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Diverse morphological, cellular and physiological changes occur during seed maturation in Bixa orellana when the seed tissues form specialized cell glands that produce reddish latex with high bixin amounts. Transcriptomic profiling during seed development in three B. orellana accessions (P12, N4 and N5) with contrasting morphologic characteristics showed enrichment in pathways of triterpenes, sesquiterpenes, and cuticular wax biosynthesis. WGCNA allows groups of all identified genes in six modules the module turquoise, the largest and highly correlated with the bixin content. The high number of genes in this module suggests a diversification of regulatory mechanisms for bixin accumulation with the genes belonging to isoprene, triterpenes and carotene pathways, being more highly correlated with the bixin content. Analysis of key genes of the mevalonate (MVA) and the 2C-methyl-D-erythritol-4-phosphate (MEP) pathways revealed specific activities of orthologs of BoHMGR, BoFFP, BoDXS, and BoHDR. This suggests that isoprenoid production is necessary for compounds included in the reddish latex of developing seeds. The carotenoid-related genes BoPSY2, BoPDS1 and BoZDS displayed a high correlation with bixin production, consistent with the requirement for carotene precursors for apocarotenoid biosynthesis. The BoCCD gene member (BoCCD4-4) and some BoALDH (ALDH2B7.2 and ALDH3I1) and BoMET (BoSABATH1 and BoSABATH8) gene members were highly correlated to bixin in the final seed development stage. This suggested a contributing role for several genes in apocarotenoid production. The results revealed high genetic complexity in the biosynthesis of reddish latex and bixin in specialized seed cell glands in different accessions of B. orellana suggesting gene expression coordination between both metabolite biosynthesis processes.

Achiote (Bixa orellana L.) is a valuable species due to the bixin content present in the aril of its seeds; this pigment is used in the food, cosmetic and medicinal industries. Germplasm characterization is the starting point to initiate breeding programs and to identify the genetic variability of a germplasm bank. Objective. This project evaluated the phenotypic and genotypic differences of 18 achiote accessions from the EARTH University Germplasm Bank. Materials and methods. Phenotype was characterized by general and botanical descriptors detailing morphological and productive characteristics of the cultivars. Genotype was characterized based on analysis of the ITS2 region of ribosomal DNA to establish phylogenetic relationships among accessions and to determine relationships between phenotypic and molecular characteristics. Self-pollination trials were also carried out to evaluate the response of the accessions to this method leading to the obtention of pure lines. Results. The results from the Maximum Parsimony dendrogram allowed to differentiate five phylogenetic groups, which are closely related to the degree of spinosity of the capsules. By condensing the dendrogram to 50%, 2 groups were obtained, differentiated by the presence or absence of spines in the capsules, which positions this trait as a determinant for the grouping of materials. Accessions with spines had a higher number of seeds per capsule, while accessions without spines had a higher bixin content and higher seed weight. Selfpollination assays revealed apparent lateacting self-incompatibility in some accessions. Conclusion. This study generated information on the identification of the accessions and is a first phase aimed at obtaining homozygous lines to preserve the genetic identity of the accessions in the EARTH University Germplasm Bank., El achiote (Bixa orellana L.) es una especie valiosa debido al contenido de bixina presente en el arilo de sus semillas, pigmento que es utilizado en la industria alimentaria, cosmética y medicinal. La caracterización de germoplasma es el punto de partida para iniciar programas de fitomejoramiento y para identificar la variabilidad genética de un banco de germoplasma. Objetivo. Evaluación de las diferencias fenotípicas y genotípicas de 18 accesiones de achiote del Banco de Germoplasma de la Universidad EARTH. Materiales y métodos. El fenotipo se caracterizó por medio de descriptores generales y botánicos que detallan características morfológicas y productivas de los cultivares. El genotipo se caracterizó a partir del análisis de la región ITS2 del ADN ribosómico para establecer relaciones filogenéticas entre las accesiones y determinar las relaciones entre características fenotípicas y moleculares. También se realizaron ensayos de autopolinización para evaluar la respuesta de las accesiones a este método que condujera a la obtención de líneas puras. Resultados. Los resultados a partir del dendrograma Maximum Parsimony permitieron diferenciar 5 grupos filogenéticos, los cuales poseen una estrecha relación con el grado de espinosidad de las cápsulas. Al condensar el dendrograma a un 50% se obtuvieron 2 grupos diferenciados por la presencia o ausencia de espinas en las cápsulas, esto posiciona a esta característica como la determinante para el agrupamiento de los materiales. Las accesiones con espinas presentan mayor número de semillas por cápsula, mientras que las accesiones sin espinas poseen mayor contenido de bixina y mayor peso de semilla. Los ensayos de autopolinización revelaron una aparente autoincompatibilidad de acción tardía en algunas accesiones. Conclusión. Esta investigación generó información sobre la identificación de las accesiones, como una primera fase encaminada a la obtención de líneas homocigotas para conservar la identidad genética de las accesio

Achiote (Bixa orellana L.) is a valuable species due to the bixin content present in the aril of its seeds; this pigment is used in the food, cosmetic and medicinal industries. Germplasm characterization is the starting point to initiate breeding programs and to identify the genetic variability of a germplasm bank. Objective. This project evaluated the phenotypic and genotypic differences of 18 achiote accessions from the EARTH University Germplasm Bank. Materials and methods. Phenotype was characterized by general and botanical descriptors detailing morphological and productive characteristics of the cultivars. Genotype was characterized based on analysis of the ITS2 region of ribosomal DNA to establish phylogenetic relationships among accessions and to determine relationships between phenotypic and molecular characteristics. Self-pollination trials were also carried out to evaluate the response of the accessions to this method leading to the obtention of pure lines. Results. The results from the Maximum Parsimony dendrogram allowed to differentiate five phylogenetic groups, which are closely related to the degree of spinosity of the capsules. By condensing the dendrogram to 50%, 2 groups were obtained, differentiated by the presence or absence of spines in the capsules, which positions this trait as a determinant for the grouping of materials. Accessions with spines had a higher number of seeds per capsule, while accessions without spines had a higher bixin content and higher seed weight. Selfpollination assays revealed apparent lateacting self-incompatibility in some accessions. Conclusion. This study generated information on the identification of the accessions and is a first phase aimed at obtaining homozygous lines to preserve the genetic identity of the accessions in the EARTH University Germplasm Bank., El achiote (Bixa orellana L.) es una especie valiosa debido al contenido de bixina presente en el arilo de sus semillas, pigmento que es utilizado en la industria alimentaria, cosmética y medicinal. La caracterización de germoplasma es el punto de partida para iniciar programas de fitomejoramiento y para identificar la variabilidad genética de un banco de germoplasma. Objetivo. Evaluación de las diferencias fenotípicas y genotípicas de 18 accesiones de achiote del Banco de Germoplasma de la Universidad EARTH. Materiales y métodos. El fenotipo se caracterizó por medio de descriptores generales y botánicos que detallan características morfológicas y productivas de los cultivares. El genotipo se caracterizó a partir del análisis de la región ITS2 del ADN ribosómico para establecer relaciones filogenéticas entre las accesiones y determinar las relaciones entre características fenotípicas y moleculares. También se realizaron ensayos de autopolinización para evaluar la respuesta de las accesiones a este método que condujera a la obtención de líneas puras. Resultados. Los resultados a partir del dendrograma Maximum Parsimony permitieron diferenciar 5 grupos filogenéticos, los cuales poseen una estrecha relación con el grado de espinosidad de las cápsulas. Al condensar el dendrograma a un 50% se obtuvieron 2 grupos diferenciados por la presencia o ausencia de espinas en las cápsulas, esto posiciona a esta característica como la determinante para el agrupamiento de los materiales. Las accesiones con espinas presentan mayor número de semillas por cápsula, mientras que las accesiones sin espinas poseen mayor contenido de bixina y mayor peso de semilla. Los ensayos de autopolinización revelaron una aparente autoincompatibilidad de acción tardía en algunas accesiones. Conclusión. Esta investigación generó información sobre la identificación de las accesiones, como una primera fase encaminada a la obtención de líneas homocigotas para conservar la identidad genética de las accesio

The aril that covers the seeds of the plant species Bixa orellana is composed of the natural colorants bixin and norbixin, phenolic components and native carbohydrates composed of hemicellulose, starch, and a small fraction of proteins. Bixin is a hydrophobic carotenoid that is water insoluble and is mostly used for coloring oil-based formulations. This thesis presents the development of different bixin formulations to be used as coloring agents in aqueous continuous-based products. Any colorant formulation needs the colorant to be available and extraction feasible. The first stage of the thesis consisted of the morphological and photochemical characterization of Bixa orellana fruits that were collected from Los Yungas of La Paz, Bolivia at different altitudes. The water content, seed weight, and the amount of bixin, norbixin and native carbohydrates were quantified. The aril of the seeds was extracted using a mild alkaline extraction method to obtain an aqueous-dispersible extract that was directly analyzed by meas of solvent dissolution and further injection into a HPLC or by means of analysis using spectroscopic techniques.In a second stage, the use of the mild alkaline base was used as a method to obtain aqueous dispersions that were mixed with an encapsulating agent. This mixture was spray dried to obtain colorant microcapsules. Micro encapsulation was applied to obtain bixin aqueous-dispersible formulations and offer protection to the colorant against light degradation. The impact of the native carbohydrates was evaluated as an encapsulating agent and as protection against bixin light degradation. The encapsulation efficiency of bixin by different wall materials was compared. Light stability experiments were performed on Bixin microcapsules (powders) and unprotected colorants. The results show that lower sensitivity towards light was obtained for microencapsulated samples. Moreover, light stability of annatto extracts applied to gummy candies was evaluated. Th

Scope: It is investigated whether at realistic dietary intake bixin and crocetin could induce peroxisome proliferator-activated receptor γ (PPARγ)-mediated gene expression in humans using a combined in vitro–in silico approach. Methods and results: Concentration–response curves obtained from in vitro PPARγ-reporter gene assays are converted to in vivo dose–response curves using physiologically based kinetic modeling-facilitated reverse dosimetry, from which the benchmark dose levels resulting in a 50% effect above background level (BMD50) are predicted and subsequently compared to dietary exposure levels. Bixin and crocetin activated PPARγ-mediated gene transcription in a concentration-dependent manner with similar potencies. Due to differences in kinetics, the predicted BMD50 values for in vivo PPARγ activation are about 30-fold different, amounting to 115 and 3505 mg kg bw−1 for crocetin and bixin, respectively. Human dietary and/or supplemental estimated daily intakes may reach these BMD50 values for crocetin but not for bixin, pointing at better possibilities for in vivo PPARγ activation by crocetin. Conclusion: Based on a combined in vitro–in silico approach, it is estimated whether at realistic dietary intakes plasma concentrations of bixin and crocetin are likely to reach concentrations that activate PPARγ-mediated gene expression, without the need for a human intervention study.

The transcription factor nuclear factor erythroid 2-related factor 2 (NRF2) is the master regulator of the cellular antioxidant response upon exposure to xenobiotics. In addition, NRF2 induces the expression of genes involved in drug metabolism, protein homeostasis, anabolic metabolism, inflammation, proliferation, and survival. Understanding the molecular mechanisms of xenobiotic-induced NRF2 activation is crucial to guiding treatments for disease prevention and intervention. On the one hand, NRF2 activation using chemopreventive compounds is an effective pharmacological strategy to protect against environmental insults, such as exposure to solar ultraviolet (UV) radiation or the metalloid arsenic, that cause toxicity and cancer. In the first part of this dissertation, the achiote-derived apocarotenoid bixin was characterized as a novel NRF2 activator. Bixin pre-treatment prevented solar UV-induced photodamage and hair graying in several mouse models. These results suggest that using bixin to activate NRF2 is a feasible strategy to prevent UV-induced skin alterations. On the other hand, prolonged activation of NRF2 has been linked to the pathogenesis of cancer and other diseases. Our laboratory previously described that the environmental toxicant and carcinogen arsenic blocked autophagy and caused non-canonical activation of NRF2. In the last part of this dissertation, the molecular alterations elicited by acute exposure to low levels of arsenic were further explored. It was determined that low-level arsenic does not generate reactive oxygen species, a paradigm shifting discovery in the arsenic field. Additionally, arsenic blocked autophagy and induced a mild and transient endoplasmic reticulum (ER) stress response, indicating that restoring protein homeostasis might be crucial for the treatment of arsenic-induced diseases.

Bixin is a carotenoid which is extracted from annatto (Bixa orellana). Its main application is as a coloring agent. It consists of conjugating double bonds with a carboxylic end, making it prone to degradation by oxygen, light and heat. This study investigates the light stability of bixin encapsulated with maltodextrin (MD), sucrose and whey by spray drying. MD was added with additives such as carboxymethylcellulose (CMC), gum arabic (GA) and pectin to examine their interaction effects. Two sets of samples were used; one with pure bixin and carbohydrates, and the other, with non-filtered bixin encapsulated with the same set of carbohydrates and a carbohydrate naturally present in the seeds named botanical tissue. The samples were illuminated using a plasma sulfur lamp for a period of 30 days. UV-Visible spectrometer was used to keep track of the degradation. Statistic tests such as Shapiro-Wilk, ANOVA and t-test were used to confirm that the degradation was linear and that they follow first order decay. Quantum yields of photo-bleaching were calculated and compared between samples. Lower the quantum yield, higher the light stability. The presence of carbohydrates significantly improved the stability of bixin in comparison with non-encapsulated samples. The non-encapsulated samples lost their color completely by day 9 while the others lasted till 30 days. Sucrose showed the highest degradation rate in the absence of botanical tissue. Although sucrose and whey showed highest stability towards light in the presence of botanical tissue, the mix of MD and CMC worked better without botanical tissue. Botanical tissue seems to play a complex role in protecting bixin as the results varied with different carbohydrates., Bixin is a coloring agent used in various industries especially food and textile industries. The color is achieved from annatto fruits found in tropical areas. It is known to produce colored products efficiently but there is an issue with its stability. The color gets bleached gradually until it gets completely lost. This is because it reacts with the oxygen in the surrounding air and gets changed in structure which will no longer have the same properties. A similar effect can be noticed when it is subjected to heat and light. Bixin can be protected from such changes by covering it with another material in a capsule form. In this case, carbohydrates were used to encapsulate bixin to provide better stability. In addition to this, bixin was found to have an additional carbohydrate (botanical tissue) cover that occurs naturally during extraction. The goal of this study was to examine how well the carbohydrates prevent degradation of color caused by light with and without the botanical tissue. Sucrose, whey and maltodextrin along with some additives were investigated as carbohydrates. The samples were placed under a lamp that provided light that was similar to daylight for 30 days. It was seen that without carbohydrates, bixin lost all of its color in 9 days, while the rest lasted for 30 days. This shows that carbohydrates do prevent color decay to a great extent. When analyzed, it was seen that the color degraded uniformly along the 30 days. Botanical tissue gave a nice color to the samples which stayed till the end of the study. Along with this, the amount of light absorbed by the samples was also calculated. Either bixin molecules or oxygen molecules were made reactive with the energy absorbed from light and this caused a reaction between them, leading to bleaching. Amongst all the samples with carbohydrates, bixin with sucrose had the highest loss of color. But this was overcome when botanical tissue was present. In terms of light stability, sucrose and whey showed

Bixin is a carotenoid which is extracted from annatto (Bixa orellana). Its main application is as a coloring agent. It consists of conjugating double bonds with a carboxylic end, making it prone to degradation by oxygen, light and heat. This study investigates the light stability of bixin encapsulated with maltodextrin (MD), sucrose and whey by spray drying. MD was added with additives such as carboxymethylcellulose (CMC), gum arabic (GA) and pectin to examine their interaction effects. Two sets of samples were used; one with pure bixin and carbohydrates, and the other, with non-filtered bixin encapsulated with the same set of carbohydrates and a carbohydrate naturally present in the seeds named botanical tissue. The samples were illuminated using a plasma sulfur lamp for a period of 30 days. UV-Visible spectrometer was used to keep track of the degradation. Statistic tests such as Shapiro-Wilk, ANOVA and t-test were used to confirm that the degradation was linear and that they follow first order decay. Quantum yields of photo-bleaching were calculated and compared between samples. Lower the quantum yield, higher the light stability. The presence of carbohydrates significantly improved the stability of bixin in comparison with non-encapsulated samples. The non-encapsulated samples lost their color completely by day 9 while the others lasted till 30 days. Sucrose showed the highest degradation rate in the absence of botanical tissue. Although sucrose and whey showed highest stability towards light in the presence of botanical tissue, the mix of MD and CMC worked better without botanical tissue. Botanical tissue seems to play a complex role in protecting bixin as the results varied with different carbohydrates., Bixin is a coloring agent used in various industries especially food and textile industries. The color is achieved from annatto fruits found in tropical areas. It is known to produce colored products efficiently but there is an issue with its stability. The color gets bleached gradually until it gets completely lost. This is because it reacts with the oxygen in the surrounding air and gets changed in structure which will no longer have the same properties. A similar effect can be noticed when it is subjected to heat and light. Bixin can be protected from such changes by covering it with another material in a capsule form. In this case, carbohydrates were used to encapsulate bixin to provide better stability. In addition to this, bixin was found to have an additional carbohydrate (botanical tissue) cover that occurs naturally during extraction. The goal of this study was to examine how well the carbohydrates prevent degradation of color caused by light with and without the botanical tissue. Sucrose, whey and maltodextrin along with some additives were investigated as carbohydrates. The samples were placed under a lamp that provided light that was similar to daylight for 30 days. It was seen that without carbohydrates, bixin lost all of its color in 9 days, while the rest lasted for 30 days. This shows that carbohydrates do prevent color decay to a great extent. When analyzed, it was seen that the color degraded uniformly along the 30 days. Botanical tissue gave a nice color to the samples which stayed till the end of the study. Along with this, the amount of light absorbed by the samples was also calculated. Either bixin molecules or oxygen molecules were made reactive with the energy absorbed from light and this caused a reaction between them, leading to bleaching. Amongst all the samples with carbohydrates, bixin with sucrose had the highest loss of color. But this was overcome when botanical tissue was present. In terms of light stability, sucrose and whey showed

Bixin is a carotenoid which is extracted from annatto (Bixa orellana). Its main application is as a coloring agent. It consists of conjugating double bonds with a carboxylic end, making it prone to degradation by oxygen, light and heat. This study investigates the light stability of bixin encapsulated with maltodextrin (MD), sucrose and whey by spray drying. MD was added with additives such as carboxymethylcellulose (CMC), gum arabic (GA) and pectin to examine their interaction effects. Two sets of samples were used; one with pure bixin and carbohydrates, and the other, with non-filtered bixin encapsulated with the same set of carbohydrates and a carbohydrate naturally present in the seeds named botanical tissue. The samples were illuminated using a plasma sulfur lamp for a period of 30 days. UV-Visible spectrometer was used to keep track of the degradation. Statistic tests such as Shapiro-Wilk, ANOVA and t-test were used to confirm that the degradation was linear and that they follow first order decay. Quantum yields of photo-bleaching were calculated and compared between samples. Lower the quantum yield, higher the light stability. The presence of carbohydrates significantly improved the stability of bixin in comparison with non-encapsulated samples. The non-encapsulated samples lost their color completely by day 9 while the others lasted till 30 days. Sucrose showed the highest degradation rate in the absence of botanical tissue. Although sucrose and whey showed highest stability towards light in the presence of botanical tissue, the mix of MD and CMC worked better without botanical tissue. Botanical tissue seems to play a complex role in protecting bixin as the results varied with different carbohydrates., Bixin is a coloring agent used in various industries especially food and textile industries. The color is achieved from annatto fruits found in tropical areas. It is known to produce colored products efficiently but there is an issue with its stability. The color gets bleached gradually until it gets completely lost. This is because it reacts with the oxygen in the surrounding air and gets changed in structure which will no longer have the same properties. A similar effect can be noticed when it is subjected to heat and light. Bixin can be protected from such changes by covering it with another material in a capsule form. In this case, carbohydrates were used to encapsulate bixin to provide better stability. In addition to this, bixin was found to have an additional carbohydrate (botanical tissue) cover that occurs naturally during extraction. The goal of this study was to examine how well the carbohydrates prevent degradation of color caused by light with and without the botanical tissue. Sucrose, whey and maltodextrin along with some additives were investigated as carbohydrates. The samples were placed under a lamp that provided light that was similar to daylight for 30 days. It was seen that without carbohydrates, bixin lost all of its color in 9 days, while the rest lasted for 30 days. This shows that carbohydrates do prevent color decay to a great extent. When analyzed, it was seen that the color degraded uniformly along the 30 days. Botanical tissue gave a nice color to the samples which stayed till the end of the study. Along with this, the amount of light absorbed by the samples was also calculated. Either bixin molecules or oxygen molecules were made reactive with the energy absorbed from light and this caused a reaction between them, leading to bleaching. Amongst all the samples with carbohydrates, bixin with sucrose had the highest loss of color. But this was overcome when botanical tissue was present. In terms of light stability, sucrose and whey showed

Bixin obtained from Annatto seeds (Bixinella orella), has been used by the food industry as a source for natural color for a long time due to its low toxicity and high colorific value. Given its potential applications in dairy and meat industry to obtain a desired reddish - orange hue, a method was sought, which would quickly extract the desired pigment from the seed coat at low cost and allow for its direct application in the desired product. The bixin mass was extracted using demineralized water (D.M.W), sunflower oil or methanol and was then re-suspended in sunflower oil to a bixin strength of 10%. A study of the extraction process, viscosity and color hue (L*a*b value) of the 10% formulated product in margarine at low doses of 0.1% was carried out. Data indicated that methanol extraction provided the desirable reddish hue in margarine and yielded a product with lowest viscosity.

Bixin obtained from Annatto seeds (Bixinella orella), has been used by the food industry as a source for natural color for a long time due to its low toxicity and high colorific value. Given its potential applications in dairy and meat industry to obtain a desired reddish - orange hue, a method was sought, which would quickly extract the desired pigment from the seed coat at low cost and allow for its direct application in the desired product. The bixin mass was extracted using demineralized water (D.M.W), sunflower oil or methanol and was then re-suspended in sunflower oil to a bixin strength of 10%. A study of the extraction process, viscosity and color hue (L*a*b value) of the 10% formulated product in margarine at low doses of 0.1% was carried out. Data indicated that methanol extraction provided the desirable reddish hue in margarine and yielded a product with lowest viscosity.

Bixin obtained from Annatto seeds (Bixinella orella), has been used by the food industry as a source for natural color for a long time due to its low toxicity and high colorific value. Given its potential applications in dairy and meat industry to obtain a desired reddish - orange hue, a method was sought, which would quickly extract the desired pigment from the seed coat at low cost and allow for its direct application in the desired product. The bixin mass was extracted using demineralized water (D.M.W), sunflower oil or methanol and was then re-suspended in sunflower oil to a bixin strength of 10%. A study of the extraction process, viscosity and color hue (L*a*b value) of the 10% formulated product in margarine at low doses of 0.1% was carried out. Data indicated that methanol extraction provided the desirable reddish hue in margarine and yielded a product with lowest viscosity.

Annatto, a naturally obtained colour is a potential and valuable Non Wood Forest Product (NWFP). It is obtained from the seeds of plant species Bixa orellana L. The present experiment was carried out to find the efficacy of different oils used for annatto colour extraction. Experiment was started with the pretreatment of seeds of B. orellana with soaking in water at 0%, 50%, 100% and 150% (v/w) for 24 hrs at room temperature and colour extraction with castor oil at 100°C showed that maximum bixin (%) was found from the pretreated seeds with 50% (v/w) water soaking i.e., 0.46% bixin. Similarly for oil soluble annatto extraction method, the maximum extraction of bixin (0.99 %) was found with castor oil at 120°C for 1 hr and least bixin (0.03 %) was recorded with mustard oil at 60oC temperature for 1 hr. Sesame oil and ground nut oil were also showed better extractability. Out of different treatments, castor oil at 120°C for 1 hr gave better result with respect to bixin extraction as compared to others.

BACKGROUND: The liver is an important organ for its ability to transform xenobiotics, making the liver tissue a prime target for toxic substances. The carotenoid bixin present in annatto is an antioxidant that can protect cells and tissues against the deleterious effects of free radicals. In this study, we evaluated the protective effect of bixin on liver damage induced by carbon tetrachloride (CCl4) in rats. RESULTS: The animals were divided into four groups with six rats in each group. CCl4 (0.125 mL kg-1 body wt.) was injected intraperitoneally, and bixin (5.0 mg kg-1 body wt.) was given by gavage 7 days before the CCl4 injection. Bixin prevented the liver damage caused by CCl4, as noted by the significant decrease in serum aminotransferases release. Bixin protected the liver against the oxidizing effects of CCl4 by preventing a decrease in glutathione reductase activity and the levels of reduced glutathione and NADPH. The peroxidation of membrane lipids and histopathological damage of the liver was significantly prevented by bixin treatment. CONCLUSION: Therefore, we can conclude that the protective effect of bixin against hepatotoxicity induced by CCl4 is related to the antioxidant activity of the compound.