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Medicinal Uses of Chlorophyll: A Critical Overview


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Reports on traditional medicinal uses of chlorophyll in alternative forms of medicine are known since ages. Now-a-days chlorophyll has been used in the field of medicine as remedy and diagnostics. Chlorophyll molecules are used in pharmacy as photosensitizer for cancer therapy. Their roles as modifier of genotoxic effects are becoming increasingly important, besides these it being known to have multiple medicinal uses. Chlorophyll has its place in modern medicine. Here, we present a review of recent developments in medicinal uses of chlorophyll. This article enumerates therapeutic claims of chlorophyll as drugs based on investigative findings of modern science. A brief overview of research and developments of medicinal uses of chlorophyll will be presented in this review along with challenges of potential applications of chlorophyll and its derivatives as chemotherapeutic agents
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Medicinal Uses of Chlorophyll: A Critical Overview
V.K. Mishra1, R.K. Bacheti2 and Azamal Husen3*
1Department of Biotechnology, Doon (P.G.) Paramedical College, Dehra Dun-248001, India
2Department of Chemistry, Graphic Era University, Dehra Dun-248001, India
3*Department of Biology, Faculty of Natural and Computational Sciences, University of Gondar
P.O. Box 196, Gondar, Ethiopia
Reports on traditional medicinal uses of chlorophyll in alternative forms of medicine are known since
ages. Now-a-days chlorophyll has been used in the field of medicine as remedy and diagnostics.
Chlorophyll molecules are used in pharmacy as photosensitizer for cancer therapy. Their roles as
modifier of genotoxic effects are becoming increasingly important, besides these it being known to
have multiple medicinal uses. Chlorophyll has its place in modern medicine. Here, we present a
review of recent developments in medicinal uses of chlorophyll. This article enumerates therapeutic
claims of chlorophyll as drugs based on investigative findings of modern science. A brief overview of
research and developments of medicinal uses of chlorophyll will be presented in this review along
with challenges of potential applications of chlorophyll and its derivatives as chemotherapeutic agents
Keywords: Chlorophyll, medicine, genotoxity, photosensitizer
CHL: Chlorophyllin
ROS: Reactive oxygen species
PDT: Photodynamic therapy
PSMA: Prostrate-specific membrane antigen
ALA: Aminolevulinic acid
CDK: Cyclin dependent kinase
Can be cited as:
V.K. Mishra, R.K. Bacheti and Azamal Husen, 2011. Medicinal Uses of Chlorophyll: a critical overview. In: Chlorophyll: Structure,
Function and Medicinal Uses, Hua Le and and Elisa Salcedo, Eds., Nova Science Publishers, Inc., Hauppauge, NY 11788 (ISBN 978-1-
62100-015-0), pp.177-196. (
1. Introduction
Natural products have been the most important source of drugs. Throughout history, these products
have been used as important source of anticancer and chemopreventive agents. Many natural products
from our daily consumption of fruits, vegetables, tea beverages whose active ingredients have
potential health benefits. Recently, their uses are becoming increasingly popular as evident from the
sales of food supplements/functional foods which is growing at an amazing proportion, $4.59 billion
for 2006 and $4.79 billion for 2007 (Knasmüller et al., 2008). Despite, growing body of epidemolgical
and investigative findings supporting health claims of dietary supplements, there is urgent need to
ensure consumer concern about their efficacy and potentiality as medicine . Among several dietary
phytochemicals, chlorophyll being most ubiquitous natural pigments with physiological effects to cure
of chronic diseases, such as some forms of cancer.
The chlorophyll and its derivatives have long history in traditional medicine ((Esten and
Dannin, 1950; Kephart, 1955), and also various therapeutic uses including wound healing (Dashwood,
1997), anti-inflammatory agent (Bower, 1947; Larato et al., 1970), internal deodorant (Young et al.,
1980). Although these applications illustrate various medicinal uses of chlorophyll but interestingly
recent research works are more focused on its role as potent anti-mutagen and anti-carcinogen
(Dashwood, 1997, 2002, Egner et al., 2001, 2003), and also as photosensitizer in photodynamic
therapy (Henderson et al., 1997; Park, 1989; Li, et al., 2005). The intent of present article is aimed at
providing better understanding of science based health claims of chlorophyll.
2. Chemotherapeutic Potential of Chlorophyll
2.1. Chlorophyll and Its derivatives
Chlorophyll has a porphyrin ring similar to that of heme in hemoglobin, although the central atom in
chlorophyll is magnesium instead of iron (Figure 1). Chlorophyllin is a semi-synthetic mixture of
sodium copper salts derived from chlorophyll. During the synthesis of chlorophyllin, the magnesium
atom at the center of the ring is replaced with copper and the phytol tail is lost. Unlike natural
chlorophyll, chlorophyllin is water-soluble. Although the content of different chlorophyllin mixtures
may vary, two compounds commonly found in commercial chlorophyllin mixtures are trisodium
copper chlorin e6 and disodium copper chlorin e4 (Figure 2).
Figure 1. Molecular structure of (a) chlorophyll and (b) red blood cell
An excellent account of structure of chlorophyll and its derivatives, stability, bioavalability and their
cancer preventing activity has been reviewed by Ferruzzi and Blakesle (2007). Chlorophyllin as been
extensively studied for its effect in animal/human, and also utilized as food grade colorant in Europe,
Asia and to a more limited and growing extent in United States (Ferruzzi and Blakesle, 2007). Some of
the important chlorophyll and its derivatives are listed in Table 1.
Table 1. Chlorophyll and its derivatives used in medicine
Natural chlorophyll
Chlorophyll a, b, c, d, e
Metal free chlorophyll derivatives
Pheophytin, Pyropheophytin
Metallochlorophyll derivatives
Cu(II)chlorin e 4
Cu-chlorin e 6
Cu-chlorin e 4 ethyl ester
Figure 2. Structure of chlorophyll and its derivatives
2.2. Potential Mechanism of action of Chlorophyll
Chlorophyll derivatives after release from the plant food matrix, natural chlorophyll (CHL) derivatives
are exposed to the acidity of the gastric digestion resulting in conversion to respective metal-free
pheophytins (PHE). Following digestive degration of commercial chlorophyll derivatives, they are
absorbed by intestinal cells and finally passes into blood circulation (Egner, 2000, Ferruzzi et al.,
2002). Chlorophyll and its derivatives act through variety of mechanisms which include: (i)
antioxidant activity; (ii) modifier of genotoxic effect; (iii) inhibition of cytochrome P450 enzymes;
(iv) induction of phase II enzymes; (v) increased level of gluta-thione S-transferase; (v) cell
differentiation, cell arrest and apoptosis.
2.2.1. Antioxidant Effect
The major source of reactive oxygen species (ROS) is electron leakage from the mitochondrial
electron transport chain, which then reacts with molecular oxygen forming ROS. ROS includes free
radical such as superoxide (O2·−) and hydroxyl radical (OH·) and non-radical species such as hydrogen
peroxide (H2O2). These free radicals set chain reaction of free radical formation when they interact
with another molecule. High concentration of ROS causes oxidative damage to bio-molecules such as
lipids, proteins and nucleic acids, leakage of electrolytes via lipid peroxidation, which results in the
disruption of the cellular metabolism. Antioxidants act as an electron sink that neutralizes free radicals
either through preventing free radical formation (preventive antioxidants) or preventing free radical
chain propagation. Free radicals have been implicated to play an important role in development several
diseases (Yoshikawa et al., 2000; Devasagayam et al., 2004; Knasmüller et al. 2008 ), which include
some forms of cancer, neurological disorders, inflammatory diseases, dermatitis, tissue damage and
sepsis, cardiovascular ailments (Elahi and Matata, 2006; Lefer and Granger 2000), and rheumatoid
arthritis , idiopathic infertility (Agarwal et al., 2006; Pasqualotto et al., 2001), decreased immune
function, several diseases of ageing (Von et al., 2004). There are contradictory views about ROS and
cancer-one suggesting increased level of ROS causes cancer formation and proliferation while other
opined that ROS may kill cancer cells (Schumacker, 2006).
Dietary chlorophyll derivative has ability to scavange long lived free radicals, such as 1,1-
diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS)
(Ferruzi et al., 2002; Lanfer-Marquez et al., 2005). Natural chlorophyll a and b exhibited lower
antioxidant activity than metal-free derivative (chlorins, pheophytins, and pyropheophytins), however
metallo-derivatives (Mg-chlorophylls, Zn-pheophytins, Zn-pyropheophytins, Cu-pheophytina, and Cu-
chlorophyllins) have highest antioxidant activity (Lanfer-Marquez et al., 2005). Chlorophyll and
derivatives have potent antioxidant and radioprotective effects in vitro and in vivo. They inhibit lipid
peroxidation (Sato et al., 1983, 1984, 1985), protein oxidation, DNA damage, membrane damage
(Kamat et al., 2000; Kumar et al., 2001). A burst of free radical formation is demonstrated during
cerebral ischaemia and reperfusion induced injury. Chlorophyll salt and the aqueous extract of
Baccopa monneria and Valeriana wallichii exerts neuroprotective effects (Rehni et al., 2007).
2.2.2. Modifier of genotoxic effect
Hartman and Shankel (1990) reviewed inhibitors that directly interact with mutagen and carcinogen
and sequester so that they may not have any harmful effect on body. These inhibitors act as interceptor
molecules against mutagen and carcinogen. Interceptors are are proficient in binding to, or reacting
with, mutagenic chemicals and free radicals, and serves as a first line of defense against mutagens and
carcinogens (Hartman and Shankel, 1990). Following interception, the defense mechanism may either
involve induction of detoxification enzyme or inhibition of carcinogen activating enzyme. Data on
activity profiles of antimutagens has been reviewed in vitro and in vivo data by Waters et al. (1996).
Among the various inhibitors reviewed, chlorophyllin (CHL) was identified as almost uniformly
protective against a broad range of direct- and indirect-acting mutagens, including aflatoxins,
polycyclic aromatic hydrocarbons, heterocyclic amines, alkylating agents and several miscellaneous
compounds (Arimoto et al., 1993; Breinholt et al., 1995; Tachino et al., 1994; Negishi et al., 1997;
Dashwood et al., 1992, 1996, 1998, Dashwood, 2000).
Although chlorophyll and its compounds has potential to act anti- mutagens in vitro (Negishi et
al., 1989, Dashwood et al., 1995) however they have shown chemopreventive properties in vivo such
as chemoprevention of aflatoxin B1 (AFB1) hepatocellular carcinoma (HCC) in rainbow trout model
(Breinholt et al., 1995, Dashwood et al., 1998; Reddy et al., 1999 Pratt et al., 2006; Simonich et al.,
2008; Castro et al., 2009) and in rodent model (Guo et al.,1995; Hasegawa et al.,1995, Simonich et
al., 2007) and human intervention (Yu, 1995; Egner et al., 2001). Chlorophyllin has strong binding
capacity to acridine, more effectively than resveratrol and xanthenes (Osowski et al., 2010), which
prevents DNA-mutagen intercalation.
2.2.3. Inhibition of Cytochrome P450 Enzymes
Cytochrome P450 enzymes are involved in the removal of carcinogenic compounds from the body.
However, in some cases they can also activate compounds consumed in food, converting
procarcinogens to carcinogens. Aflatoxin B1 is not carcinogenic until converted to the electrophilic 8,9
epoxide, which can form adduct with DNA. The metabolic activation of AFB1 is mediated by
cytochrome p450 (Tachino et al., 1994; Yun et al., 1995). Dietary supplementation of chlorophyllin
has significantly reduced AFB-1 induced DNA damage in the liver of rainbow trout and rats
(Breinholt et al., 1995). The major pathway in metabolism of aflatoxin B1 in human is presented in
Figure 3 (Guengerich et al., 2002, Guengerich, 2008).
CYP1B1 is also implicated in tobacco smoke-related cancers in several organs. Tobacco
smoke contains several procarcinogens, including polycyclic aromatic hydrocarbons (PAHs),
nitrosamines and arylamines. PAHs can be activated into carcinogens by CYP1A1, CYP1A2 and
CYP1B1. Benzo[a]pyrene (BP) is a potent pro-carcinogen and ubiquitous environmental pollutant.
John et al. (2010) observed the induction and modulation of CYP1A1 and CYP1B1 and 10-
(deoxyguanosin-N2-yl)-7,8,9-trihydroxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPdG) adduct formation
in DNA from primary normal human mammary epithelial cell (NHMEC) strains. Maximum percent
reductions of CYP1A1 and CYP1B1 gene expression and BPdG adduct formation were observed
when cells were pre-dosed with chlorophyllin followed by administration of the carcinogen.
Chlorophyllin is likely to be a good chemoprotective agent for a large proportion of the human
Figure 3. Metabolism of aflatoxin B1 in human (Guengerich et al., 2002, Guengerich, 2008)
2.2.4. Induction of phase II Enzymes
Induction of phase II response is recognized as an effective strategy for protecting cells against
oxidants, electrophiles.Phase II enzyme include glutathione-S-transferase, UDP glucoronosyl
transferase, sulfotransferase, and oxidoreductase. Phase II enzymes bind to oxygenated carcinogens
making highly polar molecule that are excreted. Phase II enzymes decrease carcinogenicity by
blocking carcinogen metabolic activation and enhancing carcinogen detoxification. Although the
Brassica vegetables have long been known to contain potent inducers of mammalian phase 2 enzymes
(Dinkova-Kostova et al., 2004), chlorophyllin may also increase the activity of the phase II enzyme,
quinone reductase (Dingley et al., 2003). Chlorophylls, chlorophyllin and related tetrapyrroles are
significant inducer of mammalian phase II cytoprotective genes, inducing the phase 2 enzyme
NAD(P) H:quinone oxidoreductase 1 (NQO1) in murine hepatoma cells (Fahey et al., 2005). The
drug metabolizing enzyme comprises phase I (oxidation, reduction and hydrolysis). Physiological
balance between Phase I and Phase II enzymes, and their level of expression and genetic
polymorphism might dictate the sensitivity or risk of individual exposed to carcinogenic species
(Kensler, 1997).
2.2.5. Effect of Chlorophyll on Cell Differentiation, Cell arrest and Apoptosis of
Cancer Cells
Generally, growth rate of pre-neoplastic or neoplastic cells is fast than normal cell. Therefore,
induction of apoptosis or cell cycle arrest can be an excellent approach to inhibit the promotion and
progression of carcinogenesis. Distinct from apoptotic events in the normal physiological process,
which are mainly mediated by interaction between death receptors and their relevant ligands (Jacks
and Weinberg, 2002), many dietery supplements appear to induce apoptosis through the mitochondria-
mediated pathways. The cytotoxic effects of chemotherapeutic compounds on neoplastic cells can be
monitored by measuring their effect on mitochondria, caspases and other apoptosis related proteins.
Chlorophyllin induced apoptosis in HCT116 human colon cancer cells, via a cytochrome c
independent pathway (Diaz et al., 2003).
Progression through cell cycle is a sequential process that directs cells to pass through G1, S.
G2 and M. There are G1-S/ or G2-M checkpoints that halts cell division whenever necessary. Cyclin
dependent kinase (CDKs) CDK inhibitors governs the progression of the cell cycle. Cell cycle arrest
induced by chemopreventive compounds potentially affects and blocks the continuous proliferation of
tumorogenic cells. Lower doses of CHL also were observed to induce cell-cycle arrest and strongly
altered markers of cell differentiation, such as E-cadherin (Carter et al., 2004).A recent study showed
that human colon cancer cells undergo cell cycle arrest after treatment with chlorophyllin (Chimploy,
2009). The mechanism involved inhibition of ribonucleotide reductase activity. Ribonucleotide
reductase plays a pivotal role in DNA synthesis and repair, and is a target of currently used cancer
therapeutic agents, such as hydroxyurea (Chimploy et al., 2009).
3. Applications in Cancer Chemotherapy
Cancer development is a long term process that involves initiation, promotion and progression that
ultimately leads to spread from one area of the body to another during the late metastasis stage.
Current clinical therapies which include surgery, radiotherapy and chemotherapy are limited to
particularly during metastasis phase. However, there is increasing body of evidences from
epidemiological and pathological studies that certain dietary substances may prevent or slow down
progression of cancer. Because advance metastasis stage cancer are almost impossible to cure,
therefore, cancer chemoprevention and containment at early stage is highly desirable. Dietary
chemopreventive agents seems to have variety of cellular and molecular mechanism that may inhibit
carcinogenesis (blocking agent) or suppress promotion and progression of carcinogenesis (suppressive
agent) or function as both. Many dietery substances such as retinoic acid, sulforaphane, curcumin,
EGCG, apigenin, qurecetin, chrysin, silibinin, silymarin and resveratrol acts through induction of
apoptosis. Potential mechanism underlying effectiveness of some of the dietery constituents is
presented in Table 2. Many dietary compounds including chlorophyll possess cancer protective
properties that include cellular detoxifying mechanism and antioxidant property that protects against
cellular damage caused by environmental carcinogens or endogenously generated reactive oxygen
species. These dietary substances can affect death signaling pathways which could prevent
proliferation of tumor cells.
Table 2: Potential Mechanism of action of some of the dietery chemopreventive compounds (modified
from Chen and Kong, 2005)
Cancer Blocking agent
of chemicals
Chlorophyll and its
Inhibit cytochrome
Vitamin E
Trap carcinogen
Suppressive agents
Cell cycle disruption
/or induce apoptosis
Chlorophyll has a potential to act as chemopreventive agent. Clinical trials with chlorophyllin have
reduced aflatoxin-DNA adducts in individuals at high risk for liver cancer (Kensler et al, 1998, Dingley
et al., 2003). In another clinical trial on patient with fibroadenomastosis of breast cancer, the drug
mamoclam- containing mega-3 polyunsaturated fatty acids, iodine and chlorophyll derivatives, produced
from the brown sea alga laminaria, was effective in pain relief and breast cyst regression (Bezpalov et
al., 2005 ).
Chlorophyll can assist with the effects of dietary and environmental exposure to carcinogens.
Notable examples are the tobacco-related carcinogens (e.g., nitrosamines and polycyclic aromatic
hydrocarbons-PAH), heterocyclic amines produced from sustained, high-temperature cooking of meats
and the fungal food contaminants aflatoxins. Research indicates that chlorophyll reduces carcinogen
binding to DNA in the target organ by inhibition of carcinogen activation enzyme or degradation of
ultimate carcinogens with the target cells. In vitro and in vivo studies further substantiated medicinal
cures offered by chlorophyll derivatives. Elucidation of the molecular mechanisms of chemical
carcinogenesis provides insight into targets for chemoprevention. Microarray and proteonomics
analysis have shown alteration at the level gene expression and protein. Recently research
investigations showing involvement of transcriptional factors and their intervention by chlorophyll and
its derivative seems to be an attractive approach showing precise action at benefits of chlorophyll at
cellular and molecular levels.
4. Photosensitizer
Photodynamic therapy (PDT) is increasingly becoming accepted as a treatment option for a variety of
cancer which is usually based on the photosensitisation of tumour cells with subsequent light exposure
leading to death of the malignant cells. The most commonly used photosensitisers, such as the
haematoporphyrin derivatives have a number of drawbacks - poor selectivity in terms of tumour drug
accumulation and low extinction coefficients so that relatively large amounts of drug and/or light are
needed in order to obtain a satisfactory phototherapeutic response. These have led to the development
of a further generation of photosensitisers based on chlorophyll derivatives, which are characterized by
increased phototoxicity and strong absorption which allows deeper light penetration into tissues, rapid
tissue clearance and minimal extravasation from the circulation (Rosenbach-Belkin, 1996).
Aminolevulinic acid (ALA), a building block of tetrapyrroles, synthesized during chlorophyll
biosynthesis, has shown photodynamic destruction of cancer cells (Reibeiz et al., 2002). It is available
as porphyric insecticides and show photodynamic property. It can be easily taken up by transformed
cells, and is rapidly cleared from the circulatory stream within 48 hr of treatment (Reibeiz et al., 2002).
Conjugating Cp 6 with histamine can help improve the effectiveness of PDT in oral cancer cells by
enhancing its intracellular delivery (Parihar et al., 2010). "Radachlorin"(®), also known in the
Bremachlorin, a composition of 3 chlorophyll a derivatives in an aqueous solution, was introduced
into the Russian Pharmacopoeia. Iand may be commercialized as a prospective second-generation
photosensitizer (Kochneva et al., 2010). Prostate-specific membrane antigen (PSMA), a validated
biomarker for prostate cancer, has attracted considerable attention as a target for imaging and
therapeutic applications for prostate cancer. PSMA inhibitor, i.e. conjugate of pyropheophorbide has
been used for targeted PDT application and the mechanism of its mediated-cell death in prostate
cancer: inducing apoptosis via activation of the caspase-8/-3 cascade pathway (Liu et al., 2010).
5. Contraindications and Safety
Natural chlorophylls are not known to be toxic, and no toxic effects have been attributed to
chlorophyllin despite more than 50 years of clinical use in humans. Although few contraindications
have been reported by some investigators ((Chernomorsky, 1988; Gogel et al., 1989; Kephart , 1995;
Egner et al., 2003 , Hendler et al., 2008) ) but much serious ill effects are less known. There is lack of
reports about the safety of chlorophyll or chlorophyllin supplements in pregnant or lactating women.
Although there is no major contraindication reported so far as but in order to be used in modern,
pharmacological aspects the way these medicine interacts with human system needs to be further
explored so as to recommend its safe, effective and widespread use of in medicine.
6. Challenges of potential application chlorophyll and its derivatives as
chemotherapeutic agents
Dietary chemoprevetive compounds offer great potential in fight against cancer. The mechanistic
insight into chemoprevention of carcinogenesis includes regulation of cell defensive and cell-death
machineries. Though progress have been made in understating apoptosis, and cell cycle arrest in
relation to chlorophyll and its derivatives, signaling pathways and gene expression events leading to
pharmacological effects require further investigation. The ultimate goal is translation of the results of
in vitro signaling and gene expression obtained in animal cell culture system /animal model to
beneficial pharmacological effects which have several challenges that need to be overcome. One of the
concern is induction of detoxifying enzymes by chemotherapeutic agents varies within human
population. Since the cancer development is a long term process, there is need to explore suitable
potency indicators to assess the effect of chemopreventive agents. Dietery chemopreventive agents
may not possess pharmacologically active properties to be used as drug. Studies on pharmacokinetics
and toxicity profile of chemopreventive agents are important in drug development. Synergistic effect
chemopreventive agent in association with other efficacious drug molecule might enhance the
efficacy. Ultimately to convert a dietery chemopreventive agents into a viable drug, a clear
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... Chlorophyll as well as its derivatives have been used in ayurvedic traditional medicine, however, chlorophyll also has a variety of therapeutic uses, including wound healing anti-inflammatory, anti-oxidative, anti-carcinogenic, and anti-mutagenic action (Mishra et al., 2011). Dietary chlorophyll derivatives possess the power to scavenge free radicals, for example, 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azinobis (3-ethylbenzothiazolin-6-sulfonate) (ABTS) (Ferruzzi et al., 2002). ...
... Chlorophyll has cancer protection properties containing the cytotoxic mechanism as well as antioxidant properties that protect against cell damage triggered by environmental carcinogenic substances and intrinsically produced reactive oxygen species. These nutrients could affect the death signal pathway, which could avert the spread of tumor cells (Mishra et al., 2011). Chlorophyll binds tightly to various carcinogens in the intestinal lumen and prevents them from being absorbed into the body (Donaldson, 2004). ...
... In addition, chlorophylls and their derivatives exhibit robust in vitro/in vivo antioxidant as well as radioprotective properties. They prevent peroxidation of lipid, protein oxidation, DNA, and membrane damage (Mishra et al., 2011). Using HPLC, a group of researchers analyzed the chlorophyll content of methanol extracts from vegetable waste. ...
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Natural colorants/pigments are nowadays gaining popularity due to increased consumer awareness and severe health problems caused by the use of synthetic pigments. The efficient extraction of natural pigments from different plant sources leads to their effective application as substitutes for synthetic pigments in the food processing sector. In the present review, authors discussed the mechanistic approach of a novel ultrasonication technique in the extraction of natural color pigments having potential bio‐functional characteristics leading to numerous health benefits. The recent advancements in the ultrasound‐assisted extraction (UAE) of plant pigments including anthocyanins, betalains, carotenoids, and chlorophyll were explored. The most significant observations were that the extraction efficiency improved due to the phenomenon of cavitation, low installation and maintenance costs, and reduced energy requirement. Further, the optimum extraction conditions, stability, and bio‐accessibility of the pigment compound significantly impact the process parameters. Moreover, recent progress in enhancing the performance of the UAE technique was addressed. Practical applications Ultrasound‐assisted extraction (UAE) is an effective way to improve the recovery of natural pigments from plants. Ultrasound treatment is a novel green extraction technology that offers a wide range of benefits. The present review conveys a better understanding of UAE mechanisms and optimum conditions of extracted pigments from different plant sources for their effective use as a natural colorant in food applications on an industrial scale.
... According to Mishra et al. (2011), chlorophyll and derivates (such as pheophytins) possess potent antioxidant and radioprotective effects in vivo and in vitro, inhibit lipid peroxidation, protein oxidation, and prevent DNA as well as membrane damage. On the other hand, carotenoids are important pigments related to immunity functions, antioxidants, and lowering the risk of developing chronic diseases (Viera et al., 2018). ...
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The functionalizing of staple and economic foods, which means adding health-promoting substances, has been visualized as a solution to reduce the concerning increase in diet-related diseases caused by bad-eating patterns. Microalgae represent an innovative way to solve this problem. Arthrospira platensis, microalgae nutritionally rich in bioactive compounds, has been used to develop hard wheat pasta (semolina) to improve its nutritional value and provide antioxidant properties, but there is scarce information about the effect of adding A. platensis in a soft wheat pasta with egg on these parameters. Given that, this work aimed to assess the effect of adding A. platensis at 1, 5, and 10% in a soft wheat pasta added with egg on the nutritional value. Besides, the total phenolic content (TPC), antioxidant capacity (by FRAP and ABTS), and the spectrophotometric estimation of chlorophylls a + b as well as total carotenoids content were determined after and before cooking. The results showed that adding A. platensis at 5% was enough to increase raw pasta's nutritional value, especially protein by 19.27%, TPC by 3.88%, antioxidant capacity by 48.54%, and 66.09% for ABTS and FRAP respectively, as well as chlorophyll a + b (5.89 mg/100g) and total carotenoids (1.31 mg/100g). After the cooking process losses of 7.40, 16.81 and 0.51% were evidenced for TPC, ABTS and FRAP assays, however, remained 10.63, 62.37 and 70.65% higher than the cooked control. Furthermore, increases of 92.19 and 54.96% for chlorophyll a+b and total carotenoids were evidenced. The addition of A. platensis to pasta represents a way to improve the nutritional value regarding protein content, increase antioxidant capacity, and the content of chlorophylls and carotenoids, without statistically significant modifications in the caloric content. Keywords: Antioxidant, Arthrospira platensis, Carotenoids, Chlorophylls, Functional Food, Pasta.
... Chlorophylls are responsible for leaf greenness, which contributes to the esthetical value of green leafy vegetables. In addition, chlorophyll has been recently found to exert a preventive action against some forms of cancer [48]. Besides the effects on quality, in plants fertilized with N and treated with the biostimulant, the improvement in plant growth was associated with the stimulation of N uptake and chlorophyll biosynthesis, which may have enhanced the photosynthetic activity and increased the translocation of photosynthates to the sinks. ...
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The nutrient and nutraceutical quality of greenhouse wild rocket is strongly influenced by the light environment and nitrogen fertilization. We investigated the effects of two cover materials, a diffuse light film (Film1) and a traditional clear film (Film2), and three nitrogen regimes, no N supply (N0) and sub-optimal (N1) and optimal (N2) doses, also in combination with a biostimulant (Stimolo Mo), on the mineral composition, antioxidant properties and chlorophyll and carotenoid content of rocket plants grown in the autumn-spring cycle. The leaf concentration of most of the minerals was higher under Film1 compared to Film2. In general, K, Ca, Mg and Na were higher, and S was lower in the presence of N supply, and the addition of the biostimulant promoted the mineral uptake. Under Film1, the hydrophilic antioxidant activity (HAA) was higher in some harvests, and the ABTS antioxidant activity (ABTS AA) in the first one, while always lower afterward, than under Film2. Nitrogen fertilization did not affect the antioxidant activity, while it reduced the content of total phenols and ascorbic acid. The biostimulant application increased ABTS AA at the optimal N dose and reduced total phenols in unfertilized plants. Both the diffuse light and the N supply inhibited the synthesis of ascorbic acid, while N fertilization and the biostimulant promoted the synthesis of chlorophylls. The experimental treatments exerted variable effects over time and significant interactions with the harvest period were found for many of the investigated parameters.
... Chlorophyll is a natural pigment widely distributed in the leaves and other parts of plants that plays an important role on the photosynthesis process. In addition, chlorophyll has multiple beneficial properties for health, nowadays has been used as remedy and diagnostics in the field of medicine, mainly as antioxidant and photosensitizer for cancer therapy [35]. ...
... Plant pigments, including chlorophylls, carotenoids and anthocyanins, determine the attractive color, play a variety of roles throughout the whole plant life cycle and are characterized by powerful antioxidant [70] and anti-inflammatory activity [11,71]. There is research confirming that chlorophyll derivatives have multiple potential health benefits [8], may be protective agents against chronic diseases and have cytostatic and cytotoxic activities against tumor cells [72], therefore it is recommended to include them in the daily diet through pigment-rich foods. At the early developmental stages, the petals of many flowering plants contain chlorophylls and, as they mature, the chlorophyll content decreases and other pigments accumulate [73]. ...
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Eating habits are changing over time and new innovative nutrient-rich foods will play a great role in the future. Awareness of the importance of a healthy diet is growing, so consumers are looking for new creative food products rich in phytochemicals, i.e., specialized metabolites (SM). The consumption of fruits, vegetables and aromatic species occupies an important place in the daily diet, but different edible flower species are still neglected and unexplored. Flowers are rich in SM, have strong antioxidant capacities and also possess significant functional and biological values with favorable impacts on human health. The main aim of this study was to evaluate the content of SM and the antioxidant capacities of the edible flower species: Calendula officinalis L. (common marigold), Tagetes erecta L. (African marigold), Tropaeolum majus L. (nasturtium), Cucurbita pepo L. convar. giromontiina (zucchini) and Centaurea cyanus L. (cornflower). The obtained results showed the highest content of ascorbic acid (129.70 mg/100 g fw) and anthocyanins (1012.09 mg/kg) recorded for cornflower, phenolic compounds (898.19 mg GAE/100 g fw) and carotenoids (0.58 mg/g) for African marigold and total chlorophylls (0.75 mg/g) for common marigold. In addition to the esthetic impression of the food, they represent an important source of SM and thus can have a significant impact if incorporated in the daily diet.
... Chlorophyll derivatives (chl a, chl b, and chl c) in microalgae and cyanobacteria are promising for health-promoting activities such as anti-inflammatory properties, anticolorectal cancer, human hygiene properties, anti-stomach disorder activities, and anti-bad breath protection [19]. A summary of previous research and development on the medical applications of chlorophyll and its derivatives has been published to provide a better understanding of science-based health claims [20]. ...
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Marine organisms are a valuable source of new compounds, many of which have remarkable biotechnological properties, such as microalgae and cyanobacteria, which have attracted special attention to develop new industrial production routes. These organisms are a source of many biologically active molecules in nature, including antioxidants, immunostimulants, antivirals, antibiotics, hemagglutinates, polyunsaturated fatty acids, peptides, proteins, biofuels, and pigments. The use of several technologies to improve biomass production, in the first step, industrial processes schemes have been addressed with different accomplishments. It is critical to consider all steps involved in producing a bioactive valuable compound, such as species and strain selection, nutrient supply required to support productivity, type of photobioreactor, downstream processes, namely extraction, recovery, and purification. In general, two product production schemes can be mentioned; one for large amounts of product, such as biodiesel or any other biofuel and the biomass for feeding purposes; the other for when the product will be used in the human health domain, such as antivirals, antibiotics, antioxidants, etc. Several applications for microalgae have been documented. In general, the usefulness of an application for each species of microalgae is determined by growth and product production. Furthermore, the use of OMICS technologies enabled the development of a new design for human therapeutic recombinant proteins, including strain selection based on previous proteomic profiles, gene cloning, and the development of expression networks. Microalgal expression systems have an advantage over traditional microbial, plant, and mammalian expression systems for new and sustainable microalga applications, for responsible production and consumption.
... Natural chlorophyll (Chl) is an essential biomolecule for photosynthesis in plants and its molecular structure is similar to the NIR fluorescent porphyrin; therefore, Chl has various imaging and therapeutic benefits. Hence, Chl may be an ideal candidate for cellular imaging, including cancer imaging and therapy, for its non-toxicity and abundance in nature [22]. ...
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The development of fluorescence dyes for near-infrared (NIR) fluorescence imaging has been a significant interest for deep tissue imaging. Among many imaging fluoroprobes, indocyanine green (ICG) and its analogues have been used in oncology and other medical applications. However, these imaging agents still experience poor imaging capabilities due to low tumor targetability, photostability, and sensitivity in the biological milieu. Thus, developing a biocompatible NIR imaging dye from natural resources holds the potential of facilitating cancer cell/tissue imaging. Chlorophyll (Chl) has been demonstrated to be a potential candidate for imaging purposes due to its natural NIR absorption qualities and its wide availability in plants and green vegetables. Therefore, it was our aim to assess the fluorescence characteristics of twelve dietary leaves as well as the fluorescence of their Chl extractions. Bay leaf extract, a high-fluorescence agent that showed the highest levels of fluorescence, was further evaluated for its tissue contrast and cellular imaging properties. Overall, this study confirms bay-leaf-associated dye as a NIR fluorescence imaging agent that may have important implications for cellular imaging and image-guided cancer surgery.
Microalgae are a source of highly valuable bioactive metabolites and a high-potential feedstock for environmentally friendly and sustainable biofuel production. Recent research has shown that microalgae benefit the environment using less water than conventional crops while increasing oxygen production and lowering CO2 emissions. Microalgae are an excellent source of value-added compounds, such as proteins, pigments, lipids, and polysaccharides, as well as a high-potential feedstock for environmentally friendly and sustainable biofuel production. Various factors, such as nutrient concentration, temperature, light, pH, and cultivation method, effect the biomass cultivation and accumulation of high-value-added compounds in microalgae. Among the aforementioned factors, light is a key and essential factor for microalgae growth. Since photoautotrophic microalgae rely on light to absorb energy and transform it into chemical energy, light has a significant impact on algal growth. During micro-algal culture, spectral quality may be tailored to improve biomass composition for use in downstream bio-refineries and boost production. The light regime, which includes changes in intensity and photoperiod, has an impact on the growth and metabolic composition of microalgae. In this review, we investigate the effects of red, blue, and UV light wavelengths, different photoperiod, and different lighting systems on micro-algal growth and their valuable compounds. It also focuses on different micro-algal growth, photosynthesis systems, cultivation methods, and current market shares.
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The rapid increase in global population and shrinkage of agricultural land necessitates the use of cost-effective renewable sources as alternative to excessive resource-demanding agricultural crops. Microalgae seem to be a potential substitute as it rapidly produces large biomass that can serve as a good source of various functional ingredients that are not produced/synthesized inside the human body and high-value nonessential bioactive compounds. Microalgae-derived bioactive metabolites possess various bioactivities including antioxidant, anti-inflammatory, antimicrobial, anti-carcinogenic, anti-hypertensive, anti-lipidemic, and anti-diabetic activities, thereof rapidly elevating their demand as interesting option in pharmaceuticals, nutraceuticals and functional foods industries for developing new products. However, their utilization in these sectors has been limited. This demands more research to explore the functionality of microalgae derived functional ingredients. Therefore, in this review, we intended to furnish up-to-date knowledge on prospects of bioactive metabolites from microalgae, their bioactivities related to health, the process of microalgae cultivation and harvesting, extraction and purification of bioactive metabolites, role as dietary supplements or functional food, their commercial applications in nutritional and pharmaceutical industries and the challenges in this area of research. Graphical abstract
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"Radachlorin"(®), also known in the EU as Bremachlorin, a composition of 3 chlorophyll a derivatives in an aqueous solution, was introduced into the Russian Pharmacopoeia. Its GMP (Good Manufacturing Practice) facility based manufacturing method was patented. Laboratory experiments and clinical phase I were performed. Protocols were designed for PDT of basal cell carcinoma of the skin to result in GCP (Good Clinical Practice)-conformed randomized phase II clinical studies. "Radachlorin"(®) solution for intravenous infusions 0.35% 10mL in the doses of 0.5-0.6 and 1.0-1.2mg/kg and a gel for topical application 0.1% 25g in the dose of 0.1g/cm(2) were photoactivated by 2.5W 662nm semiconductor laser "LAKHTA-MILON(®)" (St. Petersburg, Russia) in light doses of 200, 300 (solution), 400, 600, 800 (gel) J/cm(2). Safety study showed no side effects and a good tolerability of "Radachlorin"(®) by patients. There was no normal skin/subdermal tissue damage after both laser and sun light exposure. The main part (98%) of the drug was excreted or metabolized in the first 48h. Drug administration at a dose of 1.0-1.2mg/kg and irradiation at 3h with 662±3nm light at a dose of 300J/cm(2) (solution) and 4 PDT sessions at an interval of 1 week with 3h gel exposure, followed by 400J/cm(2) light exposure (gel) were found to be the optimal treatment regimes. Having successfully passed clinical trials, "Radachlorin"(®) achieved marketing authorization in Russia in 2009 and a conditional approval in South Korea in 2008. It is a candidate for phase III clinical trials in the EC and may be commercialized as a prospective second-generation photosensitizer.
Many people ask why one needs to take vitamins or minerals in dosages exceeding the recommended daily allowances? Among the reasons are depleted soil, low-nutrient crops, food processing and the use of pesticides and herbicides on our crops. Others include stress, special circumstances like pregnancy, and that we use nutrients for therapy, not just to avoid deficiency diseases. Let us discuss the topic of nutritional supplements in detail. FALLACIES OF THE RDAs AND MDRs Government bureaucrats meet periodically to decide the levels of the recommended daily allowances (RDA) or minimum daily allowances (MDRs) of common nutrients. The main criterion is the amount needed to prevent deficiency diseases. Vitamin C deficiency causes scurvy, vitamin A deficiency causes night blindness, and B1 deficiency causes pellagra. While this approach is okay as far as it goes, these recommendations have little to with optimum health. One of every two Americans will contract cancer and 50% of the population by age 40 have a chronic illness. In theory, most of these people meet or exceed the RDAs for nutrients. The theory of the RDAs ignores more subtle aspects of nutrition such as the effect of vitamin C or zinc on tumor inhibition. This is much harder to measure, so it is ignored. The government requires that companies print the percentage of the RDA on each supplement bottle. This can cause alarm until one understands why the RDAs are inappropriate and often irrelevent.
In this review, retinol, chlorophyllin, and N-acetylcysteine are examined and compared with regard to their antimutagenic activity against some promutagens and a group of direct-acting alkylating agents. The promutagens included aflatoxin B1, certain polycyclic aromatic hydrocarbons (e.g., benzo[a]pyrene), and certain heterocyclic amines (e.g., food pyrolysates). Results of antimutagenicity testing selected from data surveyed in the published literature are displayed graphically as activity profiles of antimutagens showing both the doses tested and the extent of inhibition or enhancement of mutagenic activity. All three antimutagens are discussed in terms of their putative mechanisms of action in vitro and in vivo with emphasis on the xenobiotic metabolizing enzyme systems.
In vitro antioxidant and antimutagenic activity of dietary chlorophyll derivatives was assessed. Antioxidant activity was determined by the ability of each compound to scavenge the long-lived free radicals 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS+). Antimutagenic activity was assayed with a modified microscreen bacterial reverse mutagenicity assay using Salmonella typhimurium TA100 and benzo[a]pyrene as the tester strain and mutagen respectively. Derivatives of chlorophyll a were found to be more effective radical quenchers than those of chlorophyll b. Furthermore, metal-free derivatives such as chlorins, pheophytins, and pyropheophytins exhibited significantly lower antiradical capacity than metallo-derivatives such as Mg-chlorophylls, Zn-pheophytins, Zn-pyropheophytins, Cu-pheophytina, andCu-chlorophyllins. Both metal-free and metallo-chlorophyll derivatives demonstrated similar dose-dependent inhibitory activity against B[a]P induced mutagenesis. These results demonstrate that dietary chlorophyll derivatives prevalent in both fresh and processed foods and dietary supplements have antioxidant and antimutagenic activities.
Hepatocellular carcinoma (HCC) is one of the major cancers in China. Accordingly, the mortality rates in 1990 (per 100 000) were 20.10 in certain cities and 24.32 in certain counties. More than 90% of HCC cases and 70% of controls were infected with the hepatitis B virus (HBV) (Odds Ratio (OR) = 10–50). In the same group of patients, 8–27% of those with HCC and 0–11% of the healthy controls were also infected with hepatitis C (HCV) (OR = 2.11–17.29). There appears to be some correlation between HBV markers and the OR. The government requires that 85% of infants be immunized with HBV vaccine. In 1992, there were 3 million infants inoculated with HB vaccines. Aflatoxins have been found as contaminants in food, particularly in corn, peanut oil, soya sauce and fermented soya beans. The intake of aflatoxin B1, (AFB1) by people of ten different villages correlated with HCC mortality rates (r =0.55; P < 0.05). The concentration of AFB1-albumin adducts is an indicator of individual exposure to aflatoxins. These adducts are higher in hyperendemic HCC areas and cases. Most people have now changed their staple food and eat rice instead of corn. Six large epidemiological studies have confirmed that people who drink pond-ditch water experience higher HCC mortality rates than people who drink deep-well water. Recent research has found that the blue-green algal toxin microcystin (MCYST) was a contaminant of pond-ditch water. MCYST is a strong promoter of HCC and will induce severe intrahepatic haemorrhages and liver necrosis. More than 30% of people in Qidong County have already changed their sources of water from pond-ditches to deep wells. Therefore, a combined strategy of the prevention of hepatitis, control of crops and control of drinking water is advocated for the primary prevention of HCC in China.
The growing body of epidemiological and experimental evidence associating diets rich in fruits and vegetables with prevention of chronic diseases such as cancer has stimulated interest in plant food phytochemicals as physiologically active dietary components. Chlorophyll and its various derivatives are believed to be among the family of phytochemical compounds that are potentially responsible for such associations. Dietary chlorophyll is predominantly composed of lipophilic derivatives including chlorophyll a and b (fresh fruits and vegetables), metal-free pheophytins and pyropheophytins (thermally processed fruits and vegetables), as well as Zn-pheophytins and Zn-pyropheophytins (thermally processed green vegetables). Water-soluble derivatives including chlorophyllides, pheophorbides, as well as a commercial-grade derivative known as sodium copper chlorophyllin (SCC) also contribute to the diversity of dietary chlorophyll derivatives. Although the use of chlorophyll derivatives, especially SCC, in traditional medical applications is well documented, it is perhaps the potential of chlorophyll as a cancer preventative agent that has drawn significant attention recently. Biological activities attributed to chlorophyll derivatives consistent with cancer prevention include antioxidant and antimutagenic activity, mutagen trapping, modulation of xenobiotic metabolism, and induction of apoptosis. Although most research has focused on commercial-grade SCC, the extent to which natural chlorophyll derivatives modulate biomarkers of cancer risk is also being explored. Recent research efforts have also included investigation of the impact of digestive factors on chlorophyll structure and bioaccessibility as a means to better understand the extent to which these pigments may be bioavailable in humans and therefore have more systemic impact in the prevention of cancer.
These products are commercially obtained in the United States to the extent of about 80,000 pounds annually by extraction of about 32,000,000 pounds of alfalfa, for the most part by only three concerns. They are finding increasing use, not only in the manufacture of popular deodorants but also in medicinal preparations for treating anemia and hypertension, as a healing agent and in oral hygiene.
The antioxidant activity of six natural isolated chlorophyll derivatives and Cu-chlorophyllin was investigated by measuring their protective action against lipid oxidation. For this, the β-carotene bleaching method and the stable radical 2,2-diphenyl-1-picryldrazyl (DPPH) scavenging assay were employed. The results obtained by the β-carotene bleaching method showed that all chlorophyll derivatives presented a dose-dependent response. Pheophorbide b and pheophytin b were the strongest natural antioxidant compounds, whose activities were comparable to BHT. The high antioxidant activity found for pheophorbide b, in comparison to pheophorbide a, demonstrated the importance of the aldehyde group for functionality. On the other hand, by the DPPH assay, all natural pigments showed low antioxidant activity when compared to Trolox. Cu-chlorophyllin, tested by both methods, presented a higher antioxidant activity than that of natural chlorophylls, showing the importance of the nature of the chelated metal in the porphyrin ring. The mechanism of antioxidant activity displayed by the natural chlorophyll derivatives does not seem to be based on the ability to donate hydrogen but maybe, on the protection of linoleic acid against oxidation and/or preventing decomposition of hydroperoxides.
Chlorophylls, chlorins, and other porphyrins have been used clinically for many years, including photodynamic therapy of tumors. More recently, the cancer chemopreventive properties of chlorophylls have come to be recognized. Chlorophylls exhibit anti-mutagenic activity in short-term genotoxicity assays, and protect against various intermediate biomarkers of cancer in vivo. The anticarcinogenic activity of sodium-copper chlorophyllin (CHL), a clinically-used water soluble salt of chlorophyll, has been studied in several species. Collectively, the results from these studies support a chemopreventive role for CHL against aromatic carcinogens (aflatoxins, polycyclic aromatic hydrocarbons, heterocyclic amines) in various target organs of rats, mice, and rainbow trout. In vivo mechanism studies indicate that inhibition is most effective when CHL is administered simultaneously with the carcinogen, thereby allowing direct interaction (molecular complex formation) between CHL and the carcinogen. Studies of post-initiation treatment with CHL have provided conflicting results, with evidence for inhibition or promotion of carcinogenesis. These findings are discussed in terms of the inhibitory and promotional mechanisms of CHL, the relevance of such mechanisms to natural chlorophylls present in the diet, and the current use of CHL as a health supplement.