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Annatto, obtained from the seeds of achiote (Bixa orellana L.), is a widely used orange pigment rich in bixin and other apocarotenoids. This work reports the optimisation of a green extraction method of pigments and antioxidant compounds from achiote as well as its integration in a one-step green extraction-cosmetic formulation process. A biphasic solvent system of water and oil was used to recover simultaneously polar polyphenols, and less polar compounds, such as δ-tocotrienol and bixin. The optimisation of the ultrasound assisted extraction is presented, as well as a comparison of different vegetable oils used as extraction solvents. The composition, physicochemical properties and antioxidant activity of the oils were studied and their extraction performance was compared. Refined sunflower oil proved to be a better solvent than virgin olive, jojoba, coconut and grapeseed oils. Both aqueous and oil phases displayed an interesting antioxidant capacity. The oil phase contained 0.9% of bixin, as well as minor apocarotenoids and δ-tocotrienol. Twelve compounds, mainly phenolics, were identified by UHPLC-DAD-HRMS/MS in the aqueous phase. Twenty-one volatile compounds were identified in the volatile fraction by SPME-GC-MS. Lastly, a one-step green process is proposed to combine the extraction and the cosmetic formulation of the bioactive compounds. [ABSTRACT FROM AUTHOR]

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.

Annatto, obtained from the seeds of achiote (Bixa orellana L.), is a widely used orange pigment rich in bixin and other apocarotenoids. This work reports the optimisation of a green extraction method of pigments and antioxidant compounds from achiote as well as its integration in a one-step green extraction-cosmetic formulation process. A biphasic solvent system of water and oil was used to recover simultaneously polar polyphenols, and less polar compounds, such as δ-tocotrienol and bixin. The optimisation of the ultrasound assisted extraction is presented, as well as a comparison of different vegetable oils used as extraction solvents. The composition, physicochemical properties and antioxidant activity of the oils were studied and their extraction performance was compared. Refined sunflower oil proved to be a better solvent than virgin olive, jojoba, coconut and grapeseed oils. Both aqueous and oil phases displayed an interesting antioxidant capacity. The oil phase contained 0.9% of bixin, as well as minor apocarotenoids and δ-tocotrienol. Twelve compounds, mainly phenolics, were identified by UHPLC-DAD-HRMS/MS in the aqueous phase. Twenty-one volatile compounds were identified in the volatile fraction by SPME-GC-MS. Lastly, a one-step green process is proposed to combine the extraction and the cosmetic formulation of the bioactive compounds.

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Annatto seed oil (ASO) and cupuassu seed fat (CSF) were combined at the ratios: 30:70, 50:50, and 70:30 (% w/w). Their fatty acid profile, nutritional quality, FTIR (Fourier Transform Infrared) spectra, and rheological behavior were evaluated. ASO increased the content of polyunsaturated fatty acids in the blends; whereas CSF conferred higher contents of monounsaturated fatty acids. The blends exhibited low atherogenicity and thrombogenicity indices, suggesting nutritional advantages. The Newtonian fluid behavior and FTIR results suggested that mixing ASO and CSF at different proportions did not affect the functional groups. ASO showed an activation energy value which indicated that this fat viscosity was more sensitive to temperature changes. The Newtonian model proved to be suitable to describe the behavior of samples, according to statistical fit parameters R², χ², and RSS. The resulting blends presented improved physicochemical properties and nutritional attributes, indicating their feasibility for the development of new products. [ABSTRACT FROM AUTHOR]

Food products with desirable sensory qualities, safety, and nutrition are in high demand in industrialized countries. Annatto (Bixa orellana L.) is a small tree cultivated in tropical and subtropical America and is appreciated for the pigmented seeds that come from which the yellow-orange achiote or annatto natural food color is made. This study investigated the effect of the annatto seeds powder on the phytochemical and physicochemical properties as well as the textural and color of the value-added cheddar cheese. Two types of cheddar cheese were produced containing different concentrations of annatto seeds powder, 0.25 and 0.5% (ASP0.25, ASP0.5), and a control (ASP0). Based on the results, annatto seed powder appeared to show good antioxidant activity (79.432 ±1.015%). The addition of annatto seed powder positively influenced the textural and color characteristics of the value-added cheese. The obtained samples highlighted the satisfactory content in phytochemicals. The sensory analysis showed that the addition of annatto seed powder had no detrimental effects on the cheese's general acceptance, the improved color being appreciated. Therefore, annatto seed powder might be a good source of natural antioxidants for the production of dairy products being a natural alternative to synthetic food coloring ingredients. [ABSTRACT FROM AUTHOR]

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.

Copyright of Revista CIENCIA UNEMI is the property of Universidad Estatal de Milagro (UNEMI) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Introducción. 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 accesiones del Banco de Germoplasma de la Universidad EARTH.

The effects of the use of solvents (96% ethanol and PEG 400), solute ratios: solvent (1: 4 and 1: 6 w/w), extraction temperatures (50 and 60 °C) and leaching techniques (conventional and assisted by ultrasound) on the concentration and time of extraction of bixin from Achiote (Bixa Orellana L.) were evaluated. The extraction kinetics were constructed, and their orders were determined. The experimental values were adjusted through the mathematical models of Peleg and Azuara. ANOVA was used based on a 23 design, blocking type of technology. The kinetics showed two stages of extraction: rapid and slow. The reaction orders were decimal numbers, which indicates the complexity of the extraction mechanism. The highest orders of reaction (2.97 and 2.075) were presented with extraction with ultrasound-assisted and ethanol as solvent, at 50 and 60 °C and solute solvent ratio 1: 4 and 1: 6, respectively. The concentration of Bixin was increased by approximately 30% by increasing the temperature. The Azuara model presented the highest adjustment with a standard error < 0.1. The conventional leaching with PEG-400 required the lowest activation energy (9.2 kJ / mol) and with this treatment in 66 min the highest concentration of bixin (2.74 mgBix / gExt) was obtained. It was concluded that ultrasound-assisted leaching facilitates the extraction of bixin pigment, and the use of PEG 400 solvent is useful to obtain higher yields. [ABSTRACT FROM AUTHOR]

The aim of this study was to characterize and identify groups of achiote (Bixa orellana L.) with characteristics related to increased yield and bixin content. A total of 149 achiote accessions were collected from the departments of Loreto, San Martin, Junín, Pasco, Huánuco, Ucayali, and Cusco, in Perú. These were then evaluated using ten quantitative and three qualitative capsule and seed descriptors. Undesirable characteristics such as high spinosity and dehiscence predominated, while none of the quantitative descriptors correlated significantly with bixin content. Principal component analysis indicates that the quantitative descriptors (except for bixin content) are statistically significant, forming four clusters. Notably, one of the clusters included accessions characterized by heavy seeds, and another cluster included accessions with high number of seeds and bixin content.

Abstract Background Full chloroplast genomes provide high resolution taxonomic discrimination between closely related plant species and are quickly replacing single and multi-locus barcoding regions as reference materials of choice for DNA based taxonomic annotation of plants. Bixa orellana, commonly known as “achiote” and “annatto” is a plant used for both human and animal foods and was thus identified for full chloroplast sequencing for the Center for Veterinary Medicine (CVM) Complete Chloroplast Animal Feed database. This work was conducted in collaboration with the Instituto de Medicina Tradicional (IMET) in Iquitos, Peru. There is a wide range of color variation in pods of Bixa orellana for which genetic loci that distinguish phenotypes have not yet been identified. Here we apply whole chloroplast genome sequencing of “red” and “yellow” individuals of Bixa orellana to provide high quality reference genomes to support kmer database development for use identifying this plant from complex mixtures using shotgun data. Additionally, we describe chloroplast gene content, synteny and phylogeny, and identify an indel and snp that may be associated with seed pod color. Results Fully assembled chloroplast genomes were produced for both red and yellow Bixa orellana accessions (158,918 and 158,823 bp respectively). Synteny and gene content was identical to the only other previously reported full chloroplast genome of Bixa orellana (NC_041550). We observed a 17 base pair deletion at position 58,399–58,415 in both accessions, relative to NC_041550 and a 6 bp deletion at position 75,531–75,526 and a snp at position 86,493 in red Bixa orellana. Conclusions Our data provide high quality reference genomes of individuals of red and yellow Bixa orellana to support kmer based identity markers for use with shotgun sequencing approaches for rapid, precise identification of Bixa orellana from complex mixtures. Kmer based phylogeny of full chloroplast genomes supports monophylly of Bixaceae consistent with alignment based approaches. A potentially discriminatory indel and snp were identified that may be correlated with the red phenotype.

To compare the encapsulation of annatto extract by external gelation (EG) and internal gelation (IG) and to maximize process yield (% Y), two central composite designs were proposed. Calcium chloride (CaCl2) concentration (0.3–3.5%), alginate to gelling solution ratio (1:2–1:6); acetic acid (CH3COOH) concentration (0.2–5.0%) and alginate to gelling solution ratio (1:2–1:6) were taken as independent variables for EG and IG respectively. Release studies were conducted under different conditions; morphology, particle size, the encapsulation efficiency (EE), and release mechanism were evaluated under optimized conditions. The optimized EG conditions were 0.3% CaCl2 and 1:1.2 alginate to gelling solution ratio, whereas a 0.3% CH3COOH and 1:5 alginate to gelling solution ratio were optimized conditions for IG. When 20% extract was employed, the highest EE was achieved, and the largest release was obtained at a pH 6.5 buffer. The Peppas–Sahlin model presented the best fit to experimental data. Polyphenol release was driven by diffusion, whereas bixin showed anomalous release. These results are promising for application as modulated release agents in food matrices. [ABSTRACT FROM AUTHOR]

Background: Full chloroplast genomes provide high resolution taxonomic discrimination between closely related plant species and are quickly replacing single and multi-locus barcoding regions as reference materials of choice for DNA based taxonomic annotation of plants. Bixa orellana, commonly known as “achiote” and “annatto” is a plant used for both human and animal foods and was thus identified for full chloroplast sequencing for the Center for Veterinary Medicine (CVM) Complete Chloroplast Animal Feed database. This work was conducted in collaboration with the Instituto de Medicina Tradicional (IMET) in Iquitos, Peru. There is a wide range of color variation in pods of Bixa orellana for which genetic loci that distinguish phenotypes have not yet been identified. Here we apply whole chloroplast genome sequencing of “red” and “yellow” individuals of Bixa orellana to provide high quality reference genomes to support kmer database development for use identifying this plant from complex mixtures using shotgun data. Additionally, we describe chloroplast gene content, synteny and phylogeny, and identify an indel and snp that may be associated with seed pod color. Results: Fully assembled chloroplast genomes were produced for both red and yellow Bixa orellana accessions (158,918 and 158,823 bp respectively). Synteny and gene content was identical to the only other previously reported full chloroplast genome of Bixa orellana (NC_041550). We observed a 17 base pair deletion at position 58,399–58,415 in both accessions, relative to NC_041550 and a 6 bp deletion at position 75,531–75,526 and a snp at position 86,493 in red Bixa orellana. Conclusions: Our data provide high quality reference genomes of individuals of red and yellow Bixa orellana to support kmer based identity markers for use with shotgun sequencing approaches for rapid, precise identification of Bixa orellana from complex mixtures. Kmer based phylogeny of full chloroplast genomes supports monophylly of Bixaceae consistent with alignment based approaches. A potentially discriminatory indel and snp were identified that may be correlated with the red phenotype. [ABSTRACT FROM AUTHOR]

Background: Residual annatto seed meal (RASM) is a byproduct after extraction of bixin in the cosmetic industry and may be used in animal diets. Objective: Two experiments were carried out to determine the nutritive value and effect of RASM inclusion levels in the diets of Japanese quails. Methods: For the digestibility experiment, 160 Japanese quails were distributed in a randomized study with two treatments and 10 replicates with eight birds each. The treatments consisted of a reference diet and a test diet containing 75% of the reference diet and 25% of RASM. For the performance assay, 192 Japanese quails were used in a randomized study with six treatments and four replicates with eight birds each. Treatments consisted of a control corn-based diet (Contr1) and another diet based on sorghum + canthaxanthin (Contr2), and four diets based on sorghum with increasing inclusion levels of RASM (0, 3, 6, and 9%). Results: Digestibility of dry matter and crude protein, metabolization coefficient of gross energy, and retention of calcium and phosphorus were 41.05, 43.12, 47.97, 25.49, and 30.35%, respectively. RASM inclusion did not influence bird productivity; however, feed conversion worsened in treatments with 0-6% RASM inclusion relative to that in Contr1. Yolk color increased due to RASM treatment. Conclusion: RASM may be included in sorghum-based diets for quails at 9%, with improvement in yolk color and no negative effects on productivity and egg quality. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate and compare the effects of co-treatment with purified annatto oil (PAO) or its granules (GRA, Chronic®) with that of testosterone on the orchiectomy-induced osteoporosis in Wistar rats. After surgery, rats were treated from day 7 until day 45 with testosterone only (TES, 7 mg/kg, IM) or TES + PAO or GRA (200 mg/kg, p.o.). The following parameters were evaluated: food/water intake, weight, HDL, LDL, glucose, triglycerides (TG), total cholesterol (TC), alkaline phosphatase levels, blood phosphorus and calcium contents, femur weight, structure (through scanning electron microscopy), and calcium content (through atomic absorption spectrophotometry). Our results show that orchiectomy could significantly change the blood lipid profile and decrease bone integrity parameters. Testosterone reposition alone could improve some endpoints, including LDL, TC, bone weight, and bone calcium concentration. However, other parameters were not significantly improved. Co-treatment with PAO or GRA improved the blood lipid profile and bone integrity more significantly and improved some endpoints not affected by testosterone reposition alone (such as TG levels and trabeculae sizes). The results suggest that co-treatment with annatto products improved the blood lipid profile and the anti-osteoporosis effects of testosterone. Overall, GRA had better results than PAO.

This study evaluated the antimicrobial activity (i.e., against Bacillus cereus and Staphylococcus aureus) and the antioxidant activity (i.e., ABTS, FRAP, and DPPH) of annatto seeds extract obtained by ultrasound-assisted extraction. A response surface design with three levels such as pH (2-11), solvent concentration (50-96 %), seed-to-solvent ratio (1:2–1:10), and treatment time (0-30 min) was employed to determine the optimal experimental conditions. Thus, a pH of 7.0, seed-to-solvent ratio of 1:7, and treatment time of 20 min were selected as optimal rendering an extract having a 0.62% of bixin, 3.81 mg gallic acid/mg equivalent of polyphenol compounds (ABTS 1035.7, FRAP 424.7, and DPPH 1161.5 μM trolox/L), and a minimal inhibitory concentration against Bacillus cereus and Staphylococcus aureus of 32 and 16 mg/L, respectively. Further, the main bioactive compounds identified by LC/ESI-MS were bixin and catechin, chlorogenic acid, chrysin, butein, hypolaetin, licochalcone A, and xanthohumol. [ABSTRACT FROM AUTHOR]

Copyright of Revista Facultad de Ciencias Basicas is the property of Revista Facultad de Ciencias Basicas and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract This work presents physicochemical, morphological, thermal, and pasting properties of a novel native starch obtained from defatted and depigmented annatto seeds for applications in food-related areas. At the end of the annatto seed processing chain, a non-conventional white starch was obtained from defatted and totally depigmented seeds. After a comparison with a commercial corn starch, such novel product presented promising characteristics. The main attributes include a null content of pigment, an amylose content (24%) similar to corn starch, thin particles (D 43 of 0.8 μm) of spherical-like shape, a typical crystalline A-type pattern with well-defined peaks at 2θ of 14.5°, 17.3°, 17.8° and 22.9°, thermal stability, and low mass loss (<10%) when heated up to 130 °C. The evaluation of pasting properties indicated a product with peak viscosity larger than corn starch, which is an important parameter for giving texture and body. Furthermore, the pasting temperature was 7.9 °C lower, enabling its use in preparations of sensible foods. Graphical abstract Image 1 Highlights • Physicochemical properties of a novel native starch are presented. • A non-conventional white starch was obtained from defatted and totally depigmented seeds. • Thin particles of spherical-like shape were obtained. • Annatto seed and corn starches showed a typical crystalline A-type pattern. [ABSTRACT FROM AUTHOR]

Abstract Cutaneous melanoma has a high capacity to metastasize and significant resistance to conventional therapeutic protocols, which makes its treatment difficult. The combination of conventional drugs with cytostatic molecules of low toxicity has been shown to be an interesting alternative for sensitization of tumor cells to chemotherapy. In this study, we evaluated the effect of bixin, an abundant apocarotenoid present in Bixa orellana , on the sensitization of human melanoma cells (A2058) to dacarbazine treatment, an anticancer agent clinically used for the therapy of metastatic melanoma. UPLC-DAD-MS/MS analyses of bioactive extracts from B. orellana seeds led to the identification of two new apocarotenoids: 6,8′-diapocarotene-6,8′-dioic acid and 6,7′-diapocarotene-6,7′-dioic acid. After being identified as its major compound, bixin (Z -bixin) was evaluated on A2058 cells expressing the oncogenic BRAF VE600 mutation and resistant to dacarbazine treatment. Bixin promoted growth inhibition, reduced cell migration, induced apoptosis and cell cycle arrest in the G2/M phase. When associated with dacarbazine, bixin restored the sensitivity of A2058 cells to chemotherapy, enhancing its antiproliferative, anti-migratory and pro-apoptotic effects. Combined treatment also induced higher ROS (reactive oxygen species) and MDA (malondialdehyde, a lipid peroxidation marker) generation than monotreatment, suggesting that the oxidative stress caused by bixin contributes significantly to its sensitizing effect. Taken together, these data suggest that bixin exerts intrinsic antimelanoma activity by mechanisms complementary to those of dacarbazine, encouraging its use in combined therapy for cutaneous melanoma treatment. Graphical abstract Image 1 Highlights • Identification of two new apocarotenoids (6,8′-diapocarotene-6,8′-dioic acid and 6,7′-diapocarotene-6,7′-dioic acid). • First detection of naringenin and β-12′-apo-carotenoic acid in B. orellana. • Purification, characterization and pharmacological evaluation of bixin (Z- bixin) on human melanoma cells. • Sensitization of melanoma cells to dacarbazine treatment by bixin (combined therapy). • Potentialization of pro-apoptotic effect of dacarbazine by increasing oxidative stress. [ABSTRACT FROM AUTHOR]

Fixed drug eruption (FDE) is a localized type IV sensitivity reaction to a systemically introduced allergen. It usually occurs as a result of new medication, making identification and avoidance of the trigger medication straightforward; however, in a rare subset of cases no pharmacological source is identified. In such cases, the causative agent is often a food or food additive. In this report we describe a case of a FDE in a 12-year-old girl recently immigrated to the United States from Ecuador who had no medication exposure over the course of her illness. Through an exhaustive patient history and literature review, we were able to hypothesize that her presentation was caused by a dietary change of the natural achiote dye used in the preparation of yellow rice to a locally available commercial dye mix containing tartrazine, or Yellow 5, which has previously been implicated in both systemic hypersensitivity reactions and specifically in FDE. This report adds to the small body of available literature on non-pharmacological fixed hypersensitivity eruptions and illustrates an effective approach to the management of such a presentation when history is not immediately revealing.

Over the years, our team has dedicated significant efforts to studying a unique natural dye-producing species, annatto (Bixa orellana L.). We have amassed knowledge and established foundations that support the applications of gene expression analysis in comprehending in vitro morphogenic regeneration processes, phase transition aspects, and bixin biosynthesis. Additionally, we have conducted gene editing associated with these processes. The advancements in this field are expected to enhance breeding practices and contribute to the overall improvement of this significant woody species. Here, we present a step-by-step protocol based on somatic embryogenesis and an optimized transformation protocol utilizing Agrobacterium tumefaciens., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Vitamin E is composed of two groups of compounds: α‐, β‐, γ‐, and δ‐tocopherols (TPs), and the corresponding unsaturated tocotrienols (TTs). TTs are found in natural sources such as red palm oil, annatto seeds, and rice bran. In the last decades, TTs (specifically, γ‐TT and δ‐TT) have gained interest due to their health benefits in chronic diseases, based on their antioxidant, neuroprotective, cholesterol‐lowering, anti‐inflammatory activities. Several in vitro and in vivo studies pointed out that TTs also exert a significant antitumor activity in a wide range of cancer cells. Specifically, TTs were shown to exert antiproliferative/proapoptotic effects and to reduce the metastatic or angiogenic properties of different cancer cells; moreover, these compounds were reported to specifically target the subpopulation of cancer stem cells, known to be deeply involved in the development of resistance to standard therapies. Interestingly, recent studies pointed out that TTs exert a synergistic antitumor effect on cancer cells when given in combination with either standard antitumor agents (i.e., chemotherapeutics, statins, "targeted" therapies) or natural compounds with anticancer activity (i.e., sesamin, epigallocatechin gallate (EGCG), resveratrol, ferulic acid). Based on these observations, different TT synthetic derivatives and formulations were recently developed and demonstrated to improve TT water solubility and to reduce TT metabolism in cancer cells, thus increasing their biological activity. These promising results, together with the safety of TT administration in healthy subjects, suggest that these compounds might represent a new chemopreventive or anticancer treatment (i.e., in combination with standard therapies) strategy. Clinical trials aimed at confirming this antitumor activity of TTs are needed. Vitamin E TTs have gained interest due to their health benefits in chronic diseases. In particular, TTs were reported to exert significant antitumor activity in a wide range of cancer cells, by counteracting cell growth or proliferation, metastasis and angiogenesis. Recent studies pointed out that TTs exert a synergistic antitumor effect on cancer cells when given in combination with either standard antitumor agents or natural compounds with anticancer activity. These results, together with the safety of TT administration in healthy subjects, suggest that these compounds might represent a new chemopreventive or anticancer treatment strategy. Clinical trials aimed at confirming this antitumor activity of TTs are needed. [ABSTRACT FROM AUTHOR]

Purpose The purpose of this study is to investigate the possibility of dyeing polyester (PET) fabric with natural dye extracted from annatto seeds using high temperature dyeing method.Design/methodology/approach PET fabric was dyed with annatto extract by varying dyeing parameters (temperature, time, pH and dye concentration) to determine the optimum dyeing conditions. The influences of KAl(SO4)2, FeSO4, gallnut mordants or a commercial UV absorber on colour yield and fastness properties were further studied.Findings Optimum results were obtained when the fabric was dyed at 130°C for 30 min in a dyebath containing 15 per cent (owf) annatto dye at pH 6. The dyed fabric had an orange shade and exhibited good to excellent wash, crock, perspiration fastness and fair light fastness. Further dyeing with mordants or UV absorber mostly resulted in lower colour yield and similar fastness properties.Research limitations/implications Although the light fastness was slightly improved to moderate level for the sample with UV absorber, a noticeable colour staining on cotton portion of multi-fibre fabric occurred when subjected to standard washing test. Compared to C.I. Disperse Orange 73, the annatto dye exhibited comparable colour fastness but had inferior light fastness when dyed at approximately the same colour strength.Originality/value Natural colourants from annatto seeds can be used to dye PET fabric at high temperature without mordants, yielding deep orange shade and satisfactory fastness properties. This study provides a promising application to reduce the environmental impact of synthetic dyes. [ABSTRACT FROM AUTHOR]

We present a study on dye-sensitized solar cells (DSCs) and we fabricate dye-sensitized solar modules (DSCMs) based on natural apocarotenoids extracted from the achiote's seeds (annatto). Use of less polar solvent such as diethyl ether improves the bixin concentration in the annatto extract which, was employed as sensitizer in the devices. We measure IPCE max (∼33%) and estimate ϕ inj  ≥ 0.438 for annatto. By accurate and progressive optimization of both TiO 2 multilayer photoanode and of electrolyte composition an efficiency (η) around 1.6%, is achieved, with an improvement of about ∼742% compare to the best performance for annatto extract, so far reported. DSCM shows stability which overcomes 1000 h (shelf-life test), under 1 sun, and produces a battery capacity of ∼46.8 Ah, the equivalent to ∼15 type AAA standard battery, in a similar time period. Although annatto based DSCMs are still below the efficiency requirements for practical applications for large scale industry, our encouraging results, testify the potentiality of this pigment in the production of non-toxic, cheap, long term stable and environmentally friendly vegetable based solar devices, as alternatives to batteries for small electronic goods market. [ABSTRACT FROM AUTHOR]

Main conclusion: Genetic transformation allows for greater bixin or norbixin production in achiote. Knowledge of genes that control the biosynthesis of these important secondary metabolites will allow for targeted amplification in transgenic plants.Annatto is a natural dye or coloring agent derived from the seeds, or their arils, of achiote (Bixa orellana L.), and is commercially known as E160b. The main active component of annatto dye is water-insoluble bixin, although water-soluble norbixin also has commercial applications. Relative to other antioxidants, bixin is light- and temperature stable and is thus safe for human consumption. Bixin is, therefore, widely applied as a dye and as an antioxidant in the medico-pharmaceutical, food, cosmetic, and dye industries. Even though bixin has also been isolated from leaves and bark, yield is lower than from seeds. More biotechnology-based research of this industrial and medicinal plant is needed. Building on provisional genetic transformation studies, it would be advantageous to transform genes that could result in greater bixin or norbixin production. Reliable protocols for the extraction of bixin and norbixin, as well as deeper knowledge of the genes that control the biosynthesis of these important secondary metabolites will allow for targeted amplification in transgenic plants. [ABSTRACT FROM AUTHOR]

Environmental exposure to solar ultraviolet (UV) radiation causes acute photodamage, premature aging, and skin cancer, attributable to UV-induced genotoxic, oxidative, and inflammatory stress. The transcription factor NRF2 [nuclear factor erythroid 2 (E2)-related factor 2] is the master regulator of the cellular antioxidant response protecting skin against various environmental stressors including UV radiation and electrophilic pollutants. NRF2 in epidermal keratinocytes can be activated using natural chemopreventive compounds such as the apocarotenoid bixin, an FDA-approved food additive and cosmetic ingredient from the seeds of the achiote tree (Bixa orellana). Here, we tested the feasibility of topical use of bixin for NRF2-dependent skin photoprotection in two genetically modified mouse models [SKH1 and C57BL/6J (Nrf2+/+ versus Nrf2 -/-)]. First, we observed that a bixin formulation optimized for topical NRF2 activation suppresses acute UV-induced photodamage in Nrf2+/+ but not Nrf2-/-) SKH1 mice, a photoprotective effect indicated by reduced epidermal hyperproliferation and oxidative DNA damage. Secondly, it was demonstrated that topical bixin suppresses PUVA (psoralen C UVA)-induced hair graying in Nrf2+/+ but not Nrf2-/-) C57BL/6J mice. Collectively, this research provides the first in vivo evidence that topical application of bixin can protect against UV-induced photodamage and PUVA-induced loss of hair pigmentation through NRF2 activation. Topical NRF2 activation using bixin may represent a novel strategy for human skin photoprotection, potentially complementing conventional sunscreen-based approaches. [ABSTRACT FROM AUTHOR]

Bixa orellana is a small tree known for its red, oil-soluble pigment contained in the seed coat that is used as a natural dye and food coloring. In this study, we assembled and characterized the complete chloroplast genome of B. orellana as a resource for future genetic studies. With a total length of 159,825 bp, the chloroplast genome comprised of a large single-copy (LSC) region of 89,476 bp, a small single-copy (SSC) region of 19,617 bp, and two inverted repeat (IR) regions of 25,356 bp each. A total of 127 genes were predicted, consisting of 83 protein-coding genes, 36 tRNA genes, and 8 rRNA genes. Phylogenetic analysis confirmed the position of B. orellana within the order Malvales.

This work reports phase equilibrium experimental data (cloud points) for the system annatto seed oil in compressed carbon dioxide + ethanol as co-solvent. The oil was obtained by supercritical fluid extraction at 40 °C and 20 MPa using carbon dioxide as solvent. The synthetic static method was employed for the measurements, which were carried out in a temperature range from 25 to 60 °C and up to 19.32 MPa. The experiments were carried out using (CO 2 + ethanol) overall mass fraction compositions ranging from 0.82 to 0.99. Experimental results were modelled using artificial neural network (ANN). Results indicate a complex phase behavior for all temperatures investigated with the occurrence of vapor-liquid, liquid-liquid and vapor-liquid-liquid transitions. ANN modelling was an efficient alternative for the thermodynamic models to accurately analysis of the phase equilibrium at high pressure. [ABSTRACT FROM AUTHOR]

The transcription factor NRF2 (nuclear factor-E2-related factor 2) orchestrates major cellular defense mechanisms including phase-II detoxification, inflammatory signaling, DNA repair, and antioxidant response. Recent studies strongly suggest a protective role of NRF2-mediated gene expression in the suppression of cutaneous photodamage induced by solar UV (ultraviolet) radiation. The apocarotenoid bixin, a Food and Drug Administration (FDA)-approved natural food colorant (referred to as 'annatto') originates from the seeds of the achiote tree native to tropical America, consumed by humans since ancient times. Use of achiote preparations for skin protection against environmental insult and for enhanced wound healing has long been documented. We have recently reported that (i) bixin is a potent canonical activator of the NRF2-dependent cytoprotective response in human skin keratinocytes; that (ii) systemic administration of bixin activates NRF2 with protective effects against solar UV-induced skin damage; and that (iii) bixin-induced suppression of photodamage is observable in Nrf2+/+ but not in Nrf2-/- SKH-1 mice confirming the NRF2-dependence of bixin-induced antioxidant and anti-inflammatory effects. In addition, bixin displays molecular activities as sacrificial antioxidant, excited state quencher, PPAR (peroxisome proliferator-activated receptor) α/γ agonist, and TLR (Toll-like receptor) 4/NFκB (nuclear factor kappa-light-chain-enhancer of activated B cells) antagonist, all of which might be relevant to the enhancement of skin barrier function and environmental stress protection. Potential skin photoprotection and photochemoprevention benefits provided by topical application or dietary consumption of this ethno-pharmacologically validated phytochemical originating from the Americas deserves further preclinical and clinical examination. [ABSTRACT FROM AUTHOR]

The current study was undertaken to analyse the effect of short-term UV-B and UV-C radiations in provoking carotenoid biosynthesis in Bixa orellana . Seeds of B . orellana were germinated and exposed to the short term UV pre-treatment under controlled environmental condition for 5 days. The UV treated young seedlings response in pigment contents; antioxidant enzyme activity and mRNA gene expression level were analysed. The pigment content such as chlorophyll was increased in both UV-B and UV-C treated seedlings, but the total carotenoid level was decreased when compared to the control seedlings this can be attributed to the plant adaptability to survive in a stressed condition. The β-carotene level was increased in UV-B, and UV-C treated young seedlings. No significant changes have occurred in the secondary pigment such as bixin and ABA. The activity of the antioxidant enzymes such as catalase, peroxidase, and superoxide dismutase was significantly increased in UV-B treated seedlings when compared to the UV-C treated seedlings and control. The mRNA expression of the genes involved in bixin biosynthesis pathways such as DXS , PSY , PDS , LCY - β , LCY - ε , CMT , LCD , ADH and CCD genes showed different expression pattern in UV-B and UV-C treated young seedlings. Further we analysed the gene co-expression network to identify the genes which are mainly involved in carotenoid/bixin biosynthesis pathway. Form our findings the CCD , LCY , PDS , ZDS and PSY showed a close interaction. The result of our study shows that the short term UV-B and UV-C radiations induce pigment content, antioxidant enzyme activity and different gene expression pattern allowing the plant to survive in the oxidative stress condition. [ABSTRACT FROM AUTHOR]

This work involves the application of physical separation methods to concentrate the pigment of semi-defatted annatto seeds, a noble vegetal biomass rich in bixin pigments. Semi-defatted annatto seeds are the residue produced after the extraction of the lipid fraction from annatto seeds using supercritical fluid extraction (SFE). Semi-defatted annatto seeds are use in this work due to three important reasons: i) previous lipid extraction is necessary to recovery the tocotrienol-rich oil present in the annatto seeds, ii) an initial removal of the oil via SFE process favors bixin separation and iii) the cost of raw material is null. Physical methods including i) the mechanical fractionation method and ii) an integrated process of mechanical fractionation method and low-pressure solvent extraction (LPSE) were studied. The integrated process was proposed for processing two different semi-defatted annatto materials denoted Batches 1 and 2. The cost of manufacture (COM) was calculated for two different production scales (5 and 50 L) considering the integrated process vs. only the mechanical fractionation method. The integrated process showed a significantly higher COM than mechanical fractionation method. This work suggests that mechanical fractionation method is an adequate and low-cost process to obtain a rich-pigment product from semi-defatted annatto seeds. [ABSTRACT FROM AUTHOR]

Achiote (Bixa orellana L.) is a plant native to tropical America. The seeds of this plant are often used in the food industry, but the leaves are underutilized. In the southeastern region of Mexico, the annatto leaves are used in the traditional Mayan medicine. While the dosage and the quality remain unclear, and we studied the chemical profile and biological activity of annatto leaves from three Mayan accessions. The qualitative analysis showed low presence of saponins and abundant anthraquinones. The annatto leaf extracts of accessions 2 and 3 had a higher content of total phenolic compounds and antioxidant capacity compared to accession 1. Accession 3 showed antiinflammatory effects like indomethacin. In conclusion, the annatto leaves of accession 3 could be used to prepare phytomedicines due to its anti-inflammatory capacity. These results add medicinal and economic value to this crop, and indicate the importance of performing detailed pharmacological studies. [ABSTRACT FROM AUTHOR]

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Los colorantes han sido ampliamente utilizados desde la antigüedad tanto para teñir telas como para colorear alimentos. Debido a las reacciones adversas que se atribuyen a algunos colorantes sintéticos, los consumidores buscan en la actualidad productos que contengan sustancias de origen natural. Por este motivo, en esta investigación se extrajo y caracterizó por espectrofotometría visible e infrarroja el colorante del achiote (Bixa orellana) que se produce y comercializa en el Ecuador. La extracción del colorante se realizó, utilizando tres solventes: acetona, hidróxido de potasio acuoso al 2 % y cloroformo, para determinar con cuál de ellos se obtiene un mayor porcentaje de rendimiento. Adicionalmente, se compararon los espectros visible e infrarrojo obtenidos con cada solvente con los espectros obtenidos en la literatura. A los resultados de porcentaje de colorante obtenidos, se les realizó un análisis de varianza de dos factores, para determinar si existen diferencias estadísticamente significativas entre las cuatro muestras obtenidas y de igual manera entre los tres solventes utilizados. Los resultados muestran que el mejor solvente para la extracción del colorante es el hidróxido de potasio al 2 % con un promedio de extracción del 9,18 %. En los espectros visibles obtenidos se aprecian dos picos representativos, a longitudes de onda de 471 y a 504 nanómetro, mientras que al analizar los espectros infrarrojos se identifican todos los grupos funcionales propios de la bixina, por lo que se puede concluir que el colorante extraído contiene dicha molécula.

This paper presents novel outcomes about the effect of degree of inulin polymerization (DP) on the technological properties of annatto seed oil powder obtained by freeze-drying. Inulins with two DP's were evaluated: GR-inulin (DP ≥ 10) and HP-inulin (DP ≥ 23). Micrographs obtained by confocal microscopy were analyzed to confirm the encapsulation of bioactive compounds using both inulins, especially the encapsulation of the natural fluorescent substance δ-tocotrienol. Microparticles formed with both inulins presented the same capacity for geranylgeraniol retention (77%). Glass transitions of microparticles formed with GR-inulin and HP-inulin succeeded at 144 °C and 169 °C, respectively. Regarding water adsorption isotherms, microparticles formed with HP-inulin and GR-inulin presented behaviors of Types II (sigmoidal) and III (non-sigmoidal), respectively. Reduction of water adsorption capacity in the matrix at high relative moistures ( > 70%) was presented when HP-inulin was used. At low relative moistures ( < 30%), the opposite behavior was observed. [ABSTRACT FROM AUTHOR]

Bixa orellana (family Bixaceae) is a neotropical fast growing perennial tree of great agroindustrial value because its seeds have a high carotenoid content, mainly bixin. It has been used since pre-colonial times as a culinary colorant and spice, and for healing purposes. It is currently used as a natural pigment in the food, in pharmaceutical, and cosmetic industries, and it is commercially known as annatto. Recently, several studies have addressed the biological and medical properties of this natural pigment, both as potential source of new drugs or because its ingestion as a condiment or diet supplement may protect against several diseases. The most documented properties are anti-oxidative; but its anti-cancer, hypoglucemic, antibiotic and antiinflammatory properties are also being studied. Bixin's pathway elucidation and its regulation mechanisms are critical to improve the produce of this important carotenoid. Even though the bixin pathway has been established, the regulation of the genes involved in bixin production remains largely unknown. Our laboratory recently published B. orellana's transcriptome and we have identified most of its MEP (methyl-D-erythritol 4- phosphate) and carotenoid pathway genes. Annatto is a potential source of new drugs and can be a valuable nutraceutical supplement. However, its nutritional and healing properties require further study. [ABSTRACT FROM AUTHOR]

The objective of this study was to compare the performance of whey protein isolate (WPI) and modified starch (SF) based on their surface properties as adsorbents in the oil–water interface aiming the encapsulation of the annatto seed oil by ultrasound-assisted emulsification. We also analyzed the influence of WPI and SF on technological quality of the microparticles formed from the respective emulsions by freeze-drying (FD) and spray-drying (SD). The effects of ultrasonication power and processing time were assessed by the droplet size of the dispersed phase and creaming stability. The finer emulsions obtained with the evaluated biopolymers were subjected to the FD and SD processes. The obtained microparticles were characterized with respect to moisture content, size distribution and average diameter, morphology, surface oil, entrapment efficiency, encapsulation efficiency, X-ray diffraction patterns and thermal stability. Ultrasound-assisted emulsification of the oil led to the formation of fine kinetically stable emulsions. Intensification of the ultrasonication process had no positive effects on reducing droplet size, demonstrating the existence of a physico-chemical limit imposed by the characteristics of WPI and SF. The FD and SD techniques resulted in different microparticle sizes and morphologies; however, the microparticles formed using both techniques exhibited an amorphous state. Micrographs obtained by confocal laser scanning microscopy (CLSM) demonstrated encapsulation of the oil in all of the treatments; however, the WPI-FD microparticles showed greater entrapment efficiency, while the SF-SD microparticles showed greater encapsulation efficiency. The type of biopolymer and method used to form the microparticles had no effect on thermal stability because both microparticles showed high stability to thermal degradation. [ABSTRACT FROM AUTHOR]

Annatto color is one of the oldest of natural carotenoids that have wide applications in food technology. Annatto color is the second natural color additives used in world industry, of economic. It is predicted that demand for it is increasing day by day. Color extraction from annatto seed is done by conventional methods, mainly. Little information is available about the use of new techniques such as response surface methodology of ultrasound technology in the extraction of annatto color. The subject of this study was to optimize the ultrasound-assistant extraction process of annatto color and determination of the best process conditions by using response surface methodology and central square design. The central composite design was used in order to evaluate the effect of extraction temperature (80-20 °C), extraction time (10-2 min), seed to solvent ratio (5-20 %) and duty cycle (0.2-0.8 s), on efficiency of dye extraction from annatto seed and to optimize the extraction process. ANOVA shows the seed to solvent ratio variable (X4) and it square effect (X4 2) have the greatest impact on dye extraction from the annatto seeds. In temperature of 72.7 °C, extraction time of 7.25 min, the seed to solvent ratio of 14 percent and duty cycle of 0.8 s were found as the optimum conditions of the ultrasound-assistant extraction process in order to obtain the maximum efficiency of color under which the absorbance value and extraction efficiency was determined as 0.865 and 6.35 percent of annatto seed, respectively. [ABSTRACT FROM AUTHOR]

Recientes evidencias sugieren que el achiote puede constituirse en una alternativa terapéutica por sus efectos directos e indirectos contra el veneno de Bothrops asper, serpiente causante de la mayoría de mordeduras en nuestro país. Considerando que la producción metabólica ex vitro de esta planta no se obtiene de manera temprana y que eventualmente puede lograrse con ayuda de cultivos celulares, el trabajo evalúa algunas combinaciones inductoras de callos en segmentos de hojas, raíces y tallos de plantas de Bixa orellana L. con el fin de inducir y masificar tejidos precursores de suspensiones celulares potencialmente productores de los metabolitos activos. Estos explantes, dispuestos en medios nutritivos sin reguladores de crecimiento, o con suplementos de algunas combinaciones de ellos, se almacenan por ocho semanas a 25± 1º C y oscuridad, verificando los porcentajes de formación de callo en las semanas dos, cinco y ocho del cultivo. La generación de callo en explantes provenientes de raíces no resulta masiva, mientras que en tallos y hojas se favorece con el tratamiento empleado y con el tiempo de exposición a los reguladores de crecimiento. Sin importar el explante utilizado, la inducción de callo se aumenta con la exposición al tratamiento que involucra NAA (5 µM) y kinetina (4,64 µM), alcanzando valores de 17.0 % (raíces), 58.9 % (tallos) y 77.5 % (hojas)Recent evidences suggest annatto can be constituted in a therapeutic alternative by its direct and indirect influences against the poison of Bothrops asper, cause of most of snake bite in our country. The ex- vitro metabolic production of this plant is not obtained in early growth stages but it is possible to obtain it by means of cell cultures. This research evaluates some inductive calli combinations of plant segments of Bixa orellana L., like leaves, roots and stems grown in vitro, in order to get high induction rates and bigger amounts of tissue precursors of cellular suspensions, potentially producers of active metabolites. These explants, located in nutritious means without growth regulators, or with supplements of some combinations of them, are stored for eight weeks (25± 1º C and darkness) and the calli formation percentage tested in second, fifth and eighth week of growing. The calli generation in explants from roots is low, whereas in those from stems and leaves is favored with the treatment and with exposition time to the growth regulators. Regardless of the kind of explant used, the calli formation percentage is increased by the treatment that uses NAA (5 µM) and kinetine (4.64 µM), and showing values of 17,0 %, 58,9 % and 77,5 % for roots, stems and leaves respectively.

Para establecer la incidencia del momento de la colección y de algunos tratamientos de desinfección e imbibición sobre la germinación seminal in vitro de Bixa orellana L, se colectan intracapsularmente o postextrusión semillas maduras, con escasa variación en peso (25.57± 0.01 mg,). Una vez desinfectadas y en algunos casos expuestos a procesos adicionales de imbibición, las semillas se transfieren a cajas de cultivo en las que se mantienen en completa oscuridad a 25 ± 1°C, por espacio de siete semanas. La germinación resulta influenciada por el tratamiento de imbibición, por las semanas de cultivo empleadas y por el momento de la colección, pues las semillas extracapsulares germinan en muy bajo porcentaje mientras que en las colectadas previamente a la ruptura de la cápsula, la germinación se manifiesta desde la primera semana de cultivo y con incrementos estadísticamente significativos en la extrusión. Asimismo, y dependiendo del tratamiento de imbibición y desinfección que se emplee, se logran aumentar los porcentajes de germinación, hasta un 78%, evitando la escasa o caprichosa tasa de germinación evidenciada por algunos autores para esta promisoria especie vegetal.To establish the incidence of the time of collection and some disinfections and imbibition treatment on the seed germination in vitro of Bixa orellana L, mature seeds are collected via intracapsule or postextrusion, with a little weight variation (25.57 ± 0.01 mg,). Once the seeds are disinfected and in some case exposed to imbibition process, they are transfered to culture medium and are kept in complete darkness to 25 ± 1 ºC, for seven weeks. The germination is influenced by the imbibition treatment, the growing time and the collection moment. On the other hand, the extracapsular seeds have a lower germination percent than those collected prior to rupture since in this the germination manifests during the first week of growing with significant statistical increases in the extrusion. Likewise, depending on the imbibition and disinfections treatment that is used, the germination percents can increase up to 78%, avoiding the poor germination rate showed by some authors from this promising vegetable species.