Chlorophyllin Research Articles

Photosensitization can be an effective risk-reduction strategy against the post-baking mold spoilage of bread

Photosensitization was developed as a risk-reduction strategy against the contamination by environmental mold spores during the bread cooling phase. Two food-grade photosensitizers -chlorophyllin (CHL) and riboflavin (RBF), were used to evaluate the effect of visible (blue) LED illumination against three common bread spoilage molds. Aided by CHL, 405 nm LEDs inactivated Rhizopus stolonifer and Penicillium expansum by 77.4 ± 3.3% and 52.1 ± 7.3% respectively in 30 min on dichloran rose bengal chloramphenicol agar. These reductions were much higher than the corresponding reductions observed with food-grade RBF and 445 nm LEDs - 22.8 ± 3.2% and 45.5 ± 5.9%, indicating that CHL-based photosensitization was more effective as an intervention than RBF-based photosensitization. When the three molds were illuminated on bread after spraying CHL and spot-inoculation, their populations were reduced by 51–58%. CHL-based photosensitization was observed to retain the texture and moisture of the bread samples, but had a statistically significant impact on their colour. The results of this study suggest that CHL-based photosensitization can be developed as a risk reduction method to prevent the spoilage of bread.

In vitro Effect of Photoactive Compounds Curcumin and Chlorophyllin Against Single Strains of Salmonella and Campylobacter

Salmonella and Campylobacter are two of the most common foodborne pathogens associated with poultry meat. Regulatory restrictions and consumer concerns have increased the interest for plant-derived antimicrobials and emerging novel technologies. The objective of this study was to determine the antimicrobial activity of photoactive compounds curcumin (CUR) and chlorophyllin (CH) followed by activating light exposure for the reduction of Salmonella and Campylobacter. Peroxyacetic acid (PAA) was also evaluated as a poultry industry standard antimicrobial processing aid. CUR and CH were evaluated in 96-well plates at concentrations of 100, 500, and 1,000 ppm, along with PAA at 100, 200, and 300 ppm, or distilled water (DW). Each well was inoculated with 105 CFU/mL of Salmonella Typhimurium or Campylobacter jejuni, and plates were exposed to activating light (430 nm) for 0 or 5 min. No detectable reductions were observed for Salmonella or Campylobacter when treated with CUR, CH, or 100 ppm PAA. However, when Salmonella was treated with 200 ppm PAA, counts were reduced from 4.57 to 2.52 log10 CFU/mL. When Salmonella was treated with 300 ppm PAA, counts were reduced to below detectable levels (5 CFU/mL). Campylobacter was reduced from 4.67 to 2.82 log10 CFU/mL when treated with 200 ppm PAA. However, no further reductions were observed when Campylobacter was treated with 300 ppm PAA (2.50 log10 CFU/mL). These results indicate that CUR and CH were not effective as antimicrobials under the evaluated conditions, particularly in comparison to the commonly used antimicrobial, PAA.

Impact of the lipid phase composition and state on the in vitro digestibility and chlorophyllin bioaccessibility of W1/O/W2 emulsions into whole milk

Water-in-oil-in-water (W1/O/W2) emulsions offer the potential to deliver hydrophilic bioactive compounds into foods, yet their application remains limited due to their instability. Thus, the impact of lipid phase composition and state on the colloidal stability, in vitro lipid digestibility and chlorophyllin (CHL) bioaccessibility of W1/O/W2 emulsions before and after incorporation into whole milk was studied. Medium-chain triglyceride oil (MCT) was used as a liquid lipid phase and MCT with glyceryl stearate (GS) or pure hydrogenated palm oil (HPO) as gelled lipid phases. The lipid phase composition was crucial to forming W1/O/W2 emulsions. MCT or MCT+GS allowed the successful formation of W1/O/W2 emulsions, being more stable upon gastric conditions those formulated with MCT+GS than pure MCT. In contrast, the use of HPO led to phase separation, which was maintained after the gastric conditions. Regarding their lipid digestibility, W1/O/W2 emulsions formulated with MCT or MCT+GS were fully digested, whereas only 40% of the lipid was digested using HPO. In accordance, the CHL bioaccessibility was higher using MCT or MCT+GS than HPO. When co-digested with whole milk, the colloidal stability and lipid digestibility of the W1/O/W2 emulsions with MCT or MCT+GS were not altered, whereas the W1/O/W2 emulsion-HPO showed enhanced colloidal stability and lipid digestibility (57.71 ± 3.06%), due to the surface-active properties of milk protein. The present study provides useful information to develop stable functional foods enriched with hydrophilic bioactive compounds by using W1/O/W2 emulsions.

The Antihyperglycemic Effect of Crude Liang (Gnetum gnemon var. tenerum) Leaves Powder on Wistar Rats.

Chlorophyll and chlorophyllin (CHL) demonstrated antidiabetic activity by inhibiting gluconeogenesis and increasing glucose uptake in rats' muscle cells. Liang leaves contain high amounts of chlorophyll and chlorophyllin and may provide an antidiabetic effect. The antidiabetic activity of chlorophyll and CHL contained in Liang leaves, Cu-chlorophyllin (CCL) Liang leaves treated with CuSO4, and untreated crude Liang leaves (CLL) were compared using commercial chlorophyllin (CHL) as a reference. Twelve Wistar male rats were separated into 4 groups (3 rats/group); the first was a normal one (based line group), the second were the diabetic rats treated with CHL, while the third and the fourth were the diabetic rats treated with 0.97 g/kg of CCL and CLL, respectively. Diabetic rats were induced by a high fructose diet, before being taken to administer commercial CHL, CCL, and CLL for 7 days. Nonfasting blood glucose and body weight were checked daily. After euthanasia, organ weight, biochemical, hematological, and histopathological properties were evaluated. CCL treatment showed no antihyperglycemic activity in the rat model but caused some biochemical abnormalities and thrombocytopenia. Commercial CHL gave a higher reduction of nonfasting blood glucose (NFBG) than Liang leaves powder CCL or CLL but also showed some signs of abnormal biochemical parameters. CLL exhibited an antihyperglycemic effect, with higher body weight and increased HDL/LDL ratio and thus could be a promising alternative natural source for diabetes treatment.

Photodynamic Activity of Chlorophyllin and Polyethylenimine on Pseudomonas aeruginosa Planktonic, Biofilm and Persister Cells.

Antimicrobial photodynamic inactivation is considered a promising antimicrobial approach that may not develop resistance in the near future. Here, we investigate the influence of the photosensitizer chlorophyllin (CHL) and the cationic permeabilizer polyethylenimine (PEI), exposed to a red light-emitting diode, on the human pathogen Pseudomonas aeruginosa free-living planktonic cells, the sessile biofilm and persister cells. The broth microdilution checkerboard method was used to test antimicrobial susceptibility. As a substrate for biofilms, the Calgary biofilm device was used, and the quantification of the biofilm biomass was carried out using a crystal violet assay. Serine hydroxamate was used for the induction of persisters. Our findings reveal that PEI ameliorates the antimicrobial activity of CHL against P. aeruginosa planktonic and biofilm states, and the concentration required to eradicate the bacteria in the biofilm is more than fourfold that is required to eradicate planktonic cells. Interestingly, the persister cells are more susceptible to CHL/PEI (31.25/100 µg mL-1) than the growing cells by 1.7 ± 0.12 and 0.4 ± 0.1 log10 reduction, respectively, after 15 min of illumination. These data demonstrate that CHL excited with red light together with PEI is promising for the eradication of P. aeruginosa, and the susceptibility of P. aeruginosa to CHL/PEI is influenced by the concentrations and the exposure time.

Plant green pigment of chlorophyllin attenuates inflammatory bowel diseases by suppressing autophagy activation in mice.

Inflammatory bowel diseases (IBD), including Crohn's disease (CD) and ulcerative colitis (UC), are intestinal complications characterized by chronic inflammation, autophagy abnormality, and lysosomal stress, which are derived from genetic predisposition and environmental risk factors. It is generally precepted that dietary green vegetable is beneficial for physiological homeostasis. In this study, we found that dextran sulfate sodium (DSS)-induced colitis and altered intestinal epithelia in mice were attenuated by oral administration of chlorophyllin (CHL), a water-soluble derivate of chlorophyll. In DSS-treated mice, autophagy was persistently activated in intestinal tissues and associated with bowel disorders. Conversely, supplement of CHL in diet or gavage suppressed intestinal inflammation, downregulated autophagy flux in intestinal tissue, and relieved endoplasmic reticulum stress. In vitro studies show that CHL could activate Akt and mTOR pathways, leading to downregulation of autophagic and lysosomal flux. Thus, consumption of green vegetables and chlorophyllin may be beneficial for IBD recovery in part through alleviation of inflammation and autolysosomal flux.NEW & NOTEWORTHY Inflammatory bowel disease (IBD) is a chronic and recurrent gastrointestinal disease, while the etiology remains poorly understood. Dietary composition and lifestyle are crucial for pathogenesis and progression of IBD. In this study, we observed that autophagy in the intestinal tissue was persistently activated in IBD mice. Chlorophyllin (CHL), a water-soluble derivate of chlorophyll, can attenuate colitis by regulating autophagy and inflammation. Thus, consumption of green vegetables and chlorophyllin may be beneficial for IBD recovery.

Phyto-chlorophyllin Prevents Food Additive Induced Genotoxicity and Mitochondrial Dysfunction via Cytochrome c Mediated Pathway in Mice Model.

The issue of food-additive-toxicity causing several health hazards needs to be therapeutically managed with an immediate effect. Alloxan, a food additive, is used for whitening and shining flour. It is capable of inducing genotoxicity, diabetes, and associated mitochondrial dysfunction. Therefore, to explore a non-toxic, phyto-based compound that can delay the onset of diabetes and prevent the multitude of damage associated, Chlorophyllin (CHL) was selected for our study, having been reported to exhibit anti-cancer, anti-diabetes, and antiinflammatory responses. Therefore, the objective of the present study is to evaluate the protective role of CHL in controlling genotoxicity, glucose imbalance, and associated cytochrome c mediated mitochondrial signaling dysfunction against food-additive-induced genotoxicity, diabetic state, and its complexities in mice model in vivo. Mice were pre-treated with CHL through oral gavage before they were exposed to alloxan. Diabetic markers, anti-oxidant enzyme profile, chromosomal study, mitochondrial functioning factors, and expression of proteins were checked against food-additive injected mice. The results revealed that CHL pre-treatment could delay the onset of diabetes, restrict alloxan-induced elevation of blood glucose, reduce DNA-damage and chromosomal aberration, optimize enzymatic profile (glucokinase, pyruvate, insulin), and modulates protein expression (insulin, IRS1, IRS2, GLUT2). Further, CHL-pre-treatment could stabilize mitochondrial-membrane-potential, intracellular calcium ion, ATP/ADP ratio, ATPase activity, thereby maintaining optimum functioning of cytochrome-c, bcl2, and caspase3 mitochondrial protein. Therefore, the present study reports, for the first time, the screening of phytobased bioactive CHL for preventing/limiting the extent of food-additive-induced genotoxicity and mitochondrial dysfunction and serves as an advanced therapeutic tool in the management of diabetes.

First report on chlorophyllin to protect mammalian and fish muscle cells from pesticide toxicity via activation of p53 and PARP

ObjectivesPesticide toxicity has become one of the major environmental menaces affecting all types of life forms of the ecosystem. Pesticides get washed off from agricultural fields into nearby water bodies and enter the aquatic organisms. Their bio-accumulated form finally reaches the human race, through consumption of pesticide infested aquatic animals, causing several physiological dysfunctions. Hence it becomes necessary to find a therapeutic cure/a preventive measure to stop the health hazard issues of pesticide. With this projection a search for a phyto-based-product was made whose primary objective would be to lower the pesticidal toxicity in fish and simultaneously in the human race. MethodsIn this study we tried to check whether the phyto-chemical, Chlorophyllin (CHL), known for its anti-genotoxic, anti-oxidant activities, could render any kind of protection against Cypermethrin (CM) induced-toxicity in fish model and mammalian cell line L6. Both the model L6 and fish were pre-treated with CHL prior to exposure of CM. Different scientific parameters like % cellular cytotoxicity, reactive oxygen species (ROS) generation, nuclear condensation, etc were checked to validate the possibility of CHL in protecting CM-induced toxicity. ResultsThe overall results revealed that pre-treatment with CHL could restrict the ROS generation leading to modulation in associated cytokine proteins expression NFkβ and IFNγ. Further, CHL lowered nuclear condensation and elevated expression of DNA repair proteins p53 and PARP, showing a kind of pre-activation of signalling cascades for overall protection against the severity of pesticidal toxicity. ConclusionThus, this phyto-based preventive approach would possibly solve many areas of human health issues related to pesticide toxicity in future.

In the Right Light: Photodynamic Inactivation of Microorganisms Using a LED-Based Illumination Device Tailored for the Antimicrobial Application

Drug-resistant bacteria threaten the health of people world-wide and cause high costs to their health systems. According to Scientific American, the number of regrettable fatalities due to the bacteria that are resistant to conventional antibiotics will sum up to 300 million until 2050 if the problem is not tackled immediately. Photodynamic Inactivation (PDI) has proven effective against microorganisms irrespective of their resistance to conventional treatment, but for the translation into clinical practice, economic, homogenous and powerful light sources holding approval as medical devices are needed. In this study we present two novel light emitting diode (LED)-based lamps (Repuls7PDI-red and Repuls7PDI-blue) tailored for application in PDI and demonstrate their photodynamic efficiency upon using either methylene blue (MB), a photoactive compound widely used in PDI, or Sodium Magnesium Chlorophyllin (CHL), a water-soluble derivative of chlorophyll, which holds approval as food additive E140, against bacteria and fungi. Gram+ Staphylococcus aureus, Gram− Escherichia coli and the yeast Candida albicans serve as model systems. Repuls7PDI-red emits a wavelength of 635 nm and an intensity of 27.6 ± 2.4 mW·cm−2 at a distance of 13.5 cm between the light source and the target, while the Repuls7PDI-blue allows an exposure at 433 nm (within the range of violet light) (6.4 ± 0.5 mW·cm−2 at 13.5 cm). Methylene blue was photoactivated with the Repuls7PDI-red at 635 nm (25.6 J·cm−2) and allows for photokilling of E. coli by more than 6 log10 steps at a concentration of 10 µM MB. Using equal parameters, more than 99.99999% of S. aureus (20 µM MB) and 99.99% of C. albicans (50 µM MB) were killed. If blue light (Repuls7PDI-blue, 433 nm, 6.6 J·cm2) is used to trigger the production of reactive oxygen species (ROS), a photoinactivation of S. aureus (5 µM CHL, CFU reduction > 7 log10) and C. albicans (>7 log10) below the detection limit is achieved. PDI based on CHL (10 µM) using red light activation reduces the number of viable S. aureus by more than 6 log10. Our data prove that both LED-based light sources are applicable for Photodynamic Inactivation. Their easy-to-use concept, high light output and well-defined wavelength might facilitate the translation of PDI into clinical practice.

Potential protective effect of chlorophyllin against the genotoxicity and oxidative stress induced by nano-TiO2 particles in mice

Wide uses of titanium dioxide (TiO2) nanoparticles in cosmetics, paints, toothpastes, sunscreens, food products, pharmaceuticals and nanomedical reagents increases human exposure and its risk. Thus this study was designed to investigate the possible protective effect of chlorophyllin (CHL) on nano-TiO2 induced genotoxicity and oxidative stress in mice. Male mice were exposed to single i.p. injection with each of three dose levels of nano-TiO2 (500, 1000 or 2000 mg/kg b.w) suspended in deionized water or CHL (200 mg/kg b.w.). Micronucleus and comet assays were carried out genotoxic endpoints. Moreover, oxidative stress was evaluated using five markers of oxidative damage to shed more light on its mechanism. A dose-dependent genotoxicity of nano TiO2 was evidenced by the observed significant elevations in micronuclei frequencies and DNA damage levels. The same end.-points indicated that TiO2 genotoxicity was significantly decreased after CHL coadministration. Moreover, CHL significantly decreased malondialdehyde level, and significantly increased reduced glutathione level and superoxide dismutase, catalase and glutathione peroxidase activities, which were significantly disrupted by TiO2 alone. In conclusion, the dose dependent nano-TiO2 induced genotoxicity was diminished by CHL co-administration via its free radicals scavenger ability.

A Single Arm Trial in Treatment of CKD Patients with Sodium Copper Chlorophyllin Formulation

In a prospective, single arm study of 34 Chronic Kidney Disease (CKD) patients in a trial of 90 days wherein patients got standard-of-care treatment and liquid composition containing Sodium Copper Chlorophyllin, the Investigational Product (IP), as add-on treatment for first 60 days and patients were ‘off’ the IP for last 30 days. Significant reduction in serum creatinine, Blood Urea and Uric acid level and significant improvement in estimated Glomerular Filtration Rate (eGFR), and Health related criteria for quality of life were observed in 60 days’ period of administration. eGFR remained significantly higher on 90th day than 0 day despite withdrawal of IP after 60 days. These observations suggest partial/limited reversal of lost kidney function as a lingering effect of the IP. Both these are first observations opening up new avenues for improving life of CKD patients. This is first clinical trial report of an orally consumed product that decreases serum creatinine and also actually improved health related quality of life within 60 days and a surprising evidence that suggests significant degree of restoration of kidney function in the progressively degenerative disease CKD. This clinical trial opens opportunities for managing CKD that were never available so far.

Homogeneous electron-transfer of chlorophyll and its derivative chlorophyllin at gold electrode

Introduction: Chlorophyll is a light harvesting pigment, which absorbs light in the visible spectrum of sunlight and promotes electron transfer, Chlorophyllin (CHL) is One of the most important derivative molecules of chlorophyll. Nowadays, chlorophyll pigment and its derivatives are utilised in organic photosynthetic solar cells for their desirable photovoltaic properties. Cyclic voltammetry (CV) is an essential technique. It is extensively used to study electroactive species to interpret the intermediates of reactions, supply information about the thermodynamics of oxidation-reduction reactions and elucidate the kinetics of electron transfer reactions. Materials and Methods: Prior to the electrochemical study, the working gold (Au) electrode surface was prepared by immersing it in the various concentrations of chlorophyllin for a period time. The electrolyte was degassed by using N2 for approximately 30 minutes inside a Faraday cage before any electrochemical experiment was performed. A three electrode system was used with, Ag/AgCl as a reference electrode, graphiteas a counter and the working electrode (Au). Results and Discussion: As a route to develop new chemical systems for artificial photosynthesis, this work reports the effectiveness of different parameters in transferring electrons between chlorophyllin (CHL) pigment and the working electrode surface (gold). These parameters such as the adsorption time, the electrolyte nature and concentration and chlorophyllin concentration are investigated. The use of chlorophyllin as a redox mediator is examined, with a gold electrode being employed. The importance of gold electrode surface preparation in determining the mechanism of redox is described, and the environment of adsorption process of the different concentrations of chlorophyllin on the surface of the gold electrode has been elucidated in this study. Conclusiones: The electrochemical method showed that the cyclic voltammetry responses of studied adsorption chlorophyllin pigment on the gold electrode were more efficient. In addition, the redox reaction was successful electrochemically in aqueous solution thanthe organic solution. It was suggested that electrons reduce to the chlorophyllin pigment by adding active species in the bulk solution homogeneous transfer. Finally, detections of chl on spinach leaves using various methods are reported.

TOXICITY OF CHLOROPHYLLIN COMPOUND ON FIELD AND SUSCEPTIBLE STRAINS OF SPODOPTERA LITTORALIS, AND ITS BIOCHEMICAL IMPACT ON Α, Β AND ACETYLCHOLIN- ESTERASES

Chlorophyllin (CHL) is water-soluble analogue of chlorophyll pigment everywhere. This is due to the presence of many useful properties and its use as food additives and in alternative medicine. The study field was investigational at station of the faculty of Agricultural, Cairo University, at Giza governorates, Egypt. Result indicated that sub-lethal concentrations of sodium copper chlorophyllin and magnesium copper chlorophyllin on field and susceptible strains Spodoptera littoralis . The concentration (10-2 M) for both sodium copper chlorophyllin and magnesium copper chlorophyllin was more effective than concentration (10-3 M) on field strains and susceptible strains of Spodoptera littoralis. The determination activity level α-esterases, βesterases and acetylcholine were determined in the total body homogenates of Spodoptera littoralis . The results show that CHL was affected both (α – β) esterase enzyme. β –esterase activity showed significant activity increase, and α-esterase showed significant activity reduction in most treatment. on the other hand the acetylcholinesterase activity show low various.

Adverse Effects of Two Kinds of Food Additive Mixtures (Sodium benzoate + Monosodium glutamate, Monosodium glutamate + Chlorophyllin and Sodium benzoate + Chlorophyllin) on Some Physiological Parameters in Male Albino Rats

Background: Food additives are substances used in food industry in order to improve the food’s taste and appearance by preserving its flavor and preventing it from souring. Materials and Methods: this study was conducted on twenty-four young male albino rats with an average 120-140 g body weight. Animals were divided into four groups (6 each). Group I: Control untreated group, Group II: it was administered sodium benzoate (SB) (5 mg/kg b.w./day) orally and monosodium glutamate (MSG) (15 mg/kg b.w./day), Group III: it was administered MSG (15 mg/kg b.w./day) and chlorophyllin (CHL) (15 mg/kg b.w./day) and Group IV administered orally with SB (5 mg/kg b.w./day) and CHL (15 mg/kg b.w./day). Blood samples were collected on the last day of experiment, for biochemical estimations. Results: There was marked elevation in levels of fasting glucose, insulin, HOMA-IR, activities of AST and ALT, urea, creatinine, total cholesterol (TC), triglycerides (TG), LDL-C , VLDL, ratios of TC/HDL-C and LDL-C/HDL-C (risk factors) in all treated group. Serum thyroid hormones (T3 and T4) increased in Group III. On the other hand, there was decline in levels of serum total proteins, albumin, testosterone and HDL-C in all treated groups in comparison with the control group. There was a highly significant reduction in the body weight, T3 and T4 in Group IV, while Group II showed highly significant increase in body weight and a highly significant reduction in globulin level as compared to control rats. Conclusion: The use of these compounds must be limited as it resulted in a vehement disturbance in the biochemical parameters.

The impact of chlorophyllin on deoxynivalenol transport across jejunum mucosa explants obtained from adult pigs

Regardless of the efforts put into preventing or reducing fungal growth, extensive mycotoxin contamination has been reported in animal feeds. In the case of pigs, one of the mycotoxins of major concern is deoxynivalenol (DON). The use of adsorbents as feed additives represents one of the strategies to control mycotoxins’ contamination in feedstuff. Therefore, the aim of the study was to verify the ability of chlorophyllin (CHL) to reduce the absorption rate of DON in swine mucosa explants. Intestine was obtained from routinely slaughtered adult pigs. The mucosa explants were studied by means of Ussing chamber technique. The effect of DON (10 and 30 μg/ml) on mucosa viability and permeability and CHL (100 μg/ml) impact on DON (30 μg/ml) absorption was verified. The results revealed that mucosa explants isolated from adult animals remained unaffected for 90 min in the presence of DON in the lower concentration (10 μg/ml). Mycotoxin in the higher dose (30 μg/ml) increased mucosa permeability (decreased transepithelial electrical resistance value) and enhanced paracellular transport of lucifer yellow and mannitol but did not affect lactate dehydrogenase leakage. The introduction of CHL neither diminished the absorption rate of DON across swine mucosa explants nor prevented the toxic effects of DON on intestine. In conclusion, the results confirm the negative effect of DON on pig jejunum mucosa. However, the toxic effect of DON was observed only when it was used in relatively high doses. A promising adsorbent agent, CHL, failed to reduce the intensity of DON transport across intestine under in vitro conditions.

Photoactivated chlorophyllin and acetylcholinesterase/ cytochrome oxidase activity in Fasciola gigantica cercaria larvae

Objective: The effect of chlorophyllin (CHL), CHL + freeze dried cow urine (FCU) and FCU on acetylcholinesterase (AChE) and cytochrome oxidase activity in the nervous tissue of the host snail Lymnaea acuminata, and parasitic cercaria larvae of Fasciola gigantica were studied. Methods: Whole cercaria larvae and snail nervous tissue was homogenized and supernatant was used as an enzyme source. Enzyme activity was measured in 10-mm path length cuvette using incubation mixtures and change in optical density was recorded. Results: Sublethal 4 h LC50 40% and 80% of these formulations in sunlight and laboratory conditions caused no significant change in AChE activity in the nervous tissue of host snail. It was noted that sublethal in vivo 8 h exposure to 40% and 80% of 4 h LC50 of single and binary combination of CHL+FCU, CHL and FCU significantly altered the AChE and cytochrome oxidase activity in the cercaria larvae. Binary combinations of CHL+FCU in sunlight were more effective in inhibiting both the enzymes. Conclusion: The present study reported that enzymes AChE and cytochrome oxidase were inhibited by chlorophyllin formulations and thus causing the larval death inside the snail body without affecting the snail itself which is an important bioindicator of aquatic ecosytem.

Detection of not allowed food-coloring additives (copper chlorophyllin; copper-sulphate) in green table olives sold on the Italian market

Table olives are a common and well-known food in the whole Mediterranean area, produced and consumed in great quantities. Many deep-green olives can be found on sale in the South of Italy. Sometimes their unnatural color could be the result of the fraudulent addition of a coloring agent (E141ii, copper chlorophyllins) during the pickling process, in spite of the European Union legislation that does not allow the addition of any colorant to table olives. The objectives of this study were to detect fraudulent color adulteration of table olives with E141ii in Italian market and to confirm the presence on the market of artificially colored table olives. We assessed the presence of compounds with chromatographic and spectral characteristics similar to the ones from the E141ii (Cu chlorin e 6 , Cu isochlorin e 4 , Cu pyropheophorbide a) in 8 samples out of 16. These result show that the fraudulent addition of colorant to table olives is a quite common practice. More pressing controls and analysis are required to ensure the complete food safety and the compliance with the current law.

Ameliorative Effect of Chlorophyllin on Oxidative Stress in Experimental Model of Diabetes

The aim of this present study was to investigate the effect of chlorophyllin (CHL) on oxidative stress in Streptozotocine (STZ) induced diabetic mice. For the study, mice were divided into Group A: normal control, Group B: diabetic control, Group C: diabetic mice treated with the ascorbic acid, and Group D: diabetic mice treated with CHL. Levels of Reactive Oxygen Species (ROS), lipid peroxidation, protein carbonyl, superoxide dismutase (CuZn SOD &Mn-SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR) activities were examined in kidney and heart tissues of different experimental groups. Histological and ultrastructuralstudies were also carried out to evaluate any changes in tissues as well as sub-cellular organelles. ROS, lipid peroxidation, and protein carbonyl levels have been significantly decreased with concomitant increased of CuZn SOD, Mn-SOD, CAT, GPx, and GR activity in CHLtreated diabetic mice. The histological and ultrastructural studies showed that CHL attenuates the detrimental effect of oxidative stress and alleviated tissue injuries in STZ induced diabetic mice. These results suggested that CHL possesses antioxidative activity and has the potential to amelioratediabetes-associated oxidative stress in mice.

Nano-encapsulated chlorophyllin significantly delays progression of lung cancer both in in vitro and in vivo models through activation of mitochondrial signaling cascades and drug-DNA interaction.

Chlorophyllin (CHL), a sodium-copper-salt derived from chlorophyll, has been widely used as a food-dye, also reportedly having some anti-cancer effect. We tested if PLGA-loaded CHL (NCHL) could have additional protective abilities through its faster and targeted drug delivery in cancer cells. Physico-chemical characterization of NCHL was done through atomic-force microscopy and UV-spectroscopy. NCHL demonstrated greater ability of drug uptake and strong anti-cancer potentials in non-small cell lung cancer cells, A549, as revealed from data of% cell viability, generation of reactive-oxygen-species and expression of bax, bcl2, caspase3, p53 and cytochrome c proteins. Circular dichroic spectral data indicated strong binding of NCHL with calf-thymus-DNA, causing a conformational/structural change in DNA. Further, NCHL could cross the blood-brain-barrier in mice and showed greater efficacy in recovery process of tissue damage, reduction in chromosomal aberrations and% of micronuclei in co-mutagens (Sodiumarsenite+Benzo[a]Pyrene)-treated mice at a much reduced dose, indicating its use in therapeutic oncology.

Phytotherapy of chlorophyllin exposed Lymnaea acuminata: A new biotechnological tool for fasciolosis control

Phytotherapy of chlorophyllin formulations against Fasciola gigantica infected Lymnaea acuminata under sunlight exposure was highly toxic against redia and cercaria larvae. Binary combinations (1:1 ratio) of chlorophyllin (CHL)+freeze dried cow urine (FCU) were more toxic against cercariae (8h LC50: 9.6mgL−1) than single treatment with chlorophyllin (8h LC50: 12.6mgL−1) in sunlight. The larvicidal activity of sunlight exposed CHL against rediae (8h LC50: 13.5mgL−1) and cercariae (8h LC50: 12.6mgL−1) was more pronounced than laboratory conditions CHL treatment (rediae- 8h LC50: 305.9mgL−1; cercariae- 8h LC50: 765.4mgL−1). Larvicidal activity of FCU was less than CHL and CHL+FCU against both redia and cercaria. Chlorophyllin and its formulations were more toxic against redia and cercaria larvae in sunlight than laboratory conditions. CHL and its different formulations may be used as potent larvicides against Fasciola gigantica larvae. Chlorophyllin formulations will be economical, ecologically sounder and their use in aquatic environment will be safe.