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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
(*Email: adroot92@yahoo.co.in)
Abstract
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
Abbreviations:
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. (https://www.novapublishers.com/catalog/product_info.php?products_id=22468)
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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).
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(a)
(b)
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
Zn-Pheophytin
Zn-pyropheophytin
Chlorophyllide
Pheophorbide
Cu(II)chlorin e 4
Cu-chlorin e 6
Cu-chlorin e 4 ethyl ester
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Figure 2. Structure of chlorophyll and its derivatives
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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
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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.
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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
population.
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Figure 3. Metabolism of aflatoxin B1 in human (Guengerich et al., 2002, Guengerich, 2008)
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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
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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.
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Table 2: Potential Mechanism of action of some of the dietery chemopreventive compounds (modified
from Chen and Kong, 2005)
Class
Function
Compounds
Cancer Blocking agent
Enhanced
detoxification
of chemicals
Indole-3-Carbinol
Chlorophyll and its
derivatives
Sulpforaphane
Curcumin
Inhibit cytochrome
P450
Isothiocyanates
Selenium
Vitamin E
Trap carcinogen
Flavonoids
Chlorophyllin
Suppressive agents
Cell cycle disruption
/or induce apoptosis
Chlorophyllin
EGCG
Quercetin
Resveratrol
Curcumin
Sulphoraphane
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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 et.al., 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
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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
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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
understanding in this area will provide impetus for future developments.
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... Chlorophylls are natural pigments present in ANS. They act as a natural antioxidant with the ability to scavenge free radicals and prevent several oxidative stress-related diseases such as cancer, neurological disorders, inflammatory diseases, dermatitis, tissue damage, and sepsis; cardiovascular disorders decreased immune function, and aging (Mishra et al., 2011;Lanfer-Marquez et al., 2005;Wang & Wink, 2016;Sangija & Wu, 2020). Chlorophyll can inhibit calcium oxalate dihydrate formation, which is the primary source of kidney stones (İnanç, 2011). ...
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African nightshades (ANS, Solanum nigrum complex) are among the most widely distributed and consumed indigenous vegetables in Tanzania. Several challenges hamper the utilization of ANS. This study sought to assess trends and constraints to ANS utilization in Kilimanjaro and Morogoro regions, Tanzania and also, to assess the effect of lactic acid fermentation on the nutritional and sensory quality of ANS. About 627 farmers' households were involved in the study. Quantitative and qualitative methods were employed to collect information. Semi-structured questionnaires, focus group discussions and Key informants’ interviews were the methods of data collection. Solanum villosum and Solanum scabrum were fermented naturally and controlled fermentation using Lactobacillus plantarum and Leuconostoc mesenteroides. The results showed that ANS is mainly used as food (96.1%), animal feed (41.3%), and medicine (38%). On average, only 5% of ANS sales contributed to family income. The main constraints to ANS utilization include; pests and diseases (92.9%), lack of knowledge (58%), shortages of fertilizer (51%), shortages of pesticides (50%), inadequate means of transport (50.4%), lack of extension services (48%), improper postharvest handling (41.4%) and inadequate storage facilities (34%). Postharvest losses accounted for a 78.4% loss of ANS. There was minimal value addition on ANS, e.g., drying (5.3%) and fermentation (1.1%). Fermentation significantly reduced pH and increased titratable acidity. Fermentation increased β-carotene 2-5 times for both pickles. Vitamin C, chlorophyll and polyphenol were significantly reduced. Fermentation increased bioavailability of minerals (P, Ca, Fe and Zn). Knowledge should be given on good agricultural practices i.e., the proper use of pesticides, fertilizers, and quality seeds. Knowledge of the processing and preservation of ANS is necessary for farmers to improve utilization, reduce losses and ensure ANS availability. Fermentation can preserve ANS, with an increase in β-carotene and reducing antinutrients. Fermentation can be recommended to small-scale farmers, processors and households to improve their nutrition and livelihood.
... Microalgal pigments in fermented foods present an intriguing intersection of nutrition, food technology, and culinary innovation. Microalgae are rich sources of diverse pigments such as chlorophyll, typically green (Chlorophyta) [99], each of which contributes not only to the color of food products but also to their nutritional and health-promoting properties. Many microalgal pigments are recognized for their potent antioxidant capabilities. ...
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Fermentation is an ancient bioprocess that harnesses the power of microorganisms to convert raw ingredients into valuable and safe food products. There has been a growing interest in using fermentation to create safe, nutritious, and sustainable foods, particularly with the incorporation of microalgae. This review highlights fermentation’s potential benefits and challenges, focusing on microalgae and its metabolites. Bioactive peptides released during microalgae fermentation have garnered attention for their potential health benefits, particularly their antidiabetic actions. Studies suggest that these peptides can help regulate blood glucose levels and improve insulin sensitivity, presenting a promising avenue for developing foods that nourish and contribute to disease prevention. As awareness grows regarding health and sustainability, there is an increasing demand for safe and sustainable food options. Fermented products, particularly those utilizing microalgae, are often perceived positively by among consumers owing to their perceived benefits. Educating consumers on the advantages of fermented foods, including their nutritional value and sustainability, can further enhance market acceptance and commercialization. Thus, the present work aims to explore the literature to study the potential and challenges of fermentation as a tool to produce sustainable foods, focusing on microalgae as an ingredient.
... According to (Belous et al., 2018), the quantitative number of chlorophylls, carotenoids, and their ratios provide significant Introduction information about the physiological state of plants. These pigments possess several health benefits too (Mishra et al., 2011). Chlorophyll possesses antioxidant activity (Ferruzzi et al., 2002;Lanfer-Marquez et al., 2005), anticancer activity (de Vogel et al., 2005), acts as an anti-inflammatory agent (Bowers, 1947;Larato & Pfau, 1970), internal deodorant (Young & Beregi Jr, 1980), potent anti-mutagen and anti-carcinogen(R. ...
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Lawsonia inermis L. (Henna) is a perennial, natural dye-producing, shrub or small tree and sole species of the genus Lawsonia in the Lythraceae family. The plant contains various compounds, including flavonoids, tannins, phenolic compounds, alkaloids, terpenoids, lawsone, and secondary metabolites is used in modern pharmacopeia and cosmetics. The plant's extracts show anti-mosquito, antimicrobial, antioxidant, wound healing, anti-inflammatory, analgesic, tuberculostatic, diuretic, hypoglycemic, and antiparasitic activities. Physical conditions affect dye properties and quality. Conventional methods for Lawsonia inermis L. propagation face problems like short viability, disease, pests, and unreliable seed propagation due to natural habitat conditions for large-scale production. Therefore, vegetative propagation via stem cuttings to multiply superior healthy plants is important for achieving increased productivity in terms of biomass. Auxins have a significant effect on the rooting and shooting of cuttings. Pigments and secondary metabolites are two good indicators of the physiological conditions, and medicinal properties. In the present study, firstly cutting of 8-10 cm each of having 3 replications were treated with 250, 500, 750, 1000, 1250, 1500, and 1750 ppm solutions of auxins (IAA, IBA, and NAA) respectively (a total of 22 treatments) following quick dip method and planted in the sand filled polybag). Vegetative growth data were collected at 30, 60, and 90 DAP (Days after planting) in 10 parameters. Secondly, the dynamics of foliar pigments (chl-a, chl-b, and carotenoids) and the qualitative presence of secondary metabolites in the leaves of Lawsonia inermis L. were assessed. Finally, the quantitative measurement of polyphenol content was done. The whole experiment was conducted following the CRD (completely randomized design) method from June 2022 to June 2023 at the Botanic Garden and Department of Botany, University of Chittagong. Statistical analysis was done using MS Excel-2021. In the field experiment of vegetative propagation through stem cutting, the maximum growth performance viz. The number of shoots (4) was observed in T20 (NAA-1250 ppm), length of shoot (26.93 cm) was observed in T7 (IAA-1500 ppm), Number of root (7.67) was observed in T14 (IBA-1500 ppm), length of root (8.37 cm) was observed in T15 (IBA-1750 ppm), Number of leaves (34) was observed in T6 (IAA-1250 ppm) at 90 DAP. In the case of the biomass of leaf study, fresh weight and dry weight of leaves highest amount (4.70g and 4.53g) was observed in T17 (NAA-500 ppm) treatment at 90 DAP. whereas minimum in control (T1). In the lab test, Maximum foliar pigments viz. Chlorophyll-a, Chlorophyll-b, Carotenoids, and total pigment content were obtained 1.22 mg g-1, 1.34 mg g-1, 1.20 mg g-1, and 3.76 mg g-1 FW in T12 (IBA-1000 ppm) treatment at 90 DAP whereas minimum in control (T1). Qualitative assessment of 10 different secondary metabolites showed positive results in the methanolic extracts of leaves. In Quantitative measurement for polyphenol contents maximum amount were observed 0.3048 %, 0.4004 %, and 0.4957 % at 30, 60, and 90 DAP respectively. The highest amount of polyphenol content (0.4957%) was found in T5 (IAA-1250 ppm) treatment at 90 DAP whereas minimum in control (T1). All the data from the above experiments were subjected to ANOVA. Treatments for most of the parameters were found significant (p<0.01 or 0.05). Therefore, the present study concludes that the above-mentioned concentrations would be considered as the most suitable treatment for producing maximum vegetative growth in stem cutting at 90 DAP. Qualitative presence of a high number of secondary metabolites approved the medicinal importance of the plant Lawsonia inermis L. Quantitative analysis of polyphenol content showed the presence of antioxidant properties in this desired medicinal plant.
... The 30 % MLE applied to the leaves of A. hybridus using foliar spraying techniques was also a standout best, recording the highest amount of 14.00 mg/100g (approximately 0.14 mg/g) total chlorophyll. This result shows the capability of MLE in enhancing the chlorophyll content of A. hybridus which is of great value to those in the food and pharmaceutical companies, where chlorophylls are being harvested and used as colorants and as a remedy to some chronic diseases respectively (Mishra et al., 2011;Sun et al., 2024). ...
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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.
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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.
Article
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.
Article
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.
Article
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.
Article
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.