the experiment extraction was carried out using CO2
and 5% solvent (chloroform): pressure – 200, 400, and
600 bar per square inch; t – 30, 40, 50, 60, and 70°C;
flow rate – 20 g/min. The control extraction of 30 g of
dried starfish was carried out using the Folch method
with a mix of chloroform-methanol (2:1) at the rate of
20 parts of the extraction mixture per one part of the
tissue at 30, 40, 50, 60, and 70°C [3].
The extracts were analyzed by high-performance liquid
chromatography (HPLC) with tandem mass spectrometry
Аннотация.
Введение. Морские звезды (Asteroidea) насчитывают более 160 видов, что делает их ценным сырьем для производства белков
и жиров. Настоящее исследование позволило определить химический состав морских звезд и доказало целесообразность
использования этого ресурса в качестве коммерческого источника жиров.
Объекты и методы исследования. В ходе исследования были определены оптимальные параметры экстракции
липидной фракции Lysastrosoma anthosticta сверхкритическим диоксидом углерода, а также описан качественный
состав полученных экстрактов.
Результаты и их обсуждение. Выход жирных кислот, полученных со сверхкритическим сорастворителем диоксида
углерода, был в 1,8 раза выше, чем при стандартной экстракции по методу Фолча. Содержание примесей оказалось ниже,
чем в образцах, где использовалась система хлороформ-метанол. Полиненасыщенные жирные кислоты, выделенные
из L. anthosticta, принадлежали к ω-3 (18,0 %), ω-6 (11,7 %), ω-7 (21,2 %), ω-9 (10,1 %) и ω-11 (6,5 %). Остальное
составляли насыщенные жирные кислоты: пальмитиновая (до 14 %) и миристиновая (до 6 %). Качественный состав
липидной фракции не отличался от метода выделения. Однако сверхкритическая экстракция увеличила выход продукта,
скорость экстракции и качество экстракционного остатка. Сверхкритический диоксид углерода оставил твердый осадок,
который не имел характерного запаха и был достаточно хрупким для дальнейшего измельчения. В будущем такой
остаток можно использовать для получения белкового концентрата.
Выводы. Сверхкритическая экстракция хлороформом может быть рекомендована для выделения жирных кислот из
морских организмов при 60°C и 400 бар.
Ключевые слова. Морская звезда, иглокожие, ненасыщенные жирные кислоты, липиды, море
Финансирование. Работа выполнена при финансовой поддержке гранта Президента Российской Федерации
(SP-3156.2019.4).
Для цитирования: Применение технологии сверхкритической экстракции для получения полиненасыщенных кислот
из морской звезды Lysastrosoma anthosticta Fisher, 1922 / А. М. Захаренко [и др.] // Техника и технология пищевых
производств. 2021. Т. 51. № 4. С. 753–758. (На англ.). https://doi.org/10.21603/2074-9414-2021-4-753-758.
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Захаренко А. М. [и др.] Техника и технология пищевых производств. 2021. Т. 51. № 4 С. 753–758
(LC-MS/MS). Reverse-phase HPLC was performed using
a Shimadzu LC-20 liquid chromatograph (Shimadzu,
Japan) equipped with a CTO-20A thermostat (Shimadzu,
Japan) and a UV-VIS SPD-20A detector (Shimadzu,
Japan). ZORBAX Eclipse XDB C18 (150×4.6 mm,
particle size: 5 microns) was used as an analytical
column at a temperature of 30°C and a total flow rate of
0.22 mL/min. Gradient elution with two mobile phases
Figure 1. Technological scheme of the supercritical extraction unit
Figure 2. HPLC and ion trap joint system with tandem mass spectrometry
50 60 70
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Zakharenko A.M. et al. Food Processing: Techniques and Technology, 2021, vol. 51, no. 4, pp. 753–758
(A – deionized water; B – acetonitrile with formic acid
0.1% v/v) was programed as follows: 0 min 0% V,
25 min 100% V, 60 min 100% V. The chromatograph
and the mass spectrometric detector were linked by the
Compass software, which made it possible to integrate
the entire system into a single complex (Fig. 2).
The mass spectra of electrospray ionization mass
spectrometry (ESI-MS) and electrospray ionization
tandem mass spectrometry (ESI-MSN) were performed
using an Amazon SL ion trap (Bruker, Germany) equipped
with an electrospray ionization source. The ESI MS
ionization parameters were optimized as follows: capillary
voltage of 4500 V, nitrogen spraying at 29 psi, dry gas
consumption of 10 l/min at 160°C. Mass spectra were
recorded in the mass range m/z 50–2000 in the mode
of negative and positive ions. The mass spectra of the
ions were recorded in the auto MS/MS mode.
Results and discussion
An analysis of yields of lipids fraction from
Lysastrosoma anthosticta under various conditions
showed that the extraction with supercritical carbon
dioxide with no co-solvent was less effective than the
standard extraction according to the Folch method
(Fig. 3). However, when extra 5% chloroform was added
to the extraction system as a co-solvent, it significantly
increased the yields of the total fatty acid fraction. The
choice of the solvent and the extraction range parameters
was based on the results of previous works where starfish
material was treated with a plant matrix [4, 5]. Despite
the fact that the main target components are soluble in
liquid CO2 only above 200 bar, it is possible to separate
significant amounts of the lipid fraction [6, 7]. The
lipid fraction obtained by supercritical extraction with
chloroform was 1.8 times higher than in the standard
Folch method. The extraction was found to be quite
effective only when a co-solvent was used. The optimal
parameters for extraction with a co-solvent included a
temperature of 60°C and a pressure of 400 bar. The data
obtained correlated with the most frequently selected
parameters. Most often, when using this technology, the
authors chose a pressure of 300–350 bar as the optimal
one [8–11]. With these parameters, a good yield of
lipids can be obtained as quickly as possible; a further
increase in the temperature and extraction pressure did
not lead to a significant increase in the yield.
Obtaining chemical profiles is an extremely important
result for any biological analysis system. In this work,
we used the HPLC-ESI-MS/MS method with additional
ionization and analysis of fragmented ions. High accuracy
mass spectrometric data were recorded on an ion trap
amaZon SL BRUKER DALTONIKS equipped with
an ESI source in the mode of negative and positive
ions. The experiment used a four-stage ion separation
mode (MS/MS mode). A qualitative analysis showed
that the ratio of polyunsaturated to saturated fatty acids
did not depend on the extraction method (Table 1). An
analysis of polyunsaturated fatty acids isolated from
L. anthosticta showed that they mainly belonged to
ω-3 (18.0%), ω-6 (11.7%), ω-7 (21.2%), ω-9 (10.1%),
and ω-11 (6.5%). The rest was saturated fatty acids,
mainly palmitic (up to 14%) and myristic (up to 6%).
Figure 3. Effect of extraction conditions on the yield of the fatty acid: a – extraction with supercritical CO2;
b – extraction with supercritical CO2 with a solvent (chloroform); c – extraction according
to the Folch method at various temperatures
0
2
4
30 40 50 60 70
a b
c d
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Захаренко А. М. [и др.] Техника и технология пищевых производств. 2021. Т. 51. № 4 С. 753–758
The predominance of polyunsaturated acids is typical
of starfish [12, 13].
A review of scientific publications showed that
starfish can be considered as a valuable source of
feed additives for agricultural animals and birds.
For instance, Danish compound feed producers are
considering the possibility of industrial use of starfish
to produce additives for compound feeds. According
to recent studies, such compound feed can reduce the
excretion of nitrogen in pigs. Scientists from the Center
for Aquatic Animals in Denmark proved that starfish
can be an effective alternative to traditional sources of
feed protein, e.g. soybeans. Starfish meal could replace
the most commonly used protein sources and increase
the weight gain in piglets. In fact, starfish-based animal
feed may be more economically rational than traditional
protein sources. In addition, starfish are being rigorously
tested as a possible protein source for bird nutrition. If
the results are confirmed, starfish will become a rich
source of protein, which will create high demand from
egg producers, who are constantly searching for new
sources of protein [14].
Conclusion
Starfish are a valuable raw material for protein
and lipid production. The chemical composition of
starfish makes it possible to use it in food and feed
industry. The content of proteins was 9.5–14.0%, lipids
– 0.5–3.5%, minerals – 1.5–32.0%. In comparison with
the integumentary tissue, the internal organs of starfish
have a higher content of potassium and iron [15, 16].
The yields of fatty acids obtained under conditions of
supercritical carbon dioxide co-solvent were 1.8 times
higher than those obtained with the standard Folch
method, while the content of impurities was lower than
when the extraction was performed using a chloroformmethanol
system. The analysis of polyunsaturated fatty
acids isolated from Lysastrosoma anthosticta showed
that they mainly belonged to ω-3 (18.0%), ω-6 (11.7%),
ω-7 (21.2%), ω-9 (10.1%), and ω-11 (6.5%). The rest
was saturated fatty acids: palmitic (14%) and myristic
(6%). Thus, isolating fatty acids from marine organisms
using supercritical extraction with chloroform can be
recommended as an effective commercial method.
In addition, supercritical carbon dioxide with a solvent
left a dry residue, brittle enough for further grinding and
without typical smell. Such a residue can presumably
be used to produce protein concentrate.
Contribution
All the authors contributed equally to the study and
bear equal responsibility for information published in
this article.
Conflict of interests
The authors declare that there is no conflict of interests
regarding the publication of this article.
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