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Файл:Ординатура / Хирургия / Библиотека им академика М.И. Перельмана / Книга_6035_Библиотеки_им_академика_М_И_Перельмана.pdf
CHAPTER 9
Strategies for Isolation and Identification of Bioactive Molecules from Natural Sources
HAMID
SHWETA SURI
1
1,*
, DEEPIKA KATHURIA2, SUNAKSHI GAUTAM3, SHAKSHI SHARMA4, and
5
2
3
4
5
*Corresponding author
ABSTRACT
Bioactive molecules nowadays are of prime interest to consumers due to their antioxidant,
stimulant, antibacterial, and other functional properties related to the food, chemical, and
pharmaceutical industries. The process of extraction, identification, purification, and characterization of these bioactive compounds from natural sources is very complex. Many
conventional and novel methods are available for the extraction of bioactive compounds
from different sources, but appropriateness with respect to single process has not been
made yet. The efficiency of any method is usually dependent upon the type of natural
source, extraction method, solvent, extract yield, preparation procedure, energy consumption, and scientific expertise. Green extraction strategies have increasingly been applied to
obtain higher extraction yields of bioactive molecules and heat-labile compounds at a lower
environmental cost in less time with effective solvent utilization. However, for effective
extraction, sample preparation processes in addition to purification (chromatographic techniques) and characterization (high-performance liquid chromatography [HPLC], Fourier
transform infrared spectroscopy [FTIR], gas chromatography [GC]) are also of prime
184
importance and are highlighted and compared in detail. This chapter will aid in acquiring
knowledge about extraction, isolation, characterization, and toxicological assessment of
bioactive molecules from natural sources.
9.1 INTRODUCTION
Bioactive compounds are secondary metabolites that are abundantly present in plants,
microbes, and animals. They manifest both pharmacological and toxicological effects on
human beings. In plants, these metabolites are produced during the development phase to
assist them in overcoming natural obstacles (Azmir et al., 2013). The bioactive compounds
in plants are polyphenols, alkaloids, terpenoids, organosulfur compounds, nitrogencontaining compounds, etc. (Altemimi et al., 2017). These metabolites engender various
functional properties such as antioxidant, anticancerous, antidiabetic, anti-inflammatory,
and antidepressive properties in humans without adding any nutritional benefits (Zhang
et al., 2016). Several conventional and novel methods are available for the extraction of
bioactive compounds from different sources. However, the selection of a suitable extraction method depends upon the maximum extract yield from tissue, preparation procedure,
and energy consumption (Azmir et al., 2013). In addition to extraction, isolation and
identification of bioactive components is also performed using a comprehensive and high
throughput screening technique that helps in providing improved quality of product. This
chapter provides details about different approaches utilized for the extraction, isolation,
and characterization of bioactive compounds from different natural sources.
9.2 BIOACTIVE COMPOUNDS IN NATURAL SOURCES AND THEIR PHARMACOLOGICAL PROPERTIES
Bioactive compounds are secondary metabolites that act as therapeutic immune stimulators
possessing antioxidant, inflammatory , and antimicrobial properties (Segneanu et al., 2017).
More than 80% of the bioactive compounds used in the food and medicine industry are
obtained from natural sources. Bioactive compounds from natural sources have gained
interest due to their lower toxicity and complete biodegradability. The pharmaceutical,
chemical, and food industries, all use bioactive compounds due to the multiple beneficial
properties they possess (Pai et al., 2022). Bioactive compounds are also used in clinical and
pharmacological studies due to the numerous health benefits associated with them (Câmara
et al., 2020). Polyphenols, vitamins, carotenoids, omega-3 fatty acids, nucleosides, organic
acids, and phytosterols are different bioactive compounds that have been in focus because
of their essential role in the prevention of certain diseases (Kamiloglu et al., 2021). Daily
consumption of food rich in bioactive compounds that possess antioxidant activity has a
positive effect on overall human health and reduces the risk of cancer , heart disease, stroke,
and Alzheimer’s disease (Santos et al., 2019; Jaime and Santoyo, 2021).
Plant biodiversity is vast and houses a wide variety of different bioactive compounds.
Medicinal plants possess therapeutic properties as they are enriched with abundant
bioactive compounds, proteins, vitamins, and minerals (Sytar and Smetanska, 2022).
Bioactive compounds are present in different parts of plants such as roots, seeds, stems,
185
leaves, and owers. Food products, whole grains, fruits, and vegetables are also found to be
enriched with different bioactive compounds. Different studies have reported the presence
of essential bioactive compounds in plants that have pharmaceutical applications (Adeleye
et al., 2022; Vaou et al., 2022). In one study, Chhikara et al. (2018) reported that Syzygium
cumini which is a traditional medicinal plant enriched with numerous bioactive compounds,
namely,, myricetin, gallic acid, oxalic acid, cyanidin diglucoside, carotenoids, hotrienol,
citronellol, avonoids phytosterols, and polyphenols. Due to these bioactive compounds,
S. cumini helps in the prevention of metabolic abnormalities and different diseases. Tran
et al. (2020) found that bioactive compounds extracted from the plant Euphorbia hirta L.
possess antibacterial, anticancerous, and antioxidant properties. Polyphenols are one of the
major bioactive compounds found in medicinal plants, fruits, vegetables, and whole grains.
Anthocyanins, avanols, phenolic acids, and stilbenes are the major polyphenols found
in grapes. These polyphenols present in grapes and their value-added product such as red
wine also provide multiple health benets and help in the prevention of certain diseases
(Buljeta et al., 2023).
Flavonoids are another large and diverse group of compounds with biological activity
commonly found in different vegetables and fruits. They also offer anticancer, antioxidant,
and antiviral properties (Ullah et al., 2020). Tannins, quercetin, catechins, and daidzein are
the members of the avonoid family that have neuroprotective and cardioprotective effects
(Samtiya et al., 2021). T erpenoids are another example of bioactive compounds obtained from
different plant sources that offer numerous health-benecial properties. Apart from plants,
bioactive molecules are also derived from animals, fungi, and microorganisms, which have also
been used for the treatment of different infectious diseases (Kim et al., 2022). Major bioactive
compounds obtained from animals include L-carnitine, taurine, alpha-lipoic acid, choline,
glutathione, and conjugated linoleic acid. These compounds also possess different health-
promoting properties such as anti-inammatory, antioxidant, and immunomodulating activities.
Fungi are also an important source of naturally occurring bioactive compounds (Table 9.1)
that offer different therapeutic properties (Suwannarach et al., 2020). Microor ganisms such as
Bacillus and Pseudomonas are well known to produce several bioactive compounds that offer
numerous health-benecial properties (Monciardini et al., 2014). Moreover, bacteria present
in marine environments also possess neuroprotective, antioxidant, and anti-inammatory
effects. The bioactive compounds obtained from marine environments also showed strong
antagonistic activity against toxic microorganisms providing antimicrobial therapeutics. They
are also used for the treatment of certain diseases, namely, Alzheimer’s, Parkinson’s, cancer,
and diabetes. Lactones, quinolines, xanthones, peptides, and terpenoids are some examples of
bioactive compounds from marine sources (Karthikeyan et al., 2022).
9.3 STRATEGIES FOR ISOLATION OF BIOACTIVE MOLECULES FROM NATURAL
SOURCES
9.3.1 SELECTION OF MATERIALS
As the emphasis on healthy living and nutritious food grows in research and lifestyle,
the globe is heading toward nutritional security. This idea is particularly pertinent to the
186
field of functional foods, which involves the extraction of bioactive/functional components
from a single food source and their application to the creation of foods that are nutritionally
enhanced. This tactic has already gained much popularity, and numerous commercially
viable items are being developed. The most important procedures for the utilization of
biologically active compounds derived from natural resources include extraction, pharmacological screening, isolating and their characterization, toxicological assessment, and
clinical trials. Among these, extraction of functional components is the most important step
(Sasidharan et al., 2011). In more depth, the general steps involved in the effective utiliza tion of functional components involve the selection of raw material or sample followed
by application of pretreatments (altering the form of raw material), extraction by suitable
techniques (conventional and nonconventional techniques), effective use of biological
assay, sample preparation for isolation of components (like liquid–liquid extraction, gel
filtration, use of macroporous resins, solid phase extraction and many more), structural
elucidation/characterization, and quantification of phytochemicals (Brusotti et al., 2013).
TABLE 9.1 Health Benefits of Different Bioactive Compounds from Plant Sources
Bioactive
Compounds
Carotenoids Carrot, squash,
Anthocyanin Maize kernels Reduce cardiovascular risk
Flavonoids Apple peal extract Anticancer effect, reduce cardiometabolic
Quercetin and
apigenin
Phenols Sorghum bran Anticancer effect Lee et al. (2020)
Lycopene Tomato Antioxidant activity,protect body from
Bioactive
peptides
Polyphenols Grapes Lower risk of cardiovascular and
Terpenes Tea tree oil, Thyme Antimicrobial, anticancer , anti-inflammatory Cox-Georgian
Tannins Legumes Antioxidant, antibacterial, antiviral activity Tong et al. (2022)
Coumarins Vegetables, fruits,
Phytosterols Vegetable oil, wheat
Source Pharmacological Properties Reference
Reduce the risk of cancer (lung, prostate
pumpkin, tomato
Onion, lettuce,
parsley
Soy, oat, pulses Lower blood pressure and hypertension Chakrabarti et al. (2018)
seeds, nuts, coffee
bran, fruits
and breast cancer)
disorders
Reduce cardiovascular risk Patil et al. (2009)
oxidative stress
neurodegenerative diseases, antidiabetic
Anticancer, neuroprotective,
anticonvulsant properties
Prevention of cardiovascular diseases Carbal and Klein (2017)
⏎
Rowles and Erdman (2020)
Toufektsian
Ullah et al. (2020)
Story et al. (2010)
Pandey and Rizvi (2009)
Venugopala et al. (2013)
et al. (2008)
et al. (2019)
Sample selection is the primary step before the pretreatment. The sample can be
selected based on the concentration of bioactive components in the natural sources which
could be plants, insects, animals, marine or land-living organisms, microorganisms like
bacteria, fungi, algae. There are some sources that are rare for the extraction of some
187
important bioactive components and some are abundant. For example, there are various
venomous snakes from which the venom is isolated to cure rare cardiovascular diseases,
and some fungi which are although toxic but their bioactive compounds in lesser quantity
can be used as medicine. The microbial and plant sources are most preferred due to the
ease of availability as well as extraction of most of the functional components. In the
case of microorganisms, wider diversity, quick growth, and presence of unique bioactive
components make them ideal among all. Plaza et al. (2010) chose the algae Himanthalia
elongate and microalgae Synechocystis sp. for the extraction of bioactive compounds
because of the abundance and novelty of these compounds. Other than the selection of
the sample, the selection of extraction technology also plays a crucial role. This depends
upon the purity of the extract required, as well as physical and chemical properties and
sensitivity of the compound of interest. The process should be cost-effective and should
yield a quality product. This selection process also depends upon the location of active
compounds such as free or in bounded forms (Patra et al., 2022). According to Choi and
Verpoorte (2019), ionic liquids and deep eutectic solvents (DES) are more applicable in
this eld rather than supercritical uids. They gave this statement due to the narrow range
of polarity of solvents used in supercritical extraction as well as the higher instrumental
cost to maintain the desired pressure. However, DES covers more solvents containing both
hydrophilic and hydrophobic with a wide range of polarity for extraction. So the use of
proper extraction techniques along with suitable sample selection leads to lesser loss of
functional components.
9.3.2 PRETREATMENTS BEFORE EXTRACTION (DRYING, MILLING, GRINDING,
HOMOGENIZATION, STANDARD PARTICLE SIZE GRANULES)
The preliminary steps before extraction are of real importance as they define the quality
and quantity of the isolated components. One of the “bottlenecks” of any analytical extraction technique is often thought to be sample preparation, which affects both the extraction
yield and the quality of extractable components. Since sample preparation is thought to be
the most challenging to accomplish, it has been the step in the analytical technique that has
been evaluated the most (Herrero et al., 2013). The main objectives of various pretreatments
such as sieving, drying, homogenization, and others are to convert the compound of interest
or analyte into a more suitable form, to ease separation, to enhance the concentration in
the solvent, and to make them easily detectable and quantifiable (Luthria, 2006). Drying
as reported in many studies can affect the bioactive molecule directly, so appropriateness
related to the raw sample with the selected drying method should be taken care of to retain
maximum bioactive molecules besides the particle size of natural material obtained for
extraction (Hamid et al., 2020a; Hamid et al., 2022). The pretreatments accelerate the
kinetics of extraction of analyte and increase the contact area of constituent with solvents
which leads to higher extraction yield (Sasidharan et al., 2011). According to Brusotti et al.
(2013), it is very important to provide pretreatment to the crude extracts as it removes the
barriers and degrades the compounds that are strongly bounded to metabolites and does not
allow them to extract. Such components are mostly tannins, pigments, lipids, and certain
188
nutrient-inhibitor components. Thus, the removal of such undesirable molecules allows
the smooth extraction as well as higher extraction yield with purity. There are several
techniques for this purpose including maceration, solvent extraction methods, Soxhlet
extraction, green methods like microwave-assisted extraction, ultrasound-assisted extraction, pulse electric field, pressurized liquid extraction (PLE), high-pressure processing,
and supercritical CO2 extraction (SC-CO2) are some of the extraction techniques. These
techniques are very target specific and use very less volume of solvent with lesser wastage,
thus gaining popularity (Wijngaard et al., 2012). Nowadays researchers are more focused
on isolations using nonthermal pretreatments. This is to ensure the retention of bioactive
components which are very prone to temperature treatment. This include the treatments like
high-pressure processing, pulse electric field, microwave-assisted technology, ultrasoundassisted techniques, vacuum techniques, mechanical disruption, freeze-thaw, and many
more (Zhao et al., 2014; Pongmalai et al., 2015). There is another technique, enzymeassisted extraction (EAE), which is very safe and effective and is mostly used to enhance
the extraction yield. The principle behind this is the weakening and degradation of cell
walls, which allows the component to escape more easily from the bounded matrix. The
prominent examples of enzymes used are pectinases, cellulases, hemicellulases, proteases,
and many more (Doria et al., 2022).
Steam blanching softened the structure of the cabbage making the extraction simpler
and improved the process of extraction of glucosinolates (Pongmalai et al., 2015). Fresh
cauliower is used for the extraction of bioactive compounds by using the bacterial extraction method. For this purpose, 0.5–1 cm diameter was obtained by grinding followed
by treatment with 500 ml bacterial culture of Bacillus subtilis supernatant for extracting
enzymes like cellulase and endo-xylanase. These enzymes are further used for enzymatic
extraction. The results show that the quantity of cellulolytic enzymes used corresponds
with the recovery of chemicals, proving that pretreatment using unpuried culture broth
efciently encourages the discharge of bioactives from the vegetable matrix (Doria et al.,
2022). See et al. (2016) extracted the bioactive component from Java tea by ultrasound,
microwave, and mechanical grinding techniques. For enhancing the extraction yield, they
applied the pretreatments such as drying at 45ºC till 15% moisture content and grinding to
attain particle size of various grades like A (above 500 µm), B (250–500 µm), C (180–250
µm), and D (<180 µm). They concluded that with a decrease in particle size, the extraction
yield of bioactive components increases. These treatments were found to be very effective
because drying reduces the moisture content and grinding plant tissues breaks the cells
which allows the diffusion of bounded bioactive particles from matrix to solution easily.
Similarly, Hiranvarachat et al. (2013) used particle size of 1.4–2.0 mm ground carrots for
the extraction of beta-carotene through a microwave-assisted technique. For the isolation
of functional components from Artemisia ower, Anibogwu et al. (2021) enlisted various
pretreatments like maceration, stirring, grinding, drying, use of chemicals, acid hydrolysis,
alkaline hydrolysis, and biological treatment. For the extraction of bioactive components
from agarwood leaves, Lee
et al. (2017) used oven drying (60°C), followed by grinding
into 215 µm particle size for better extraction. For the extraction of proanthocyanidin from
grape extract, Da Porto et al. (2014) air-dried the grapes till 14.30% moisture content,
followed by grinding to attain a particle size of 2.0 mm.
189
9.3.3 TYPES AND PROPERTIES OF SOLVENT FOR EXTRACTION
Extraction is the core step for the isolation of any of the functional components. In this
step, solvents of different polarity are preferred for the extraction of bioactive compounds
from plant material. Solvents act as a medium to diffuse/leach out bioactive components
from any sample. Several studies have shown that polar solvents (methanol, ethanol,
hexane, ethyl alcohol, etc.) are mostly recommended for better extraction of antioxidants
(Altemimi et al., 2017). In order to choose a good solvent, various properties should be
kept in mind, such as low boiling point, quick mass transfer of solutes to solvent, low
toxicity, and should not produce complexity or dissociation. Thus, for easy and effective
handling of active ingredients, solvent selection is a step of utmost priority (Da Silva et al.,
2015). For the extraction of esters, oils, hydrocarbons, and fatty acids, nonpolar solvents
such as petroleum ether and hexane are preferred, whereas highly polar solvents (ethanol,
methanol) are effective for the extraction of phenolics as well as organic acids (SaucedoPompa et al., 2018). The solubility of phenolics varies on a number of factors, including
size, polarity, and the properties of the extraction medium, which is to be considered for
any substance. By altering the level of solvents used, for instance, the temperature, the
properties of the fluid (supercritical fluids and subcritical water) can be modified. The
solvent selected has a significant impact on the success of extraction (Wijngaard et al.,
2012). For instance, a limited number of solvents, such as water (with a mixture of base
or acid), volatile food components, and a few extremely volatile solvents like butane,
propane, ethanol, ethyl acetate, N2O, CO2, and acetone are permitted in the manufacturing
of food under European Union rules (Choi and Verpoorte, 2019). Based on the properties
of extractable components, one can use the solvents. For example, for the extraction of
hydrophilic compounds, polar solvents (ethyl acetate, methanol) are on the preferable side
(Table 9.2). For the removal of chlorophyll pigments from plants or algae, hexane can be
used; however, for hydrophobic or lipophilic ones, dichloromethane or its combination
with methanol in an equal ratio can be used (Sasidharan et al., 2011).
Wijngaard et al. (2012) used supercritical CO2 and 100% water in pressurized
uid extraction for extracting carotenoids from tomato paste waste, seeds, and skin;
polyphenols from apple and peach pomace, grape skin, guava seeds, and pistachio seeds
and anthocyanins from liquid grape seed extract using solvents like ethanol, methanol
under different conditions. Supercritical uids are known as the rst-generation green
solvents, having properties between gas and liquid. These uids have a density lying
between 0.1 and 1.0 g/ml and optimum dissolving power suitable for the extraction of
a wide range of functional components (Choi and Verpoorte, 2019). The supercritical
uid extraction (SCFE) technique is environment-friendly and gives excellent results
due to specic characteristics and critical points of solvents. Thus, it is generally used
to isolate alcohols, aromatic compounds, hydrocarbons, and most gases (Uwineza and
Waskiewicz, 2020). The main advantage of this method is that important extraction
conditions like temperature and pressure can be maintained according to demand, and
there are almost no solvent traces left in extractable components (Ingle et al., 2017).
Other than this, supercritical uids are low in surface tension and viscosity and high in
diffusivity, thus slight changes in temperature and pressure conditions lead to changes
TABLE 9.2 Utilization of Solvents for the Extraction of Bioactive Compounds
⏎
Solvent Used Extraction Method Raw Material Used Bioactive Compound Extracted Source
Hexane, ethanol, and water Pressurized liquid
extraction
Dried algae Himanthalia
elongate and microalgae
Phytol, neophyte diene, fecosterol, or
palmitic, and oleic acids palmitoleic
Plaza et al. (2010)
Synechocystis sp.
Water Supercritical water
extraction
Haematococcus pluvialis
microalga
Carvacrol, chlorogenic acid, ferulic
acid, gallic acid, p- hydroxybenzoic
Rodríguez-Meizoso
et al. (2010)
acid, sinapic acid, protocatechuic acid
Ethanol (25%), acetone (25%), and hexane
(50%)
Microwave-assisted
extraction and Soxhlet
Baby carrots (Daucus
carota)
Beta-carotenoids Hiranvarachat et al.
(2013)
extraction
Methanol, ethanol, and a mixture of these
alcohols with water
CO
, water, and ethanol SCFE Grape extract Proanthocyanidins, catechin Da Porto et al.
2
Water, ethanol, hexane, and isopropanol Maceration method
DES, used various DES benzyl triethyl
ammonium chloride, benzyl trimethyl
Soxhlet extraction,
sonication, and
maceration method
followed by Soxhlet
extraction for 6 h at 60°C
Ultrasound-assisted
extraction
Different plant extracts Alkaloids, anthraquinone, cardiac
glycosides, flavonoids, pyrrolizidine
alkaloids, steroids
Agarwood leaves Agarwood oil, squalene, phytol,
n-hexadecenoic acid, octadecatrienoic
acid
Natural and agricultural
waste by-products
Alkaloids, essential oils, phenolics,
proteins, and carotenoids
Sasidharan et al.
(2011)
(2014)
Lee et al. (2017)
Socas-Rodriguez
et al. (2021)
ammonium chloride, benzyl tributyl
ammonium bromide, benzyl tributyl
ammonium chloride, and choline chloride in
combination with water with different ratios
Methanol, petroleum ether, ethyl acetate,
chloroform
Soxhlet extraction
Euphoria cotinifolia
leaves
Presence of terpenoids steroids,
glycosides confirmed with the
Jayalakshmi et al.
(2021)
antimicrobial activity against Bacillus
cereous, Klebsiella pneumonia,
Eschercia coli, Salmonella typhi.
Methanol Fractionation Marine extracts from
sponges Lendenfeldia,
Tertiary amines or quaternary amines Dinarvand and
Spain (2021)
Ircinia and Dysidea sp.
190
191
in density which can be altered according to the desired component (Da Porto et al.,
2014). Newly reported ionic uids popularly known as DES are gaining more focus due
to their desirable physical and chemical properties. This includes a wider solubility and
miscibility range, optimum thermal properties, negligible vapor pressure, and very good
recycling properties (Lvanovic et al., 2020). Besides this, DES contains a homogenous
mixture of different solvents and shows lower melting points and greater versatility than
any other individually used solvent (Socas-Rodriguez et al., 2021).
9.4 EXTRACTION METHODS (CONVENTIONAL AND MODERN)
Different techniques are available for the extraction of bioactive components that play
an important role in developing quality products for the consumer. Furthermore, separation, identification, and characterization of bioactive compounds are possible only after
following proper extraction methods. Various conventional methods such as maceration,
decoction, percolation, hydrodistillation, infusion, and Soxhlet extraction are being
followed for the extraction. In these techniques, the extraction proceeds with the following
steps such as solvent diffusion into the solid matrix; mixing and dissolution of solute and
solvent; diffusion of solution from the solid material; and collection of extracted solute.
These techniques facilitate extraction with enhanced diffusivity and solubility properties.
Furthermore, various novel extraction techniques have also been developed to address
the drawbacks of conventional methods. Novel techniques such as microwave-assisted,
ultrasound-assisted, pulsed electric field-assisted, pressurized liquid enzyme-assisted,
supercritical fluid, and natural deep eutectic solvent-assisted techniques are gaining popularity due to the improved quality and yield of the extract as well as possessing shorter
operational time. Furthermore, the novel techniques are less detrimental to the environment
leading to the reduced usage of chemically developed organic chemicals. The food and
pharmaceutical industries are interested in assisted novel extraction techniques for rapid
and efficient extraction of bioactive components (Table 9.3). To understand the selectivity
of the extraction method, a brief discussion on various conventional and novel methods is
given below.
9.4.1 CONVENTIONAL METHODS
It is the process of molecular diffusion which is generally followed at the household
level for a very long time. This process consists of soaking the solid material in a liquid
medium (solvent/menstruum) in a closed container at ambient temperature for 2–3 days
with continuous stirring. The reason for this is to soften and rupture the protective cell
wall around the plant tissues for easy release of soluble components in a solvent which is
then separated by filtration or decantation process (Azwanida, 2015, Handa et al., 2008).
The efficiency of extraction is higher using ground solid material as it facilitates higher
TABLE 9.3 Comparison of Conventional and Modern Extraction Methods
Extraction Methods Principle Advantages Disadvantages Reference
Maceration Diffusion Minimum cost of investment, simple operation
procedure, easy to modulate process based on
solvent choice
Decoction Diffusion Simple operating procedure Requirement of huge amount of
Percolation Diffusion Increases yield of extraction by improved
contact time of solvent through herbal material
Hydrodistillation Decomposition of cell wall Automatic separation of bioactive compounds
Conventional
Infusion Diffusion Time taking operation, takes longer time to
Soxhlet Simplicity, boosted process kinetics due to
SCFE Extraction at a critical point
where solvent diffuses like
gas and dissolves like liquid
Microwaveassisted extraction
Ultrasoundassisted extraction
Novel
Pressurized liquid
extraction
Pulsed electric
field
Cell destruction by heat
generation caused by polar
molecules
Cavitation causes collapse
of cell structure
Molecular diffusion under
pressure
Electroporation Reduction of solvent use and processing time Activity of enzymes requires
from the water
form concentrated extract
applicability at high temperature
Higher extraction yield, reduced extraction
temperature, desirable for less polar
compound, no residual solvent in extract
Require minimum solvent and time for
extraction, higher yield and desirable for
thermolabile components
Minimum extraction temperature and time Not effective in a highly viscous
Application of pressure increases solubility
and mass transfer rate, higher extraction yield,
reduces solvent use
⏎
Longer processing time Azwanida (2015),
Jha and Sit (2022)
Azwanida (2015)
solvent
Longer operating time and require
large amount of solvent
Loss of volatile compound on
using high extraction temperature
Requires large amount of solvent
Poor extraction efficiency, timetaking procedure, not desirable for
thermolabile compound
Higher cost of investment Al Khawli et al.
Require small sample size,
effective using polar solvents, high
extraction temperature
fluid, swelling of the plant material
Higher cost of investment, high
temperature affects structural and
functional properties, undesirable
for thermolabile components
conductivity
Hussain et al.
(2019)
Omeroglu
(2019)
Hussain et al.
(2019)
Xiao
et al. (2008)
(2019), Jha and Sit
(2022)
Chan et al. (2011),
Jha and Sit (2022)
Tiwari (2015), Jha
and Sit (2022)
Nieto et al. (2010)
Heinz et al. (2003),
Jha and Sit (2022)
192
et al.