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Microalgae are a source of a wide variety of commodities, including particularly valuable pigments. The typical pigments present in microalgae are the chlorophylls, carotenoids, and phycobiliproteins. However, other types of pigments, of the family of water-soluble polyphenols, usually encountered in terrestrial plants, have been recently reported in microalgae. Among such microalgal polyphenols, many flavonoids have a yellowish hue, and are used as natural textile dyes. Besides being used as natural colorants, for example in the food or cosmetic industry, microalgal pigments also possess many bioactive properties, making them functional as nutraceutical or pharmaceutical agents. Each type of pigment, with its own chemical structure, fulfills particular biological functions. Considering both eukaryotes and prokaryotes, some species within the four most promising microalgae groups (Cyanobacteria, Rhodophyta, Chlorophyta and Heterokontophyta) are distinguished by their high contents of specific added-value pigments. To further enhance microalgae pigment contents during autotrophic cultivation, a review is made of the main related strategies adopted during the last decade, including light adjustments (quantity and quality, and the duration of the photoperiod cycle), and regard to mineral medium characteristics (salinity, nutrients concentrations, presence of inductive chemicals). In contrast to what is usually observed for growth-related pigments, accumulation of non-photosynthetic pigments (polyphenols and secondary carotenoids) requires particularly stressful conditions. Finally, pigment enrichment is also made possible with two new cutting-edge technologies, via the application of metallic nanoparticles or magnetic fields.

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The method describes pigment analysis from microalgae/phytoplankton extracts using electron-transfer Matrix-Assisted Laser Desorption Ionization Mass Spectrometry (ET MALDI MS). Current microalgae/phytoplankton pigment analysis requires resource- and time-intensive chromatographic methods due to the broad polarity range of the target analytes. On the other hand, traditional MALDI MS chlorophyll analysis, using proton-transfer matrices such as 2,5-dihydroxybenzoic acid (DHB) or α-cyano-4-hydroxycinnamic acid (CHCA), results in central metal loss and phytol-ester cleavage. ET MALDI MS is an alternative for the rapid screening and detection of pigments in microalgae extracts.•MALDI matrices with ionization energies above 8.0 eV guarantee electron-transfer processes from photosynthetic and photoprotective pigments whose ionization energies lay below 7.5 eV.•ET MALDI MS pigment analysis agrees with data gathered from conventional chromatographic techniques (HPLC) and optical microscopy for pigment extracts from C. vulgaris cultures and freshwater phytoplankton samples.•The ET MALDI MS method allows fast and reliable detection of pigments in microalgae cultures and freshwater phytoplankton samples.

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Chronic wound healing represents an impactful financial burden on healthcare systems. In this context, the use of natural products as an alternative therapy reduces costs and maintains effectiveness. Phytotherapeutic gels applied in photodynamic therapy (PDT) have been developed to act as topical healing medicines and antibiotics. The bioactive system is composed of Spirulina sp. (source of chlorophylls) and Copaifera reticulata oil microdroplets, both incorporated into a polymeric blend constituted by kappa-carrageenan (k-car) and F127 copolymer, constituting a system in which all components are bioactive agents. The flow behavior and viscoelasticity of the formulations were investigated. The photodynamic activity was accessed from studies of the inactivation of Staphylococcus aureus bacteria, the main pathogen of hospital relevance. Furthermore, in vivo studies were conducted using eighteen rabbits with dermatitis (grade III and IV) in both paws. The gels showed significant antibiotic potential in vitro, eliminating up to 100% of S. aureus colonies in the presence or absence of light. The k-car reduced 41% of the viable cells; however, its benefits were enhanced by adding chlorophyll and copaiba oil. The animals treated with the phytotherapeutic medicine showed a reduction in lesion size, with healing and re-epithelialization verified in the histological analyses. The animals submitted to PDT displayed noticeable improvement, indicating this therapy's viability for ulcerative and infected wounds. This behavior was not observed in the iodine control treatment, which worsened the animals' condition. Therefore, gel formulations were a viable alternative for future pharmaceutical applications, aiming at topical healing.

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Recently, the number of new cases of cutaneous leishmaniasis has been of concern among health agencies. Research that offers new therapeutic alternatives is advantageous, especially those that develop innovative drugs. Therefore, this paper presents the incorporation of Copaifera reticulata Ducke and chlorophyll extract into Pluronic®® F127 and Carbopol gels, under optimized polymer quantities. The chlorophyll extract (rich in photosensitizing compounds) was obtained by continuous-flow pressurized liquid extraction (PLE), a clean, environmentally friendly method. The system aims to act as as a leishmanicidal, cicatrizant, and antibiotic agent, with reinforcement of the photodynamic therapy (PDT) action. Rheological and mechanical analyses, permeation studies and bioadhesiveness analyses on human skin, and PDT-mediated activation of Staphylococcus aureus were performed. The emulgels showed gelation between 13° and 15 °C, besides pseudoplastic and viscoelastic properties. Furthermore, the systems showed transdermal potential, by releasing chlorophylls and C. reticulata Ducke into the deep layers of human skin, with good bioadhesive performance. The application of PDT reduced three logarithmic colony-forming units of S. aureus bacteria. The results support the potential of the natural drug for future clinical trials in treating wounds and cutaneous leishmania.

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Green or purple lettuce varieties produce many secondary metabolites, such as chlorophylls, carotenoids, anthocyanins, flavonoids, and phenolic compounds, which is an emergent search in the field of biomolecule research. The main objective of this study was to use multivariate and machine learning algorithms on Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR)-based spectra to classify, predict, and categorize chemometric attributes. The cluster heatmap showed the highest efficiency in grouping similar lettuce varieties based on pigment profiles. The relationship among pigments was more significant than the absolute contents. Other results allow classification based on ATR-FTIR fingerprints of inflections associated with structural and chemical components present in lettuce, obtaining high accuracy and precision (>97%) by using principal component analysis and discriminant analysis (PCA-LDA)-associated linear LDA and SVM machine learning algorithms. In addition, PLSR models were capable of predicting Chla, Chlb, Chla+b, Car, AnC, Flv, and Phe contents, with R2P and RPDP values considered very good (0.81−0.88) for Car, Anc, and Flv and excellent (0.91−0.93) for Phe. According to the RPDP metric, the models were considered excellent (>2.10) for all variables estimated. Thus, this research shows the potential of machine learning solutions for ATR-FTIR spectroscopy analysis to classify, estimate, and characterize the biomolecules associated with secondary metabolites in lettuce.

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Coloring compounds are widely applied to manufacturing foods and beverages. The worldwide food market is replacing artificial colorants with natural alternatives, given the increased consumer demand for natural products. However, these substitutes are still an issue due to their high production cost and low chemical and physical stability. Furthermore, natural pigments are highly sensitive to processes applied in conventional extraction techniques, such as thermal, mechanical, and chemical stresses. In this regard, pulsed electric field (PEF) technology has emerged as a promising non-thermal alternative for recovering and producing natural colorings from food matrices. Its action mechanism on cell structures through the electroporation effect is a smart alternative to overcoming the challenging issues associated with producing natural colorants. In this scenario, this review provides an overview of the PEF assisted extraction of natural pigments and colorants, such as anthocyanins (red-blue-purple), betalains (red), carotenoids (yellow-orange-red), and chlorophylls (green) from plant sources. Moreover, the potential and limitations of this emerging technology to integrate the extraction process of natural colorants were discussed.

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Microalgae are photosynthetic microorganisms that stand out from conventional food sources and ingredients due to their high growth rate and adaptability. In addition to being highly sustainable, significant concentrations of proteins, lipids, and pigments accumulate in their cell structures from photosynthesis. Hence, this study sought to evaluate the food potential of Scenedesmus obliquus biomasses obtained from photosynthetic cultures enriched with 3, 5, 10, 15, 20, and 25% carbon dioxide (CO2) (v/v). Cultivations with 3, 5, and 10% CO2 showed greater amino acids and proteins synthesis; the protein content reached values above 56% of the dry biomass and high protein quality, due to the presence of most essential amino acids at recommended levels for the human diet. The highest concentrations of chlorophylls were found in cultures with 15, 20, and 25% CO2 (24.2, 23.1 and 30.8 mg g-1, respectively), although the profiles showed higher percentages of degradation compounds. Carotenoid concentrations were three times higher in cultures with 3, 5, and 10% CO2 (25.3, 22.7 and 18.1 mg g-1, respectively) and all-trans-ß-carotene was the major compound. Lipid synthesis was intensified at higher CO2 enrichment; the percentages obtained were 14.8% of lipids in the culture with 15% CO2, 15.0% with 20% CO2, and 13.7% with 25% CO2. In addition, greater polyunsaturated fatty acids accumulation and a significant reduction in the n6/n3 ratio were also observed at the highest CO2 concentrations. Our findings showed that CO2 treatments significantly altered all compounds concentrations in S. obliquus biomasses, which presented satisfactory composition for application in foods and as ingredients.

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This study aimed to investigate the bioaccessibility of carotenoids and chlorophylls from the biomass of microalgae Chaetoceros calcitrans. The samples were submitted to an in vitro digestion protocol, and the compounds were determined by HPLC-PDA-MS/MS. A total of 13 compounds were identified in all tests. After in vitro digestion, the relative bioaccessibility of carotenoids and chlorophylls ranged from 4 to 58%. The qualitative profile of carotenoids reflected the initial sample, with all-E-zeaxanthin (57.2%) being the most bioaccessible compound, followed by all-E-neochrome (31.26%), the latter being reported for the first time in the micellar fraction. On the other hand, among the chlorophylls only pheophytin a (15.01%) was bioaccessible. Furthermore, a chlorophyll derivative (Hydroxypheophytin a') was formed after in vitro digestion. Considering all compounds, xanthophylls (12.03%) and chlorophylls (12.22%) were significantly (p < 0.05) more bioaccessible than carotenes (11.22%). Finally, the considerable individual bioaccessibilities found, especially for zeaxanthin, demonstrate the bioactive potential of this bioresource. However, the large reduction in the totality of compounds after in vitro digestion suggests that additional technological strategies should be explored in the future to increase the efficiency of micellarization and enhance its bioactive effects.

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The sinami palm (Oenocarpus mapora H. Karst) is a plant from the South American Amazonia that has great potential for industrial applications in the development of functional foods, nutraceuticals and cosmeceuticals. In this manuscript, the physicochemical properties, total polyphenol content and antioxidant activity of sinami oil that was obtained using four extraction systems, namely expeller press extraction (EPE), cold press extraction (CPE), ultrasound-assisted extraction (UAE) and supercritical fluid extraction (SFE), were studied and compared. The oxidative stability (OSI) was statistically non-significant in EPE and SFE. The chromatic properties (CIELab) were influenced by the extraction methods and SFE presented high values of L* and a lower content of plant pigments. Ultrasound-assisted extraction showed a higher content of polyphenols and higher antioxidant activity. Different analyses for the evaluation of the physicochemical properties, the content of total polyphenols and antioxidant activity were used to classify sinami oil according to chemometrics using principal component analysis (PCA). For example, the sinami oil that was obtained using each extraction method was in a different part of the plot. In summary, sinami oil is an excellent resource for plant pigments. Additionally, the information that was obtained on the quality parameters in this study provided a good foundation for further studies on the characterization of major and minor compounds.

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BACKGROUND: Wild edible herbs have historically been used as local nutritional and medicinal sources. These plants grow spontaneously, depending on the season. They adapt well to different edaphoclimatic conditions, generating a diversity constituent beneficial to health. They impart compounds needed in the human diet in regard to macro and micronutrients. When consumed raw, they keep their properties intact and provide health benefits. Five undervalued edible plants: Stellaria media (L.) Vill, Tropaeolum majus L., Sonchus oleraceus L., Chenopodium album L. and Diplotaxis erucoides (L.) DC are characteristic of the autumn-winter season in the Valencian coastal region and could have new sustainable agro-ecological potential for the local commercial sector. However, little information is available from the nutritional quality and bioactive composition viewpoint for these species. Concurrently, the volatiles compounds profiles describing the characteristic flavors are unknown. METHODS: Nutritional characteristics, bioactive compounds, and other chemical components of the fresh leaves were analyzed. In addition, the volatiles composite profile was performed. The analyzed species come from the soil reservoir; their wild growth is adjusted to the autumn season. The proximate analysis was carried out by Association of Official Analytical Chemists methods. Total antioxidants were measured as 2.2-diphenyl-1-picrylhydrzyl hydrate (DPPH) and total polyphenols content via the Folin-Ciocalteu procedure. Volatiles profile was determined by gas chromatography-mass spectrometry. The vegetative part analyzed was the tender leaves with edible potential. RESULTS: A high variability has been obtained in the composition of the species studied. The proximate analysis found a considerable content of fiber (1.22-5.4 g·100 g-1), potassium (157.7-1,250.6 mg·100 g-1), iron (0.6-2.0 mg·100 g-1), and a low caloric value (16.1-43.02 kcal·mg·100 g-1). In bioactive compounds analysis, a high level of antioxidants was highlighted (1,604.3-4,874.6 µmol·100 g-1), followed by chlorophylls. Volatiles profile revealed that the species were rich in benzenoids (33.8-89.9%) as the majority family. The pyrazines class was characteristic only in D. erucoides L. DISCUSSION: Fresh edible leaves of the undervalued plants show considerable nutritional potential and a high bioactive components level, which highlight the antioxidant capacity. Leaves of C. album L. stand out due to their higher concentration of nutritional compounds, while D. erucoides L. is noted for its higher antioxidant capacity. Aromatic descriptor of pyrazines detected in the leaves of D. erucoides L. is associated with the slightly spicy flavors that characterize this species. Results suggest that studied species could be of great relevance in introducing these five edible herbs as a source of new grown material, postulating them as healthy food ingredients with attractive flavors for the gourmet cuisine industry.

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ABSTRACT Cacao has high social importance in Colombia but requires cultivation techniques that improve its productivity, especially in the production of organic cacao. Pruning and organic fertilizers were assessed as treatments to increase the bean yield in an old cacao crop in Cundinamarca. The positive effects of organic fertilizer application and pruning on the pod morphometry and bean yield were established. The application of organic fertilizers with pruning, in general, increased the length and circumference of cacao pods. A negative correlation (R=-0.618) was found among the relative chlorophyll contents in leaves at the end of the harvest period and with the number of pods per tree. A positive correlation (R=0.748) was obtained between the fresh weight of pods and the dry weight of beans. No correlation was established between the relative chlorophyll contents in leaves and the bean dry weight. Pruning plus organic fertilization increased the total dry weight of beans per tree at almost 15 %, which makes pruning with organic fertilizer application the best tool for increasing yield in organic cacao production in old plantations in Cundinamarca.

RESUMEN El cultivo de cacao tiene una alta importancia social en Colombia, pero requiere la implementación de técnicas de cultivo que mejoren su productividad, principalmente, en la producción de cacao orgánico. La poda y los fertilizantes orgánicos fueron evaluados, como los tratamientos para incrementar el rendimiento de cacao, en una plantación antigua en el departamento Cundinamarca. La aplicación de fertilizantes orgánicos y la poda tuvieron efectos positivos sobre la morfometría del fruto y el rendimiento del cacao. La aplicación de fertilizantes orgánicos con poda, en general, aumentó la longitud y el diámetro de frutos de cacao. Se encontró una correlación negativa (R=-0,618) entre el contenido relativo de clorofila en hojas al final del período de cosecha y el número de frutos por árbol. Se obtuvo una correlación positiva (R=0,748) entre el peso fresco de frutos y el peso seco de semillas. No se estableció correlación entre el contenido relativo de clorofila en hojas y el peso seco de semillas. La poda más fertilización orgánica aumentó el peso seco total de semillas por árbol, en casi un 15 %, lo que convierte la poda con aplicación de fertilizante orgánicos en una herramienta para aumentar el rendimiento de cacao orgánico en antiguas plantaciones, en el departamento Cundinamarca.

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The aim of this work was to study the evolution of fruit size and weight together with the soluble solid and total titratable acidity contents during development of Hexachlamys edulis fruit. Also, the patterns of accumulation of chlorophylls, carotenoids, phenols and antioxidant activity were analysed to define the optimal time for harvesting to obtain maximum nutraceutical characteristics. Fruits were harvested from H. edulis plants growing at the experimental field of the University of Morón (Moreno, Buenos Aires, 34°35'4.98″ SL, 58°48'52.09″ WL, 14 m.a.s.l.). Fresh fruit weight was significantly higher in Medium ripe, Ripe and Overripe fruits (40.1, 39.6 and 38.5 g, respectively) than in Unripe fruits (19.5 g). Soluble solids/total titratable acidity was significantly higher in Overripe fruits (7.3) than in Unripe, Medium ripe and Ripe fruits (3.7-4.5). Total polyphenols were maximum in Unripe fruits (905.8 mg tannic acid/100 g fresh fruit weight) decreasing during the fruit development (426.2-130.4 mg tannic acid/100 g fresh fruit weight). Also, DPPH radical scavenging activity was significantly higher in Unripe fruits (75.7%) compared with Medium ripe, Ripe and Overripe fruits (64.1-17.0%). Positive and significant correlations were observed between total polyphenol content and DPPH radical scavenging activity at each extract concentration (r = 0.74, 0.87, 0.74 and 0.60 for 1.25; 2.50; 5.00 and 12.50 mg/mL, respectively). Total carotenoid content increased during fruit development while at the same time decreased chlorophyll content. Chlorophyll b is the main chlorophyll found. Chromatographic analysis showed that lutein is the main carotenoid found in H. edulis fruits, followed by ß-cryptoxanthin and ß-carotene. As shown by the chromatograms at 280 nm, the concentration of biophenols and the complexity of the biophenol profile decreases during fruit development. Levels of polyphenols and pigments together with the antioxidant activity allow us to consider H. edulis fruit as a functional food.

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Different cultivation strategies have been developed with the aim of increasing the production rate of microalgal pigments. Specifically, biotechnological approaches are designed to increase antioxidant metabolites as chlorophyll and carotenoids. However, although significant advances have been built up, available information regarding both the chlorophyll metabolism and their oxidative reactions in photobioreactors is scarce. To unravel such processes, the detailed chlorophyll and carotenoid fraction of Scenedesmus obliquus has been studied by HPLC-ESI/APCI-hrTOF-MS from phototrophic and heterotrophic cultures. Scenedesmus is provided with a controlled strategy of interconversion between chlorophyll a and b to avoid the formation of reactive oxygen species (ROS) at high irradiances in addition to the photoacclimation of carotenoids. Indeed, precise kinetics of 132-hydroxy- and 151-hydroxy-lactone chlorophyll metabolites shows the existence of a chlorophyll oxidative metabolism as a tool to manage the excess of energy at high light conditions. Unexpectedly, the oxidation under phototrophy favored chlorophyll b metabolites over the chlorophyll a series, while the heterotrophic conditions exclusively induced the formation of 132-hydroxy-chlorophyll a. In parallel, during the first 48 h of growth in the dark, the chlorophyll fraction maintained a promising steady state. Although future studies are required to resolve the biochemical reactions implied in the chlorophyll oxidative metabolism, the present results agree with phytoplankton metabolism.

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Researches about light emitting diodes (LEDs) as energy source in microalgae cultivations has been growing in recent years due to its spectral quality, durability and reduced energy consumption. In this study, green, red and yellow LEDs were evaluated as energy source in Synechococcus nidulans LEB 115 cultures. Productivities and specific growth rates were up to 2.5 times greater than in cultures using fluorescent light. The different LED colors evaluated did not influence the chlorophyll, carotenoid or lipid productions. Biomass cultivated with LEDs showed high amounts of saturated fatty acids (above 48%), which is desirable for biodiesel production. In addition to the Synechococcus nidulans LEB 115 growth stimulation, the application of green, red and yellow LEDs in the cultivations produces potential biomass for biodiesel synthesis and other industrial interest biomolecules utilization.

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Extra-virgin olive oil (EVOO) from Mediterranean were analyzed by Ultraviolet-Visible (UV-Vis) spectroscopy and Independent Component Analysis (ICA). The use of ICA resolution provided information over dienes (primary oxidation compound), polyphenolic compounds, tocopherol, carotenoids and chlorophylls when EVOO was evaluated by UV-Vis spectroscopy. Based on these data, ICA could be used to determine the contribution of chemical compounds to the composition of EVOO produced in different regions from Mediterranean. The results suggest that the combination of UV-Vis measurements and ICA makes possible the EVOO evaluation, and can contribute to suggesting that a foodstuff comes from an alleged origin. The proposed methodology is a low cost, fast and sample preparation free methodology to highlights the EVOO characteristics produced in the Mediterranean region.

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LEDs have narrow wavelength bands, which can influence microalgae biomass. This study pioneers the evaluation of blue LEDs as an energy source in Chlorella fusca and Synechococcus nidulans cultures. Blue LEDs increased the specific growth rate in Synechococcus nidulans LEB 115 cultures by 80% compared to the standard light used in indoor cultivations. Moreover, blue LEDs also induced lipid accumulation in Chlorella fusca LEB 111 cells, yielding concentrations of this bioproduct of up to 23% (ww-1). The chlorophylls and carotenoids were photostimulated proportionally to the LED light intensity. When the intensity of the blue LEDs was increased from 50 to 150µmolm-2s-1, the biomass accumulated up to 4.5 and 2.4 times more chlorophylls and carotenoids, respectively. The potential of blue LEDs as an alternative environmentally friendly light source to stimulate biomass and metabolite production for different purposes was demonstrated.

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The identification of chlorophyll molecules with peroxyl radical scavenger capacity in microalgae Phormidium autumnale was determined. The ultrasound-assisted extraction was utilized for obtaining the chlorophyll compounds from biomass. A total of eleven molecules were separated in microalgae chlorophyll extract, with pheophytin a' (371µg·g-1) and chlorophyll a (159.3µg·g-1) as the major ones. The chlorophyll extract was shown to be a potent scavenger of peroxyl radical, being almost 200 times more potent than α-tocopherol. These facts suggest the microalgae Phormidium autumnale as potential source of bioactive tetrapyrrole compounds.

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This study aimed at applying different intensities (30 and 60mT) and exposure times (24 and 1hd-1) of MF to cultures of Chlorella kessleri and evaluated their effects on cell growth, proximate composition of biomass, pigment production and antioxidant activity. The condition of 60mT for 1hd-1 stimulated biomass concentration of 83.2% by comparison with control culture (CC). Besides stimulated 13.7% lipid synthesis, 38.9% chlorophyll a and 59.1% chlorophyll b, 25.0% total carotenoids and antioxidants up to 185.7%. Thus MF application is an excellent alternative to stimulate cell growth and high biotechnological interest biocompounds.

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The spectrophotometric properties of chlorophylls' derivatives (Chls) formulated in the Pluronics® F-127 and P-123 were evaluated and the results have shown that the Chls were efficiently solubilized in these drug delivery systems as monomers. The relative location of the Chls in the Pluronics® was estimated from the Stokes shift and micropolarity of the micellar environment. Chls with phytyl chain were located in the micellar core, where the micropolarity is similar to ethanol, while phorbides' derivatives (without phytyl chain) were located in the outer shell of the micelle, i.e., more polar environment. In addition, the thermal stability of the micellar formulations was evaluated through electronic absorption, fluorescence emission and resonance light scattering with lowering the temperature. The Chls promote the stability of the micelles at temperatures below the Critical Micellar Temperature (CMT) of these surfactants. For F-127 formulations, the water molecules drive through inside the nano-structure at temperatures below the CMT, which increased the polarity of this microenvironment and directly affected the spectrophotometric properties of the Chls with phytyl chain. The properties of the micellar microenvironment of P-123, with more hydrophobic core due to the small PEO/PPO fraction, were less affected by lowering the temperature than for F-127. These results enable us to better understand the Chls behavior in micellar copolymers and allowed us to design new drug delivery system that maintains the photosensitizer's properties for photodynamic applications.

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