Molecules 28 05344

molecules
Review
Enhancing Health Benefits through Chlorophylls and
Chlorophyll-Rich Agro-Food: A Comprehensive Review
Tânia Martins 1,2,† , Ana Novo Barros 1,2, *,† , Eduardo Rosa 1,2 and Luís Antunes 1,2
1 Centre for the Research and Technology of Agro-Environmental and Biological Sciences,
University of Trás-os-Montes and Alto Douro (CITAB), 5000-801 Vila Real, Portugal; taniam@utad.pt (T.M.);
erosa@utad.pt (E.R.); labanimalsgroup@gmail.com (L.A.)
2 Institute for Innovation, Capacity Building and Sustainability of Agri-Food Production (Inov4Agro),
5000-801 Vila Real, Portugal
* Correspondence: abarros@utad.pt
† These authors contributed equally to this work.
Abstract: Chlorophylls play a crucial role in photosynthesis and are abundantly found in green fruits
and vegetables that form an integral part of our diet. Although limited, existing studies suggest
that these photosynthetic pigments and their derivatives possess therapeutic properties. These
bioactive molecules exhibit a wide range of beneficial effects, including antioxidant, antimutagenic,
antigenotoxic, anti-cancer, and anti-obesogenic activities. However, it is unfortunate that leafy
materials and fruit peels often go to waste in the food supply chain, contributing to the prevailing
issue of food waste in modern societies. Nevertheless, these overlooked materials contain valuable
bioactive compounds, including chlorophylls, which offer significant health benefits. Consequently,
exploring the potential of these discarded resources, such as utilizing them as functional food
ingredients, aligns with the principles of a circular economy and presents exciting opportunities
for exploitation.
Keywords: chlorophylls; chlorophyllin; health; biological activity

Citation: Martins, T.; Barros, A.N.;
Rosa, E.; Antunes, L. Enhancing
Health Benefits through Chlorophylls
and Chlorophyll-Rich Agro-Food: A
Comprehensive Review. Molecules
1. Introduction
2023, 28, 5344. https://doi.org/ Chlorophyll, Figure 1, a complex green pigment found in plants, algae, and certain
10.3390/molecules28145344 bacteria, plays a crucial role in the process of photosynthesis by absorbing light energy and
converting it into chemical energy [1]. While early beliefs about the bioavailability and
Academic Editors: Alexandra
stability of chlorophyll under differaent conditions limited research on its health effects, re-
Christine Helena F. Sawaya, Maria
Cristina Marcucci and
cent studies have shed light on the potential benefits of chlorophyllin as a chemopreventive
Riccardo Petrelli
agent [2]. Nagini et al. have provided insights into its molecular mechanisms [2]. Although
in vitro and in vivo studies suggest its anticancer effects, evidence of its efficacy in humans
Received: 3 June 2023 remains scarce [2]. Dietary supplements containing chlorophyll and chlorophyllin are
Revised: 23 June 2023 available and generally considered safe, with no reported adverse side effects over several
Accepted: 10 July 2023
decades of human use [3]. However, skepticism about their effectiveness persists due to
Published: 11 July 2023
the lack of robust scientific evidence supporting their claimed health benefits [3].
Despite the potential health benefits associated with chlorophyll, a significant number
of chlorophyll-rich vegetables, leafy materials, and fruits are lost throughout the food
Copyright: © 2023 by the authors.
supply chain [4]. This loss occurs despite the underutilized potential of these agro-food
Licensee MDPI, Basel, Switzerland.
residues [5,6]. Harnessing and utilizing this discarded material could contribute to the
This article is an open access article transition towards a more sustainable circular economy.
distributed under the terms and Due to the poor bioavailability and stability of chlorophylls, studies on chlorophylls
conditions of the Creative Commons are scarce until now. In summary, the main purpose of this review is to provide a com-
Attribution (CC BY) license (https:// prehensive understanding of the current knowledge on chlorophylls and their potential
creativecommons.org/licenses/by/ applications in the context of bioavailability, stability, antioxidant activity, and antimuta-
4.0/). genic properties. The results and conclusions can be applied by readers and researchers to
Molecules 2023, 28, 5344. https://doi.org/10.3390/molecules28145344 https://www.mdpi.com/journal/molecules

Molecules 2023, 28, 5344 2 of 21
Molecules 2023, 28, x FOR PEER REVIEW 2 of 21
deepen their knowledge, guide future studies, explore applications in food and pharma-
ceutical industries, and stimulate further research in this field.
Figure 1. Structure from Chlorophyll a and Chlorophyll b.

Figure 1. Structure from Chlorophyll a and Chlorophyll b.
2. Chlorophyll: Chemical Properties and Metabolism
Due to the poor
Chlorophyll is abioavailability
complex molecule and stability
made upofofchlorophylls,
a porphyrinstudies
ring, a on chlorophylls
magnesium ion,
are scarce until now. In summary, the main purpose of this review
and an attached hydrocarbon tail. The porphyrin ring is responsible for absorbing is to provide a
light
comprehensive understanding of the current knowledge on chlorophylls
energy and the magnesium ion acts as an electron acceptor. Chlorophyll has many forms and their
potential applications
such as chlorophyll a, in the context
chlorophyll of bioavailability,
b, chlorophyll stability,d antioxidant
c, chlorophyll and chlorophyllactivity,
e [1].and
antimutagenic properties.
The most common Theofresults
form and conclusions
chlorophyll found in plantscan isbechlorophyll
applied bya.readers and
Its chemical
researchers to deepen their knowledge, guide future studies, explore applications
structure includes a porphyrin ring with a central magnesium ion, and an attached hydro- in food
and pharmaceutical
carbon tail known asindustries, andporphyrin
a phytol. The stimulate further research
ring is made in four
up of this field.
nitrogen-containing
groups called pyrrole, and the phytol tail is composed of isoprenoid units [7]. Chlorophyll
2. Chlorophyll:
a absorbs light Chemical Properties
most efficiently in theandredMetabolism
and blue regions of the spectrum, with peak
absorption at around
Chlorophyll 430 and 662
is a complex nanometers,
molecule made uprespectively.
of a porphyrin ring, a magnesium ion,
and an Chlorophyll b is another form
attached hydrocarbon tail. of
The chlorophyll
porphyrinfound ring isin responsible
plants, algae,for and some bacteria.
absorbing light
Its chemical structure is similar to that of chlorophyll a, but
energy and the magnesium ion acts as an electron acceptor. Chlorophyll has many formsit has a slightly different
porphyrin
such ring. This
as chlorophyll a, difference
chlorophyll results in chlorophyll
b, chlorophyll b absorbing
c, chlorophyll d andlight in the blue-green
chlorophyll e [1].
region
Theofmost
the spectrum,
common form with of
peak absorption
chlorophyll at around
found in plants453isnanometers. Its chemicalb
chlorophyll a.Chlorophyll
also has aincludes
structure role in photosynthesis,
a porphyrin ring butwith
its main function
a central is to protect
magnesium ion,chlorophyll a from
and an attached
excess light. tail
hydrocarbon In addition phytol. The aporphyrin
known astoa chlorophyll and b, there ringare other forms
is made of chlorophyll
up of four nitrogen-
such as chlorophyll
containing c, chlorophyll
groups called pyrrole, and d, and
the phytol tail is e.
chlorophyll These are
composed offound in a variety
isoprenoid units [7].of
organisms such as algae, and they have different absorption spectra
Chlorophyll a absorbs light most efficiently in the red and blue regions of the spectrum,and different functions.
Chlorophyll
with c absorbsat
peak absorption light in the430
around blue-green spectrum, chlorophyll d absorbs
region of the respectively.
and 662 nanometers,
lightChlorophyll
in the red region
b is of the spectrum,
another form ofand chlorophyllfound
chlorophyll e absorbs light in algae,
in plants, the far-red
and region
some
of the spectrum
bacteria. [7]. structure is similar to that of chlorophyll a, but it has a slightly
Its chemical
Photosynthesis
different porphyrin ring.is the process
This by which
difference plants,
results algae, andb some
in chlorophyll absorbingbacteria
lightconvert
in the
light energy into chemical energy. The light energy is absorbed
blue-green region of the spectrum, with peak absorption at around 453 nanometers. by chlorophyll and other
pigments, which
Chlorophyll excite
b also has electrons
a role ininphotosynthesis,
the pigment molecules.but its mainAfter function
absorbingislight energy,
to protect
the excited electrons in chlorophyll are utilized to facilitate the synthesis
chlorophyll a from excess light. In addition to chlorophyll a and b, there are other forms of ATP (adenosine
triphosphate)
of chlorophyll suchand NADPH (nicotinamide
as chlorophyll adenine
c, chlorophyll dinucleotide
d, and chlorophyllphosphate),
e. These arewhich
foundare in
essential components for the subsequent phases of photosynthesis.
a variety of organisms such as algae, and they have different absorption spectra and These energy-rich
molecules play a crucial role in the production of glucose and the release of oxygen as
different functions. Chlorophyll c absorbs light in the blue-green region of the spectrum,
byproducts. The first stage of photosynthesis is known as the light-dependent reactions
chlorophyll d absorbs light in the red region of the spectrum, and chlorophyll e absorbs
which take place in the thylakoid membrane of chloroplasts. The light energy absorbed by
light in the far-red region of the spectrum [7].
chlorophyll and other pigments is used to drive the transfer of electrons, which results in
Photosynthesis is the process by which plants, algae, and some bacteria convert light
the production of ATP and NADPH. The second stage of photosynthesis is known as the
energy into chemical energy. The light energy is absorbed by chlorophyll and other
light-independent reactions, also called the Calvin cycle, which takes place in the stroma of
pigments, which excite electrons in the pigment molecules. After absorbing light energy,
chloroplasts. In this stage, the ATP and NADPH produced in the light-dependent reactions
the excited electrons in chlorophyll are utilized to facilitate the synthesis of ATP
are used to drive the production of glucose and oxygen [8].
(adenosine triphosphate) and NADPH (nicotinamide adenine dinucleotide phosphate),
which are essential components for the subsequent phases of photosynthesis. These
energy-rich molecules play a crucial role in the production of glucose and the release of
oxygen as byproducts. The first stage of photosynthesis is known as the light-dependent
reactions which take place in the thylakoid membrane of chloroplasts. The light energy
absorbed by chlorophyll and other pigments is used to drive the transfer of electrons,
which results in the production of ATP and NADPH. The second stage of photosynthesis
is known as the light-independent reactions, also called the Calvin cycle, which takes
Molecules 2023, 28, 5344 place in the stroma of chloroplasts. In this stage, the ATP and NADPH produced in 3 of 21
the
light-dependent reactions are used to drive the production of glucose and oxygen [8].
Chlorophyll, chlorophyllides, and phycobiliproteins such as phycoerythrin and
phycocyanin,
Chlorophyll, are chlorophyllides,
all pigments that and are involved in the
phycobiliproteins suchprocess of photosynthesis.
as phycoerythrin and phy-
Chlorophyll
cocyanin, areisallthe primarythat
pigments pigment found ininplants
are involved and algae,
the process while chlorophyllides
of photosynthesis. Chlorophylland
phycobiliproteins
is the primary pigment are found
found in plants
smaller andquantities.
algae, whileThechlorophyllides
main difference andbetween these
phycobilipro-
pigments
teins is their
are found chemical
in smaller structure,The
quantities. which
mainresults in different
difference betweenabsorption spectra
these pigments and
is their
therefore structure,
chemical different functions in photosynthesis.
which results in different absorption spectra and therefore different
The process
functions of obtaining pheophorbides from chlorophyll is called chlorophyll
in photosynthesis.
The process
degradation, which of obtaining
is a process pheophorbides from chlorophyll
that occurs naturally in plants and is called chlorophyll
algae. This processdegra-
can
dation, whichbyisdifferent
be triggered a processenvironmental
that occurs naturally in plants
factors such andintensity,
as light algae. This process canand
temperature, be
triggered by different
water availability. environmental
During chlorophyllfactors such as light
degradation, intensity, temperature,
chlorophyll is broken downand water
into
availability. During including
different pigments, chlorophyll degradation,
pheophytin, chlorophyll
which is a form is broken down into
of chlorophyll thatdifferent
lacks a
pigments,
magnesium including
ion, andpheophytin,
pheophorbide, which is a form
which of chlorophyll
is a form of chlorophyll that that
lackshas
a magnesium
been modified ion,
and pheophorbide, which is
by the removal of the phytol tail [8]. a form of chlorophyll that has been modified by the removal
of theChlorophyllides
phytol tail [8]. are pigments that are closely related to chlorophyll, and they differ
in theChlorophyllides
arrangement ofare pigments
atoms. Theirthat are closely
chemical relatedisto
structure chlorophyll,
similar and theybut
to chlorophyll differ
theyin
the
have arrangement
a different ofcentral
atoms.atomTheirsuch
chemical structure
as zinc, iron, isorsimilar
copper, to chlorophyll
and they have but they have
different
a different central
absorption spectra.atomTheysuch as zinc,iniron,
are found or copper,
prokaryotic and theysuch
organisms haveas different absorption
cyanobacteria and
spectra. They are found in prokaryotic organisms such
they have a role in photosynthesis similar to chlorophyll [9], Figure 2. as cyanobacteria and they have a
role in photosynthesis similar to chlorophyll [9], Figure 2.
Figure 2. Conversion of Chlorophyll in phaeophytin, chlorophyllide and pheophorbide.

Figure 2. Conversion of Chlorophyll in phaeophytin, chlorophyllide and pheophorbide.
Phycobiliproteins such as phycoerythrin and phycocyanin are found in cyanobacteria
and red algae. They are water-soluble
Phycobiliproteins pigments that are
such as phycoerythrin andcomposed of a protein
phycocyanin are component
found in
and
cyanobacteria and red algae. They are water-soluble pigments that are composed than
a pigment component. They absorb light in different regions of the spectrum of a
chlorophyll, they transfer
protein component and athe energycomponent.
pigment to chlorophyll andabsorb
They they are important
light in photosynthe-
in different regions of
sis
theby increasing
spectrum the chlorophyll,
than efficiency of light
they harvesting
transfer the[10].
energy to chlorophyll and they are
important in photosynthesis by increasing the efficiency of light harvesting [10].
3. Effects of Chlorophyll in Health
3.1. Historic
3. Effects ofPerspective/Herbal Ethnomedicines
Chlorophyll in Health
The utilization
3.1. Historic of medicinal
Perspective/Herbal plants has a long history, dating back to ancient times, with
Ethnomedicines
its practice spanning across different regions worldwide [11,12]. Even in the present era, herbal
The utilization of medicinal plants has a long history, dating back to ancient times,
ethnomedicines continue to be of fundamental importance in primary healthcare, particularly
with its practice spanning across different regions worldwide [11,12]. Even in the present
for impoverished populations in remote areas [13] and developing countries [14]. These
era, herbal ethnomedicines continue to be of fundamental importance in primary
medicinal plants are employed to address a wide array of illnesses, including cancer, skin
diseases, cardiovascular disorders, endocrinal imbalances, gastrointestinal ailments, genitouri-
nary conditions, respiratory system disorders, musculoskeletal disorders, liver diseases, and
even treatment for poisonous bites, among others [12,13,15–18]. While the therapeutic effects
of medicinal plants are often attributed to their secondary metabolites [18–21], it is worth
noting that photosynthetic pigments may also play a significant role [5,6], with chlorophyll
being the most well-known pigment associated with these plants.
Among the various plant parts used for ethnomedicinal purposes, leaves stand out
as the most utilized [22–26]. Leaves possess several advantages, such as easy accessibility,
Molecules 2023, 28, 5344 4 of 21
straightforward processing, prolonged availability, and therapeutic properties resulting

from the accumulation of photosynthates and phytochemicals [18,20,24,27]. Notably, har-
vesting leaves does not pose a threat to the survival of the plant, unlike the harvesting
of whole plants or roots, which can endanger the survival of medicinal plant species and
contribute to a decline in plant biodiversity within specific regions [22,23].
3.2. Chlorophyll Bioavailability

Chlorophylls, the most abundant pigments on Earth, are present in photosynthetic
organisms such as bacteria, algae, and higher plants. In plants, chlorophyll a and b are the
predominant pigments, with their ratio varying depending on the species, environmental
conditions, and ripening stage [6]. However, chlorophylls are highly sensitive to physical
and chemical changes and exhibit instability when isolated or consumed outside of their
biological context. This has led to the development of chlorophyll derivatives [28]. Con-
sequently, natural chlorophylls are not commonly used in experimental research, as their
purification is challenging and expensive.
Semi-synthetic sodium copper-chlorophyllins (SCC) have been commercially available
as food colorants and supplements. SCC is produced through the saponification of natural
chlorophylls, where the magnesium ion in the tetrapyrrole is substituted by copper and the
hydrophobic side chain is eliminated. These modifications enhance the stability, solubility
in water, and accessibility of SCC [29,30]. Commercial SCC products may vary in compo-
sition, although two primary components, Cu-chlorin e4 and Cu-chlorin e6, are typically
present [31–33]. Due to these advantageous properties, SCC has found widespread use in
biological experiments. However, it is important to note that the bioavailability of SCC
may not match that of natural chlorophylls obtained from our diet.
Chlorophylls can be obtained from the human diet through the consumption of
green fruits and vegetables. However, their content varies significantly depending on
factors such as cultivar, maturity stage, growing conditions, harvest time, plant parts
used, storage conditions, food processing methods, and extraction and quantification
techniques [6,28]. While a diet rich in vegetables and green fruits may provide a substantial
amount of chlorophylls, their bioavailability, metabolism, and the effects of food processing
influence their potential impact on human health. Early studies assumed that humans
did not absorb chlorophylls, resulting in limited research on their absorption through the
gastrointestinal tract. However, a few studies have demonstrated that native chlorophylls
undergo significant transformation during the digestive process, and the absorption of
different chlorophyll derivatives may differ based on their molecular structure.
In vitro studies have provided valuable insights into the digestion, metabolism, and
absorption of natural chlorophylls. Using an in vitro model simulating the gastric and
small intestinal digestive processes, Ferruzi et al. [34] demonstrated that native chloro-
phylls obtained from fresh spinach puree undergo various transformations. The highly
acidic gastric phase led to the conversion of chlorophylls into their metal-free pheophytin
derivatives, while Zn-pheophytins treated with ZnCl2 remained stable. Furthermore, the
micellarization of chlorophyll a series was found to be more efficient compared to the b
series. The uptake of micellarized chlorophyll derivatives by Caco-2 human intestinal cells
predominantly consisted of pheophytins and their epimers, comprising around 5–10% of
the absorbed compounds. Another study utilizing the same in vitro approach revealed
that Cu-chlorin e4 in SCC remained stable during digestion, whereas 90% of Cu-chlorin
e6 underwent degradation. However, incorporating SCC into a food matrix reduced the
degradation of Cu-chlorin e6 [32]. Additionally, SCC derivatives were taken up by Caco-2
cells and transported to the basolateral compartment, suggesting their potential absorption
and transport to peripheral tissues [32,34].
Furthermore, a study using chlorophylls from pea puree demonstrated that native
chlorophylls were completely transformed into their magnesium-free derivatives during
gastric digestion in vitro, primarily due to the acidic conditions [35]. Pheophorbide a,
the most micellarized chlorophyll derivative, exhibited the highest absorption by Caco-2
Molecules 2023, 28, 5344 5 of 21
cells [35]. Another investigation by Gandul-Rojas et al. [36], using standard pigments such
as chlorophyll a, chlorophyll b, pheophytin a, pheophytin b, pyropheophytin a, pheophor-
bide a, and pyropheophorbide a, indicated that the de-esterification of the phytol group
enhanced the transfer efficiency of dephytylated derivatives (pheophorbide a and py-
ropheophorbide a) to the aqueous micellar fraction and their transport to Caco-2 intestinal
cells, thereby increasing their bioaccessibility. Moreover, the cellular uptake of phytylated
chlorophyll derivatives (pheophytin a and pyropheophytin a) by Caco-2 cells occurred
through simple diffusion, whereas the uptake of dephytylated derivatives was mediated
by facilitated diffusion at lower concentrations tested [36].
A recent study investigated the in vitro digestion of chlorophyll pigments derived from
three types of edible dried seaweeds: Nori (red algae, containing only chlorophyll a series),
Sea lettuce (green algae, containing a and b series), and Kombu (brown algae, presenting a
and c series). The findings revealed that chlorophylls from a series were more susceptible to
pheophytinization compared to the b and c series. The formation of pheophorbides during
digestion occurred when the initial chlorophyll profile contained significant amounts of
pheophytins, as observed in the Kombu algae. Furthermore, the digestive stability and
recovery of chlorophyll derivatives after in vitro digestion appeared to depend on the
chemical structure and the food matrix [37]. The same authors demonstrated that the
percentage of micellization and the uptake by Caco-2 cells were higher for dephytylated
chlorophylls compared to phytylated derivatives. Additionally, chlorophylls from Nori
algae exhibited higher bioaccessibility than those from Sea lettuce and Kombu [38].
Regarding studies demonstrating in vivo absorption of chlorophyll derivatives, a
chemoprevention trial in humans using SCC revealed that daily ingestion of SCC tablets
(300 mg/day) resulted in the absorption of Cu-chlorin e4 ethyl ester into the bloodstream,
and to a lesser extent, Cu-chlorin e4 [31]. This provided the initial evidence that chlorophyll
derivatives can be absorbed by the human gastrointestinal tract. Another study conducted
on human volunteers detected the presence of pheophytin and pheophorbide derivatives in
the blood three hours after the ingestion of 1.2 kg of freshly boiled spinach [39]. In a study by
Gomes et al. [33], rats fed a diet supplemented with 10 or 30 g/kg SCC showed absorption
of Cu-chlorin e4, which was detected in the serum, liver, and kidneys. The absorption was
macroscopically visible as a green color in these biological materials. However, Cu-chlorin
e6 was not found in the serum or organs, suggesting degradation during passage through
the gastrointestinal tract or interaction with other food components [33].
Regarding chlorophylls from natural sources, Fernandes et al. [40] studied the appar-
ent absorption of chlorophylls in dogs by analyzing ingested and excreted chlorophylls.
The results showed that after supplementation with 0.8% freeze-dried ground spinach
leaves (18 mg chlorophyll/day) for 10 days, the average apparent absorption of chlorophyll
derivatives was 3.4%. By analyzing the excreta, the authors inferred that pheophytiniza-
tion, the major degradation process occurring in the acidic gastrointestinal tract of dogs,
transformed chlorophylls a and b into their respective magnesium-free pheophytins a and
b. However, they were unable to detect chlorophyll derivatives in the plasma of dogs
after consuming a diet containing 10% freeze-dried spinach [40]. In a biodistribution study
conducted in rabbits, where a single dose of 100 g of freeze-dried spinach powder (prepared
from fresh spinach) was administered after 24 h of fasting, and rabbits were sacrificed after
2, 4, 8, 12, and 24 h, the major metabolites observed were chlorophyllides and pheophor-
bides [41]. The occurrence and levels of chlorophyll derivatives were organ-specific, found
in the plasma, liver, gallbladder, and kidney. In the feces, the major metabolites detected
were native chlorophylls and pheophytins [41]. More recently, Vieira et al. [42] analyzed
the livers of mice fed a diet containing 15% spirulina powder, a blue-green microalgae
primarily composed of chlorophyll a. The results demonstrated that the formation and
absorption of pheophorbides, pyro-derivatives, and phytyl-chlorin e6 required first-pass
metabolism. The absorption and accumulation of pheophorbide a in the liver may be
partially protein-mediated through the scavenger receptor B type I, while the presence of
phytol in the liver may occur due to the de-esterification of pheophytin a, leading to the
Molecules 2023, 28, 5344 6 of 21
formation of pheophorbide a and phytol [42]. In the feces, the percentage of pheophorbide
derivatives and allomerized derivatives was similar to that in the supplemented feed, while
pheophytin derivatives and pyro-derivatives exhibited increased content compared to
the supplemented feed. Additionally, native chlorophyll a was detected in the feces of
mice [42].
In summary, the collective findings from both in vitro and in vivo studies using native
chlorophylls indicate that the potential health benefits associated with chlorophyll a and b
are likely attributed to their metal-free derivatives. While these studies provide valuable
insights into the bioavailability of chlorophylls, there is still a need for a comprehensive
characterization of the chlorophyll derivatives formed in the gastrointestinal tract and a
better understanding of their pharmacokinetic parameters. Unfortunately, as of now, there
is a lack of published in vivo studies involving omnivorous species, which poses challenges
in translating this information directly to humans. Further research in this area is warranted
to bridge the gap in our understanding of chlorophyll metabolism and absorption in the
human body.
3.3. Bioactive Properties of Chlorophyll Compounds

The chemical structure of chlorophylls is a key determinant of their bioactivity, influ-
encing their potential health benefits [5]. Understanding the relationship between chemical
structure and bioactivity is crucial for unraveling the therapeutic properties of chloro-
phylls and their derivatives [43–45]. The chemical structure of chlorophylls consists of a
porphyrin ring, which serves as the core framework, and a long hydrophobic side chain.
This unique structure confers distinctive physicochemical and biological properties to
chlorophylls. For instance, the presence of magnesium at the center of the porphyrin ring
enables chlorophylls to efficiently capture light energy during photosynthesis.
Moreover, the chemical structure of chlorophylls contributes to their bioactive proper-
ties. Studies have shown that chlorophyll derivatives, which undergo structural modifica-
tions, exhibit enhanced bioactivity compared to native chlorophylls. These modifications
can involve alterations in the porphyrin ring, such as the addition of functional groups or
substitutions. These structural changes lead to variations in the solubility, stability, and
interaction capabilities of chlorophyll compounds [7,46].
The bioactivity of chlorophylls is attributed to their ability to act as antioxidants,
antimutagens, and anticarcinogens. The unique chemical structure allows chlorophylls to
scavenge harmful free radicals, mitigate DNA damage, and modulate cellular processes
involved in disease development. Furthermore, their hydrophobic side chains facilitate
interactions with biological membranes, influencing cellular uptake and signaling path-
ways [5,47].
Despite advances in understanding the relationship between the chemical structure
of chlorophyll compounds and their bioactivity, further research is warranted. Investi-
gations into the specific structural features responsible for different bioactive properties
are necessary to unlock the full potential of chlorophylls as therapeutic agents. Moreover,
exploring the interactions between chlorophylls and other bioactive compounds in a syner-
gistic manner can provide insights into the holistic benefits of consuming chlorophyll-rich
foods [48].
In conclusion, the bioactive properties of chlorophyll compounds are intricately linked
to their chemical structure. By deciphering the structural determinants of their bioactivity,
we can uncover new opportunities for utilizing chlorophylls and their derivatives in
promoting human health [47]. Continued research in this field holds great promise for
harnessing the full potential of chlorophylls as functional ingredients and contributing to
the development of novel therapeutic approaches (Figure 3) [49–51].
consuming chlorophyll-rich foods [48].
In conclusion, the bioactive properties of chlorophyll compounds are intricately
linked to their chemical structure. By deciphering the structural determinants of their
bioactivity, we can uncover new opportunities for utilizing chlorophylls and their
derivatives in promoting human health [47]. Continued research in this field holds great
Molecules 2023, 28, 5344 promise for harnessing the full potential of chlorophylls as functional ingredients
7 of 21 and
contributing to the development of novel therapeutic approaches (Figure 3) [49–51].
Figure 3. Bioactive properties and health benefits of chlorophyll compounds.

Figure 3. Bioactive properties and health benefits of chlorophyll compounds.
3.3.1. Antioxidant Activity
3.3.1. Antioxidant Activity
Oxidative stressOxidative
plays a significant
stress plays rolea in the development
significant role in theofdevelopment
various diseases. Natural
of various diseases.
chlorophylls possess
Natural antioxidant
chlorophylls properties [52], which
possess antioxidant make them
properties [52], promising
which makecandidates
them promising
for preventing or mitigating
candidates for the formation
preventing of reactive
or mitigating the species.
formationIn ofvitro studies
reactive conducted
species. In vitro studies
by Ferruzi et al. conducted
[32] demonstrated
by Ferruzithat standard
et al. chlorophyllthat
[32] demonstrated a derivatives exhibited ahigher
standard chlorophyll derivatives
exhibited
antioxidant capacity higher to
compared antioxidant
chlorophyll capacity comparedHowever,
b derivatives. to chlorophyll b derivatives. et
Lanfer-Marquez However,
al.
Lanfer-Marquez
reported contrasting findings, et al. reported
showing thatcontrasting
pheophorbide findings,
b andshowing that pheophorbide
pheophytin b were theb and
most potent naturalpheophytin
antioxidantb were the mostcompared
derivatives potent natural antioxidant
to chlorophyll derivatives[45].
a derivatives compared
Fer- to
chlorophyll a derivatives [45]. Ferruzi et al. [32] also observed that metallo-chlorophyll
ruzi et al. [32] also observed that metallo-chlorophyll derivatives (such as Mg-chlorophylls,
derivatives (such as Mg-chlorophylls, Zn-pheophytins, Zn-pyropheophytins, Cu-
Zn-pheophytins, Zn-pyropheophytins, Cu-pheophytin a, and Cu-chlorophyllins) exhibited
pheophytin a, and Cu-chlorophyllins) exhibited higher antiradical capacity than metal-
higher antiradical freecapacity than
derivatives metal-free
(such derivatives
as chlorins, (such
pheophytins, andaspyropheophytins).
chlorins, pheophytins, and that
This suggests
pyropheophytins). This suggests that in addition to the fundamental porphyrin
in addition to the fundamental porphyrin structure contributing to free radical reductionstructure
contributing to free
[53],radical reduction
metal chelation [53], metal
enhances chelation
antioxidant enhances
activity antioxidant
[32]. Moreover, activity [32]. et al.
Lanfer-Marquez
Moreover, Lanfer-Marquez et al. [45]
[45] demonstrated demonstrated that
that Cu-chlorophyllin Cu-chlorophyllin
displayed displayed
higher antioxidant higher
activity compared
to natural
antioxidant activity chlorophylls,
compared highlighting
to natural the influence
chlorophylls, of the chelated
highlighting metal in the
the influence ofporphyrin
the
chelated metal in ring onporphyrin
the the strengthringof theonantioxidant
the strengthcapacity. In an
of the in vitro study
antioxidant conducted
capacity. by Kang
In an
et al., Zn-pheophytins exhibited the highest radical scavenging
in vitro study conducted by Kang et al., Zn-pheophytins exhibited the highest radical capacity and β-carotene
scavenging capacity and β-carotene bleaching activities, surpassing the antioxidant activity
of chlorophylls and pheophytins [54]. Zn-pheophytins also demonstrated inhibitory activ-
ity against lipopolysaccharide (LPS)-induced nitric oxide production and suppressed the
expression of inducible nitric oxide synthase in LPS-stimulated macrophage cells without
exerting cytotoxic effects [54]. Regarding in vivo investigations, Gomes et al. [33] reported
that a diet supplemented with SCC protected against lipid peroxidation in the brain of
rats, but not in the liver. Additionally, pretreatment with chlorophyll b in mice resulted in
reduced oxidative stress and lipid peroxidation induced by cisplatin, although the antiox-
idant effects were inconsistent [55]. Despite the potential antioxidant activity of natural
chlorophylls, research in this area has been relatively limited in recent years. This may
be attributed to the perception that chlorophylls are less important compared to other
phytochemicals found in fruits and vegetables, as well as the belief that chlorophylls are
poorly absorbed by the human gastrointestinal tract. However, it is worth noting that the
understanding of the antioxidant properties of natural chlorophylls is an evolving field,
and further investigations are necessary to fully elucidate their potential and establish their
significance relative to other phytochemicals.
Molecules 2023, 28, 5344 8 of 21
3.3.2. Antimutagenic and Antigenotoxic Properties

Mutagenic and genotoxic agents are ubiquitous in our environment and food, and
some are even used as chemotherapy agents such as cisplatin. In a pioneering study by
Lai et al. [56], the chlorophyll content of fruits and vegetables was correlated with their
antimutagenic activity. Using the gene reversion mutagen test with Salmonella strain
TA100, the researchers found that higher chlorophyll content in both aqueous and acetone
extracts corresponded to decreased mutagenic activity induced by 3-methylcholanthrene
or benzo[a]pyrene [56]. Interestingly, the study also revealed that SCC had a more potent
effect than vegetable extracts. In another study conducted in Drosophila melanogaster, the
effects of SCC and natural chlorophylls from spinach and chlorella were evaluated against
the genotoxicity of 4-nitroquinoline 1-oxide (4NQO) in the flies [57]. All the chlorophyll
preparations demonstrated a reduction in wing spot formation caused by 4NQO. The
authors proposed that the mechanisms of action involve the formation of complexes be-
tween chlorophyll and 4NQO, as well as the inhibition of metabolic activation of this
mutagen through enzyme inhibition or degradation of active metabolites [57]. Similarly,
Kocaoğlu Cenkci and Kaya [58] demonstrated the effectiveness of chlorophyll a and b
in reducing the genotoxic effects of 2-amino-3,8-dimethylimidazo [4,5-f]quinoxaline in
Drosophila melanogaster. An additional study evaluated the antimutagenic effect of stan-
dard chlorophyll derivatives in Salmonella typhimurium TA100 exposed to the mutagen
benzo[a]pyrene. The results indicated that both metallo and metal-free derivatives ex-
hibited similar dose-dependent antimutagenic activity, suggesting that the tetrapyrrole
macrocycle is essential for chlorophyll’s antimutagenic activity rather than the presence of
a central metal atom [32]. In mice, pretreatment with chlorophyll b was found to mitigate
chromosomal breakage and micronucleus formation induced by cisplatin in peripheral
blood and bone marrow cells [55]. Similar protective effects were observed in the liver
and kidneys, where DNA damage was reduced [59]. Despite these promising findings,
the potential antimutagenic and antigenotoxic activity of natural chlorophylls remains
largely unexplored, similar to their antioxidant activity mentioned earlier. Further research
is needed to comprehensively investigate and understand the extent of these properties in
natural chlorophylls.
3.3.3. Anticancer Activity

Cancer, as the second leading cause of death worldwide, necessitates the development
of new therapeutic agents with minimal side effects. Studies have demonstrated the
potential anticarcinogenic action of chlorophylls and sodium copper chlorophyllin (SCC)
against various types of cancer. For instance, topical application of pheophorbide a inhibited
skin tumor promotion induced by 7,12-dimethylbenz[a]anthracene (DMBA) and 12-O-
tetradecanoyl-phorbol-13-acetate (TPA) in ICR mice [60]. Similarly, chlorophyll a and b
extracted from green tea leaves suppressed skin tumorigenesis and edema formation in
BALB/c mice when applied prior to treatment with the tumor promoter TPA, initiated by
DMBA [61].
In rainbow trout exposed to the potent environmental carcinogen dibenzo[a,l]pyrene
(DBP), concurrent exposure to native chlorophyll preparations or SCC significantly reduced
DBP-DNA adduct levels in the liver [62]. Dietary intake of natural chlorophyll and SCC
also reduced DBP-DNA adducts in the liver, inhibited tumor incidence and multiplicity
in the liver, and reduced tumor incidence in the stomach of rainbow trout when co-fed
with DBP [63,64]. Aflatoxins, potent inducers of hepatocellular carcinoma (HCC), are food
contaminants produced by fungi. Chlorophyllin forms a strong noncovalent complex with
aflatoxin-B1 (AFB1), inhibiting hepatic AFB1-DNA adduction and hepatocarcinogenesis
in rainbow trout [65–67]. In rats administered with AFB1, chlorophylls derived from
spinach and chlorophyllin protected against early biochemical and late pathophysiologic
biomarkers of AFB1 carcinogenesis in the liver and colon [68]. This protection is thought
to occur through the inhibition of AFB1 intestinal uptake involving complex formation,
thereby reducing its bioavailability [68]. In a study involving human subjects from Qidong,
Molecules 2023, 28, 5344 9 of 21
People’s Republic of China, where aflatoxin contamination in food is linked to a high risk of
HCC, the consumption of 100 mg of chlorophyllin three times a day for 4 months reduced
the urinary excretion of aflatoxin-DNA adducts by 55%, indicating a reduced biologically
effective dose of aflatoxin [50].
High consumption of red meat is associated with an increased risk of colon cancer.
However, studies in rats subjected to dietary heme, which mimics red meat ingestion,
demonstrated that natural chlorophylls inhibit colonic cytotoxicity, proliferation of colonic
epithelial cells, epithelial cell turnover, and the formation of lipid radicals induced by
heme [69]. In contrast, Na-chlorophyllin and Cu-chlorophyllin were unable to prevent these
heme-induced effects [69]. Diets supplemented with freeze-dried spinach also decreased
colonic cytotoxicity and colonic hyperproliferation induced by heme [49]. These protective
effects are attributed to the chlorophyll content, which prevents heme degradation and
blocks the formation of cytotoxic heme metabolites [49].
Aside from skin, liver, stomach, and colon cancer, SCC has been reported to decrease
the proliferation of human pancreatic cancer cell lines in vitro [70] and delay the progression
of lung cancer in vitro and in vivo [71].
The anticarcinogenic activity of chlorophylls and SCC is suggested to occur through
various mechanisms, including the formation of a molecular complex with aromatic car-
cinogens. This complex formation reduces carcinogen uptake and bioavailability, enhances
elimination of the unmetabolized carcinogen, inhibits carcinogen activation, and promotes
antioxidant activity and induction of apoptosis in cancer cells [2,51,72]. The protective
effect through complex formation mainly occurs when chlorophylls are administered si-
multaneously with the carcinogen, highlighting the importance of consuming green and
leafy vegetables and fruits to combat dietary carcinogens and mutagens [72]. Moreover,
high-fiber diets have been associated with better prognosis in oncology [73], emphasizing
the need for targeted policies to promote the consumption of healthier foods over processed
ones, thereby improving public health.
Photodynamic therapy (PDT) is a two-stage treatment involving a photosensitizing
drug and activating light, combined with molecular oxygen to induce cell death (phototoxi-
city). PDT has been increasingly used in cancer treatment [74]. However, the prolonged
half-life of photosensitizers often leads to prolonged generalized cutaneous photosensitivity,
causing a phototoxic reaction when the treated lesion is exposed to sunlight. Chlorophyll
acts as a photosensitizer due to its natural ability to absorb light. Notably, it loses its
photosensitizing activity within a few hours and requires only a relatively short incubation
period [75], making it a promising candidate for PDT. In a clinical trial, cancer patients
with basal cell carcinoma, squamous cell cancer, and papillary carcinoma received a single
intravenous injection of mono-L-aspartyl chlorin e6 (NPe6), a photosensitizer derived from
chlorophyll a. The study showed that NPe6 PDT had some efficacy against cutaneous
tumors with minimal phototoxic side effects [76]. Previous research demonstrated that
NPe6 persisted in the plasma of cancer patients for 6 weeks, although no persistent skin
photosensitization was observed [77]. NPe6 PDT has also been used for the treatment of
lung cancer [74]. Recently, Zhuo et al. [78] demonstrated that chlorophyllin e6-mediated
PDT induces apoptosis in human bladder cancer cells, possibly through the inhibition of
superoxide dismutase activity and the generation of reactive oxygen species. Although
the use of chlorophyll or NPe6 in PDT shows promising results, further clinical trials with
robust and compelling positive outcomes are needed to confirm their efficacy.
3.3.4. Anti-Obesity Effects

The prevalence of obesity has significantly increased in recent decades, becoming a
major concern in developed countries. The imbalance between energy intake and energy
expenditure, coupled with reduced physical activity, is one of the primary factors contribut-
ing to the development of obesity [79]. To combat obesity and its associated diseases such
as diabetes, cardiovascular diseases, and atherosclerosis [80], it is crucial to make dietary
Molecules 2023, 28, 5344 10 of 21
changes that prioritize the consumption of healthier foods. Recent studies have suggested
that chlorophylls could have a positive impact on obesity control.
In an in vitro study, chlorophyll a extracted from the aquatic plant Ludwigia octovalvis
exhibited an antiproliferative effect on 3T3-L1 adipose cells. It induced cell apoptosis by
activating the CD95 (APO/CD95) death receptor and pro-caspase-3 proteins. The anti-
adipogenic activity of chlorophyll a appeared to be mediated through the activation of the
AMPK signaling transduction pathway [81]. Another study by Wang et al. [82] investigated
the effects of chlorophyll and its derivatives on the digestion of soybean oil under simulated
human gastrointestinal conditions. The researchers discovered that chlorophyll reduced the
release rate of free fatty acids, altered the fatty acid composition, and increased the particle
size of oil droplets. These changes could decrease the uptake of fatty acids by intestinal
epithelial cells. Furthermore, the study found that pheophytin bound to and inhibited
pancreatic lipase activity during intestinal digestion, thereby affecting lipid digestion
in vitro [82].
Thylakoids, which are membranes within plant chloroplasts where photosynthesis
occurs, consist of galactolipids, proteins, vitamins, antioxidants, chlorophylls, carotenoids,
and other pigments [83]. Spinach baby leaves, for example, may contain around 3000 mg of
chlorophyll per 100 g of thylakoids [84]. In vivo studies have demonstrated that thylakoid
supplementation can suppress appetite, reduce body weight gain and body fat, lower serum
glucose, triglyceride, and free fatty acid levels, and modulate appetite-regulating hormones
in animals fed a high-fat diet (HFD) [85,86]. Supplementation with thylakoids has also
been shown to induce weight loss, decrease total and LDL cholesterol levels, suppress
the appetite for palatable food in overweight women [84], and exert a prebiotic effect that
influences microbiota composition, thus improving lipid and glucose homeostasis [83].
A study by Seo et al. [87] investigated the anti-obesity and browning effects of
Spirulina maxima extract, a microalga rich in chlorophyll a. The researchers found that
Spirulina extract suppressed lipid accumulation by reducing the expression of adipogenic
and lipogenic proteins in vitro. Furthermore, supplementation with Spirulina extract de-
creased body weight gain, fat mass, triglyceride and total cholesterol serum levels, and also
reduced the expression of adipogenic proteins while increasing thermogenic factors in mice
fed an HFD [87]. In the same study, chlorophyll a alone was found to inhibit adipogenesis
and lipogenesis in vitro [87].
Other recent studies reported that supplementation with chlorophyll-rich spinach
extract significantly reduced body weight gain, low-grade inflammation, and improved
glucose tolerance in mice fed an HFD [88,89]. Moreover, this chlorophyll-rich extract
supplementation also alleviated gut microbiota dysbiosis induced by the HFD [88,89].
Additional studies have demonstrated that chlorophyll-rich thylakoid supplementation
or chlorophyllin can modulate the diversity and composition of gut microbiota in human
subjects and BALB/c mice, respectively [90,91]. The composition of the gut microbiota is
now recognized to be related to the state of health or disease, including obesity [92]. It
has been observed that dietary substances can influence the composition of the gut micro-
biota [93,94]. Furthermore, there is evidence of an interaction between herbal medicines
and gut microbiota. It is suggested that the gut microbiota biotransforms components of
herbal medicines into bioactive small molecules that are absorbed into the bloodstream.
Simultaneously, herbal medicines can modify the composition of gut microbiota by pro-
moting the growth of beneficial bacteria and inhibiting harmful ones, leading to favorable
physiological changes [95].
3.3.5. Protection against Endocrine Disruptors

Research suggests that chlorophyll has the ability to mitigate the harmful effects of
endocrine disruptors through several mechanisms. Firstly, chlorophyll has been found to
possess antioxidant properties. It can scavenge reactive oxygen species (ROS) and reduce
oxidative stress, which is known to be associated with endocrine disruption. By reducing
oxidative stress, chlorophyll helps to protect endocrine organs, such as the ovaries, testes,
Molecules 2023, 28, 5344 11 of 21
and thyroid gland, from damage caused by endocrine disruptors. Secondly, chlorophyll has
been shown to exhibit detoxification properties. It can bind to and effectively remove certain
endocrine-disrupting chemicals from the body. This binding ability, known as chelation,
helps to prevent the chemicals from interacting with hormone receptors and disrupting
normal endocrine function. By facilitating the elimination of these harmful compounds,
chlorophyll aids in reducing their detrimental effects on the endocrine system [96,97]. A
study from Okai et al. [98] showed that, compared to SCC, chlorophyll a and pheophytin a
derived from green tea showed strong preventive effects against the endocrine disruptor
p-nonylphenol (NP)-induced inhibition of cell growth and cellular respiration in yeast
Saccharomyces cerevisiae. These protective effects of natural chlorophylls were linked to their
ability to attenuate oxygen radical formation induced by NP in yeast cells.
Furthermore, chlorophyll has been found to possess anti-inflammatory properties.
Endocrine disruptors can induce chronic inflammation, which can further exacerbate their
disruptive effects on hormone regulation. Chlorophyll has been shown to modulate inflam-
matory pathways and reduce the production of inflammatory mediators, thereby mitigating
the inflammatory response triggered by endocrine disruptors [99]. A water-extract from
the leaves of the red algae Dulse, constituted by phycobiliproteins and chlorophyll a, atten-
uated the secretion of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and nitric oxide
induced by LPS in vitro [100]. Furthermore, the acid-insoluble fraction of the water-extract,
containing concentrated pheophorbide a and pheophorbide a derivatives, was able to
reduce the secretion of proinflammatory mediators. Additionally, the oral administration of
the Dulse water-extract to mice decreased the acute inflammation in carrageenan-induced
paw edema [100]. Moreover, chlorophyll has been observed to support liver health and
enhance detoxification processes. The liver plays a crucial role in metabolizing and elimi-
nating endocrine-disrupting chemicals from the body. Chlorophyll promotes liver function
and aids in the detoxification of these compounds, thereby reducing their accumulation
and potential impact on the endocrine system. While more research is needed to fully
understand the extent of chlorophyll’s protective effects against endocrine disruptors, the
available evidence suggests its potential as a natural defense mechanism. Incorporating
chlorophyll-rich foods, such as leafy green vegetables, into the diet may contribute to
minimizing the adverse effects of endocrine disruptors on hormone balance and overall
health [51,101].
3.3.6. Neuroprotective and Anti-Inflammatory Effects

One way chlorophyll may exert its neuroprotective effects is through its antioxidant
properties. Oxidative stress, which occurs when there is an imbalance between the produc-
tion of harmful free radicals and the body’s antioxidant defenses, has been implicated in the
development and progression of neurodegenerative diseases such as Alzheimer’s, Parkin-
son’s, and Huntington’s diseases. Chlorophyll’s antioxidant activity allows it to neutralize
free radicals and reduce oxidative damage to brain cells. By reducing oxidative stress,
chlorophyll may help preserve the structure and function of neurons, potentially slowing
down the onset or progression of neurodegenerative disorders [102]. Rehni et al. [103]
demonstrated that a pre-treatment with a chlorophyll salt had neuroprotective effects in
mice subjected to cerebral ischemia followed by reperfusion. Results showed that chloro-
phyll was able to decrease cerebral infarct size, increase short-term memory, attenuate
motor incoordination and increase lateral push response. These protective effects were
attributed to the antioxidant activity of chlorophylls. Additionally, chlorophyll has been
shown to possess anti-inflammatory properties. Chronic inflammation is a common feature
of many neurological conditions, and it can contribute to neuronal damage and degener-
ation. By modulating inflammatory pathways, chlorophyll may help dampen excessive
inflammation in the brain, thereby protecting neurons from inflammatory damage. Further-
more, chlorophyll has been found to enhance the body’s natural detoxification processes.
Toxins and environmental pollutants can accumulate in the brain and contribute to neu-
ronal dysfunction and neurodegenerative processes [104,105]. Chlorophyll’s detoxifying
Molecules 2023, 28, 5344 12 of 21
properties may aid in the elimination of these harmful substances, reducing their impact on
brain health and potentially preventing or delaying neurodegenerative conditions. More-
over, chlorophyll-rich foods have been associated with improved cognitive function and
brain health. Leafy green vegetables, which are abundant sources of chlorophyll, have
been linked to better cognitive performance and a lower risk of cognitive decline [47].
While the exact mechanisms underlying these associations are not fully understood, the
neuroprotective properties of chlorophyll may play a role. It is important to note that
the research on chlorophyll’s neuroprotective effects is still in its early stages, and more
studies are needed to fully elucidate the extent and mechanisms of its action. Nonetheless,
the available evidence suggests that incorporating chlorophyll-rich foods into the diet
may contribute to brain health and potentially offer protection against neurodegenerative
disorders. As always, maintaining a balanced and nutritious diet, along with a healthy
lifestyle, is crucial for overall brain health and well-being [106].
In general, the bibliography from the last decades has shown essentially the potential
of chlorophyll and chlorophyllin as anticancer and anti-obesogenic agents, but this evi-
dence comes basically from a few in vitro and in vivo studies. The same applies for the
other health benefits mentioned above. This lack of reliable clinical trials and follow-up
studies in humans also translates into doubts regarding the real beneficial properties of
chlorophylls. Nevertheless, these gaps should not divert attention from the potential use of
these pigments, and these questions should continue to be addressed in future studies.
4. Chlorophyll Content in Fruits and Vegetables

Modern societies are currently facing food waste problems that are increasing as the
world’s population also increases, leading to economic and environmental issues. Food
losses and waste occur at all stages of the food supply chain: agricultural production,
post-harvest handling and storage, processing, distribution, and consumption stages [4].
Simultaneously with these difficulties, changes in eating habits, increased consumption of
more processed foods, and less variety in diets have contributed to the increase in modern
societies’ diseases such as obesity, diabetes, cardiovascular diseases, and atherosclerosis.
The discarded material can be a valuable resource to answer these problems. For instance,
leafy material or fruit peels, which are discarded in these first stages, are usually rich in
bioactive compounds beneficial to health [107–109]. The use of these currently discarded
products may represent a return to past eating habits, with the use of more diverse foods,
sometimes not so appealing, but with less caloric concentrations and rich in a high variety
of bioactive compounds. For example, broccoli is one of the most produced crops world-
wide, where only the inflorescence part is used, while the stem and leaves are discarded.
Nevertheless, this discarded material, in addition to glucosinolates, is also extremely rich
in chlorophylls, especially the leaves. Our group evaluated the chlorophyll and carotenoid
contents in the broccoli plant in two different harvest years (Supplementary Methods).
Table 1 shows the contents of chlorophylls a and b, total chlorophylls, and total carotenoids
in the inflorescences, stalks and leaves of broccoli plants, harvested in October 2018 and
July 2019. For both crops, the leaves are the part of the broccoli plant that contains sig-
nificantly (p < 0.05) more total chlorophyll content and carotenoids compared to stems
and inflorescences. Comparing the two harvests, only the leaves registered significant
differences (p < 0.0001), whereas the 2018 crop plants showed higher contents of pigments
than the 2019 crop. This difference may be due to the fact that plants harvested in July
were subjected to greater environmental stress [110]. These results can also be correlated
with the high antioxidant activity of the leaves compared to stalks and inflorescences [111].
These results suggest that the leaves can be a broccoli by-product with high interest and
potential for exploitation as a functional food ingredient due to their high content of chloro-
phylls. Nevertheless, the stalks can also be a value-added product of great potential since
chlorophylls are also present in high amounts.
Molecules 2023, 28, 5344 13 of 21
Table 1. Contents of chlorophyll a, chlorophyll b, total chlorophylls, and total carotenoids (mg/g
DW), in the inflorescences, stalks and leaves of broccoli plants.
Chlorophyll a Chlorophyll b Total Chlorophylls Total Carotenoids

Broccoli Part 2018 2019 2018 2019 2018 2019 2018 2019
Inflorescence 0.75 ± 0.05 aA 0.91 ± 0.09 bA 0.30 ± 0.03 aA 0.34 ± 0.03 aA 1.05 ± 0.24 bA 1.25 ± 0.24 bA 0.18 ± 0.02 bA 0.26 ± 0.02 bA
Stalk 0.30 ± 0.03 aA 0.14 ± 0.03 aA 0.14 ± 0.01 aA 0.05 ± 0.01 aA 0.44 ± 0.12 bA 0.19 ± 0.081 cA 0.07 ± 0.004 aA 0.05 ± 0.01 aA
Leaves 12.43 ± 0.14 bA 4.96 ± 0.36 cB 4.74 ± 0.06 bA 1.85 ± 0.16 bB 17.17 ± 0.59 aA 6.81 ± 0.36 aB 2.49 ± 0.03 cA 1.15 ± 0.08 cB
Data presented as mean ± SD (n = 5). For each year, values for the same parameter evaluated (within the same
column) followed by different lowercase letters are significantly different at p < 0.05, according to Tukey’s test.
Uppercase letters indicate differences between the harvest years (p < 0.0001). DW, dry weight.
When it comes to green plants and vegetables, storage and processing conditions
greatly impact the green color of these foods conferred by chlorophyll, whose degradation
can be delayed or accelerated by these conditions [112–116]. This, in turn, has a great
influence on the behavior of the final consumer, that is, not consuming them if the products
do not have an attractive green color, thus further contributing to food waste.
Table 2 lists chlorophyll-rich fruits and vegetables. Compared to fruits, vegetables are
particularly rich in chlorophyll, among which spinach, broccoli, garden rocket and wild
rocket have the highest amount of this pigment. On the contrary, fruits and vegetables that
are lower in chlorophyll include: kiwifruit purée, cabbage, celery and cucumber. For some
fruits and vegetables, the extent of chlorophyll loss after food processing is also indicated
(Table 2). This is particularly important since several processing food techniques may have
a large contribution to final chlorophyll content, affecting the nutritional and commercial
value. The green color, conferred by a high proportion of chlorophylls a and b, is often used
as a quality measurement [117]. The post-harvest storage time per se leads to the loss of
green color due to chlorophyll degradation, triggered by ROS and ethylene formation [118].
The use of exogenous diacetyl to inhibit ethylene and ROS generation, has been proposed
to retard the yellowing and nutrient loss of postharvest broccoli, thus maintaining its
nutritional and economic value [119]. Postharvest treatment of broccoli with melatonin has
also been shown to prolong shelf life [120]. Table 2 shows the chlorophyll content in fruits
and vegetables before and after processing. However, this information has to take into
account the concepts previously described regarding its bioavailability. Furthermore, recent
data shows that food composition is also a variable that greatly influences the bioavailability
of chlorophylls. Viera et al. [121] have shown that, during the in vitro digestion, the stability
of total chlorophylls ranges from 15% to 85%, which is influenced by the amount of salt
present in the food.
Based on the provided information, here are some observations regarding the process-
ing methods and conditions that can help retain higher chlorophyll content in certain fruits
and vegetables: (i) Boiling: Boiling for a shorter duration appears to be more effective in
retaining chlorophyll content. For example, in the case of green beans, boiling for 5 min
resulted in higher chlorophyll content compared to longer boiling times; (ii) Steaming:
Steaming for a moderate duration seems to be beneficial for maintaining chlorophyll lev-
els. In the case of spinach, steaming for 7.5 min resulted in higher chlorophyll content
compared to both shorter and longer steaming times; (iii) Microwaving: Microwaving
for a shorter duration tends to preserve chlorophyll content. For instance, in the case of
peas, microwaving for 1.5 min resulted in higher chlorophyll content compared to longer
microwaving times; (iv) Storage conditions: Some vegetables, such as celery and leek,
demonstrated a decrease in chlorophyll content after storage at low temperatures (0 ◦ C) for
an extended period. Therefore, it is advisable to minimize storage time at low temperatures
to maintain higher chlorophyll levels.
It is important to note that the optimal method and conditions for preserving chloro-
phyll content may vary depending on the specific fruit or vegetable being processed.
Additionally, other factors such as the desired texture, taste, and nutrient retention should
also be considered when determining the best processing method for a particular food
Molecules 2023, 28, 5344 14 of 21
item. Further research and experimentation may be necessary to obtain more specific and
comprehensive guidelines for maximizing chlorophyll retention during food processing.
Table 2. Chlorophyll content in fruits and vegetables before and after processing.
Total Chlorophyll
Chlorophyll Content
Fruit/Vegetable Content (Fresh/Raw Food Processing References
after Processing
Material)
Broccoli 19.1 mg/kg (d.w.) [122]
21 mg/kg (f.w.) [123]
72.6 mg/kg (f.w.) [124]
128.2 mg/kg (f.w.) [112]
6940 mg/kg (d.w.) Boiling for 5 min 4140 mg/kg (d.w.)
Steaming for 7.5 min 2800 mg/kg (d.w.) [115]
Microwaving for 1.5 min 4100 mg/kg (d.w.)
Brussels sprouts 31.8 mg/kg (f.w.) [125]
57.5 mg/kg (f.w.) Microwaving for 6 min 30.4 mg/kg (f.w.) [124]
Cabbage 15.3 mg/kg (f.w.) [124]
Celery 23 mg/kg (f.w.) [123]
34.5 mg/kg (f.w.) Stored at 0 ◦ C for 21 days 15 mg/kg (f.w.)
Thermally treated by [116]
immersion (50 ◦ C for 90 s)
30 mg/kg (f.w.)
and stored at 0 ◦ C for
21 days
Thermally treated by
heated air (48 ◦ C for 1 h)
14 mg/kg (f.w.)
and stored at 0 ◦ C for
21 days
Cucumber 36 mg/kg (f.w.) [123]
Dandelion 2482.5 mg/kg (f.w.) [126]
Garden rocket 3596.2 mg/kg (f.w.) [126]
Green beans 71–133 mg/kg (f.w.) [127]
75 mg/kg (f.w.) [123]
Green paprika 38 mg/kg (f.w.) [123]
Green peas 50 mg/kg (f.w.) [123]
Green pepper 86.1 mg/kg (f.w.) [112]
797.8 mg/kg (d.w.) [122]
Heat steam sterilization at
Kale 282 mg/kg (f.w.) n.d. [128]
121 ◦ C for 5 min
1834 mg/kg (f.w.) Microwaving for 6 min 1142 mg/kg (f.w.) [124]
Puréed fruit after boiling
Kiwifruit purée 6.8 mg/kg (f.w.) n.d.
for 5 min
Frozen purée at −18 ◦ C
[129]
stored for:
1 day 3.9 mg/kg (f.w.)
36 days 1.9 mg/kg (f.w.)
68 days 0.9 mg/kg (f.w.)
Leek 1800 mg/kg (d.w.) Boiling for 5 min 480 mg/kg (d.w.)
Lettuce 2888.1 mg/kg (d.w.) [122]
Parsley 632 mg/kg (f.w.) [123]
Heat steam sterilization at
995 mg/kg (f.w.) n.d. [128]
121 ◦ C for 5 min
Peas 87 mg/kg (f.w.) [123]
Molecules 2023, 28, 5344 15 of 21
Table 2. Cont.
Total Chlorophyll
Chlorophyll Content
Fruit/Vegetable Content (Fresh/Raw Food Processing References
after Processing
Material)
639.1 mg/kg (f.w.)
Spinach [128]
(spinach purée)
791 mg/kg (f.w.) [123]
1083.4 mg/kg (f.w.) [112]
1148 mg/kg (f.w.) [124]
9470 mg/kg (d.w.) Blanched 9250 mg/kg (d.w.)
Processed at 121 ◦ C for:
4 min 6800 mg/kg (d.w.) [113]
15 min 1480 mg/kg (d.w.)
30 min n.d.
20330 mg/kg (d.w.) Baking at 105 ◦ C for:
20 min 14000 mg/kg (d.w.)
40 min 9580 mg/kg (d.w.)
80 min 7380 mg/kg (d.w.)
Blanching for:
6 min 9670 mg/kg (d.w.)
9 min 8270 mg/kg (d.w.)
15 min 5270 mg/kg (d.w.)
[114]
Steaming for:
7.5 min 4990 mg/kg (d.w.)
30 min 3510 mg/kg (d.w.)
60 min 1140 mg/kg (d.w.)
Microwave cooking for:
1 min 16,770 mg/kg (d.w.)
5 min 11,850 mg/kg (d.w.)
9 min 7980 mg/kg (d.w.)
39,090 mg/kg (d.w.) Boiling for 5 min 25,770 mg/kg (d.w.)
Steaming for 7.5 min 25,380 mg/kg (d.w.) [115]
Microwaving for 1 min 24,740 mg/kg (d.w.)
Squash 1660 mg/kg (d.w.) Boiling for 5 min 890 mg/kg (d.w.)
Wild rocket 3032.3 mg/kg (f.w.) [126]
Zucchini 68 mg/kg (f.w.) [123]
f.w., fresh weight; d.w., dry weight; n.d., not detectable.
5. Conclusions and Future Perspective

The rise in obesity, diabetes, cardiovascular diseases, and atherosclerosis can be at-
tributed to significant changes in human dietary patterns. However, the precise phar-
macokinetics of dietary chlorophylls and their derivatives, which play a crucial role in
conferring health benefits, remain poorly understood, needing further investigation. While
native chlorophylls have been the focus of limited research, both in vitro and in vivo studies
have shed light on the therapeutic potential of chlorophyll derivatives. These derivatives
exhibit a range of beneficial effects, including antioxidant, antimutagenic, antigenotoxic,
anticarcinogenic, and anti-obesogenic properties. Nevertheless, additional research is
required to validate the efficacy of dietary chlorophylls in treating the aforementioned
diseases and to explore their therapeutic potential in other medical conditions.
It is well-established that a high intake of vegetables and fruits promotes improved
health outcomes. The synergistic interplay among various bioactive compounds, rather
than the action of a single component, primarily underlies these benefits. In light of this,
investigating the potential health advantages and circular economy value of discarded
agri-food materials, such as broccoli leaves, which contain chlorophyll and other phy-
tochemicals, presents an intriguing avenue for research. Such exploration can provide
valuable insights into harnessing the health-promoting properties of these wasted resources
and their potential contribution to a sustainable and holistic approach to nutrition.
Molecules 2023, 28, 5344 16 of 21
Author Contributions: Conceptualization, T.M., L.A., A.N.B. and E.R.; writing—original draft prepara-
tion, T.M. and A.N.B.; writing—review and editing, T.M., A.N.B., L.A. and E.R.; funding acquisition,
A.N.B. and E.R. All authors have read and agreed to the published version of the manuscript.
Funding: This work was funded by the Portuguese Foundation for Science and Technology (FCT) and co-
financed by the European Regional Development Fund (FEDER) through COMPETE 2020—Operational
Competitiveness and Internationalization Programme (POCI), grant PTDC/ASP-HOR/29152/2017,
POCI-01-0145-FEDER-029152 (VALORIZEBYPRODUCTS). This work was also supported by National
Funds by FCT—Portuguese Foundation for Science and Technology, under the project UIDB/04033/2020.
Conflicts of Interest: The authors declare no conflict of interest.
References
1. Björn, L.O.; Papageorgiou, G.C.; Blankenship, R.E.; Govindjee, A. Viewpoint: Why chlorophyll a? Photosynth. Res. 2009, 99, 85–98.
[CrossRef] [PubMed]
2. Nagini, S.; Palitti, F.; Natarajan, A.T. Chemopreventive Potential of Chlorophyllin: A Review of the Mechanisms of Action and
Molecular Targets. Nutr. Cancer. 2015, 67, 203–211. [CrossRef] [PubMed]
3. MacKeen, D. Influencers Are Drinking Chlorophyll Water. But Why? The New York Times: New York, NY, USA, 2 July 2021.
4. FAO. Global Food Losses and Food Waste: Extent, Causes and Prevention; FAO: Rome, Italy, 2011.
5. Queiroz Zepka, L.; Jacob-Lopes, E.; Roca, M. Catabolism and bioactive properties of chlorophylls. Curr. Opi. Food Sci. 2019, 26,
94–100. [CrossRef]
6. Roca, M.; Chen, K.; Pérez-Gálvez, A. 6-Chlorophylls. In Handbook on Natural Pigments in Food and Beverages; Carle, R., Schweiggert,
R.M., Eds.; Woodhead Publishing: Cambridge, UK, 2016; pp. 125–158.
7. Durrett, T.P.; Welti, R. The tail of chlorophyll: Fates for phytol. J. Biol. Chem. 2021, 296, 100802. [CrossRef]
8. Pareek, S.; Sagar, N.A.; Sharma, S.; Kumar, V.; Agarwal, T.; González-Aguilar, G.A.; Yahia, E.M. Chlorophylls: Chemistry and
Biological Functions. In Fruit and Vegetable Phytochemicals; Yahia, E.M., Ed.; Wiley: Hoboken, NJ, USA, 2017; pp. 269–284.
9. Wang, Y.-T.; Yang, C.-H.; Huang, K.-S.; Shaw, J.-F. Chlorophyllides: Preparation, Purification, and Application. Biomolecules 2021,
11, 1115. [CrossRef]
10. Hsieh-Lo, M.; Castillo, G.; Ochoa-Becerra, M.A.; Mojica, L. Phycocyanin and phycoerythrin: Strategies to improve production
yield and chemical stability. Algal Res. 2019, 42, 101600. [CrossRef]
11. Halberstein, R.A. Medicinal Plants: Historical and Cross-Cultural Usage Patterns. Ann. Epidemiol. 2005, 15, 686–699. [CrossRef]
12. Soelberg, J.; Davis, O.; Jäger, A.K. Historical versus contemporary medicinal plant uses in the US Virgin Islands. J. Ethnopharmacol.
2016, 192, 74–89. [CrossRef]
13. Weckerle, C.S.; Ineichen, R.; Huber, F.K.; Yang, Y. Mao’s heritage: Medicinal plant knowledge among the Bai in Shaxi, China, at a
crossroads between distinct local and common widespread practice. J. Ethnopharmacol. 2009, 123, 213–228. [CrossRef]
14. Calixto, J.B. Twenty-five years of research on medicinal plants in Latin America: A personal view. J. Ethnopharmacol. 2005, 100,
15. Ayyanar, M.; Ignacimuthu, S. Ethnobotanical survey of medicinal plants commonly used by Kani tribals in Tirunelveli hills of
Western Ghats, India. J. Ethnopharmacol. 2011, 134, 851–864. [CrossRef] [PubMed]
16. Boy, H.I.A.; Rutilla, A.J.H.; Santos, K.A.; Ty, A.M.T.; Yu, A.I.; Mahboob, T.; Tangpoong, J.; Nissapatorn, V. Recommended
Medicinal Plants as Source of Natural Products: A Review. Digit. Chin. Med. 2018, 1, 131–142. [CrossRef]
17. Menendez-Baceta, G.; Aceituno-Mata, L.; Molina, M.; Reyes-García, V.; Tardío, J.; Pardo-de-Santayana, M. Medicinal plants
traditionally used in the northwest of the Basque Country (Biscay and Alava), Iberian Peninsula. J. Ethnopharmacol. 2014, 152,
113–134. [CrossRef] [PubMed]
18. Zorofchian Moghadamtousi, S.; Rouhollahi, E.; Karimian, H.; Fadaeinasab, M.; Firoozinia, M.; Ameen Abdulla, M.; Abdul Kadir, H.
The Chemopotential Effect of Annona muricata Leaves against Azoxymethane-Induced Colonic Aberrant Crypt Foci in Rats and
the Apoptotic Effect of Acetogenin Annomuricin E in HT-29 Cells: A Bioassay-Guided Approach. PLoS ONE 2015, 10, e0122288.
[CrossRef]
19. Chassagne, F.; Huang, X.; Lyles, J.T.; Quave, C.L. Validation of a 16th Century Traditional Chinese Medicine Use of Ginkgo biloba
as a Topical Antimicrobial. Front. Microbiol. 2019, 10, 775. [CrossRef]
20. Ibrahim, M.H.; Jaafar, H.Z.; Karimi, E.; Ghasemzadeh, A. Primary, secondary metabolites, photosynthetic capacity and antioxidant
activity of the Malaysian Herb Kacip Fatimah (Labisia Pumila Benth) exposed to potassium fertilization under greenhouse
conditions. Int. J. Mol. Sci. 2012, 13, 15321–15342. [CrossRef]
21. Rastogi, S.; Pandey, M.M.; Kumar Singh Rawat, A. Medicinal plants of the genus Betula--traditional uses and a phytochemical-
pharmacological review. J. Ethnopharmacol. 2015, 159, 62–83. [CrossRef]
22. Alamgeer; Sharif, A.; Asif, H.; Younis, W.; Riaz, H.; Bukhari, I.A.; Assiri, A.M. Indigenous medicinal plants of Pakistan used to
treat skin diseases: A review. Chin. Med. 2018, 13, 52. [CrossRef]
Molecules 2023, 28, 5344 17 of 21
23. Bhat, J.A.; Kumar, M.; Bussmann, R.W. Ecological status and traditional knowledge of medicinal plants in Kedarnath Wildlife
Sanctuary of Garhwal Himalaya, India. J. Ethnobiol. Ethnomed. 2013, 9, 1. [CrossRef]
24. Ghorbani, A. Studies on pharmaceutical ethnobotany in the region of Turkmen Sahra, north of Iran: (Part 1): General results.
J. Ethnopharmacol. 2005, 102, 58–68. [CrossRef]
25. González, J.A.; García-Barriuso, M.; Amich, F. Ethnobotanical study of medicinal plants traditionally used in the Arribes del
Duero, western Spain. J. Ethnopharmacol. 2010, 131, 343–355. [CrossRef] [PubMed]
26. Tariq, A.; Mussarat, S.; Adnan, M. Review on ethnomedicinal, phytochemical and pharmacological evidence of Himalayan
anticancer plants. J. Ethnopharmacol. 2015, 164, 96–119. [CrossRef]
27. Tajidin, N.E.; Shaari, K.; Maulidiani, M.; Salleh, N.S.; Ketaren, B.R.; Mohamad, M. Metabolite profiling of Andrographis paniculata
(Burm. f.) Nees. young and mature leaves at different harvest ages using 1H NMR-based metabolomics approach. Sci. Rep. 2019,
9, 16766. [CrossRef] [PubMed]
28. Yilmaz, C.; Gökmen, V. Chlorophyll. In Encyclopedia of Food and Health; Caballero, B., Finglas, P.M., Toldrá, F., Eds.; Academic
Press: Oxford, UK, 2016; pp. 37–41.
29. Lanfer-Marquez, U.M.; Sinnecker, P. Chlorophylls: Properties, Biosynthesis, Degradation and Functions. In Food Colorants:
Chemical And Functional Properties; Socaciu, C., Ed.; CRC Press: Boca Raton, FL, USA, 2007; pp. 195–211.
30. Viera, I.; Pérez-Gálvez, A.; Roca, M. Green Natural Colorants. Molecules 2019, 24, 154. [CrossRef] [PubMed]
31. Egner, P.A.; Stansbury, K.H.; Snyder, E.P.; Rogers, M.E.; Hintz, P.A.; Kensler, T.W. Identification and characterization of chlorin e(4)
ethyl ester in sera of individuals participating in the chlorophyllin chemoprevention trial. Chem. Res. Toxicol. 2000, 13, 900–906.
[CrossRef]
32. Ferruzzi, M.G.; Failla, M.L.; Schwartz, S.J. Sodium copper chlorophyllin: In vitro digestive stability and accumulation by Caco-2
human intestinal cells. J. Agric. Food Chem. 2002, 50, 2173–2179. [CrossRef] [PubMed]
33. Gomes, B.B.; Barros, S.B.; Andrade-Wartha, E.R.; Silva, A.M.; Silva, V.V.; Lanfer-Marquez, U.M. Bioavailability of dietary sodium
copper chlorophyllin and its effect on antioxidant defence parameters of Wistar rats. J. Sci. Food Agric. 2009, 89, 2003–2010.
[CrossRef]
34. Ferruzzi, M.G.; Failla, M.L.; Schwartz, S.J. Assessment of degradation and intestinal cell uptake of carotenoids and chlorophyll
derivatives from spinach puree using an in vitro digestion and Caco-2 human cell model. J. Agric. Food Chem. 2001, 49, 2082–2089.
[CrossRef]
35. Gallardo-Guerrero, L.; Gandul-Rojas, B.; Mínguez-Mosquera, M.I. Digestive stability, micellarization, and uptake by Caco-2
human intestinal cell of chlorophyll derivatives from different preparations of pea (Pisum sativum L.). J. Agric. Food Chem. 2008,
56, 8379–8386. [CrossRef]
36. Gandul-Rojas, B.; Gallardo-Guerrero, L.; Mínguez-Mosquera, M.I. Influence of the chlorophyll pigment structure on its transfer
from an oily food matrix to intestinal epithelium cells. J. Agric. Food Chem. 2009, 57, 5306–5314. [CrossRef]
37. Chen, K.; Roca, M. In vitro digestion of chlorophyll pigments from edible seaweeds. J. Funct. Foods 2018, 40, 400–407. [CrossRef]
38. Chen, K.; Roca, M. In vitro bioavailability of chlorophyll pigments from edible seaweeds. J. Funct. Foods 2018, 41, 25–33. [CrossRef]
39. Chao, P.-Y.; Huang, M.-Y.; Huang, W.-D.; Lin, K.-H.R.; Chen, S.-Y.; Yang, C.-M. Study of Chlorophyll-related Compounds from
Dietary Spinach in Human Blood. Not. Bot. Horti. Agrobo. 2018, 46, 309–316. [CrossRef]
40. Fernandes, T.M.; Gomes, B.B.; Lanfer-Marquez, U.M. Apparent absorption of chlorophyll from spinach in an assay with dogs.
Innov. Food Sci. Emerg. Technol. 2007, 8, 426–432. [CrossRef]
41. Hsu, C.Y.; Yeh, T.H.; Huang, M.Y.; Hu, S.P.; Chao, P.Y.; Yang, C.M. Organ-specific distribution of chlorophyll-related compounds
from dietary spinach in rabbits. Indian J. Biochem. Biophys. 2014, 51, 388–395. [PubMed]
42. Viera, I.; Chen, K.; Ríos, J.J.; Benito, I.; Pérez-Gálvez, A.; Roca, M. First-Pass Metabolism of Chlorophylls in Mice. Mol. Nutr. Food
Res. 2018, 62, e1800562. [CrossRef]
43. Fasakin, C.F.; Udenigwe, C.C.; Aluko, R.E. Antioxidant properties of chlorophyll-enriched and chlorophyll-depleted polyphenolic
fractions from leaves of Vernonia amygdalina and Gongronema latifolium. Food Res. Int. 2011, 44, 2435–2441. [CrossRef]
44. Ferruzzi, M.G.; Böhm, V.; Courtney, P.D.; Schwartz, S.J. Antioxidant and Antimutagenic Activity of Dietary Chlorophyll
Derivatives Determined by Radical Scavenging and Bacterial Reverse Mutagenesis Assays. J. Food Sci. 2002, 67, 2589–2595.
[CrossRef]
45. Lanfer-Marquez, U.M.; Barros, R.M.C.; Sinnecker, P. Antioxidant activity of chlorophylls and their derivatives. Food Res. Int. 2005,
38, 885–891. [CrossRef]
46. Yahia, E.M. Fruit and Vegetable Phytochemicals: Chemistry and Human Health, 2nd ed.; John Wiley & Sons Ltd.: Hoboken, NJ, USA,
2018; Volume I.
47. Perez-Galvez, A.; Viera, I.; Roca, M. Chemistry in the Bioactivity of Chlorophylls: An Overview. Curr. Med. Chem. 2017, 24,
4515–4536. [CrossRef]
48. Cai, J.-Q.; Liu, X.-M.; Gao, Z.-J.; Li, L.-L.; Wang, H. Chlorophylls derivatives: Photophysical properties, assemblies, nanostructures
and biomedical applications. Mater. Today 2021, 45, 77–92. [CrossRef]
49. De Vogel, J.; Jonker-Termont, D.S.; van Lieshout, E.M.; Katan, M.B.; van der Meer, R. Green vegetables, red meat and colon cancer:
Chlorophyll prevents the cytotoxic and hyperproliferative effects of haem in rat colon. Carcinogenesis 2005, 26, 387–393. [CrossRef]
[PubMed]
Molecules 2023, 28, 5344 18 of 21
50. Egner, P.A.; Wang, J.B.; Zhu, Y.R.; Zhang, B.C.; Wu, Y.; Zhang, Q.N.; Qian, G.S.; Kuang, S.Y.; Gange, S.J.; Jacobson, L.P.; et al.
Chlorophyllin intervention reduces aflatoxin-DNA adducts in individuals at high risk for liver cancer. Proc. Natl. Acad. Sci. USA
2001, 98, 14601–14606. [CrossRef]
51. Hayes, M.; Ferruzzi, M.G. Update on the bioavailability and chemopreventative mechanisms of dietary chlorophyll derivatives.
Nutr. Res. 2020, 81, 19–37. [CrossRef]
52. Khattab, R.; Goldberg, E.; Lin, L.; Thiyam, U. Quantitative analysis and free-radical-scavenging activity of chlorophyll, phytic
acid, and condensed tannins in canola. Food Chem. 2010, 122, 1266–1272. [CrossRef]
53. Endo, Y.; Usuki, R.; Kaneda, T. Antioxidant effects of chlorophyll and pheophytin on the autoxidation of oils in the dark. II. The
mechanism of antioxidative action of chlorophyll. J. Am. Oil Chem. Soc. 1985, 62, 1387–1390. [CrossRef]
54. Kang, Y.-R.; Park, J.; Jung, S.K.; Chang, Y.H. Synthesis, characterization, and functional properties of chlorophylls, pheophytins,
and Zn-pheophytins. Food Chem. 2018, 245, 943–950. [CrossRef]
55. Serpeloni, J.M.; Grotto, D.; Aissa, A.F.; Mercadante, A.Z.; Bianchi Mde, L.; Antunes, L.M. An evaluation, using the comet assay
and the micronucleus test, of the antigenotoxic effects of chlorophyll b in mice. Mutat. Res. 2011, 725, 50–56. [CrossRef]
56. Lai, C.-N.; Butler, M.A.; Matney, T.S. Antimutagenic activities of common vegetables and their chlorophyll content. Mut. Res.
1980, 77, 245–250. [CrossRef]
57. Negishi, T.; Rai, H.; Hayatsu, H. Antigenotoxic activity of natural chlorophylls. Mutat. Res. 1997, 376, 97–100. [CrossRef]
58. Kocaoğlu Cenkci, S.; Kaya, B. Effects of Chlorophyll a and b in Reducing Genotoxicity of 2-Amino-3,8-dimethylimidazo[4,5-
F]quinoxaline (MeIQx). Biology 2022, 11, 602. [CrossRef] [PubMed]
59. Serpeloni, J.M.; Batista, B.L.; Angeli, J.P.; Barcelos, G.R.; Bianchi Mde, L.; Barbosa, F., Jr.; Antunes, L.M. Antigenotoxic properties
of chlorophyll b against cisplatin-induced DNA damage and its relationship with distribution of platinum and magnesium
in vivo. J. Toxicol. Environ. Health A 2013, 76, 345–353. [CrossRef] [PubMed]
60. Nakamura, Y.; Murakami, A.; Koshimizu, K.; Ohigashi, H. Inhibitory effect of pheophorbide a, a chlorophyll-related compound,
on skin tumor promotion in ICR mouse. Cancer Lett. 1996, 108, 247–255. [CrossRef] [PubMed]
61. Higashi-Okai, K.; Okai, Y. Potent suppressive activity of chlorophyll a and b from green tea (Camellia sinensis) against tumor
promotion in mouse skin. J. UOEH 1998, 20, 181–188. [CrossRef]
62. Harttig, U.; Bailey, G.S. Chemoprotection by natural chlorophylls in vivo: Inhibition of dibenzo[a,l]pyrene-DNA adducts in
rainbow trout liver. Carcinogenesis 1998, 19, 1323–1326. [CrossRef] [PubMed]
63. McQuistan, T.J.; Simonich, M.T.; Pratt, M.M.; Pereira, C.B.; Hendricks, J.D.; Dashwood, R.H.; Williams, D.E.; Bailey, G.S. Cancer
chemoprevention by dietary chlorophylls: A 12,000-animal dose-dose matrix biomarker and tumor study. Food Chem. Toxicol.
2012, 50, 341–352. [CrossRef]
64. Simonich, M.T.; McQuistan, T.; Jubert, C.; Pereira, C.; Hendricks, J.D.; Schimerlik, M.; Zhu, B.; Dashwood, R.H.; Williams, D.E.;
Bailey, G.S. Low-dose dietary chlorophyll inhibits multi-organ carcinogenesis in the rainbow trout. Food Chem. Toxicol. 2008, 46,
1014–1024. [CrossRef]
65. Breinholt, V.; Arbogast, D.; Loveland, P.; Pereira, C.; Dashwood, R.; Hendricks, J.; Bailey, G. Chlorophyllin chemoprevention in
trout initiated by aflatoxin B(1) bath treatment: An evaluation of reduced bioavailability vs. target organ protective mechanisms.
Toxicol. Appl. Pharmacol. 1999, 158, 141–151. [CrossRef]
66. Breinholt, V.; Hendricks, J.; Pereira, C.; Arbogast, D.; Bailey, G. Dietary chlorophyllin is a potent inhibitor of aflatoxin B1
hepatocarcinogenesis in rainbow trout. Cancer Res. 1995, 55, 57–62.
67. Breinholt, V.; Schimerlik, M.; Dashwood, R.; Bailey, G. Mechanisms of chlorophyllin anticarcinogenesis against aflatoxin B1:
Complex formation with the carcinogen. Chem. Res. Toxicol. 1995, 8, 506–514. [CrossRef]
68. Simonich, M.T.; Egner, P.A.; Roebuck, B.D.; Orner, G.A.; Jubert, C.; Pereira, C.; Groopman, J.D.; Kensler, T.W.; Dashwood, R.H.;
Williams, D.E.; et al. Natural chlorophyll inhibits aflatoxin B1-induced multi-organ carcinogenesis in the rat. Carcinogenesis 2007,
28, 1294–1302. [CrossRef] [PubMed]
69. De Vogel, J.; Jonker-Termont, D.S.; Katan, M.B.; van der Meer, R. Natural chlorophyll but not chlorophyllin prevents heme-induced
cytotoxic and hyperproliferative effects in rat colon. J. Nutr. 2005, 135, 1995–2000. [CrossRef] [PubMed]
70. Koníčková, R.; Vaňková, K.; Vaníková, J.; Váňová, K.; Muchová, L.; Subhanová, I.; Zadinová, M.; Zelenka, J.; Dvořák, A.; Kolář,
M.; et al. Anti-cancer effects of blue-green alga Spirulina platensis, a natural source of bilirubin-like tetrapyrrolic compounds.
Ann. Hepatol. 2014, 13, 273–283. [CrossRef] [PubMed]
71. Das, J.; Samadder, A.; Mondal, J.; Abraham, S.K.; Khuda-Bukhsh, A.R. Nano-encapsulated chlorophyllin significantly delays
progression of lung cancer both in in vitro and in vivo models through activation of mitochondrial signaling cascades and
drug-DNA interaction. Environ. Toxicol. Pharmacol. 2016, 46, 147–157. [CrossRef] [PubMed]
72. Dashwood, R. Chlorophylls as anticarcinogens (review). Int. J. Oncol. 1997, 10, 721–727. [CrossRef]
73. McRae, M.P. The Benefits of Dietary Fiber Intake on Reducing the Risk of Cancer: An Umbrella Review of Meta-analyses.
J. Chiropr. Med. 2018, 17, 90–96. [CrossRef] [PubMed]
74. O’Connor, A.E.; Gallagher, W.M.; Byrne, A.T. Porphyrin and nonporphyrin photosensitizers in oncology: Preclinical and clinical
advances in photodynamic therapy. Photochem. Photobiol. 2009, 85, 1053–1074. [CrossRef]
Molecules 2023, 28, 5344 19 of 21
75. Kim, J.E.; Hwang, J.I.; Lee, J.I.; Cho, B.K.; Park, H.J. Pilot study on photodynamic therapy for acne using chlorophyll: Evaluator-
blinded, split-face study. J. Dermatolog. Treat. 2012, 23, 35–36. [CrossRef]
76. Chan, A.L.; Juarez, M.; Allen, R.; Volz, W.; Albertson, T. Pharmacokinetics and clinical effects of mono-L-aspartyl chlorin e6
(NPe6) photodynamic therapy in adult patients with primary or secondary cancer of the skin and mucosal surfaces. Photodermatol.
Photoimmunol. Photomed. 2005, 21, 72–78. [CrossRef]
77. Kessel, D. Pharmacokinetics of N-aspartyl chlorin e6 in cancer patients. J. Photochem. Photobiol. B 1997, 39, 81–83. [CrossRef]
78. Zhuo, Z.; Song, Z.; Ma, Z.; Zhang, Y.; Xu, G.; Chen, G. Chlorophyllin e6-mediated photodynamic therapy inhibits proliferation
and induces apoptosis in human bladder cancer cells. Oncol. Rep. 2019, 41, 2181–2193. [CrossRef]
79. Galgani, J.; Ravussin, E. Energy metabolism, fuel selection and body weight regulation. Int. J. Obes. 2008, 32 (Suppl. S7),
S109–S119. [CrossRef]
80. Jahangir, E.; De Schutter, A.; Lavie, C.J. The relationship between obesity and coronary artery disease. Transl. Res. 2014, 164,
81. Wu, S.J.; Ng, L.T.; Wang, G.H.; Huang, Y.J.; Chen, J.L.; Sun, F.M. Chlorophyll a, an active anti-proliferative compound of Ludwigia
octovalvis, activates the CD95 (APO-1/CD95) system and AMPK pathway in 3T3-L1 cells. Food Chem. Toxicol. 2010, 48, 716–721.
[CrossRef] [PubMed]
82. Wang, X.; Li, Y.; Shen, S.; Yang, Z.; Zhang, H.; Zhang, Y. Chlorophyll Inhibits the Digestion of Soybean Oil in Simulated Human
Gastrointestinal System. Nutrients 2022, 14, 1749. [CrossRef] [PubMed]
83. Erlanson-Albertsson, C.; Albertsson, P. The Use of Green Leaf Membranes to Promote Appetite Control, Suppress Hedonic
Hunger and Loose Body Weight. Plant Foods Hum. Nutr. 2015, 70, 281–290. [CrossRef] [PubMed]
84. Montelius, C.; Erlandsson, D.; Vitija, E.; Stenblom, E.L.; Egecioglu, E.; Erlanson-Albertsson, C. Body weight loss, reduced urge for
palatable food and increased release of GLP-1 through daily supplementation with green-plant membranes for three months in
overweight women. Appetite 2014, 81, 295–304. [CrossRef] [PubMed]
85. Köhnke, R.; Lindqvist, A.; Göransson, N.; Emek, S.C.; Albertsson, P.A.; Rehfeld, J.F.; Hultgårdh-Nilsson, A.; Erlanson-Albertsson,
C. Thylakoids suppress appetite by increasing cholecystokinin resulting in lower food intake and body weight in high-fat fed
mice. Phytother. Res. 2009, 23, 1778–1783. [CrossRef]
86. Montelius, C.; Szwiec, K.; Kardas, M.; Lozinska, L.; Erlanson-Albertsson, C.; Pierzynowski, S.; Rehfeld, J.F.; Weström, B. Dietary
thylakoids suppress blood glucose and modulate appetite-regulating hormones in pigs exposed to oral glucose tolerance test.
Clin. Nutr. 2014, 33, 1122–1126. [CrossRef]
87. Seo, Y.J.; Kim, K.J.; Choi, J.; Koh, E.J.; Lee, B.Y. Spirulina maxima Extract Reduces Obesity through Suppression of Adipogenesis
and Activation of Browning in 3T3-L1 Cells and High-Fat Diet-Induced Obese Mice. Nutrients 2018, 10, 712. [CrossRef]
88. Li, Y.; Cui, Y.; Hu, X.; Liao, X.; Zhang, Y. Chlorophyll Supplementation in Early Life Prevents Diet-Induced Obesity and Modulates
Gut Microbiota in Mice. Mol. Nutr. Food Res. 2019, 63, e1801219. [CrossRef] [PubMed]
89. Li, Y.; Cui, Y.; Lu, F.; Wang, X.; Liao, X.; Hu, X.; Zhang, Y. Beneficial effects of a chlorophyll-rich spinach extract supplementation
on prevention of obesity and modulation of gut microbiota in high-fat diet-fed mice. J. Funct. Foods 2019, 60, 103436. [CrossRef]
90. Stenblom, E.L.; Weström, B.; Linninge, C.; Bonn, P.; Farrell, M.; Rehfeld, J.F.; Montelius, C. Dietary green-plant thylakoids decrease
gastric emptying and gut transit, promote changes in the gut microbial flora, but does not cause steatorrhea. Nutr. Metab. 2016,
13, 67. [CrossRef] [PubMed]
91. Zheng, H.; You, Y.; Hua, M.; Wu, P.; Liu, Y.; Chen, Z.; Zhang, L.; Wei, H.; Li, Y.; Luo, M.; et al. Chlorophyllin Modulates Gut
Microbiota and Inhibits Intestinal Inflammation to Ameliorate Hepatic Fibrosis in Mice. Front. Physiol. 2018, 9, 1671. [CrossRef]
92. Sanmiguel, C.; Gupta, A.; Mayer, E.A. Gut Microbiome and Obesity: A Plausible Explanation for Obesity. Curr. Obes. Rep. 2015, 4,
250–261. [CrossRef] [PubMed]
93. Kau, A.L.; Ahern, P.P.; Griffin, N.W.; Goodman, A.L.; Gordon, J.I. Human nutrition, the gut microbiome and the immune system.
Nature 2011, 474, 327–336. [CrossRef]
94. Sonnenburg, J.L.; Bäckhed, F. Diet-microbiota interactions as moderators of human metabolism. Nature 2016, 535, 56–64.
[CrossRef]
95. An, X.; Bao, Q.; Di, S.; Zhao, Y.; Zhao, S.; Zhang, H.; Lian, F.; Tong, X. The interaction between the gut Microbiota and herbal
medicines. Biomed. Pharmacother. 2019, 118, 109252. [CrossRef]
96. Hasanuzzaman, M.; Bhuyan, M.; Parvin, K.; Bhuiyan, T.F.; Anee, T.I.; Nahar, K.; Hossen, M.S.; Zulfiqar, F.; Alam, M.M.; Fujita, M.
Regulation of ROS Metabolism in Plants under Environmental Stress: A Review of Recent Experimental Evidence. Int. J. Mol. Sci.
2020, 21, 8695. [CrossRef]
97. Liu, H.; Xiao, C.; Qiu, T.; Deng, J.; Cheng, H.; Cong, X.; Cheng, S.; Rao, S.; Zhang, Y. Selenium Regulates Antioxidant,
Photosynthesis, and Cell Permeability in Plants under Various Abiotic Stresses: A Review. Plants 2022, 12, 44. [CrossRef]
98. Okai, Y.; Higashi-Okai, K. Protective effects of chlorophyll a and pheophytin a derived from green tea (Camellia sinensis) on
p-nonylphenol-induced cell growth inhibition and oxygen radical generation in yeast (Saccharomyces cerevisiae). J. Sci. Food
Agric. 2001, 81, 1443–1446. [CrossRef]
99. Wu, Z.; Ren, Y.; Liang, Y.; Huang, L.; Yang, Y.; Zafar, A.; Hasan, M.; Yang, F.; Shu, X. Synthesis, Characterization, Immune
Regulation, and Antioxidative Assessment of Yeast-Derived Selenium Nanoparticles in Cyclophosphamide-Induced Rats. Acs
Omega 2021, 6, 24585–24594. [CrossRef] [PubMed]
Molecules 2023, 28, 5344 20 of 21
100. Lee, D.; Nishizawa, M.; Shimizu, Y.; Saeki, H. Anti-inflammatory effects of dulse (Palmaria palmata) resulting from the
simultaneous water-extraction of phycobiliproteins and chlorophyll a. Food Res. Int. 2017, 100, 514–521. [CrossRef] [PubMed]
101. Sies, H.; Jones, D.P. Reactive oxygen species (ROS) as pleiotropic physiological signalling agents. Nat. Rev. Mol. Cell Biol. 2020, 21,
102. Hannan, M.A.; Dash, R.; Sohag, A.A.M.; Haque, M.N.; Moon, I.S. Neuroprotection Against Oxidative Stress: Phytochemicals
Targeting TrkB Signaling and the Nrf2-ARE Antioxidant System. Front. Mol. Neurosci. 2020, 13, 116. [CrossRef]
103. Rehni, A.K.; Pantlya, H.S.; Shri, R.; Singh, M. Effect of chlorophyll and aqueous extracts of Bacopa monniera and Valeriana
wallichii on ischaemia and reperfusion-induced cerebral injury in mice. Indian J. Exp. Biol. 2007, 45, 764–769.
104. Saleh, H.A.; Yousef, M.H.; Abdelnaser, A. The Anti-Inflammatory Properties of Phytochemicals and Their Effects on Epigenetic
Mechanisms Involved in TLR4/NF-κB-Mediated Inflammation. Front. Immunol. 2021, 12, 606069. [CrossRef]
105. Subramoniam, A.; Asha, V.V.; Nair, S.A.; Sasidharan, S.P.; Sureshkumar, P.K.; Rajendran, K.N.; Karunagaran, D.; Ramalingam, K.
Chlorophyll revisited: Anti-inflammatory activities of chlorophyll a and inhibition of expression of TNF-α gene by the same.
Inflammation 2012, 35, 959–966. [CrossRef]
106. Canosa-Carro, L.; Bravo-Aguilar, M.; Abuín-Porras, V.; Almazán-Polo, J.; García-Pérez-de-Sevilla, G.; Rodríguez-Costa, I.; López-
López, D.; Navarro-Flores, E.; Romero-Morales, C. Current understanding of the diagnosis and management of the tendinopathy:
An update from the lab to the clinical practice. Dis. Mon. 2022, 68, 101314. [CrossRef]
107. Castelao-Baptista, J.P.; Barros, A.; Martins, T.; Rosa, E.; Sardao, V.A. Three in One: The Potential of Brassica By-Products against
Economic Waste, Environmental Hazard, and Metabolic Disruption in Obesity. Nutrients 2021, 13, 4194. [CrossRef]
108. Esparza, I.; Jiménez-Moreno, N.; Bimbela, F.; Ancín-Azpilicueta, C.; Gandía, L.M. Fruit and vegetable waste management:
Conventional and emerging approaches. J. Environ. Manag. 2020, 265, 110510. [CrossRef] [PubMed]
109. Martins, T.; Colaco, B.; Venancio, C.; Pires, M.J.; Oliveira, P.A.; Rosa, E.; Antunes, L.M. Potential effects of sulforaphane to fight
obesity. J. Sci. Food Agric. 2018, 98, 2837–2844. [CrossRef]
110. Talebzadeh, F.; Valeo, C. Evaluating the Effects of Environmental Stress on Leaf Chlorophyll Content as an Index for Tree Health.
IOP Conf. Ser. Earth Environ. Sci. 2022, 1006, 012007. [CrossRef]
111. Castelão-Baptista, J.P.; Valente, S.A.; Canário, S.; Oppolzer, D.; Barros, A.; Venâncio, C.; Martins, T.; Antunes, L.; Sardão, V.A.;
Rosa, E.; et al. Brassica oleracea Var italica by-Products Prevent Lipid Accumulation and Cell Death in a Liver Cell Model of
Lipid Toxicity. Nutrients 2023, 15, 924. [CrossRef] [PubMed]
112. Sánchez, C.; Baranda, A.B.; Martínez de Marañón, I. The effect of High Pressure and High Temperature processing on carotenoids
and chlorophylls content in some vegetables. Food Chem. 2014, 163, 37–45. [CrossRef]
113. Schwartz, S.J.; Lorenzo, T.V. Chlorophylls in foods. Crit. Rev. Food Sci. Nutr. 1990, 29, 1–17. [CrossRef]
114. Teng, S.S.; Chen, B.H. Formation of pyrochlorophylls and their derivatives in spinach leaves during heating. Food Chem. 1999, 65,
115. Turkmen, N.; Poyrazoglu, E.S.; Sari, F.; Sedat Velioglu, Y. Effects of cooking methods on chlorophylls, pheophytins and colour of
selected green vegetables. Int. J. Food Sci. Techn. 2006, 41, 281–288. [CrossRef]
116. Viña, S.Z.; Osornio, M.M.L.; Chaves, A.R. Quality changes in fresh-cut celery as affected by heat treatment and storage. J. Sci.
Food Agric. 2007, 87, 1400–1407. [CrossRef]
117. Herrera, M.; Viera, I.; Roca, M. HPLC-MS(2) Analysis of Chlorophylls in Green Teas Establishes Differences among Varieties.
Molecules 2022, 27, 6171. [CrossRef]
118. Fang, H.; Zhou, Q.; Cheng, S.; Zhou, X.; Wei, B.; Zhao, Y.; Ji, S. 24-epibrassinolide alleviates postharvest yellowing of broccoli via
improving its antioxidant capacity. Food Chem. 2021, 365, 130529. [CrossRef] [PubMed]
119. Li, X.; Meng, Z.; Malik, A.U.; Zhang, S.; Wang, Q. Maintaining the quality of postharvest broccoli by inhibiting ethylene
accumulation using diacetyl. Front. Nutr. 2022, 9, 1055651. [CrossRef] [PubMed]
120. Cano, A.; Giraldo-Acosta, M.; García-Sánchez, S.; Hernández-Ruiz, J.; Arnao, M.B. Effect of Melatonin in Broccoli Postharvest and
Possible Melatonin Ingestion Level. Plants 2022, 11, 2000. [CrossRef]
121. Viera, I.; Herrera, M.; Roca, M. Influence of food composition on chlorophyll bioaccessibility. Food Chem. 2022, 386, 132805.
[CrossRef]
122. Burns, J.; Fraser, P.D.; Bramley, P.M. Identification and quantification of carotenoids, tocopherols and chlorophylls in commonly
consumed fruits and vegetables. Phytochemistry 2003, 62, 939–947. [CrossRef] [PubMed]
123. Bohn, T.; Walczyk, T.; Leisibach, S.; Hurrell, R.F. Chlorophyll-bound Magnesium in Commonly Consumed Vegetables and Fruits:
Relevance to Magnesium Nutrition. J. Food Sci. 2004, 69, S347–S350. [CrossRef]
124. Khachik, F.; Beecher, G.R.; Whittaker, N.F. Separation, identification, and quantification of the major carotenoid and chlorophyll
constituents in extracts of several green vegetables by liquid chromatography. J. Agri. Food Chem. 1986, 34, 603–616. [CrossRef]
125. Olivera, D.F.; Viña, S.Z.; Marani, C.M.; Ferreyra, R.M.; Mugridge, A.; Chaves, A.R.; Mascheroni, R.H. Effect of blanching on the
quality of Brussels sprouts (Brassica oleracea L. gemmifera DC) after frozen storage. J. Food Eng. 2008, 84, 148–155. [CrossRef]
126. Žnidarčič, D.; Ban, D.; Šircelj, H. Carotenoid and chlorophyll composition of commonly consumed leafy vegetables in Mediter-
ranean countries. Food Chem. 2011, 129, 1164–1168. [CrossRef]
127. Cubas, C.; Gloria Lobo, M.; González, M. Optimization of the extraction of chlorophylls in green beans (Phaseolus vulgaris L.) by
N,N-dimethylformamide using response surface methodology. J. Food Compost. Anal. 2008, 21, 125–133. [CrossRef]
Molecules 2023, 28, 5344 21 of 21
128. Arnold, C.; Schwarzenbolz, U.; Böhm, V. Carotenoids and chlorophylls in processed xanthophyll-rich food. Food Sci. Technol.
2014, 57, 442–445. [CrossRef]
129. Robertson, G.L. Changes in the chlorophyll and pheophytin concentrations of kiwifruit during processing and storage. Food
Chem. 1985, 17, 25–32. [CrossRef]
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molecules

Keywords: chlorophylls; chlorophyllin; health; biological activity

Molecules 2023, 28, 5344. https://doi.org/10.3390/molecules28145344 https://www.mdpi.com/journal/molecules

Molecules 2023, 28, x FOR PEER REVIEW 2 of 21

Figure 1. Structure from Chlorophyll a and Chlorophyll b.

Figure 2. Conversion of Chlorophyll in phaeophytin, chlorophyllide and pheophorbide.

straightforward processing, prolonged availability, and therapeutic properties resulting

3.2. Chlorophyll Bioavailability

3.3. Bioactive Properties of Chlorophyll Compounds

Figure 3. Bioactive properties and health benefits of chlorophyll compounds.

3.3.2. Antimutagenic and Antigenotoxic Properties

3.3.3. Anticancer Activity

3.3.4. Anti-Obesity Effects

3.3.5. Protection against Endocrine Disruptors

3.3.6. Neuroprotective and Anti-Inflammatory Effects

4. Chlorophyll Content in Fruits and Vegetables

Chlorophyll a Chlorophyll b Total Chlorophylls Total Carotenoids

5. Conclusions and Future Perspective

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