Environmental and Climate Technologies

2023, vol. 27, no. 1, pp. 422–437

https://doi.org/10.2478/rtuect-2023-0031
https://content.sciendo.com

Methods for Extraction of Bioactive Compounds

from Products: A Review
Ilze LUKSTA1*, Kriss SPALVINS2
1,2Institute of Energy Systems and Environment, Riga Technical University, Āzenes street 12/1, Riga,
LV-1048, Latvia
Received 06.01.2023; accepted 08.06.2023

Abstract – Most agricultural by-products are not recycled or fully utilized. Most of them end
up in waste and cause economic, social and environmental problems. Today, much of the by-
products are considered waste, although they can be appropriately recycled to produce new
value-added products. By-products have great potential to create, for example, food additives.
Many of these byproducts are a source of valuable compounds such as proteins, lipids,
starches, trace elements, bioactive compounds and dietary fiber. A wide variety of extraction
methods are currently available, but several factors must be considered to determine which
of these methods can efficiently and environmentally produce high-value-added products
from by-products. The article examines, summarizes and compares various extraction
methods, as well as products that are currently obtained using specific methods.

Keywords – Agriculture; by-products; extraction; value added products.

1. INTRODUCTION
Agricultural product residues (by-products) can be defined as primary or secondary
depending on their origin. Primary residues include the production or cultivation of plant
products, such as grains, fruits, vegetables and herbs, as well as the transportation, storage
and processing of said products (if the products do not change significantly) on the farm and
onward delivery to businesses. Secondary residues are obtained in the food processing
industry and become residues during the processing stage (peels, pulps, stones, nut shells) [1].
The generation of agricultural waste varies from country to country, for example in Europe
more than 40 % of the weight of the original agricultural product is thrown away, while in
North Africa, East Central Asia and Latin America this percentage is higher. For example,
the amount of by-products produced for an artichoke is about 50–60 % of the original weight,
and for an orange – about 60 % [1], [2].
Agricultural by-products such as fruit and vegetable pomace and grain bran contain
bioactive compounds that can act as free radical and antioxidant scavengers as well as
health-promoting agents. Formative by-products also include substances such as phenol,
catechin, epicatechin, anthraquinones, as well as essential oils. Methods involving their
extraction using organic solvents such as methanol, ethanol, and acetone have been used to
recover these substances. However, these methods are harmful to human health and the
environment, as the process uses a large amount of organic solvents, and the methods are
time-consuming. Therefore, alternative methods that are effective, cost-effective,

* Corresponding author.
E-mail address: [email protected]

©2023 Author(s). This is an open access article licensed under the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/4.0). 422
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environmentally friendly, safe and fast are being studied and sought to reduce the
disadvantages of using organic solvents [3]. Therefore, in this article, the most popular
extraction methods have been collected, with both the advantages and disadvantages of their
use, as well as the principle of operation of the methods themselves and the type of solvent
(environmentally friendly or unfriendly) required. The research conducted in recent years
reveals the possibilities of using by-products in the production of products with high added
value, however, there are also various disadvantages, such as the lack of appropriate methods
for the extraction of these compounds. New extraction methods are emerging and being used
from time to time to optimize the extraction of bioactive compounds from byproducts.
However, given that progress has been made in this area, research has not been renewed in
recent years, despite significant advances in extraction methods. Solvents also play an
important role in many chemical and chemical engineering unit operations. Solvents usually
need to be used for a limited period of time during the process, as they are most often intended
as a solvent, diluent, dispersant or extractant, and then must be separated from the sample [9].
The correct choice of solvent is important and several methods of solvent selection have been
developed. Many years ago, choosing the most suitable solvent was purely empirical and was
done through error experiments or trials. This approach to solvent selection generally follows
the alchemist principle of similia similibus solventur [10]. Green extraction methods, which
do not cause pollution and use environmentally friendly solvents in their processes, are
becoming more and more popular [6]. For example, pressurized liquid extraction and
supercritical CO2 extraction use heat in combination with other parameters to improve the
extraction result. Several other non-thermal approaches are also available, such as pulsed
electric field extraction and ultrasound extraction, which can be particularly effective for
thermally stable compounds. Microwave extraction method is also used to improve the
extraction of biologically active compounds of plant origin [7], [8].
By-products are increasingly being used in various industries. As, for example, there are
currently no production processes in the pharmaceutical, perfumery, cosmetic, food
ingredient, biofuel, or chemical industries that do not use solvents in the extraction process,
such as maceration, percolation, steam or hydrodistillation, decoction and infusion. Mainly,
the latest trends in extraction methods are focused on finding solutions that replace or reduce
the use of petroleum solvents. This should be achieved by ensuring that the quality of the
extract does not decrease and that there is less impact on the environment. The article
combined the more commonly used extraction methods. The extraction methods reviewed
indicated their advantages and disadvantages, the products obtained by these methods, and
the extracts that can be obtained by extracting specific products.

2. EXTRACTION METHODS
In most analytical situations, the sample to be analyzed is a complex mixture consisting of
many components. Usually only one or two components of a sample are of interest for
research or production purposes, but these compounds are present in the matrix along with
many other compounds. In order to quantify the compounds of interest, they must first be
isolated from the other compounds in the sample [4], [5].
Extraction is the step to separate the desired natural products from the raw materials.
Extraction methods include solvent extraction, distillation method, pressing and sublimation
according to the extraction principle. Solvent extraction is the most widely used method. The
extraction of natural products takes place in the following steps: (1) the solvent penetrates the
solid matrix; (2) solute dissolves in solvents; (3) the solute is separated from the solid matrix;

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(4) the extracted solutes are collected. Any factor that improves diffusion and solubility in
the above steps will facilitate extraction. Extraction solvent properties, raw material particle
size, solvent-to-solid ratio, extraction temperature, and extraction duration affect extraction
efficiency [14], [15]. These are the most important aspects that must be taken into account
when performing the extraction, because the specific by-products are effectively extracted
only when the necessary parameters are specified in the process.
Refer to Table 1. the extraction methods, the products that can be obtained by the specific
methods, the extracts obtained, as well as the efficiency of the extraction methods. Of course,
traditional methods are more environmentally friendly because they use large amounts of
solvents, however, as the methods develop and come out, they are more environmentally
friendly and do not harm the environment during their processes, and researchers have worked
to ensure that the efficiency of the technology is not lost and is improved.
TABLE 1. COMPOUNDS USED IN EXTRACTION METHODS, BIOACTIVE SUBSTANCES FOUND AND
EFFICIENCY
Method Compounds Bioactives Efficiency
Percolation Medicinal plants [6], [4] Essential oils 35 % to 50 %
Plants with bioactive constituents [7] [8]
Reflux extraction Pepper [6] Spice oleoresin
Bamboo [9] polyphenols 70 % [9]
Chinese ginseng [10]
Medicinal herbs [11]
Soxhlet extraction algal biomass [12] Lipids 80–90 % [14]
Plants
Soybeans [5] Proteins and carbohydrates
Soils, sediments, and animal and plant
tissues [13]
Pressurized liquid Medicinal plants [12] Essential oils 48 % [16]
extraction Fruits skin(ex. grapes) [15]

Pulsed electric field Microalgae [17] Lipids 50 % [21]

extraction Flower seeds and olive by-products [18] Oils [18] [19]
Pomace (ex. apple, carrot) [19] [20]
Waste (ex. berries, vegetable) [20]
Maceration Coarsely powdered drug material, either Alkaloids, flavonoids, terpenes, 91 % [23]
leaves or stem bark or root bark [22] saponins, steroids, and glycosides [22]
Superciritical CO2 Sources - plant parts, such as leaves, Natural substances, aromas, fats, oils, 95 % [27]
extraction stem, roots, seeds, flowers, and fruits waxes, polymers, enzymes and
[24] [25] colourants [26]
flavonoids, essential oils, seed oils,
Meat, meat products and fish carotenoids, and fatty acids [24]
lipids and cholesterol
Enzyme assisted Sources - plants, bacteria, fungi, algae, Oils, proteins, carbohydrates, and 90 % [30]
extraction and animals [28] phenolics [28]
Tuna and mussel [29] Enzymes (pronase E and subtilisin) [29]

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Ultrasound-assisted Sources - animal and plant protein Vitamins, carotenoids and phenolic 90 % [35]
extraction sources [31] compounds [31]
Polyphenols, aromas, polysaccharides
brown seaweed species [32] from plant [32]
Alkaloids, flavonoids, glycosides [34]
microalgae, yeasts, food and plant Phenolics, phlorotannins, flavonoids
materials [33] [32]
Antioxidants, oils [33]
Heat reflux Source – plants [36][37] Essential oils, polysaccharide, 60 % [40]
extraction polyphenolics and flavonoids [39]
Pleioblastus amarus (Keng) Shell [38]
Mechanochemical- Source – plant [41] (bamboo leaves, Flavonoids, magnolol, rutin and 50 % [42]
assisted extraction Eleutherococcus senticosus, polysaccharides [43]
Ginkgo leaves,
Camellia oleifera Abel. Meal, Magnolia
officinalis, Hibiscus mutabilis L.,
Siberian fir needles) [42]
Raw tea, animal material (ex., Shark
cartilage [43])
Microwave-assisted Source – plants (ex. grape skin, Anthocyanins, 28 % [45]
extraction citrus peel, phenolic acids,
apple pomace, pectin,
soybean, tea leaves, vitamin E: γ-tocopherol and α-
tomatoes, tocopherol,
basil, garden mint, thyme, lycopene, essential oil,
green pepper, yellow pepper, sea Ascorbic acid, vitamin C [44]
buckthorn) [44]
Instant controlled Fruits peels Essential oil and antioxidants [46] 86 % [47]
pressure drop Plants (ex. green cardamom) [46]
extraction

The Table 1 describes the extraction methods as well as the products that can be extracted
by these methods. Next, each method will be discussed and the advantages and disadvantages
of these methods will be indicated.

2.1. Percolation
Given that the process is continuous, this method is more efficient than the maceration
method, and the saturated solvent in the process is independently replaced by fresh solvent.
However, it should be noted that during the extraction, high concentrations and larger
amounts of ethanol are required, in which the sample is soaked (about 12 - 24 hours) [48].
Percolation extraction is a traditional extraction method used in traditional Chinese medicine
processing. After placing the medicinal material powder in the percolation tank, the extraction
solvent is continuously added and the percolation extract is collected at the same time. The
percolation equipment and operation is simple. This applies to a wide range of medicinal
materials. Components that are unstable under thermal conditions can be effectively extracted
by the percolation process. However, there are also disadvantages, such as high solvent
consumption, long extraction times, and high energy consumption in subsequent
concentration processes [49].

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2.2. Reflux extraction
Reflux extraction is a solid-liquid extraction process that takes place at a constant
temperature with repeated evaporation and condensation of the solvent over a period of time
so that no solvent is lost. The system is easy to use, efficient and cost effective [50]. This
process is useful in industrial and laboratory settings where distillation is used. In addition,
this technique is useful in chemistry for supplying energy to sustain reactions for long periods
of time. In industrial distillation processes, reflux is useful for bulk distillation columns and
fractionators, including oil refineries, petrochemical plants, chemical plants, and natural gas
processing plants. In laboratory applications, the reagents and solvent mixture are placed in
a round-bottomed flask. It is then connected to a water-cooled condenser, which is usually
open to the atmosphere above. The round-bottomed flask is then heated, allowing the reaction
mixture to boil. The vapors formed from the mixture are condensed through the condenser,
thus returning by gravity to the round-bottomed flask. This technique is important because it
can thermally accelerate the reaction by carrying out the process at an elevated and controlled
temperature at ambient pressure, instead of losing a large amount of the mixture [51].
Extraction has the advantage of being more efficient than, for example, maceration or
percolation extraction, as less solvent is needed and the extraction time is shorter. However,
it cannot be used to obtain thermolabile natural products [52].

2.3. Soxhlet extraction

In this method, a finely ground sample is placed in a porous bag or “thimble” made of
cellulose or filter paper and placed in the thimble chamber of a Soxhlet apparatus. The
extraction solvents are heated in the lower flask, the sample evaporates in the “thimble”,
condenses in the condenser and drips back to the starting position. When the liquid content
reaches the siphon arm, the liquid content is again emptied into the lower flask and the process
continues [4]. This method requires a smaller amount of solvent compared to, for example,
maceration. Soxhlet extraction also has disadvantages, such as exposure to hazardous and
flammable liquid organic solvents that can produce toxic emissions during extraction. The
solvents used in the extraction system must be of high purity, which can increase the cost of
the extraction method. This procedure is not environmentally friendly and can cause pollution
compared to, for example, supercritical fluid extraction. The ideal sample for Soxhlet
extraction is a dry and finely divided solid, and the method requires consideration of many
factors such as temperature, solvent-to-sample ratio, and stirring speed [4].

2.4. Pressurized liquid extraction

Pressurized fluid extraction is considered an advanced extraction technology because of its
advantages over traditional extraction methods. Pressurized liquid extraction uses solvent
extraction at high temperatures and pressures that are always below this critical point so that
the solvent remains in a liquid state throughout the extraction process. This technique is also
known as accelerated solvent extraction, pressurized liquid extraction, pressurized hot solvent
extraction, high pressure solvent extraction, high pressure high temperature solvent extraction
and subcritical solvent extraction [53]. The use of these special pressure and temperature
conditions causes changes in the physicochemical properties of the solvent. For example, the
rate of mass transfer is increased while reducing the surface tension and viscosity of the
solvent and increasing the solubility of the analyte. This allows the solvent to penetrate more
easily and deeply into the sample to be extracted. As a result, the extraction process is more
efficient and it is possible to obtain a larger amount of extract compared to conventional
extraction methods. Low-pressure liquid extraction provides not only faster extraction

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processes, but also less solvent consumption for solid sample preparation. In addition, most
instruments used for pressurized fluid extraction are automated, allowing for less
labor-intensive methods and improved reproducibility [53]. This method has many
advantages, for example, it is not time-consuming (starting from 20 minutes), more
environmentally friendly and less toxic compared to traditional extraction methods, and this
technology is mainly used in the pharmaceutical and food industries. Various applications of
pressurized liquid extraction have been developed for the extraction of contaminants from
various food products; for organic pollutants from various solid and semi-solid environmental
samples, such as soil matrices, sediments and sewage sludge; and for the extraction of
bioactive compounds from natural matrices. In the field of natural products, the most studied
specimens are plants. However, various studies have described the extraction of bioactive
compounds from seaweed, microalgae and other food by-products. Pressurized liquid
extraction is mainly used to extract antioxidants (such as phenolic compounds and
carotenoids) and other bioactive compounds with anti-inflammatory, antimicrobial and
antiviral properties [53]. However, a drawback of pressurized fluid extraction is that the
presence of relatively high water percentages in the samples to be treated strongly decreases
analyte extraction efficiency when using hydrophobic organic solvents as water hinders
contact between the solvent and the analyte [54].

2.5. Pulsed electric field extraction

The pulsed electric field method uses moderate to high electric field strengths ranging from
100–300 V/cm in discontinuous mode and 20–80 kV/cm in continuous mode. Food or other
materials (residues, by-products, plants) are placed between the electrodes and the high
voltage electric field. The cell membrane is punctured, creating hydrophilic pores that open
protein channels. A force per unit charge, called the electric field, is applied to the sample
when high-voltage electrical pulses are applied through the electrodes. The membrane loses
its structural functionality and is extracted like plant material [18]. The electric field can be
applied in exponentially decaying, oscillating square waves, unipolar triangular or bipolar
pulses. The electroporation that occurs is either reversible or irreversible, but depending on
the application, this effect can be controlled. Generally, low specific energy (1–10 kJ/kg) and
pulse cycle time (nanoseconds to milliseconds) are effective in the extraction process. High
electric field strength increases cell membrane permeability, which depends on cell size and
cell geometry. For fragile plant tissue, an electric field strength of 0.1–10 kV/cm is sufficient;
however, resistant materials such as seeds require high intensities (i.e., 10–20 kV/cm) for
efficient extraction. It also provides some additional benefits such as maintaining the
nutritional value and sensory properties of liquid food [18].

2.6. Maceration
Very simple extraction method with low extraction efficiency and long extraction time
(three days). This method can be used to extract thermolabile components. This is an
extraction procedure in which coarsely powdered medicinal material, either leaf or stem bark
or root bark, is placed in a container; menstruum is poured on top until the medicinal material
is completely covered. After that, the container is closed and kept for at least three days. The
contents are stirred periodically and, if bottled, must be shaken from time to time to ensure
complete extraction. At the end of the extraction, the micelle is separated from the chaga by
filtration or decantation. The micelle is then separated from the menstruum by evaporation in
an oven or over a water bath. This method is convenient and very suitable for thermolabile
plant material [22]. Studies have shown that phenolics and total anthocyanins can be extracted

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from fruits under optimized conditions of 50 % ethanol, solids to solvent ratio of 1:20 and
particle size of 0.75 mm. The efficiency of the method can be improved by combining it, for
example, with microwave and ultrasonic extraction methods, and the combination of these
methods brings economic benefits, as it is possible to reduce the process temperature during
extraction. However, when the maceration method is combined with other extraction
methods, the efficiency is not high enough to obtain valuable compounds. When studying the
application of this method, it can be concluded that this method is effective only in
combination with other extraction methods [55].

2.7. Supercritical fluid extraction

Supercritical fluid extraction is a technique that uses a supercritical fluid, a substance whose
physical properties as both a gas and a liquid exceed the critical point. CO2 with a critical
point above 31.1 °C and 7380 kPa is the most commonly used supercritical fluid in
supercritical fluid extraction. It is flammable, relatively non-toxic, chemically stable and
inexpensive. Its critical temperature is suitable for the extraction of thermolabile compounds.
However, since it is non-polar, it is recommended to add polar modifiers such as ethanol,
methanol, ethyl acetate or acetone to extract the polar phenolic compounds. A pressure of 50–
600 bar, a temperature of 20–35 °C and a time of 5–180 min are considered to be the
parameters that provide the highest yield of phenolic compounds extracted from various
samples (e.g., plant materials) by the supercritical method. Although CO2 is the preferred
extraction solvent (for extraction of non-polar compounds), the polarity of supercritical
carbon dioxide extraction can be increased by adding a miscible polar compound (e.g.,
ethanol) as a modifier. Due to the selectivity of the supercritical fluid extraction process, the
extracts obtained by this method have a low concentration of undesirable compounds.
Additionally, supercritical carbon dioxide becomes gaseous after depressurization and can be
easily removed from the flow system [56]. Supercritical fluid extraction has many advantages
over traditional extraction methods, such as lower organic solvent consumption during the
process, high extract selectivity and separation, and shorter extraction time compared to other
extraction methods. The main advantage of this method is the reduced possibility of sample
contamination by solvent impurities and the avoidance of decomposition and oxidation of the
extracted compounds, as it is performed in a closed system without air and light. However,
the initial cost of supercritical fluid extraction equipment is very high, and the cost of using
it on an industrial scale often exceeds the technical benefits [49], [56].
Advantages of this method: (a) improved transport properties – the solute diffuses faster
through a supercritical solvent than through a liquid solvent; (b) the equilibrium ratio and
separation coefficients are usually quite high; and (c) the solvent can be regenerated with gas
at reduced pressure, resulting in significant energy savings. Small changes in temperature and
pressure at the critical point result in extremely large changes in solvent density and
solubility, which is an advantage of supercritical fluid extraction. Carbon dioxide has a low
cost, no harmful chemical properties and a low critical temperature [57]. The properties of
supercritical fluid can be changed by varying the pressure and temperature, allowing for the
selective extraction of actives, giving it an advantage over other extraction processes [58].
Supercritical CO2 is cheap and readily available in bulk with high purity, ensuring minimal
changes in bioactive compounds and maintaining their therapeutic or functional properties,
able to dissolve lipophilic substances and easily separate from end products [49]. The
disadvantage of supercritical fluid extraction is that the capital cost of supercritical fluid
extraction equipment is significantly higher (at least 50 %) than conventional extraction
equipment [57].

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2.8. Enzyme assisted extraction
Enzyme-assisted extraction is suitable for extracting pectins from waste and by-products
by increasing the permeability of cell walls. Enzymes are for the extraction of many phenolic
compounds, additional flavonoids and anthocyanidins. Enzyme activity, processing time,
substrate ratio, and particle size are all aromatized to achieve maximum efficiency during
enzymatic processing [59]. Enzyme-assisted extraction has high efficiency and easy
extraction conditions while maintaining extraction properties and stability. This method is
widely used in the extraction of biologically active components from plants [60].
One of the advantages of using enzymes is that they can be added to hydrophilic and multi-
step lipophilic extractions, especially for by-product valorization. In Europe, for example,
postharvest to distribution food losses of grains, fruits, and vegetables range from 20, 41, and
46 %, respectively. Supplementing them with the help of enzymes increases the phenolic
content of the lipophilic extracts, which is perfectly applicable in the production of nutritional
or pharmaceutical products [61]. However, by-product safety restrictions are increasing and
the topic is receiving more attention. In comparison, for hydrophilic extracts, enzymes
effectively increase the water-soluble content of new derivatives used in the food industry. In
addition, enzymatic extraction methods are characterized by easy reaction conditions,
substrate specificity, industrial applicability and many other advantages. These extracts can
be used continuously in many fields and, surprisingly, in the development of green
synthesis [62].

2.9. Ultrasound-assisted extraction

An extraction method that uses ultrasound to create rapid solvent movement, resulting in
increased mass transfer rates that speed up extraction times [63]. This extraction method has
become more popular due to its several advantages, such as the short extraction time (10 – 30
minutes) [64], low power consumption (approx. 0–100 W) [65], less damage to the active
compound and high extraction efficiency compared to traditional extraction methods [34]. In
the ultrasonic treatment process, acoustic vibrations or mechanical waves with a frequency
from 20 kHz to 100 MHz are applied to the sample. Ultrasonic extraction is based on
cavitation, mechanical and thermal properties and is a very efficient method for the extraction
of compounds of interest. These combined properties result in a decrease in particle size, cell
wall ruptures, and an increase in reaction rates as mass moves across the cell wall. Ultrasound
treatment creates a mechanical effect on the cell wall. This improves the penetration of the
solvent into the cell. Thus, the maximum amount of intracellular compounds dissolves in the
solvent, which is collected and purified. The ultrasonic method can accelerate the extraction
of bioactive compounds. The extraction is carried out in a gaseous or liquid solvent by
cavitation, which relies on liquid-liquid or gas-liquid interfaces. During the extraction,
mechanical fragmentation of the cell wall occurs, which helps to easily remove the extract
from the cell. The ultrasonic method is used for the extraction of bioactive compounds with
reduced energy consumption, less amount of solvent addition, shorter processing time. This
method is very fast and effective, because the cell wall is ruptured as a result of the ultrasound
action. When ultrasound waves pass through a liquid, it causes cycles of compression and
expansion. The expansion part forms bubbles or cavities in the liquid. The process of bubble
formation, growth and explosive disruption is called cavitation. In a pure liquid system, the
bubble maintains its circular structure because the environment around the bubble is
homogeneous. When a circular bubble bursts on a solid surface, its shape changes and creates
a high-velocity solvent jet against the cell wall and enhances solvent penetration into the cell
wall. It improves the interaction between solid and liquid. In addition, in the solid material,

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the ultrasound wave causes swelling, wetting and dilation of the pores in the cell wall. This
increases diffusion and thus improves mass transfer. However, cavitation properties can also
introduce new reaction mechanisms to form different types of free radicals. Sometimes highly
reactive free radicals can be released, which can modify other compounds such as proteins as
well as speed up the extraction of bioactive compounds [31].
The ultrasound method has various advantages. Technology with the help of ultrasound
improves food processing to improve the physicochemical and quality of food products.
Today, the ultrasonic extraction method is chosen mainly because high extraction yields of
plant bioactive compounds can be obtained with this method. Ultrasound-assisted extraction
can also increase the availability of bioactive compounds in food. It is likely that ultrasound
assistance is a good alternative method to improve the yield and bioavailability of compounds
compared to the heat reflux method [66].
The ultrasonic wave in this method could also improve the physical properties of
compounds such as suspension, particle size, viscosity, color, pH, acidity and stability. The
stability of bioactive compounds is an important criterion for bioavailability. Studies have
observed that sonicated juice had lower oxygen levels, which was a favorable environment
for carotenoid retention [66]. Ultrasonic technology is also useful in inhibiting the enzymatic
browning process. Studies have shown that sonication with an aqueous extract of purslane
can help delay browning of potatoes. This could be due to the “sponge effect” created by the
ultrasonic wave with a frequency of 40 kHz during cavitation, although the treatment is only
10 min. Ultrasound can repeatedly squeeze and release the purslane extract into the cells,
which is called the “sponge effect”. This phenomenon created an external low-concentration
solution to achieve an effective anti-browning concentration in the cells. In addition,
cavitation can also change the partial pressure of microbial cells, thereby promoting cell death
caused by cell stress. Reducing the microbial load in products would extend shelf life, as
microbes are always a major contributor to product spoilage [66]. Disadvantages could be
limited recovery, efficiency, time and solvent consumption [67].

2.10. Heat reflux extraction

The process is simple and only in combination with other extraction methods is more
effective. The sample is placed in a container of distilled water, then filtered and concentrated
using a rotary evaporator [68]. The reflux apparatus allows the solution to be easily heated,
but without the loss of solvent that occurs when heating in an open vessel. In a reflux setup,
solvent vapors are trapped by a condenser and the concentration of reactants remains constant
throughout the process. Heat reflux extraction is a more efficient method compared to
percolation and decoction because it requires a shorter extraction time and less solvent. The
technology involves heating the matrix for a certain period of time, causing a complex
chemical reaction. Since the process uses a reflux extractor as the main reactor, better mass
transfer and contact efficiency between the solvent and the treated matrix is achieved. The
vapor flows to the flask, thereby controlling the temperature of the reaction. This technology
is preferred due to its simplicity and convenient operation. The technology has found
application in the extraction of many natural, phytochemical compounds and essential
oils [69].
This method has both advantages and disadvantages. Advantages include low investment
costs, for example compared to the supercritical CO2 extraction method, and the method
provides a very high extraction efficiency at the end of the process [10]. However, the
disadvantages include the high consumption of the solution, which is environmentally

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unfriendly, and the high temperature required for the process (compared to other methods and
similar to the Soxhlet method) [70].

2.11. Mechanochemical-assisted extraction

Highly efficient, economical and environmentally friendly extraction of target bioactive
components from complex matrices of dietary supplements or natural medicines has always
been a challenge for both bioactivity determination, batch preparation and, more importantly,
quality assessment of natural products. Mechanochemical extraction, which is a green
technology, shows a clear superiority in the application of high-energy mechanical force
acting on raw materials to separate target components from natural products in an aqueous
environment at normal temperature and without using harmful, dangerous and expensive
substances. organic solvents. In addition, mechanical chemical extraction in a one-pot
extraction provides a rapid process, good reproducibility, and direct solution transfer to
instruments such as reverse-phase high-performance liquid chromatography systems for
further analysis [71]. This technology is based on the study of the physicochemical and
chemical transformation of compounds caused by mechanical force, such as grinding in a ball
mill. It consists of mechanically treating a sample (e.g., plant material) under very insensitive
mechanical pressure in a ball mill with a solid reagent (usually carbonated salts) prior to
solvent extraction. During the process, the cell walls are ruptured, allowing the target
compounds to be obtained with improved water solubility. This property allows the process
to use water instead of other common solvents, reducing extraction costs and simplifying the
purification process. The most commonly used reagents have been solid alkali reagents such
as Na2CO3, NaHCO3, and NaOH, depending on their alkali strength and the chemical
properties of the target compounds. Studies have shown that the extraction process provides
a more efficient time and temperature reduction and does not use organic solvents. However,
since the technique is quite new, the effect of different extraction parameters is still
inconsistent and a complete understanding is lacking, which is essential for the scale-up
process and future applications [55]. Of course, this method has both advantages and
disadvantages. Mechano-chemical extraction can have a significant impact on the
environmentally friendly processing of medicinal plants [72]. Mechanical chemical extraction
is considered a good alternative method to traditional methods for the extraction of bioactive
compounds from plant and animal by-product raw materials. The main advantages of
mechanical chemical extraction are: (1) selectivity and increased extraction yield, (2) low
operating temperature, thus allowing the extraction of thermolabile compounds, (3) in most
cases, water is used as a solvent instead of conventional. organic solvent, (4) short extraction
time [73], (5) simplified process. Thus, it is possible to obtain bioactive compounds from
plant and animal materials by mechanochemical extraction method [56]. Studies have proven
the increased efficiency and selectivity of mechanical chemical extraction as a new extraction
method, the absence of consumption of organic solvents, simplification of manipulations and
shorter extraction time. The mechanochemical extraction process can avoid the loss of heat-
sensitive compounds due to its relatively low extraction temperature (~40 °C). Thus, the
mechanical chemical extraction technique has a wide application in the pharmaceutical and
food industries in the future. Although the performance of the mechanical chemical extraction
technique is impressive, various problems arise when using this method. Not applicable to
hydrophobic and neutral compounds [52]. Further research and application of mechanical
chemical extraction is important, including the extraction of other plant materials, especially
water-insoluble substances and heat-sensitive compounds. In addition, further development
of extraction equipment is also required. Available ball mills with different weights can allow

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mechanical chemical extraction to be used on a larger scale. The mechanism and kinetics of
mechanochemical extraction are still debated and require further investigation. A deeper
understanding of the mechanism is essential for the further development of mechanical
chemical extraction. For example, the physicochemical and biological changes induced by
mechanochemical pretreatment should be further investigated. Although mechanical
chemical extraction is not yet widely used in the modern food and pharmaceutical industry,
we believe that it will be a competitive tool for the extraction of bioactive substances in the
future. Further development of the extraction process and equipment will promote the
application of mechanical chemical extraction technology [73].

2.12. Microwave-assisted extraction

Microwave extraction is a method that uses non-ionizing electromagnetic wave radiation
with a frequency of 300 MHz to 300 GHz to induce motion of molecules in polar or
polarizable materials or solvents by acting on dipoles. The molecular motions heat the sample,
which causes moisture to evaporate from the sample, creating pressure that causes the cell
wall to rupture and the necessary compounds to be released. During radiation, the solvent
molecules in the normal phase are aligned with the electric field. In the rapid electric field
changes that occur in microwave extraction, the solvent molecules are unable to rearrange
and begin to vibrate, causing the solvent to heat up due to frictional forces. This allows the
solvent to easily penetrate the sample matrix and facilitates the extraction of the desired
compounds. Solvents should be selected based on their boiling points, dispersion and
dielectric properties. Based on these properties, ethanol, acetone, or their combination with
water are often used to extract phenolic compounds using microwave-assisted extraction.
Since microwave energy is transferred only by dielectric absorption, non-polar solvents with
lower dielectric constants can absorb much less energy, which can lead to lower extraction
efficiency. Therefore, microwave-assisted extraction is considered a selective method for
polar molecules and solvents with high dielectric constants. Microwave extraction has many
advantages similar to ultrasonic extraction, including lower solvent usage, shorter extraction
time and processing costs, and high extraction efficiency. However, this method is limited to
low molecular weight phenolic compounds such as phenolic acids, quercetin, isoflavone and
trans-resveratrol, which were shown to be stable under microwave heating conditions up to
100 °C for 20 min. Phenolic compounds with a higher number of hydroxyl substituents, such
as tannins, or thermosensitive compounds such as anthocyanins, may not be suitable for
microwave extraction [56].
The advantage of microwave extraction is that it is an automated green extraction method
that has a short extraction time (can be a few seconds [23]) and low solvent consumption
during the process, as well as the ability to obtain several (up to 40) samples at the same time,
significantly improving the permeability of the samples [74]. Although the small sample size
used in many analytical extraction processes has some advantages and is more
environmentally friendly, it also has at least one disadvantage, such as the amount of
microwave energy absorbed decreases as the sample size decreases. If the sample is small in
size, a large amount of energy is not absorbed but reflected. Reflected energy can damage the
magnetron, so when using small samples for analytical work, it is recommended to use
microwave systems designed to protect the magnetron from reflected power. Commercially
available applicators are not compatible with microextraction, as the minimum amount of
solvent required is usually around 10 mL. Sample size is directly related to the amount of
solvent required for extraction. The nature of the sample and the dielectric properties of the
sample or its components can be very important in microwave extraction. For example, water

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added to or naturally present in the sample is a key factor because water molecules have a
high dipole moment, resulting in a high heating efficiency for the sample. A related
inconvenience is the requirement to control the water content of the matrix to obtain
reproducible results. Other possible matrix components (such as ferrous material) may cause
buckling due to absorption of microwave energy. The organic carbon content of the matrix is
known to hinder extraction due to the many strong analyte-matrix interactions that are
difficult to break. In this case, microwave extraction is a very effective technique [75].

2.13. Instant controlled pressure drop extraction

This process firstly involves a short heating stage (10–60 s) including the injection of
saturated steam under high pressure (up to 1 MPa) applied to the product initially placed
under vacuum. This step involves condensation of the vapor and heating of the product, which
increases the moisture content of the product by 0.1 g H2O/g dry matter. The initial vacuum
ensures rapid contact between the vapor and the sample and thus improves heat transfer.
Compressed air can sometimes be used as a pressurizing agent, such as for multi-cycle
instantaneous controlled pressure drop extraction treatments. After the first heating stage, a
sudden pressure drop (0.5 MPa s−1) towards vacuum (3–5 kPa) in just 10–60 ms causes
automatic evaporation of water in the product, which creates steam and significant
mechanical stress. This allows the product to expand. In addition, automatic water
evaporation ensures rapid cooling, which prevents thermal degradation of sensitive
compounds and thus ensures the high quality of processed products. The cooling rate can
reach an extraordinary 1500–2000 kW m 2 [47].
Instantaneous controlled pressure drop extraction technology is also currently being used
at research and industrial levels. Depending on the application, products can be collected from
the processing tank, while extracts and liquids can be collected from the vacuum tank.
Furthermore, it is important to emphasize that the processing conditions defined at the
laboratory scale can be directly applied at the industrial scale, which becomes an additional
advantage of the instantaneous controlled pressure drop technology [76]. Research at
laboratory and industrial scales has led to the development of many applications of
instantaneous controlled pressure drop technology in food processing. This process is well
guaranteed to maintain functional properties, organoleptic quality, effective
microbiological/fungal decontamination and reduced energy consumption and shorter drying
time. Instant controlled pressure drop extraction treatment is a high temperature/short time
process that improves both drying process performance and high quality functional food [77].
The efficiency of obtaining essential oils can be improved by combining this method with
other types of methods (for example, the hydrodistillation method) [46]. On the other hand,
it also has disadvantages, such as it can be energy-intensive and often has a long processing
time, which can cause negative chemical changes in the essential oil [46].

3. CONCLUSIONS
The main purpose of extraction is to separate the components from a mixture using a
solvent. The application of the extraction method is important in almost all industries, such
as pharmaceuticals, chemical industry, wastewater treatment processes, food and cosmetic
production processes, etc. Depending on the properties of the materials (samples) to be
extracted, it is important to study in detail and find the best and most efficient extraction
methods according to the processed sample (by-product).

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Over time, mining methods have evolved and become greener and more environmentally
friendly. New mining methods have been sought, as well as improved ones used so far. For
extraction, reflux extraction is the most common product separation method. Modern
extraction methods, which are also considered green extraction methods, including ultrasonic
extraction, microwave-assisted extraction, supercritical fluid extraction, and pressurized fluid
extraction, have received increased attention in recent years due to their high extraction yield,
selectivity, and stability. target extracts and process safety benefits. Some of these green
methods have become common sample preparation methods for analytical purposes.
There is a clear and growing interest in obtaining and isolating natural products and their
beneficial uses. These specific applications also include the extraction techniques used for
conditioning and the new stationary and mobile phases to be used in these techniques.
Consequently, these trends are expected to continue in the near future as they are primarily
driven by consumer demands and safety, environmental and regulatory issues.
In conclusion, advanced methods have some advantages over traditional methods in terms
of extraction yield and time. However, traditional methods still have their values. Traditional
press extraction is more suitable for extracting crude oil or other high quality oils, but it is
time consuming. Solvent extraction may have the highest extraction yield, but may result in
solvent contamination. Supercritical fluid extraction can be considered a kind of green
technique if the green solvent is chosen, but it has higher operating costs due to the complexity
of the process. In addition, microwave and ultrasonic extraction can increase the extraction
yield, but it is difficult to use for large sample sizes. These extraction methods need more
research to be able to obtain larger sized material. In general, each of the methods discussed
above can be modified or combined with other extraction methods to improve the efficiency
of the process. Each method has its pros and cons, but it is also possible to find and adapt the
best extraction technology for each material. As well as continuing to develop the extraction
methods used so far, they must be more and more efficient, as well as better in terms of time
and cost, in order to continue to focus on environmentally friendly processes, taking into
account the problem of environmental pollution.

ACKNOWLEDGEMENT
This work has been supported by the European Social Fund within the Project No 8.2.2.0/20/I/008 “Strengthening of
PhD students and academic personnel of Riga Technical University and BA School of Business and Finance in the strategic
fields of specialization” of the Specific Objective 8.2.2 “To Strengthen Academic Staff of Higher Education Institutions
in Strategic Specialization Areas” of the Operational Programme “Growth and Employment”.
This research/publication was supported by Riga Technical University's Doctoral Grant programme.

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