Food and Bioprocess Technology (2025) 18:7646–7660

[Link]

RESEARCH

Rational Design of Eco‑Friendly Deep Eutectic Solvent Systems

for the Recovery of High‑Value Phytosterol Compounds
María Antonieta Escobedo‑Monge1 · Sergio de‑la‑Huerta‑Sainz1 · Pedro A. Marcos2 · José Antonio Esteban‑Ollo3 ·
Laura Montejo‑Gil3 · María Conde‑Rioll3 · Mert Atilhan4 · Alfredo Bol2,5 · Santiago Aparicio1,5

Received: 8 January 2025 / Accepted: 10 May 2025 / Published online: 2 June 2025
© The Author(s) 2025

Abstract
The extraction and purification of high-value phytosterol compounds from plant-derived feedstocks remains an important
challenge in the fields of nutraceuticals, functional foods, and pharmaceutical applications. Conventional solvent-based
extraction methods often rely on volatile, toxic, petroleum-derived solvents that raise sustainability concerns. Conventional
solvent-based extraction methods often rely on volatile, toxic, petroleum-derived solvents such as n-hexane, ethanol, petro-
leum ether, or dichloromethane, which are considered volatile organic compounds (VOCs) and raise sustainability concerns.
Natural Deep Eutectic Solvents (NADES) have emerged as alternative and promising green solvents for the selective recovery
of phytosterols. In this work, we employ COSMO-RS (COnductor-like Screening MOdel for Real Solvents) modelling to
systematically design and screen a large library of NADES formulations, with natural compounds as components, for the
efficient extraction from vegetable sources of 19 typically occurring phytosterols. The COSMO-RS methodology allows the
simulation of each NADES property, including their melting point and composition. It also calculates the activity coefficients
with the phytosterols that will be used to rule out unsuitable combinations and evaluates toxicological parameters to assess
their safety. After conducting nearly 2000 solubility tests, NADES demonstrated 1.5 to 5 times higher extractive capacity
than common solvents for all phytosterols, with significantly lower volatilities. Finally, the use of NADES, especially the
carvacrol-menthol mixture, significantly improves the extraction of phytosterols compared to conventional solvents, offering
a more efficient, safe, and sustainable alternative, capable of dissolving a wide range of phytosterols.

Keywords Natural Deep Eutectic Solvents · Phytosterol extraction · COSMO-RS modelling · Sustainable separation ·
Plant-derived bioactives

Introduction

Phytosterols (PTS) are natural triterpenes of plant ori-

gin, which are mainly present in oilseed plants. They are
characterized by a chemical structure very similar to cho-
* Santiago Aparicio
sapar@[Link] lesterol (Bai et al., 2021; Othman et al., 2013), having a
fused polycyclic structure with varying carbon side chains,
1
Department of Chemistry, University of Burgos, with or without a double bond. PTS esterified with fatty
09001 Burgos, Spain acids or conjugated with glucosides play an essential role
2
Department of Physics, University of Burgos, 09001 Burgos, in regulating membrane fluidity and permeability (Brufau
Spain et al., 2008; Plat et al., 2019; Roche et al., 2008). Regarding
3
Research, Innovation and Development Center, Soria Natural, technological applications of PTS, they are used in the cos-
42162 Garay, Spain metic and pharmaceutical industries (Evtyugin et al., 2023;
4
Department of Chemical and Paper Engineering, Western Nattagh‐Eshtivani et al., 2022; Younas et al., 2024) and in
Michigan University, Kalamazoo, MI 49008‑5462, USA the food industry as additives, natural ingredients, and nutra-
5
International Research Center in Critical Raw Materials ceutical products with therapeutic effects (Ostlund, 2002;
for Advanced Industrial Technologies (ICCRAM), University Witter et al., 2018). In medicine, their cholesterol- and, in
of Burgos, 09001 Burgos, Spain

Vol:.(1234567890)
Food and Bioprocess Technology (2025) 18:7646–7660 7647

general, lipid-lowering properties are of interest, with antia- et al., 2020; MS et al., 2018). Conventional extraction meth-
therogenic and immunomodulatory properties (Morand & ods such as maceration, percolation, reflux, hydrodistilla-
Tomás-Barberán, 2019; Nashed et al., 2005; Nes, 2011; Oth- tion, and Soxhlet extraction (Fernández-Cuesta et al., 2011),
man et al., 2013; Trautwein et al., 2018; Witter et al., 2018), all of which involve organic solvents, heat, and mechanical
especially showing changes in cholesterol metabolism in agitation to extract PTS, have been developed (Grosso et al.,
patients with heart disease, degenerative diseases, deficiency 2015; Jha & Sit, 2022; Kozłowska et al., 2016; Mustapa
of essential neurotransmitters, and neuronal damage induced et al., 2015; Péres et al., 2006; Vilela et al., 2013).
by oxidative stress, among other chronic conditions (Feng Therefore, solvents play a fundamental role in the PTS
et al., 2020; Jansen et al., 2006; Moreau et al., 2018; Yan extraction and separation processes. The most commonly
et al., 2021). PTS have hepatoprotective properties (Feng used solvents for the extraction of phytosterols are n-hex-
et al., 2020), anti-inflammatory, anti-atherosclerotic, antioxi- ane (Kozłowska et al., 2016), petroleum ether (Péres et al.,
dant, and anti-cancer effects (Awad et al., 2003; Bae et al., 2006), ethanol (Mustapa et al., 2015), or dichloromethane
2021; Blanco-Vaca et al., 2019; Müller & Bracher, 2015; (Abdel-Aal et al., 2015; Fernández-Cuesta et al., 2011), all
Sierksma et al., 1999). of which are volatile organic compounds (VOCs) (Garcia-
It has been identified 250 different PTS structures (Maran- Llatas et al., 2021) with relevant environmental and safety
goni & Poli, 2010), being the most common β-sitosterol, problems (Morand & Tomás-Barberán, 2019; Nashed et al.,
campesterol, and stigmasterol (Brufau et al., 2008; Gajewski 2005) or high material loss due to their volatility. Likewise,
et al., 2009; Han et al., 2008; Shin et al., 2010; Nes, 2011; these extraction techniques use high temperatures and long
Yuan et al., 2017). However, there is considerable variability extraction times that can result in the thermal degradation of
in PTS content among plant species (Plat et al., 2019; Zhang bioactive compounds (Jha & Sit, 2022) as well as processes
et al., 2020). Among fruits and vegetables, the lowest con- being technoeconomically non-viable, thus hindering their
tents are found in watermelon (2 mg/100 g) and eggplant (3 industrial scaling up.
mg/100 g) and the highest in peas at 54 mg/100 g (Han et al., This situation has prompted industry to develop new
2008). On the contrary, the highest values are recorded in green extraction techniques that are more reliable, sustain-
oilseeds, such as canola, soybean, and sunflower, with 399, able, energy-efficient, faster, and safer for humans and the
353, and 225 mg/100 g, respectively (Amar et al., 2009b; environment (Kaderides et al., 2019; Cannavacciuolo et al.,
Costa et al., 2011; Fernández-Cuesta et al., 2011; Fernán- 2024), employing supercritical fluids (Hrabovski et al.,
dez-Cuesta et al., 2012). In cereal seeds, values fluctuate 2012; Nyam et al., 2010; Shi et al., 2015; Sperotto et al.,
between 44 and 110 mg/100 g (Piironen et al., 2000; Ryan 2022; Tolcha et al., 2021; Tuhanioglu & Ubeyitogullari,
et al., 2007; Shao et al., 2021; Singh et al., 2003; Zangenberg 2022), pressurized liquids (Alvarez-Rivera et al., 2020;
et al., 2004). In the group of pseudocereals, quinoa stands Nieto et al., 2010; Péres et al., 2006; Santos et al., 2021;
out with values of 118 mg/100 g (Hernández-Ledesma, Setyaningsih et al., 2016), ultrasounds (Péres et al., 2006; J.
2019; Ryan et al., 2007; Villacrés et al., 2013a, 2013b) fol- Wang et al., 2025), pulsed electric fields, microwaves (Bel-
lowed by buckwheat (up to 139 mg/100 g) (Dziedzic et al., wal et al., 2020), enzymes, and even high-voltage electrical
2018b; Raguindin et al., 2021a, 2021b) and amaranth (178 discharges to extract various bioactive compounds (Gowda
mg/100 g) (Fernando and Bean, 1985; Marcone et al., 2003; et al., 2022; Soquetta et al., 2018). All these methods, how-
Ogrodowska et al., 2014). ever, have appeared in the search for a radically different
Considering the availability of PTS in vegetal sources, it extraction procedure, rather than looking for a suitable sub-
is essential to develop suitable extraction techniques, which stitute for the traditional solvents themselves, at the expense
are largely dependent on the nature of the matrix and the of complex experimental setups, expensive operation costs,
form of the PTS, either as free, esterified, or glycosylated or losses of product.
compounds (Lagarda et al., 2006; MS et al., 2018). The qual- As a straightforward alternative, the extraction of PTS
ity of PTS extracts is the result of a combination of adequate is possible using Natural Deep Eutectic Solvents (NADES)
raw materials (quality of the plant material: species, variety, (Hansen et al., 2021; Makris & Lalas, 2020; Serna-Vázquez
maturity of the plant, environmental factors where it has et al., 2021; Švarc-Gajić & Morais, 2022; Zhou et al.,
been developed), the extraction methods and processes used 2023; Singh et al., 2024), a category of solvents aligned
(parameters such as temperature and extraction time impact with the priorities for the reduction of hazardous solvents in
the efficiency of the process), the type of solvents used, the industrial processes. Any DES consists of a combination of
purification and refining processes, and the storage condi- two different compounds interacting via strong intermolecu-
tions (Gupta et al., 2012; Jha & Sit, 2022). The first step lar forces, resulting in a mixture with a melting point lower
in most of the methods for obtaining PTS is their extrac- than that of its constituents due to the existence of a eutec-
tion from vegetable oil, separating them from the rest of tic point, hence their name. The components of a DES are
the matrix, followed by analysis and identification (Dikshit commonly known as the hydrogen bond donor (HBD) and
7648 Food and Bioprocess Technology (2025) 18:7646–7660

hydrogen bond acceptor (HBA) in the case such interaction calculate the chemical potential of a given molecule (Gau-
is produced via hydrogen bonding, and for a NADES, both din & Aubry, 2025), which further allows a full range of
compounds are organic and can be found in natural sources. thermodynamic properties like activity coefficients, phase
One of the most relevant characteristics of NADES is that equilibria, solubilities, and many more, with very low com-
they can solubilize polar and nonpolar compounds depend- putational cost. This has allowed the design and tuning of
ing on their composition, making them attractive substitutes DES for particular molecules (Norhidzam et al., 2025; G.
for conventional extraction methods (Zhou et al., 2023; Zou Oliveira et al., 2021; D. Wang et al., 2024) as well as mass
et al., 2024) and suitable for organic molecules of inter- screening for a wide range of solutes in a previous study
est such as polyphenols (I. L. de Oliveira et al., 2025; Ly (de-la-Huerta-Sainz et al., 2025) and with promising per-
& Sothornvit, 2024), anthocyanins (Kaur & Qadri, 2025; spectives in food and cosmetic research fields (Gaudin &
Kuasnei et al., 2022), and saponins (Deng et al., 2025). The Aubry, 2025).
physicochemical properties of the NADES are tunable by For this work, a similar strategy will be used for phytos-
modifying its composition, which enables the creation of terols: a selected group of PTS will be optimized through
a NADES that fits each process, maximizing the positive DFT calculations and later tested against a large library
effect of the solvent. of previously computed HBA/HBD combinations via the
There is a clear research gap in this field. Although COSMO-RS methodology. The obtained values in activity
NADES have been studied for the extraction of various bio- coefficients, solubilities, and thermodynamic properties of
active compounds, there is little research on their affinity and the NADES will allow us to easily discard the less promising
efficacy in the extraction of specific phytosterols and how cases and, with toxicological and environmental information
they may impact the environment upon their use. Molecular- as a complement, select the best NADES for phytosterol
level interactions between NADES and solutes have not been extraction from an efficient, safe, and sustainable point of
thoroughly explored through molecular dynamics (MD) view.
simulations, quantum mechanical computations (DFT), or
thermodynamic modelling, so a systematic approach to the
design of optimal NADES, while also considering their
effect on human health and the environment, is lacking. No Materials and Methods
systematic strategy has been developed to design and tune
NADES composition based on properties such as polarity, Natural Compounds Selection
solubility, viscosity, and thermal stability. The impact of
the physicochemical properties of NADES on extraction As established before, the main point of interest of this work
and their safety is poorly understood; computational mod- is the analysis/extraction of PTS. To study this, 19 PTS were
elling is needed to predict how different combinations of selected based on their availability in vegetal sources as well
hydrogen bond donors and acceptors in NADES influence as their suitability for industrial uses. The molecular struc-
phytosterol solubility. There is a marked limitation in the ture of the vast majority is fairly similar (Fig. 1), although a
integration of experimental and computational data, and couple of them–daucosterol and saringosterol–have an addi-
few theoretical and predictive studies have been conducted tional polar group attached to the general carbon skeleton,
that also include potential toxicological and environmen- so a certain difference in behaviour is expected.
tal hazards, which hinders the understanding and efficient On the other hand, the NADES library used in this study
design of these solvents. This study will address this gap by is complex and extensive. A total of 66 naturally occur-
applying advanced computational tools to design, optimize, ring compounds with various chemical natures–terpenoids,
and predict the behaviour of NADES for more efficient and alcohols, carboxylic acids, ketones, and lactones, to name
sustainable phytosterol extraction. a few–were selected. All compounds have an oxygen atom
The main objective of this study is to design, using com- able to donate an electron pair, thus qualifying them to act as
putational chemistry methods, suitable NADES for PTS a hydrogen bond acceptor (HBA). Of these 66 compounds,
extraction, via the careful and detailed optimization of the 45 of them also have a protic hydrogen, mainly in the form
solvent properties for the correct and efficient extraction of an alcohol or acid group, allowing them to also act as a
of PTS. For this purpose, a large in silico NADES library hydrogen bond donor (HBD). By combining pairs of com-
was developed and screened using the COSMO-RS meth- pounds, one as HBA and the other as HBD, a potential
odology (Eckert & Klamt, 2002; Klamt, 1995; Klamt et al., NADES is conceived, resulting in a total of 2970 mixtures
1998). This methodology is based on the elaboration of a to test.
density charge surface, called the σ-profile, calculated with Additionally, a selection of other organic compounds typ-
quantum chemistry methods such as the density functional ically used as extractants for phytosterols was also gathered
theory (DFT). The σ-profile is used as the starting point to for comparison. All of them are considered volatile organic
Food and Bioprocess Technology (2025) 18:7646–7660 7649

Fig. 1  Chemical structures of all 19 phytosterols used as target compounds

compounds (VOCs) and will be treated as solvents, just as suitable for typically occurring organic compounds such as
the NADES. All compounds–HBA, HBD, phytosterols, and the considered molecules. Due to the restrictions imposed
VOCs–are gathered in the [Link] file of the Sup- by other software, specifically the COSMO-RS methodol-
porting Information. ogy and the 2021 version of the COSMOTherm software
(Arroyo-Avirama et al., 2023; BIOVIA COSMOtherm, n.d.-
Structural Optimization b; Chagnoleau et al., 2024), the BP86 functional with a def-
TZVP basis set is used in order to generate the proper files
All selected compounds, both PTS and NADES component for further processing and thermodynamic calculations. This
candidates, require fully optimized geometries to ensure will allow importing all the molecules involved to COSMO-
accurate simulation at their most stable configuration, with Therm using the BP_TZVP_21.ctd parametrization (Chag-
the lowest energy (Arroyo-Avirama et al., 2023; Santiago noleau et al., 2024) for all later calculations.
et al., 2022). For that purpose, a structural optimization was
carried out by means of the TURBOMOLE software pack- Activity Coefficients
age (Abranches et al., 2019; Balasubramani et al., 2020;
Chagnoleau et al., 2024; TURBOMOLE V7.8, 2023, n.d.) Taking into account all compounds and mixtures consid-
on all molecules individually, without considering any mix- ered–2970 DES candidates and 19 phytosterols–the total
tures or combinations to begin with, as that could restrict the amount of combinations is impractically huge, easily sur-
potential intermolecular orientations when using COSMO- passing the 55,000 possible tests to compute. Although all
RS. The main calculation method is the density functional compounds could technically be simulated with enough time
theory (DFT) (Moreno et al., 2018) due to its good balance and computational resources, a more smart and approachable
between accuracy and computational cost, making it very method has been used. This method eliminates cases where a
7650 Food and Bioprocess Technology (2025) 18:7646–7660

favourable interaction between compounds is not observed, n.d.), PubChem (Kim et al., 2023), ChemSpider (ChemSpi-
which will be quantified through the activity coefficients. der, n.d.) or CRC Handbooks (Haynes, 2014), and, when
Since the main phenomenon considered is the solubiliza- no other data were available, the contribution method from
tion of a PTS in an NADES candidate, an attractive inter- Joback and Reid (Joback & Reid, 1987).
action between it and each NADES component must be
present; otherwise, the PTS would not be properly solved. NADES Safety: Volatility and Toxicity
The activity coefficients between each PTS and HBA or
HBD–each one isolated and not mixed–have been calcu- Besides the operating conditions of the extractive process,
lated with the COSMO-RS methodology with an infinite the safety, toxicity hazards, and sustainability must also be
solution approach at 25 °C (Arroyo-Avirama et al., 2023). considered. One major concern with solvent extraction is the
The result is yielded as the decimal logarithm of the activity volatility of commonly used solvents, which are typically
coefficient, log(γ), in which a negative value indicates an VOCs and pose inherent toxicity and pollution risks. One
attractive interaction, while positive values show a repulsion. of the main concerns about solvent extraction is the volatil-
COSMOTherm (BIOVIA COSMOtherm, n.d.-a) also ity of commonly used solvents, which are typically VOCs
allows calculating the activity coefficients for all PTS within and pose inherent toxicity and pollution hazards. For that
a liquid mixture; however, since the main purpose of the purpose, two additional thermodynamic calculations were
result only concerns its sign and not its actual numerical performed for each DES candidate: vapour pressure at 25
value, it was preferred the quicker one-to-one approach to °C and boiling point at 1013.25 hPa, as representative of the
setting all possible PTS-HBA-HBD combinations. relevant working conditions. Again, this has been performed
with COSMO-RS in a very similar way to the SLE case,
needing now the vaporization enthalpy as external data and
NADES Solid–Liquid Equilibria: Melting Temperature using the eutectic composition as the input.
For the toxicity analysis, a few representative properties
Another possible selection criterion is the usability of the were selected for each DES component and VOC, these
NADES candidates, as they would determine the working being the vapour pressure, as a measure of their volatility;
conditions in which the extraction would take place. These the octanol/water partition coefficient, as a sample of their
properties are the melting point and the composition at bioaccumulation potential; and the Cramer toxicological
which it is lowest. The melting point is the main condition- level, which identifies chemical structures more prone to
ing factor, as the NADES needs to be in a liquid state in produce harmful effects on humans (Cramer et al., 1978).
order to be used as an extractive solvent. For that, liquid These three parameters were obtained through the QSAR
phase at room temperature would be the ideal case, although Toolbox software suite (QSAR Toolbox, n.d.), developed
those mixtures that only need mild heating could be also and validated in collaboration with the ECHA (European
employed. Composition also plays a role in its usability. Chemicals Agency, n.d.).
Mixtures dominated by a single component would offer little All three safety parameters were fetched from QSAR
benefit over using the pure compound. In contrast, mixtures Toolbox experimental data databases whenever possible, the
with simple ratios, such as 1:1, 1:2, or 1:3, are easier to ECHA REACH (Lulei, 2008) being the most used and exter-
prepare and work with. Combining both properties, the best nal sources like PubChem (Kim et al., 2023) as a comple-
conditions would be a melting point considerably lower than ment. For those cases where no data were available, QSAR
25 °C and a molar fraction–of either component–between models provided an estimation: KOWWIN for the partition
0.45 and 0.55, as it would be quite close to equimolarity. coefficient and MPBPWIN for the vapour pressure.
These NADES properties will be calculated, once
again, using the COSMO-RS methodology and perform- Solubility Tests
ing a full solid–liquid equilibrium (SLE) calculation for
each of the 2970 DES candidates. This kind of calculation Once the formerly discussed discarding criteria have been
performs a temperature scan, ranging from 100 to 0 °C, chosen and their relevant data gathered, the main step is
and yields the melting point and eutectic composition, if the solubility tests once more with COSMOTherm. These
one is found. However, while COSMO is able to perform tests consider the NADES candidate, the extractant, the
such duties just from the original files, including external solute, the PTS, and the working temperature, which was
data, such as the melting point or fusion enthalpy, is rec- set to 25 °C. Only such HBA-HBD (NADES)-PTS combi-
ommended; otherwise, it could assume any compound is nations that fulfil all criteria–NADES liquid at room tem-
liquid, which can lead to false results. An extensive search perature and nearly equimolar composition, and negative
was conducted, gathering both data from databases such as log(γ) between each PTS and both HBA and HBD–will be
the NIST (National Institute of Standards and Technology, calculated. As a complement, the solubility of each PTS
Food and Bioprocess Technology (2025) 18:7646–7660 7651

on relevant VOCs, at the same temperature as before, has are in Tm, with some anomalous cases sparely found for
been calculated as well in order to perform a compara- very different compounds.
tive analysis and choose the best NADES. A flow diagram
showcasing the process can be seen in Fig. 2. Vapour Pressure and Boiling Point

The vapour pressure and boiling point results are very

promising. All valid mixtures computed showcase the non-
Results and Discussion volatile nature of a typical DES, with the vapour pressure
never surpassing 1 hPa, with the highest values found in
DES Properties combinations involving naturally fragrant compounds such
as geraniol, linalool, or citral, to name a few. The boiling
Almost all 2970 HBA-HBD combinations formed eutec- points obtained match this non-volatility, as they easily span
tic mixtures, with few exceptions involving high melting a range between 180 and 470 °C, the higher the lower the
temperature compounds like santonin and matricin, likely vapour pressure, as expected. While these criteria are not as
due to their melting points falling outside the temperature important from an operative point of view, from a toxico-
range considered in the scan. Results from identical com- logical and environmental one, they present DES as a much
pounds, where all HBDs are also HBAs, and duplicates, safer alternative to VOCs, with lower solvent losses and haz-
where both compounds can act as both donor and acceptor, ards of intoxication by inhalation.
were discarded–half of them in the latter case. The remain-
ing amount of remaining DES candidates was 1929. Solubility Analysis
Upon comparing the DES melting point Tm with those
of its components–as observed in Fig. 3–a clear trend can A few remarks will be done about the activity coefficients,
be observed, with the majority of the DES candidates as their calculation was merely a simple decision criterion
being liquid at room temperature and the highest melting in order to reduce the workload to the point that only its
points appearing when both components have a high Tm sign was considered and not its actual value. No remark-
as well. This is expected, as by definition a eutectic must able trends were observed, as even between similar com-
have a lower Tm than its constituents, so if one of them has pounds–either PTS or HBA/HBD–the results were fairly
a low one, so will the eutectic. inconsistent.
The molar fraction results are, on the other hand, con- Once all selection conditions have been applied, a total of
siderably more erratic than those of the melting points, and 1286 solubility calculations involving the DES candidates
a trend regarding the components Tm, while observable, is were conducted, and another 152 for the VOCs solubility
not as clear as before (Fig. 4). Most of the mixtures yield tests. Each calculation gave out three results, with the final
a ratio way too distant from equimolarity to be considered phytosterol solubility expressed as molar fraction, weight
eligible by our former criteria, although the most balanced fraction, and volumetric solubility, with the first and the last
ratios occur more easily the more similar HBA and HBD one in decimal logarithmic form. For the VOCs, no selection

Fig. 2  Flowchart diagram of the general computational workflow of this paper

7652 Food and Bioprocess Technology (2025) 18:7646–7660

Fig. 3  Melting point 2D scatter

plot for all tested HBA/HBD
combinations, coloured as per
their melting point. Green, Tm <
25 °C; yellow, 25 °C < Tm < 40
°C; orange, Tm > 40 °C

rules were applied, as they are all single compounds, liquid many interactions present, so it is not surprising that such
at room temperature, and their number is much lower than trends are not seen.
for HBA/HBD, so the calculation of all solubility combina- It is interesting to note that, in the VOCs resulting data,
tions was more than affordable. the effect of the solvent polarity is clear. Saringosterol and
At first glance, the solubility of phytosterols in either DES daucosterol, though slightly soluble overall, show far more
or VOCs is consistently low, with molar fractions barely sur- solubility with polar VOCs like isopropanol, ethanol, or
passing 0.1 and going as low as − 10 in the logarithm. Due dichloromethane, as expected due to their additional alco-
to the high molecular weight of the phytosterols, the cor- hol groups. On the other hand, some phytosterols seem to
responding weight fraction is considerably larger, although have a greater affinity for apolar VOCs and others for the
no combination manages to reach a 0.5 value either way. A polar ones.
few tendencies are observed, however. For example, the top
solubilities are predominantly reached by two specific phy- Global Analysis and NADES Selection
tosterols–cycloartenol and β-amyrin–while the lower end of
the values is quite populated by the two more dissimilar sol- In order to do a further analysis on a more large-scale basis
utes, saringosterol and daucosterol, with a polar residue and and considering a future industrial application, an additional
a whole sugar residue, respectively, so the solubility values parameter was calculated, which we will call the S:F ratio,
are mostly solute-driven. In these two later cases, the higher a representation of the mass of solute (S) extracted per mass
hydrophilicity explains the lower solubility in a hydrophobic unit of feed solvent (F). This amount will be used as an
solvent such as a NADES. Other than that, there have not extractive power value and can be calculated easily from the
been any other observed tendencies, unlike some observa- weight fraction w, defined as the mass ratio between solute
tions addressing the different solubilities between phytos- and solution, as follows:
terols and phytostanols (Översti et al., 2025) due to the dif-
S
ferent grade of unsaturation. However, the solubility within w= (1)
S+F
a NADES is a much more complex phenomenon due to the
Food and Bioprocess Technology (2025) 18:7646–7660 7653

Fig. 4  Molar fraction scatter

plot for all tested HBA/HBD
combinations, coloured as per
their molar fraction. Green, 0.45
< x < 0.55; yellow, 0.33 < x <
0.45 or 0.55 < x < 0.67; orange,
x < 0.33 or x > 0.67

The vapour pressure values confirm the very low vola-

w(S + F) = S (2)
tilities of the DES components, with none of them reach-
ing 1 hPa, except for cineole, matching its fragrant nature.
wF = S(1 − w) (3) The VOCs, on the other hand, as expected, show very high
vapour pressures, with the least volatile compound being
S
=
w
(4) nearly 30 times more than cineole. Therefore, even when the
F 1−w solvent is safe, considerable solvent losses by evaporation
For the solubility tests conducted for both the NADES are a risk to have in mind.
and the VOCs, we considered that a satisfactory separation The partition coefficients for all compounds are in line
was achieved when the solvent is able to extract at least 10% with their chemical structure, so it is not a surprise to find
of its weight in the process, thus selecting an S:F of 0.1 as a that the most bioaccumulable compounds are hydrocar-
threshold. Seventy-three of the NADES tests and 15 of the bons or fatty acids. As a norm, it can be considered that
VOC tests fulfilled this requirement. a compound accumulates when the partition coefficient is
Table 1 shows the toxicological parameters and number of 5 or higher (STOCKHOLM CONVENTION ON PERSIS-
successful extractions for every VOC and DES component–for TENT ORGANIC POLLUTANTS (POPS) TEXT AND
the latter case, only the components involved in at least three ANNEXES REVISED IN 2023, n.d.), so with this criterion,
successful extractions are shown. The full solubility table can all DES components are safe except the few fatty acids.
be found in the Supporting Information, in the Solubility Tests. The Cramer toxicological level also shows a simi-
xlsx file. The VOCs are very balanced in versatility, except for lar picture for both compound categories, with most of
the ethanol, which had no successful tests; this is understanda- them being on the low level and a few of the DES com-
ble, considering its polar nature clashes with the inherent hydro- ponents on the intermediate. Two compounds are on the
phobicity of PTS. On the DES side, the most prevalent com- high level, however, dichloromethane and cineole, which
pounds are menthol and carvacrol, appearing in nearly a third of is not surprising despite being a simple and structure-
the cases, with camphor and pulegone following behind. based method (Cramer et al., 1978). Being a halogenated
compound, dichloromethane is already a hazard for both
7654 Food and Bioprocess Technology (2025) 18:7646–7660

Table 1  Toxicological Compound Frequency Cramer category log ­KOW PVap (hPa)
parameters and number of
successful extractions for Menthol 27 Low (Class I) 3.3812 0.085
different compounds proposed
Carvacrol 25 Low (Class I) 3.4900 0.000
as HBA or HBD. Values in bold
are extracted from databases, Camphor 13 Intermediate (Class II) 2.3800 0.867
the rest calculated with QSAR Pulegone 10 Intermediate (Class II) 3.2028 0.164
models α-Bisabolol 8 Low (Class I) 5.6285 0.000
cis-Pinocarveol 7 Low (Class I) 2.8089 0.000
Linoleic acid 7 Low (Class I) 7.5144 0.000
Capric acid 6 Low (Class I) 4.0900 0.001
Cubebol 4 Low (Class I) 5.3162 0.000
Geraniol 4 Low (Class I) 3.4700 0.040
Palmitoleic acid 4 Low (Class I) 6.7472 0.000
Verbenone 4 Intermediate (Class II) 3.2119 0.000
Cineole 3 High (Class III) 2.7400 2.533
Menthone 3 Intermediate (Class II) 2.8698 0.360
Myristoleic acid 3 Low (Class I) 5.7650 0.000
Thymol 3 Low (Class I) 3.3000 0.021
Zingiberol 3 Low (Class I) 4.8847 0.000
Hexane 1 Low (Class I) 3.9000 202.000
Ethanol 2 Low (Class I) −0.3500 58.000
Dichloromethane 2 High (Class III) 1.2500 465.000
Pentane 2 Low (Class I) 3.3900 685.277
2-Methylpentane 2 Low (Class I) 3.2100 281.310
3-Methylpentane 2 Low (Class I) 3.6000 253.310
Isopropanol 2 Low (Class I) 0.0500 60.200
Isopentane 2 Low (Class I) 4.0000 918.590

the environment and human health (Cordes et al., 1988; however, the difference is clear: for all PTS, NADES consist-
Fagin et al., 1980; Kobayashi et al., 2008; Wells & Wal- ently reach higher extraction yields than the best VOC tested,
dron, 1984), and cineole, as many other monoterpenes and with some cases achieving more than five times better.
related ketones and ethers, are also prone to cause respira- Lastly, although a complete comparison is not possible given
tory issues or even delirium and dementia in high quanti- that not all NADES were tested for all PTS, these results are
ties (Burkhard et al., 1999; Woolf, 1999). consistent with former work on PTS solubility via NADES, as
Lastly, while we have considered the DES components seen in Resende et al. (2024), with solubilities ranging from
on their own so far, the main purpose of this study is to use 10 to 100 mg/mL, depending on the DES used, and consistent
them in a mixture. Table 2 shows the number of successful with the calculated average values from Table 4, further validat-
extractions for each DES that managed to do at least two, ing the accuracy of the computational approach employed. The
with their boiling point and vapour pressure, and, given extraction yields themselves are expected to be lower due to
that each DES is unique, this is also how many PTS they the complexity of the vegetal matrix and the presence of many
are suitable for. It can be seen that, with no doubt, the other compounds the NADES may interact with.
best combination is the carvacrol–menthol one; not only
can it extract more than half of the PTS considered, it is
also composed of two low-level compounds with very low Conclusions
volatilities. The DES itself has a low volatility as well.
However, the yield for each PTS can be analysed on The main objective of this research was to apply a systematic
its own as well, as can be seen in Table 3, where the best computational methodology to design NADES suitable for
NADES and VOC are displayed as well as their respective the efficient extraction of phytosterols from plant sources.
S:F ratio. The carvacrol–menthol appears many times, as An extensive analysis of nearly 2000 different DES was con-
seen before, and isopropanol and isopentane are also quite ducted, choosing only the best suitable in terms of operation
predominant on the VOC side. Comparing the S:F values, conditions and production costs.
Food and Bioprocess Technology (2025) 18:7646–7660 7655

Table 2  NADES with the highest number of different PTS success- Table 4  Comparison of PTS average volume solubility in mg/mL
fully extracted NADES and VOCs
NADES NPTS PVap (hPa) TB (ºC) PTS SNADES (mg/mL) SVOCs (mg/mL)
Carvacrol Menthol 10 0.086 224.87 α-Amyrin 36.95 32.12
Carvacrol cis-Pinocarveol 4 0.055 234.02 Avenasterol 2.50 1.08
α-Bisabolol Pulegone 3 0.075 263.56 β-Amyrin 109.85 96.78
Cineole Menthol 3 0.285 215.72 Brassicasterol 36.43 19.80
Linoleic acid Palmitoleic acid 2 0.000 344.00 Campesterol 28.13 19.64
Capric acid Cubebol 2 0.011 240.50 Cycloartenol 214.70 248.79
Capric acid Patchoulol 2 0.018 237.23 Δ−7-Stigmasterol 14.62 9.78
Carvacrol Piperitone 2 0.020 253.63 Daucosterol 0.00 0.00
α-Bisabolol Carvacrol 2 0.022 255.53 Ergosterol 23.09 12.92
Carvacrol Pulegone 2 0.044 241.24 γ-Sitosterol 31.20 16.47
α-Bisabolol Menthol 2 0.046 245.77 Gramisterol 12.63 9.01
Linoleic acid Menthol 2 0.050 249.05 Isofucosterol 53.07 31.59
Menthol Verbenone 2 0.051 242.57 Lanosterol 57.05 49.57
Menthol Palmitoleic acid 2 0.052 251.43 Lupeol 39.34 26.70
Carvacrol Menthone 2 0.073 236.33 Saringosterol 1.89 1.11
Menthol Pulegone 2 0.098 231.99 β-Sitoesterol 36.92 18.32
Camphor Carvacrol 2 0.116 231.97 Stigmastanol 36.69 17.39
Camphor Linoleic acid 2 0.117 259.02 Stigmasterol 16.06 10.97
Camphor Capric acid 2 0.143 222.55 Taraxerol 47.40 35.69
Camphor Zingiberol 2 0.151 252.53
Camphor Thymol 2 0.188 226.48
Geraniol Pulegone 2 0.232 204.38

Table 3  Comparison of S:F PTS NADES VOCs S:FNADES/VOC

values from the best NADES
and VOC for each of the PTS Solvents S:F Solvent S:F
tested, as well as the ratio
NADES:VOC α-Amyrin α-Bisabolol Pulegone 0.095 Isopentane 0.062 1.52
Avenasterol Carvacrol Menthol 0.010 Isopropanol 0.003 3.78
β-Amyrin Carvacrol Menthol 0.452 Isopentane 0.207 2.18
Brassicasterol Carvacrol Menthol 0.116 Isopropanol 0.044 2.63
Campesterol α-Bisabolol Pulegone 0.057 Isopentane 0.038 1.49
Cycloartenol Carvacrol Menthol 0.885 Isopentane 0.668 1.32
Δ−7-Stigmasterol α-Bisabolol Pulegone 0.032 Isopropanol 0.017 1.87
Daucosterol Carvacrol Menthol 0.000 Isopropanol 0.000 5.71
Ergosterol Carvacrol Menthol 0.075 Isopropanol 0.035 2.16
γ-Sitosterol Carvacrol Menthol 0.115 Isopropanol 0.030 3.78
Gramisterol Carvacrol Menthol 0.026 Isopentane 0.016 1.65
Isofucosterol Carvacrol Menthol 0.173 Isopropanol 0.065 2.66
Lanosterol α-Bisabolol Pulegone 0.162 Isopentane 0.101 1.60
Lupeol Carvacrol Menthol 0.181 Dichloromethane 0.063 2.87
Saringosterol Carvacrol Menthol 0.010 Isopropanol 0.005 1.99
β-Sitoesterol Carvacrol Menthol 0.125 Isopropanol 0.041 3.07
Stigmastanol Carvacrol Menthol 0.161 Isopentane 0.031 5.26
Stigmasterol Carvacrol Menthol 0.045 Isopropanol 0.028 1.64
Taraxerol Carvacrol Menthol 0.170 Isopentane 0.068 2.50
7656 Food and Bioprocess Technology (2025) 18:7646–7660

A similarly large amount of solubility tests was performed 101092347) and the Junta de Castilla y León (Spain, Project NADES-
through the COSMO-RS methodology, confirming the typi- forNature, Ref.: BU047P23). We also acknowledge SCAYLE (Super-
computación Castilla y León, Spain) for providing supercomputing
cally low solubility of phytosterols on usual conditions. A facilities. The statements made herein are solely the responsibility of
remarkably low solubility was seen for dauctosterol, whose the authors.
sugar residue impedes even more the dissolution in such a
hydrophobic medium. Data Availability Data is provided within the manuscript or supplemen-
tary information files. Additional information may be obtained from
From all NADES tested, the increased performance the corresponding author upon request.
upon comparison with the commonly used VOCs is clear,
increasing the solute extracted in a considerable amount Declarations
for all 19 phytosterols considered. The carvacrol–menthol
mixture appears as a highly versatile solvent, able to suc- Competing interests The authors declare no competing interests.
cessfully extract 10 of the 19 PTS, and it is the best option
for almost all of them. Its components are also non-volatile Open Access This article is licensed under a Creative Commons Attri-
bution 4.0 International License, which permits use, sharing, adapta-
and non-toxic, so it is also a safer alternative to commonly tion, distribution and reproduction in any medium or format, as long
used VOCs. as you give appropriate credit to the original author(s) and the source,
We conclude that the objectives of this study have been provide a link to the Creative Commons licence, and indicate if changes
achieved. Further research will follow, with experimental were made. The images or other third party material in this article are
included in the article’s Creative Commons licence, unless indicated
testing of these findings and their potential application in otherwise in a credit line to the material. If material is not included in
a more industrial setting, with more research on differ- the article’s Creative Commons licence and your intended use is not
ent approaches to isolate the phytosterols and recycle the permitted by statutory regulation or exceeds the permitted use, you will
DES. Given the extremely low vapour pressure observed in need to obtain permission directly from the copyright holder. To view a
copy of this licence, visit [Link]
the NADES compendium, traditional separation methods,
such as distillation, are discarded due to the high energetic
cost that would imply and the potential degradation of PTS
with high temperatures (Barriuso et al., 2012; Lin et al.,
References
2017). The low solubility of PTS, on the other hand, could
be exploited to attempt a separation by precipitation if the Abdel-Aal, E. I., Haroon, A. M., & Mofeed, J. (2015). Successive sol-
PTS solubility shows noticeable changes with temperature vent extraction and GC–MS analysis for the evaluation of the
(Kong et al., 2025; Översti et al., 2025). By combining an phytochemical constituents of the filamentous green algae Spi-
extraction unit at mild heating and a precipitation unit at rogyra longata. Egyptian Journal of Aquatic Research, 41(3),
233–246. [Link]
room temperature or cooler, a closed loop with NADES Abranches, D. O., Larriba, M., Silva, L. P., Melle-Franco, M., Palomar,
recirculation could be achieved. With this in mind, the J. F., Pinho, S. P., & Coutinho, J. A. P. (2019). Using COSMO-
next logical research steps would imply the temperature RS to design choline chloride pharmaceutical eutectic solvents.
dependence for PTS solubility for the carvacrol–menthol Fluid Phase Equilibria, 497, 71–78. [Link]
fluid.​2019.​06.​005
mixture, as well as small-scale experimental setups to con- Alvarez-Rivera, G., Bueno, M., Ballesteros-Vivas, D., Mendiola, J. A.,
firm the computational results. & Ibañez, E. (2020). Pressurized liquid extraction. In Liquid-
Phase Extraction (pp. 375–398). Elsevier. [Link]
Supplementary Information The online version contains supplemen- 1016/​B978-0-​12-​816911-​7.​00013-X
tary material available at [Link] oi.o​ rg/1​ 0.1​ 007/s​ 11947-0​ 25-0​ 3896-5. Amar, S., Becker, H. C., & Möllers, C. (2009a). Genetic variation in
phytosterol content of winter rapeseed ( Brassica napus L.) and
Author Contribution - María Antonieta Escobedo-Monge: Investiga- development of NIRS calibration equations. Plant Breeding,
tion, Writing – Original Draft, Writing – Review & Editing–Sergio 128(1), 78–83. [Link]
de-la-Huerta-Sainz: Software, Formal analysis, Investigation, Writing Arroyo-Avirama, A. F., Carreño-Guzmán, S., Lorenzo-Llanes, J.,
– Original Draft, Writing – Review & Editing, Visualization–Pedro A. Gajardo-Parra, N. F., Santiago, R., Held, C., Palomar, J., &
Marcos: Writing – Review & Editing–José Antonio Esteban – Ollo: Canales, R. I. (2023). In situ product recovery of β-ionone from
Conceptualization, Writing – Review & Editing–Laura Montejo-Gil: a fermentation broth: Computational solvent selection and pro-
Conceptualization, Writing – Review & Editing–María Conde-Rioll: cess design of its extraction and purification. ACS Sustainable
Conceptualization, Writing – Review & Editing–Mert Atilhan: Writing Chemistry & Engineering, 11(24), 9065–9076. [Link]
– Review & Editing–Alfredo Bol: Writing – Review & Editing–San- 10.​1021/​acssu​schem​eng.​3c017​39
tiago Aparicio: Conceptualization, Methodology, Supervision, Project Awad, A., Roy, R., & Fink, C. (2003). β-sitosterol, a plant sterol,
Administration, Funding Acquisition, Writing – Review & Editing. induces apoptosis and activates key caspases in MDA-MB-231
human breast cancer cells. Oncology Reports. [Link]
Funding Open access funding provided by FEDER European Funds 3892/​or.​10.2.​497
and the Junta de Castilla y León under the Research and Innovation Bae, H., Park, S., Yang, C., Song, G., & Lim, W. (2021). Disruption of
Strategy for Smart Specialization (RIS3) of Castilla y León 2021-2027. endoplasmic reticulum and ROS production in human ovarian
This work was funded by the European Union (Horizon Program, cancer by campesterol. Antioxidants, 10(3), 379. [Link]
Ref: HORIZON-CL4-2022-RESILIENCE-01-Convert2 green-GA 10.​3390/​antio​x1003​0379
Food and Bioprocess Technology (2025) 18:7646–7660 7657

Bai, G., Ma, C., & Chen, X. (2021). Phytosterols in edible oil: Distri- Bol, A., & Aparicio, S. (2025). Nature’s tool kit: Designing bio-
bution, analysis and variation during processing. Grain & Oil compatible and affordable NADES for sustainable extraction of
Science and Technology, 4(1), 33–44. [Link] plant bioactives. Sustainable Chemistry One World, 5, 100043.
gaost.​2020.​12.​003 [Link]
Balasubramani, S. G., Chen, G. P., Coriani, S., Diedenhofen, M., Frank, Deng, Y., Wang, X., Zhang, C., Xie, P., & Huang, L. (2025). Enhanced
M. S., Franzke, Y. J., Furche, F., Grotjahn, R., Harding, M. E., and green extraction of saponins from Gleditsia sinensis Lam.
Hättig, C., Hellweg, A., Helmich-Paris, B., Holzer, C., Huniar, Pods by ultrasound-assisted deep eutectic solvents: Optimi-
U., Kaupp, M., Marefat Khah, A., Karbalaei Khani, S., Müller, zation and comprehensive characterization. Food and Bio-
T., Mack, F., … Yu, J. M. (2020). TURBOMOLE: Modular pro- process Technology, 18(2), 1919–1938. [Link]
gram suite for ab initio quantum-chemical and condensed-matter s11947-​024-​03577-9
simulations. The Journal of Chemical Physics, 152(18). [Link] Dikshit, S., Bubna, S., Gupta, A., & Kumar, P. (2020). Advances in
doi.​org/​10.​1063/5.​00046​35 various techniques for isolation and purification of sterols. Jour-
Barriuso, B., Otaegui-Arrazola, A., Menéndez-Carreño, M., Astiasarán, I., nal of Food Science and Technology, 57(7), 2393–2403. [Link]
& Ansorena, D. (2012). Sterols heating: Degradation and formation doi.​org/​10.​1007/​s13197-​019-​04209-3
of their ring-structure polar oxidation products. Food Chemistry, Dziedzic, K., Górecka, D., Szwengiel, A., Sulewska, H., Kreft, I.,
135(2), 706–712. [Link] Gujska, E., & Walkowiak, J. (2018). The content of dietary fibre
Belwal, T., Pandey, A., Bhatt, I. D., & Rawal, R. S. (2020). Opti- and polyphenols in morphological parts of buckwheat (Fag-
mized microwave assisted extraction (MAE) of alkaloids and opyrum tataricum). Plant Foods for Human Nutrition, 73(1),
polyphenols from Berberis roots using multiple-component 82–88. [Link]
analysis. Scientific Reports, 10(1), 917. [Link] Eckert, F., & Klamt, A. (2002). Fast solvent screening via quantum
s41598-​020-​57585-8 chemistry: COSMO-RS approach. AIChE Journal, 48(2), 369–
BIOVIA COSMOtherm (Release 2021). (n.d.-b). Dassault Systèmes. 385. [Link]
Retrieved October 23, 2024, from [Link] European Chemicals Agency. (n.d.). www.​echa.​europa.​eu
BIOVIA COSMOtherm. (n.d.-a). Dassault Systèmes. [Link] Evtyugin, D. D., Evtuguin, D. V., Casal, S., & Domingues, M. R.
Blanco-Vaca, F., Cedó, L., & Julve, J. (2019). Phytosterols in cancer: (2023). Advances and challenges in plant sterol research: Fun-
From molecular mechanisms to preventive and therapeutic poten- damentals, analysis, applications and production. Molecules,
tialS. Current Medicinal Chemistry, 26(37), 6735–6749. [Link] 28(18), 6526. [Link]
doi.​org/​10.​2174/​09298​67325​66618​06070​93111 Fagin, J., Bradley, J., & Williams, D. (1980). Carbon monoxide poison-
Brufau, G., Canela, M. A., & Rafecas, M. (2008). Phytosterols: Physi- ing secondary to inhaling methylene chloride. BMJ, 281(6253),
ologic and metabolic aspects related to cholesterol-lowering 1461–1461. [Link]
properties. Nutrition Research, 28(4), 217–225. [Link] Feng, S., Belwal, T., Li, L., Limwachiranon, J., Liu, X., & Luo, Z.
10.​1016/j.​nutres.​2008.​02.​003 (2020). Phytosterols and their derivatives: Potential health-pro-
Burkhard, P. R., Burkhardt, K., Haenggeli, C.-A., Landis, T., Burkhard, moting uses against lipid metabolism and associated diseases,
P. R., Burkhardt, · K, Landis, · T, & Haenggeli, C.-A. (1999). mechanism, and safety issues. Comprehensive Reviews in Food
Plant-induced seizures: Reappearance of an old problem. In J Science and Food Safety, 19(4), 1243–1267. [Link]
Neurol (Vol. 246). 1111/​1541-​4337.​12560
Cannavacciuolo, C., Pagliari, S., Celano, R., Campone, L., & Rastrelli, Fernández-Cuesta, Á., Velasco, L., & Fernández-Martínez, J. M.
L. (2024). Critical analysis of green extraction techniques used (2011). Phytosterols in the seeds of wild sunflower species.
for botanicals: Trends, priorities, and optimization strategies- Helia, 34(55), 31–38. [Link]
A review. TrAC Trends in Analytical Chemistry, 173, 117627. Fernández-Cuesta, Á., Aguirre-González, M. R., Ruiz-Méndez, M. V.,
[Link] & Velasco, L. (2012). Validation of a method for the analysis of
Chagnoleau, J.-B., Papaiconomou, N., Fernandez, X., & Coutinho, phytosterols in sunflower seeds. European Journal of Lipid Sci-
J. A. P. (2024). Using COSMO-RS to design organic biphasic ence and Technology, 114(3), 325–331. [Link]
systems containing deep eutectic solvents for the separation of ejlt.​20110​0138
natural compounds. Journal of Molecular Liquids, 393, 123601. Fernando, T., & Bean, G. (1985). A comparison of the fatty acids
[Link] and sterols of seeds of weedy and vegetable species of Amaran-
ChemSpider. (n.d.). [Link] thus spp. Journal of the American Oil Chemists’ Society, 62(1),
Cordes, D. H., Brown, W. D., & Quinn, K. M. (1988). Chemically 89–91. [Link]
induced hepatitis after inhaling organic solvents. The Western Gajewski, M., Przybyå, J. L., Kosakowska, O., & Szymczak, P. (2009).
Journal of Medicine, 148(4), 458–460. Some factors influencing free sterols content in broccoli (Bras-
Costa, J., Amaral, J. S., Mafra, I., & Oliveira, M. B. P. P. (2011). Refining sica oleracea L. var. botrytis italica plenck.). Journal of Food
of Roundup Ready® soya bean oil: Effect on the fatty acid, phytos- Biochemistry, 33(6), 881–894. [Link]
terol and tocopherol profiles. European Journal of Lipid Science and 4514.​2009.​00262.x
Technology, 113(4), 528–535. [Link] Garcia-Llatas, G., Alegría, A., Barberá, R., & Cilla, A. (2021). Current
Cramer, G. M., Ford, R. A., & Hall, R. L. (1978). Review Section methodologies for phytosterol analysis in foods. Microchemical
Estimation of toxic hazard-a decision tree approach (Vol. 16). Journal, 168, 106377. [Link]
Pergamon Press. Gaudin, T., & Aubry, J.-M. (2025). Driving the future of cosmetics,
de Oliveira, I. L., Strieder, M. M., Bragagnolo, F. S., Sanches, V. L., de fragrances, and foods with COSMO-RS. Part 1—Bibliometric
Souza Mesquita, L. M., Viganó, J., & Rostagno, M. A. (2025). analysis and introductory framework. Current Opinion in Col-
Selective separation of α-punicalagin, β-punicalagin, and ellagic loid & Interface Science, 75, 101874. [Link]
acid from pomegranate husk aqueous extract by an optimized cocis.​2024.​101874
adsorption process employing a (deep) eutectic solvent. Food Gowda, N. A. N., Gurikar, C., Anusha, M. B., & Gupta, S. (2022).
and Bioprocess Technology, 18(3), 2519–2530. [Link] Ultrasound-assisted and microwave-assisted extraction, GC-MS
10.​1007/​s11947-​024-​03608-5 characterization and antimicrobial potential of Freeze-dried L.
de-la-Huerta-Sainz, S., Escobedo-Monge, M. A., Marcos, P. A., Este- camara flower. Journal of Pure and Applied Microbiology, 16(1),
ban-Ollo, J. A., Montejo-Gil, L., Conde-Rioll, M., Atilhan, M., 526–539. [Link]
7658 Food and Bioprocess Technology (2025) 18:7646–7660

Grosso, C., Valentão, P., Ferreres, F., & Andrade, P. (2015). Alternative and & Matsumura, M. (2008). Severe optic neuropathy caused by
efficient extraction methods for marine-derived compounds. Marine dichloromethane inhalation. Journal of Ocular Pharmacology
Drugs, 13(5), 3182–3230. [Link] and Therapeutics, 24(6), 607–612. [Link]
Gupta, A., Naraniwal, M., & Kothari, V. (2012). Modern extraction 2007.​0100
methods for preparation of bioactive plant extracts. International Kong, F., Lin, Z., Zhang, Y., Suo, H., Yan, L., & Zou, B. (2025). Effi-
Journal of Applied and Natural Sciences. cient cooling crystallization of phytosterol assisted by solubil-
Han, J.-H., Yang, Y.-X., & Feng, M.-Y. (2008). Contents of phytos- ity and thermodynamics analysis in organic solvents. Journal
terols in vegetables and fruits commonly consumed in China. of Molecular Liquids, 420, 126840. [Link]
Biomedical and Environmental Sciences, 21(6), 449–453. [Link] molliq.​2024.​126840
doi.​org/​10.​1016/​S0895-​3988(09)​60001-5 Kozłowska, M., Gruczyńska, E., Ścibisz, I., & Rudzińska, M. (2016).
Hansen, B. B., Spittle, S., Chen, B., Poe, D., Zhang, Y., Klein, J. M., Fatty acids and sterols composition, and antioxidant activity of
Horton, A., Adhikari, L., Zelovich, T., Doherty, B. W., Gurkan, oils extracted from plant seeds. Food Chemistry, 213, 450–456.
B., Maginn, E. J., Ragauskas, A., Dadmun, M., Zawodzinski, T. [Link]
A., Baker, G. A., Tuckerman, M. E., Savinell, R. F., & Sangoro, Kuasnei, M., Wojeicchowski, J. P., Santos, N. H., Pinto, V. Z., Ferreira,
J. R. (2021). Deep eutectic solvents: A review of fundamentals S. R. S., & Zielinski, A. A. F. (2022). Modifiers based on deep
and applications. Chemical Reviews, 121(3), 1232–1285. [Link] eutectic mixtures: A case study for the extraction of anthocyanins
doi.​org/​10.​1021/​acs.​chemr​ev.​0c003​85 from black bean hulls using high pressure fluid technology. The
Haynes, W. M. (Ed.). (2014). CRC Handbook of chemistry and physics. Journal of Supercritical Fluids, 191, Article 105761. [Link]
CRC Press. [Link] org/​10.​1016/j.​supflu.​2022.​105761
Hernández-Ledesma, B. (2019a). Quinoa (Chenopodium quinoa Lagarda, M. J., García-Llatas, G., & Farré, R. (2006). Analysis of phy-
Willd.) as source of bioactive compounds: A review. Bioactive tosterols in foods. Journal of Pharmaceutical and Biomedical
Compounds in Health and Disease, 2(3), 27. [Link] Analysis, 41(5), 1486–1496. [Link]
31989/​bchd.​v2i3.​556 02.​052
Hrabovski, N., Sinadinović-Fišer, S., Nikolovski, B., Sovilj, M., & Lin, Y., Knol, D., Valk, I., van Andel, V., Friedrichs, S., Lütjohann,
Borota, O. (2012). Phytosterols in pumpkin seed oil extracted D., Hrncirik, K., & Trautwein, E. A. (2017). Thermal stability
by organic solvents and supercritical CO 2. European Journal of of plant sterols and formation of their oxidation products in veg-
Lipid Science and Technology, 114(10), 1204–1211. [Link] etable oils and margarines upon controlled heating. Chemistry
org/​10.​1002/​ejlt.​20120​0009 and Physics of Lipids, 207(1), 99–107. [Link]
Jansen, P. J., Lütjohann, D., Abildayeva, K., Vanmierlo, T., Plösch, T., chemp​hyslip.​2017.​01.​007
Plat, J., von Bergmann, K., Groen, A. K., Ramaekers, F. C. S., Lulei, M. (2008). REACH: Zu den Leitlinien der Europäischen Agen-
Kuipers, F., & Mulder, M. (2006). Dietary plant sterols accumu- tur für chemische Stoffe (ECHA)1. Bundesgesundheitsblatt -
late in the brain. Biochimica et Biophysica Acta (BBA) - Molecu- Gesundheitsforschung - Gesundheitsschutz, 51(12), 1444–1452.
lar and Cell Biology of Lipids, 1761(4), 445–453. [Link] oi.o​ rg/​ [Link]
10.​1016/j.​bbalip.​2006.​03.​015 Ly, L., & Sothornvit, R. (2024). Novel sequential microwave-ultra-
Jha, A. K., & Sit, N. (2022). Extraction of bioactive compounds from sonic-assisted extraction of phenolic compounds from jack-
plant materials using combination of various novel methods: A fruit (Artocarpus heterophyllus Lam.) Peel waste using choline
review. Trends in Food Science & Technology, 119, 579–591. chloride-lactic acid deep eutectic solvent. Food and Bioprocess
[Link] Technology, 17(11), 4249–4261. https:// ​ d oi. ​ o rg/ ​ 1 0. ​ 1 007/​
Joback, K. G., & Reid, R. C. (1987). Estimation of pure-component s11947-​024-​03393-1
properties from group-contributions. Chemical Engineering Makris, D. P., & Lalas, S. (2020). Glycerol and glycerol-based deep
Communications, 57(1–6), 233–243. [Link] eutectic mixtures as emerging green solvents for polyphenol
00986​44870​89604​87 extraction: The evidence so far. Molecules, 25(24), 5842. [Link]
Kaderides, K., Papaoikonomou, L., Serafim, M., & Goula, A. M. doi.​org/​10.​3390/​molec​ules2​52458​42
(2019). Microwave-assisted extraction of phenolics from pome- Marangoni, F., & Poli, A. (2010). Phytosterols and cardiovascular
granate peels: Optimization, kinetics, and comparison with health. Pharmacological Research, 61(3), 193–199. [Link]
ultrasounds extraction. Chemical Engineering and Processing org/​10.​1016/j.​phrs.​2010.​01.​001
- Process Intensification, 137, 1–11. [Link] Marcone, M. F., Kakuda, Y., & Yada, R. Y. (2003). Amaranth as a rich
cep.​2019.​01.​006 dietary source of β-sitosterol and other phytosterols. Plant Foods
Kaur, D., & Qadri, O. S. (2025). Ultrasound-assisted extraction of for Human Nutrition, 58(3), 207–211. [Link] oi.o​ rg/1​ 0.1​ 023/B:​
Syzygium cumini anthocyanins using natural deep eutectic QUAL.​00000​40334.​99070.​3e
solvents. Food and Bioprocess Technology, 18(2), 1821–1833. Morand, C., & Tomás-Barberán, F. A. (2019). Contribution of plant
[Link] food bioactives in promoting health effects of plant foods: Why
Kim, S., Chen, J., Cheng, T., Gindulyte, A., He, J., He, S., Li, Q., look at interindividual variability? European Journal of Nutri-
Shoemaker, B. A., Thiessen, P. A., Yu, B., Zaslavsky, L., Zhang, tion, 58(S2), 13–19. [Link] oi.o​ rg/1​ 0.1​ 007/s​ 00394-0​ 19-0​ 2096-0
J., & Bolton, E. E. (2023). PubChem 2023 update. Nucleic Acids Moreau, R. A., Nyström, L., Whitaker, B. D., Winkler-Moser, J. K., Baer,
Research, 51(D1), D1373–D1380. [Link] D. J., Gebauer, S. K., & Hicks, K. B. (2018). Phytosterols and their
gkac9​56 derivatives: Structural diversity, distribution, metabolism, analysis,
Klamt, A. (1995). Conductor-like screening model for real solvents: and health-promoting uses. Progress in Lipid Research, 70, 35–61.
A new approach to the quantitative calculation of solvation phe- [Link]
nomena. The Journal of Physical Chemistry, 99(7), 2224–2235. Moreno, D., Gonzalez-Miquel, M., Ferro, V. R., & Palomar, J. (2018).
[Link] Molecular and thermodynamic properties of zwitterions ver-
Klamt, A., Jonas, V., Bu, T., & Lohrenz, J. C. W. (1998). Refinement sus ionic liquids: A comprehensive computational analysis to
and Parametrization of COSMO-RS. [Link] develop advanced separation processes. ChemPhysChem, 19(7),
nggui​delin​es 801–815. [Link]
Kobayashi, A., Ando, A., Tagami, N., Kitagawa, M., Kawai, E., MS, U., Ferdosh, S., Haque Akanda, Md. J., Ghafoor, K., A. H., R.,
Akioka, M., Arai, E., Nakatani, T., Nakano, S., Matsui, Y., Ali, Md. E., Kamaruzzaman, B. Y., M. B., F., S., H., Shaarani,
Food and Bioprocess Technology (2025) 18:7646–7660 7659

S., & Islam Sarker, Md. Z. (2018). Techniques for the extraction Péres, V. F., Saffi, J., Melecchi, M. I. S., Abad, F. C., de Assis Jacques,
of phytosterols and their benefits in human health: A review. R., Martinez, M. M., Oliveira, E. C., & Caramão, E. B. (2006).
Separation Science and Technology, 53(14), 2206–2223. [Link] Comparison of soxhlet, ultrasound-assisted and pressurized
doi.​org/​10.​1080/​01496​395.​2018.​14544​72 liquid extraction of terpenes, fatty acids and Vitamin E from
Müller, C., & Bracher, F. (2015). Determination by GC-IT/MS of phytos- Piper gaudichaudianum Kunth. Journal of Chromatography A,
terols in herbal medicinal products for the treatment of lower uri- 1105(1–2), 115–118. [Link]
nary tract symptoms and food products marketed in Europe. Planta 113
Medica, 81(07), 613–620. [Link] Piironen, V., Lindsay, D. G., Miettinen, T. A., Toivo, J., & Lampi,
Mustapa, A. N., Martin, Á., Mato, R. B., & Cocero, M. J. (2015). Extrac- A.-M. (2000). Plant sterols: Biosynthesis, biological function
tion of phytocompounds from the medicinal plant Clinacanthus and their importance to human nutrition. Journal of the Science
nutans Lindau by microwave-assisted extraction and supercriti- of Food and Agriculture, 80(7), 939–966. [Link] oi.o​ rg/1​ 0.1​ 002/​
cal carbon dioxide extraction. Industrial Crops and Products, 74, (SICI)1​ 097-0​ 010(200005​ 15)8​ 0:7%3​ c939::A ​ ID-J​ SFA64​ 4%3​ e3.0.​
83–94. [Link] CO;2-C
Nashed, B., Yeganeh, B., HayGlass, K. T., & Moghadasian, M. H. Plat, J., Baumgartner, S., Vanmierlo, T., Lütjohann, D., Calkins, K. L.,
(2005). Antiatherogenic effects of dietary plant sterols are associ- Burrin, D. G., Guthrie, G., Thijs, C., Te Velde, A. A., Vreugden-
ated with inhibition of proinflammatory cytokine production in hil, A. C. E., Sverdlov, R., Garssen, J., Wouters, K., Trautwein,
Apo E-KO mice. The Journal of Nutrition, 135(10), 2438–2444. E. A., Wolfs, T. G., van Gorp, C., Mulder, M. T., Riksen, N. P.,
[Link] Groen, A. K., & Mensink, R. P. (2019). Plant-based sterols and
National Institute of Standards and Technology. (n.d.). [Link] stanols in health & disease: “Consequences of human develop-
ok.​nist.​gov/​chemi​stry/ ment in a plant-based environment?” Progress in Lipid Research,
Nattagh-Eshtivani, E., Barghchi, H., Pahlavani, N., Barati, M., Amiri, 74, 87–102. [Link]
Y., Fadel, A., Khosravi, M., Talebi, S., Arzhang, P., Ziaei, R., QSAR Toolbox (4.7). (n.d.). OECD.
& Ghavami, A. (2022). Biological and pharmacological effects Raguindin, P. F., Adam Itodo, O., Stoyanov, J., Dejanovic, G. M.,
and nutritional impact of phytosterols: A comprehensive review. Gamba, M., Asllanaj, E., Minder, B., Bussler, W., Metzger, B.,
Phytotherapy Research, 36(1), 299–322. [Link] oi.o​ rg/1​ 0.1​ 002/​ Muka, T., Glisic, M., & Kern, H. (2021a). A systematic review
ptr.​7312 of phytochemicals in oat and buckwheat. Food Chemistry, 338,
Nes, W. D. (2011). Biosynthesis of cholesterol and other sterols. Chem- 127982. [Link]
ical Reviews, 111(10), 6423–6451. [Link] oi.o​ rg/1​ 0.1​ 021/c​ r200​ Resende, J., Sosa, F. H. B., Coutinho, J. A. P., Rocha, J., Silvestre, A. J.
021m D., & Santos, S. A. O. (2024). Sustainable phytosterol extraction
Nieto, A., Borrull, F., Pocurull, E., & Marcé, R. M. (2010). Pressurized from codium tomentosum using eutectic solvents. ACS Sustain-
liquid extraction: A useful technique to extract pharmaceuticals able Chemistry & Engineering, 12(24), 9037–9044. [Link]
and personal-care products from sewage sludge. TrAC Trends in org/​10.​1021/​acssu​schem​eng.​4c001​45
Analytical Chemistry, 29(7), 752–764. [Link] Roche, Y., Gerbeau-Pissot, P., Buhot, B., Thomas, D., Bonneau, L.,
trac.​2010.​03.​014 Gresti, J., Mongrand, S., Perrier-Cornet, J., & Simon-Plas, F.
Norhidzam, M. S., Kai, C. W., Hong, S. L., Chuah, J. H., Yusoff, R., (2008). Depletion of phytosterols from the plant plasma membrane
Goh, B. H., & Teoh, W. H. (2025). The Design of natural deep provides evidence for disruption of lipid rafts. The FASEB Journal,
eutectic solvent for the extraction of acetyl-11-keto-β-boswellic 22(11), 3980–3991. [Link]
acid using COSMO-RS. Microchemical Journal, 208, 112425. Ryan, E., Galvin, K., O’Connor, T. P., Maguire, A. R., & O’Brien, N.
[Link] M. (2007). Phytosterol, squalene, tocopherol content and fatty
Nyam, K. L., Tan, C. P., Lai, O. M., Long, K., & Che Man, Y. B. acid profile of selected seeds, grains, and legumes. Plant Foods
(2010). Optimization of supercritical fluid extraction of phytos- for Human Nutrition, 62(3), 85–91. [Link]
terol from roselle seeds with a central composite design model. s11130-​007-​0046-8
Food and Bioproducts Processing, 88(2–3), 239–246. [Link] oi.​ Santiago, R., Díaz, I., González-Miquel, M., Navarro, P., & Palomar, J.
org/​10.​1016/j.​fbp.​2009.​11.​002 (2022). Assessment of bio-ionic liquids as promising solvents in
Ogrodowska, D., Zadernowski, R., Czaplicki, S., Derewiaka, D., & industrial separation processes: Computational screening using
Wronowska, B. (2014). Amaranth seeds and products – The COSMO-RS method. Fluid Phase Equilibria, 560, 113495.
source of bioactive compounds. Polish Journal of Food and [Link]
Nutrition Sciences, 64(3), 165–170. [Link] Santos, P. H., Kammers, J. C., Silva, A. P., Oliveira, J. V., & Hense, H.
v10222-​012-​0095-z (2021). Antioxidant and antibacterial compounds from feijoa leaf
Oliveira, G., Farias, F. O., Sosa, F. H. B., Igarashi-Mafra, L., & Mafra, extracts obtained by pressurized liquid extraction and supercriti-
M. R. (2021). Green solvents to tune the biomolecules’ solubili- cal fluid extraction. Food Chemistry, 344, 128620. [Link]
zation in aqueous media: An experimental and in silico approach org/​10.​1016/j.​foodc​hem.​2020.​128620
by COSMO-RS. Journal of Molecular Liquids, 341, 117314. Serna-Vázquez, J., Ahmad, M. Z., Boczkaj, G., & Castro-Muñoz, R. (2021).
[Link] Latest insights on novel deep eutectic solvents (DES) for sustainable
Ostlund, R. E. (2002). Phytosterols in human nutrition. Annual Review extraction of phenolic compounds from natural sources. Molecules,
of Nutrition, 22(1), 533–549. [Link] 26(16), 5037. [Link]
nutr.​22.​020702.​075220 Setyaningsih, W., Saputro, I. E., Palma, M., & Barroso, C. G. (2016).
Othman, R. A., Myrie, S. B., & Jones, P. J. H. (2013). Non-cholesterol Pressurized liquid extraction of phenolic compounds from rice
sterols and cholesterol metabolism in sitosterolemia. Atheroscle- (Oryza sativa) grains. Food Chemistry, 192, 452–459. [Link] oi.​
rosis, 231(2), 291–299. [Link] org/​10.​1016/j.​foodc​hem.​2015.​06.​102
2013.​09.​038 Shao, Y., Hu, Z., Liu, C., Xu, Q., Zhang, H., Yan, Q., Zhu, D., & Zhu, Z.
Översti, P., Han, B., Kavakka, J., Torssell, S., Tirronen, E., Louhi- (2021). Phenolic acids and phytosterols in rice grains and wheat
Kultanen, M., & Oinas, P. (2025). The effect of water on the crys- flours consumed in five regions of China. Journal of Food Science,
tallization of phytosterols and phytostanols in organic solutions. 86(5), 1878–1892. [Link]
European Journal of Pharmaceutical Sciences, 204, 106956. Shi, Y., Ma, Y., Zhang, R., Ma, H., & Liu, B. (2015). Preparation
[Link] and characterization of foxtail millet bran oil using subcritical
7660 Food and Bioprocess Technology (2025) 18:7646–7660

propane and supercritical carbon dioxide extraction. Journal of Global Advanced Research Journal of Food Science and Tech-
Food Science and Technology, 52(5), 3099–3104. [Link] oi.o​ rg/​ nology, 2(4), 44–53.
10.​1007/​s13197-​014-​1311-0 Villacrés, E., Pástor, G., Quelal, M. B., Zambrano Villavicencio, I., &
Shin, E.-C., Pegg, R. B., Phillips, R. D., & Eitenmiller, R. R. (2010). Morales, S. H. (2013b). Effect of processing on the content of
Commercial peanut (Arachis hypogaea L.) cultivars in the United fatty acids, tocopherols and sterols in the oils of quinoa (Cheno-
States: Phytosterol composition. Journal of Agricultural and Food podium quinoa Willd), lupine (Lupinus mutabilis Sweet), ama-
Chemistry, 58(16), 9137–9146. [Link] ranth (Amaranthus caudatus L.) and sangorache (Amaranthus
Sierksma, A., Weststrate, J. A., & Meijer, G. W. (1999). Spreads quitensis L.). Global Advanced Research Journal of Food Sci-
enriched with plant sterols, either esterified 4,4-dimethylsterols ence and Technology, 2(4), 044–053.
or free 4-desmethylsterols, and plasma total- and LDL-choles- Wang, D., Zhang, M., Law, C. L., & Zhang, L. (2024). Natural deep
terol concentrations. British Journal of Nutrition, 82(4), 273– eutectic solvents for the extraction of lentinan from shiitake
282. [Link] mushroom: COSMO-RS screening and ANN-GA optimizing
Singh, V., Moreau, R. A., & Hicks, K. B. (2003). Yield and phytosterol conditions. Food Chemistry, 430, Article 136990. [Link]
composition of oil extracted from grain sorghum and its wet- org/​10.​1016/j.​foodc​hem.​2023.​136990
milled fractions. Cereal Chemistry, 80(2), 126–129. [Link] Wang, J., Du, S., Li, H., Wang, S., & Ling, B. (2025). Efficient extrac-
org/​10.​1094/​CCHEM.​2003.​80.2.​126 tion and characterization of pectin from pomelo peel by sequen-
Singh, L., Singh, B., & Bhatt, I. D. (2024). NADES-based extraction tial ultrasonic and radio frequency treatment. Food and Bio-
optimization and enrichment of cyanidin-3-O-galactoside from process Technology, 18(2), 1431–1445. [Link]
Rhododendron arboreum Sm.: Kinetics and thermodynamics s11947-​024-​03538-2
insights. Food Chemistry, 455, 139793. [Link] oi.o​ rg/1​ 0.1​ 016/j.​ Wells, G. G., & Waldron, H. A. (1984). Methylene chloride burns.
foodc​hem.​2024.​139793 Occupational and Environmental Medicine, 41(3), 420–420.
Soquetta, M. B., de Terra, L. M., & Bastos, C. P. (2018). Green tech- [Link]
nologies for the extraction of bioactive compounds in fruits and Witter, S., Witter, R., Vilu, R., & Samoson, A. (2018). Medical plants
vegetables. CyTA - Journal of Food, 16(1), 400–412. [Link] and nutraceuticals for amyloid-β fibrillation inhibition. Journal
org/​10.​1080/​19476​337.​2017.​14119​78 of Alzheimer’s Disease Reports, 2(1), 239–252. [Link]
Sperotto, F., Yang, J., Isom, L., Weller, C., & Ciftci, O. N. (2022). 10.​3233/​ADR-​180066
Supercritical carbon dioxide extraction, purification, and char- Woolf, A. (1999). Essential oil poisoning. In Clinical Toxicology (Vol.
acterization of wax from sorghum and sorghum by-products as 37, Issue 6). www.​dekker.​com
an alternative natural wax. Journal of the American Oil Chem- Yan, Y., Niu, Z., Wang, B., Zhao, S., Sun, C., Wu, Y., Li, Y., Ying,
ists’ Society, 99(5), 433–441. [Link] oi.o​ rg/1​ 0.1​ 002/a​ ocs.1​ 2575 H., & Liu, H. (2021). Saringosterol from Sargassum fusiforme
STOCKHOLM CONVENTION ON PERSISTENT ORGANIC POLLUT- modulates cholesterol metabolism and alleviates atherosclerosis
ANTS (POPS) TEXT AND ANNEXES REVISED IN 2023. (n.d.). in ApoE-deficient mice. Marine Drugs, 19(9), 485. [Link]
Švarc-Gajić, J., & Morais, S. (2022). Recent advances in the devel- org/​10.​3390/​md190​90485
opment and application of green extraction techniques. Applied Younas, R., Sahar, A., Sameen, A., Khan, M. I., Ali, M. A., Tahir,
Sciences, 12(20), 10510. [Link] M. A., Mohsin, M., Usman, M., & Aadil, R. M. (2024). A
Tolcha, T., Gemechu, T., Al-Hamimi, S., Megersa, N., & Turner, C. narrative review on extraction techniques of phytosterols and
(2021). Multivariate optimization of a combined static and their applications in food industry. Biomass Conversion and
dynamic supercritical fluid extraction method for trace analysis of Biorefinery, 14(24), 30897–30911. [Link]
pesticides pollutants in organic honey. Journal of Separation Sci- s13399-​023-​05007-w
ence, 44(8), 1716–1726. [Link] Yuan, C., Xie, Y., Jin, R., Ren, L., Zhou, L., Zhu, M., & Ju, Y. (2017).
Trautwein, E. A., Vermeer, M. A., Hiemstra, H., & Ras, R. T. (2018). Simultaneous analysis of tocopherols, phytosterols, and squalene
LDL-cholesterol lowering of plant sterols and stanols—Which in vegetable oils by high-performance liquid chromatography.
factors influence their efficacy? Nutrients, 10(9), 1262. [Link] Food Analytical Methods, 10(11), 3716–3722. [Link]
doi.​org/​10.​3390/​nu100​91262 10.​1007/​s12161-​017-​0927-x
Tuhanioglu, A., & Ubeyitogullari, A. (2022). Extraction of high-value Zangenberg, M., Hansen, H. B., Jørgensen, J. R., & Hellgren, L.
lipids and phenolic compounds from sorghum bran via a sequen- I. (2004a). Cultivar and year-to-year variation of phytosterol
tial supercritical carbon dioxide approach. ACS Food Science content in rye (Secale cereale L.). Journal of Agricultural and
& Technology, 2(12), 1879–1887. [Link] Food Chemistry, 52(9), 2593–2597. [Link]
odsci​tech.​2c002​66 jf035​1873
TURBOMOLE V7.8 2023. (n.d.). University of Karlsruhe, Forschung- Zhang, X., Lin, K., & Li, Y. (2020). Highlights to phytosterols accu-
szentrum Karlsruhe GmbH. mulation and equilibrium in plants: Biosynthetic pathway and
Verleyen, T., Sosinska, U., Ioannidou, S., Verhe, R., Dewettinck, K., feedback regulation. Plant Physiology and Biochemistry, 155,
Huyghebaert, A., & De Greyt, W. (2002). Influence of the veg- 637–649. [Link]
etable oil refining process on free and esterified sterols. Journal Zhou, M., Fakayode, O. A., & Li, H. (2023). Green extraction of poly-
of the American Oil Chemists’ Society, 79(10), 947–953. [Link] phenols via deep eutectic solvents and assisted technologies from
doi.​org/​10.​1007/​s11746-​002-​0585-4 agri-food by-products. Molecules, 28(19), 6852. [Link]
Vilela, C., Santos, S. A. O., Oliveira, L., Camacho, J. F., Cordeiro, 10.​3390/​molec​ules2​81968​52
N., Freire, C. S. R., & Silvestre, A. J. D. (2013). The ripe pulp Zou, R., Zhou, X., Qian, M., Wang, C., Boldor, D., Lei, H., & Zhang,
of Mangifera indica L.: A rich source of phytosterols and other X. (2024). Advancements and applications of microwave-assisted
lipophilic phytochemicals. Food Research International, 54(2), deep eutectic solvent (MW-DES) lignin extraction: A compre-
1535–1540. [Link] hensive review. Green Chemistry, 26(3), 1153–1169. [Link]
Villacrés, E., Pástor, G., Quelal, M. B., Villavicencio, I. Z., & Morales, org/​10.​1039/​D3GC0​4501C
S. H. (2013a). Effect of processing on the content of fatty acids,
tocopherols and sterols in the oils of quinoa (Chenopodium qui-
noa Willd), lupine (Lupinus mutabilis Sweet), amaranth (Ama- Publisher's Note Springer Nature remains neutral with regard to
ranthus caudatus L.) and sangorache (Amaranthus quitensis L.). jurisdictional claims in published maps and institutional affiliations.