From Extraction to Stabilization: Employing a 22 Experimental Design in Developing Nutraceutical-Grade Bixin from Bixa orellana L.

Bixin is the main carotenoid found in the outer portion of the seeds of Bixa orellana L., commercially known as annatto. This compound is industrially employed in pharmaceutical, cosmetic, and food formulations as a natural dye to replace chemical additives. This study aimed to extract bixin from annatto seeds and obtain encapsulated bixin in a powder form, using freeze-drying encapsulation and maltodextrin as encapsulating agent. Bixin was extracted from annatto seeds employing successive washing with organic solvents, specifically hexane and methanol (1:1 v/v), followed by ethyl acetate and dichloromethane for subsequent washes, to effectively remove impurities and enhance bixin purity, and subsequent purification by crystallization, reaching 1.5 ± 0.2% yield (or approximately 15 mg of bixin per gram of seeds). Bixin was analyzed spectrophotometrically in different organic solvents (ethanol, isopropyl alcohol, dimethylsulfoxide, chloroform, hexane), and the solvents chosen were chloroform (used to solubilize bixin during microencapsulation) and hexane (used for spectrophotometric determination of bixin). Bixin was encapsulated according to a 22 experimental design to investigate the influence of the concentration of maltodextrin (20 to 40%) and bixin-to-matrix ratio (1:20 to 1:40) on the encapsulation efficiency (EE%) and solubility of the encapsulated powder. Higher encapsulation efficiency was obtained at a maltodextrin concentration of 40% w/v and a bixin/maltodextrin ratio of 1:20, while higher solubility was observed at a maltodextrin concentration of 20% w/v for the same bixin/maltodextrin ratio. The encapsulation of this carotenoid by means of freeze-drying is thus recognized as an innovative and promising approach to improve its stability for further processing in pharmaceutical and food applications.

The role of bixin as antioxidant, anti-inflammatory, anticancer, and skin protecting natural product extracted from Bixa orellana L.

Since long, medicinal plants or herbs are being used in different traditional treatment systems as therapeutic agents to treat a variety of illnesses. Bixa orellana L., an medicinal plant (family: Bixaceae), is an Ayurvedic herb used to treat dyslipidemia, diarrhoea, and hepatitis since ancient times. B. orellana L., seeds contain an orange-red coloured component known as bixin (C25H30O4), which constitutes 80% of the extract.Chemically, bixin is a natural apocarotenoid, biosynthesized through the oxidative degradation of C40 carotenoids. Bixin helps to regulate the Nrf2/MyD88/TLR4 and TGF-1/PPAR-/Smad3 pathways, which further give it antifibrosis, antioxidant, and anti-inflammatory properties. This current review article presents a comprehensive review of bixin as an anti-inflammatory, antioxidant, anticancer,and skin protecting natural product. In addition, the biosynthesis and molecular target of bixin, along with bixin extraction techniques, are also presented.

Hybrid Lipid/Clay Carrier Systems Containing Annatto Oil for Topical Formulations.

Nanocomposites formed by clay and lipid carriers (NLCs) show a high potential for providing controlled release and specific delivery of bioactive molecules and have recently gained attention in the pharmaceutical sector due to their ability to transport hydrophilic and hydrophobic drugs. Recent studies have recognized the biological activity of the oil of Bixa orellana L. (AO) with regards to its healing, antioxidant, antibacterial, and anti-leishmanial properties. Therefore, the purpose of this study is the preparation and characterization of hybrid systems based on lipid nanocarriers and laponite for the delivery of AO. NLCs were prepared by the fusion-emulsification method, using cetyl palmitate (CP) or myristyl myristate (MM), AO, and Poloxamer 188. The morphology, hydrodynamic diameters, zeta potential (ZP), polydispersity index (PDI), thermal analysis, X-ray diffraction analysis (XRD), viscosity behavior, and cytotoxicity testing of the hybrid systems were performed. The thermal study and X-ray diffraction analyses (XRD) revealed polymorphic structural changes compatible with the amorphization of the material. Rheological assays highlighted a typical pseudoplastic behavior in all systems (MM and CP with LAP). The hybrid systems' morphology, size diameters, and PDIs were similar, preset spherical and monodisperse structures (≈200 nm; <0.3), without significant change up to sixty days. The ZP values differed from each other, becoming higher with increasing AO concentration. XEDS spectra and elemental X-ray maps show peaks of lipids (organic components, C and O) and inorganic components O, Mg, and Si. All samples showed cell viability above 60%. The results indicated a stable, biocompatible hybrid system that can be an alternative for topical application.

Identifying Bixa orellana L. New Carotenoid Cleavage Dioxygenases 1 and 4 Potentially Involved in Bixin Biosynthesis.

Carotene cleavage dioxygenases (CCDs) are a large family of Fe2+ dependent enzymes responsible for the production of a wide variety of apocarotenoids, such as bixin. Among the natural apocarotenoids, bixin is second in economic importance. It has a red-orange color and is produced mainly in the seeds of B. orellana. The biosynthesis of bixin aldehyde from the oxidative cleavage of lycopene at 5,6/5',6' bonds by a CCD is considered the first step of bixin biosynthesis. Eight BoCCD (BoCCD1-1, BoCCD1-3, BoCCD1-4, CCD4-1, BoCCD4-2, BoCCD4-3 and BoCCD4-4) genes potentially involved in the first step of B. orellana bixin biosynthesis have been identified. However, the cleavage activity upon lycopene to produce bixin aldehyde has only been demonstrated for BoCCD1-1 and BoCCD4-3. Using in vivo (Escherichia coli) and in vitro approaches, we determined that the other identified BoCCDs enzymes (BoCCD1-3, BoCCD1-4, BoCCD4-1, BoCCD4-2, and BoCCD4-4) also participate in the biosynthesis of bixin aldehyde from lycopene. The LC-ESI-QTOF-MS/MS analysis showed a peak corresponding to bixin aldehyde (m/z 349.1) in pACCRT-EIB E. coli cells that express the BoCCD1 and BoCCD4 proteins, which was confirmed by in vitro enzymatic assay. Interestingly, in the in vivo assay of BoCCD1-4, BoCCD4-1, BoCCD4-2, and BoCCD4-4, bixin aldehyde was oxidized to norbixin (m/z 380.2), the second product of the bixin biosynthesis pathway. In silico analysis also showed that BoCCD1 and BoCCD4 proteins encode functional dioxygenases that can use lycopene as substrate. The production of bixin aldehyde and norbixin was corroborated based on their ion fragmentation pattern, as well as by Fourier transform infrared (FTIR) spectroscopy. This work made it possible to clarify at the same time the first and second steps of the bixin biosynthesis pathway that had not been evaluated for a long time.

Bixa orellana L. carotenoids: antiproliferative activity on human lung cancer, breast cancer, and cervical cancer cells in vitro.

Emerging evidence on the potential pro-oxidant effect of carotenoids provokes apoptosis of cancer cells. Bixa orellana L. is native to Central and South America, interestingly, is also cultivated worldwide. Apo-carotenoids present in B. orellana L. are mainly dominated by bixin and norbixin and demonstrate fundamental antioxidant activity. Anti-proliferative activity on human cancer cells is rarely investigated. We isolated bixin from B. orellana L. found in the island of Java using Ultra-Fast Liquid Chromatography and confirmed the isolated compound using Liquid Chromatography-MS/MS. Bixin and crude extract were examined on human lung cancer (A549), cervical cancer (HeLa), and breast cancer (MCF-7). Anti-proliferative activity revealed to be promising on both, the isolated pigment and crude extract. Further investigation on the mechanism of action and effect on other cell lines, both in vitro and in vivo, are required before clinical translation.

Treating leishmaniasis in Amazonia, part 2: Multi-target evaluation of widely used plants to understand medicinal practices.

ETHNOPHARMACOLOGICAL RELEVANCE: Leishmaniasis are widely distributed among tropical and subtropical countries, and remains a crucial health issue in Amazonia. Indigenous groups across Amazonia have developed abundant knowledge about medicinal plants related to this pathology. AIM OF THE STUDY: We intent to explore the weight of different pharmacological activities driving taxa selection for medicinal use in Amazonian communities. Our hypothesis is that specific activity against Leishmania parasites is only one factor along other (anti-inflammatory, wound healing, immunomodulating, antimicrobial) activities. MATERIALS AND METHODS: The twelve most widespread plant species used against leishmaniasis in Amazonia, according to their cultural and biogeographical importance determined through a wide bibliographical survey (475 use reports), were selected for this study. Plant extracts were prepared to mimic their traditional preparations. Antiparasitic activity was evaluated against promastigotes of reference and clinical New-World strains of Leishmania (L. guyanensis, L. braziliensis and L. amazonensis) and L. amazonensis intracellular amastigotes. We concurrently assessed the extracts immunomodulatory properties on PHA-stimulated human PBMCs and RAW264.7 cells, and on L. guyanensis antigens-stimulated PBMCs obtained from Leishmania-infected patients, as well as antifungal activity and wound healing properties (human keratinocyte migration assay) of the selected extracts. The cytotoxicity of the extracts against various cell lines (HFF1, THP-1, HepG2, PBMCs, RAW264.7 and HaCaT cells) was also considered. The biological activity pattern of the extracts was represented through PCA analysis, and a correlation matrix was calculated. RESULTS: Spondias mombin L. bark and Anacardium occidentale L. stem and leaves extracts displayed high anti-promatigotes activity, with IC50 ≤ 32 µg/mL against L. guyanensis promastigotes for S. mombin and IC50 of 67 and 47 µg/mL against L. braziliensis and L. guyanensis promastigotes, respectively, for A. occidentale. In addition to the antiparasitic effect, antifungal activity measured against C. albicans and T. rubrum (MIC in the 16-64 µg/mL range) was observed. However, in the case of Leishmania amastigotes, the most active species were Bixa orellana L. (seeds), Chelonantus alatus (Aubl.) Pulle (leaves), Jacaranda copaia (Aubl.) D. Don. (leaves) and Plantago major L. (leaves) with IC50 < 20 µg/mL and infection rates of 14-25% compared to the control. Concerning immunomodulatory activity, P. major and B. orellana were highlighted as the most potent species for the wider range of cytokines in all tested conditions despite overall contrasting results depending on the model. Most of the species led to moderate to low cytotoxic extracts except for C. alatus, which exhibited strong cytotoxic activity in almost all models. None of the tested extracts displayed wound healing properties. CONCLUSIONS: We highlighted pharmacologically active extracts either on the parasite or on associated pathophysiological aspects, thus supporting the hypothesis that antiparasitic activities are not the only biological factor useful for antileishmanial evaluation. This result should however be supplemented by in vivo studies, and attracts once again the attention on the importance of the choice of biological models for an ethnophamacologically consistent study. Moreover, plant cultural importance, ecological status and availability were discussed in relation with biological results, thus contributing to link ethnobotany, medical anthropology and biology.

Annatto Oil Loaded Nanostructured Lipid Carriers: A Potential New Treatment for Cutaneous Leishmaniasis.

Annatto (Bixa orellana L.) is extensively used as food pigment worldwide. Recently, several studies have found it to have healing and antioxidant properties, as well as effective action against leishmaniasis. Therefore, the purpose of this study was to incorporate the oil obtained from annatto seeds into a nanostructured lipid carrier (NLC) and evaluate its physicochemical properties and biological activity against Leishmania major. Nanoparticles were prepared by the fusion-emulsification and ultrasonication method, with the components Synperonic™ PE (PL) as the surfactant, cetyl palmitate (CP) or myristyl myristate (MM) as solid lipids, annatto oil (AO) (2% and 4%, w/w) as liquid lipid and active ingredient, and ultra-pure water. Physicochemical and biological characterizations were carried out to describe the NLCs, including particle size, polydispersity index (PDI), and zeta potential (ZP) by dynamic light scattering (DLS), encapsulation efficiency (EE%), thermal behavior, X-ray diffraction (XRD), transmission electron microscopy (TEM), Electron Paramagnetic Resonance (EPR), cytotoxicity on BALB/c 3T3 fibroblasts and immortalized human keratinocyte cells, and anti-leishmaniasis activity in vitro. Nanoparticles presented an average diameter of ~200 nm (confirmed by TEM results), a PDI of less than 0.30, ZP between -12.6 and -31.2 mV, and more than 50% of AO encapsulated in NLCs. Thermal analyses demonstrated that the systems were stable at high temperatures with a decrease in crystalline structure due to the presence of AOs (confirmed by XRD). In vitro, the anti-leishmania test displayed good activity in encapsulating AO against L. major. The results indicate that the oily fraction of Bixa orellana L. in NLC systems should be evaluated as a potential therapeutic agent against leishmaniasis.

Bixa orellana L. by-products' fractions from an industrial process: antiproliferative activity on tumor cells and chemical profile.

This study evaluated the phytochemical characterization of Bixa orellana (BO extract) unsaponifiable extract and resulting fractions (F fraction - FF, Geranyl fraction - GF and R fraction- RF) obtained as by-products of an industrial process investigating in vitro antiproliferative activities in human tumoral cells. The main compounds identified by GC-MS for BO extract were Geranylgeraniol (61.51%); for FF: Geranylgeraniol (70.23%); for GF: Geranylgeraniol (78.92%) and for RF: ß-cubebene (27.75%). Quantifications of geranylgeraniol by GC-FID presented the percentage content: BO 27.52%; FF 38.52%; GF 51.44% and RF 1.81%. BO extract showed a significant antiproliferative activity, with GI50 up to 4 µg/mL. All fractions had a remarkably similar antiproliferative activity profile (GI50 27-47 µg/mL). Data reported herein showed an important cytostatic effect for BO extract, nevertheless this activity is not attributed exclusively to geranylgeraniol. In conclusion, this by-product becomes of great value, being a potential candidate for development of new anti-tumor ingredients.

Thiodiketopiperazines with two spirocyclic centers extracted from Botryosphaeria mamane, an endophytic fungus isolated from Bixa orellana L.

Three thiodiketopiperazines, botryosulfuranols A-C (1-3) were isolated from the endophytic fungus Botryosphaeria mamane. The three compounds present sulfur atoms on α- and ß-positions of phenylalanine derived residues and unprecedented two spirocyclic centers at C-4 and C-2'. Their planar structures were determined by spectroscopic analysis and absolute configurations were achieved by X-ray diffraction analysis and ECD and NMR chemical shifts calculations. Botryosulfuranol A (1) was the most cytotoxic compound against four cancer cell lines (HT-29, HepG2, Caco-2, HeLa) and two healthy cell lines (IEC6, Vero) highlighting the importance of an electrophilic center for cell growth inhibition.

Obtaining bixin from semi-defatted annatto seeds by a mechanical method and solvent extraction: Process integration and economic evaluation.

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 50L) 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.

Extracción de compuestos fenólicos y actividad antioxidante de hojas de Bixa orellana L. (achiote) / Extraction of phenolic compounds and antioxidant activity from leaves of Bixa orellana L. (achiote)

Introducción: Bixa orellana L. también conocida como achiote, es una planta con un rico depósito de compuestos bioactivos, tales como, saponinas, alcaloides y flavonoides, a los cuales se les atribuyen propiedades fisiológicas entre las que se encuentran: actividad antimicrobiana, antiinflamatoria, antialérgica, antiviral, anticancerígena, antioxidante, entre otras. Objetivos: evaluar el efecto del tiempo de extracción y la relación solvente/ sobre el contenido de fenoles totales, así como el efecto del contenido de sólidos y el pH de la solución, sobre la actividad antioxidante del extracto de hojas de B. orellana. Métodos: el contenido total de fenoles fue evaluado por el método de Folin-Ciocalteu. La actividad antioxidante se determinó por los métodos espectrofotométricos de reacción con el radical 2,2´-azino-bis (3-etilbenzotiazolin-6- ácido sulfónico) y la medida de la capacidad reductora sobre el Fe+3, los resultados se expresaron como micromoles de equivalentes Trolox por gramo de extracto (µmol ET.g-1). Resultados: las condiciones del proceso que más favorecen la extracción de compuestos fenólicos desde las hojas de Bixa orellana L. son: tiempo de extracción de 60 h y relación solvente/ hojas (v/p) de 4/1. El contenido máximo de fenoles totales fue de 144,77 ± 9,66 mgAT.g-1, que al someterlo a una solución de pH de 8 y 11,7 °Brix, presenta una actividad antioxidante de 4406,83 ± 43,30 µmol ET.g-1 por el método de reacción con el radical 2,2´-azino-bis (3-etilbenzotiazolin-6- ácido sulfónico) y 4547,22 ± 53,19 µmolET.g-1 por el método de medida de la capacidad reductora sobre el Fe+3. Conclusiones: se demostró que la cantidad de fenoles totales extraídos de hojas de B. orellana dependen de la relación solvente/material vegetal y del tiempo de extracción; asimismo, se encontró que el pH tiene efecto sobre la actividad antioxidante determinada por la reacción con el radical 2,2´-azino-bis (3-etilbenzotiazolin-6- ácido sulfónico) y la medida de la capacidad reductora sobre el Fe+3(AU)

Introduction: Bixa orellana L. also known as annatto, is a plant with a high content of bioactive compounds, such as saponins, alkaloids and flavonoids that have physiological properties as antimicrobial, anti-inflammatory, anti-allergenic, antiviral, anticarcinogenic and antioxidant properties, among others. Objectives: To evaluate the effect of extraction time and the solvent/ leaf ratio over the total phenolic content and the effect of solids and pH of the solution on the antioxidant activity in extracts from Bixa orellana leaves. Methods : The ethanol extract of leaves of Bixa orellana was obtained by percolation and filtering. Total phenol content was evaluated by the Folin-Ciocalteu. The antioxidant activity was determined by spectrophotometric methods as reaction with 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) and ferric reducing antioxidant power, the results were expressed as micromoles of Trolox equivalents per gram of extract (µmolTE.g -1). Results: Process conditions that improve the extraction of phenolic compounds from Bixa. orellana extract are: extraction time of 60 h and solvent/ leaves ratio (v/w) of 4/1. The maximum total phenol content was 144.77 ± 9.66mgAT.g-1, which when subject to a solution of pH 8 and 11.7 °Brix, has an antioxidant activity of 4406.83 ± 43.30 µmolTE.g-1 by the 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) reaction and 4547.22 ± 53.19 µmolET.g-1 by ferric reducing antioxidant power. Conclusion: It showed that the amount of TF extracted from leaves of Bixa orellana depend on the solvent/ leaves ratio and the extraction time. In addition, it found that the pH has an effect on the antioxidant activity determined by the methods 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) reaction and ferric reducing antioxidant power(AU)

Economic Analysis of an Integrated Annatto Seeds-Sugarcane Biorefinery Using Supercritical CO2 Extraction as a First Step.

Recently, supercritical fluid extraction (SFE) has been indicated to be utilized as part of a biorefinery, rather than as a stand-alone technology, since besides extracting added value compounds selectively it has been shown to have a positive effect on the downstream processing of biomass. To this extent, this work evaluates economically the encouraging experimental results regarding the use of SFE during annatto seeds valorization. Additionally, other features were discussed such as the benefits of enhancing the bioactive compounds concentration through physical processes and of integrating the proposed annatto seeds biorefinery to a hypothetical sugarcane biorefinery, which produces its essential inputs, e.g., CO2, ethanol, heat and electricity. For this, first, different configurations were modeled and simulated using the commercial simulator Aspen Plus® to determine the mass and energy balances. Next, each configuration was economically assessed using MATLAB. SFE proved to be decisive to the economic feasibility of the proposed annatto seeds-sugarcane biorefinery concept. SFE pretreatment associated with sequential fine particles separation process enabled higher bixin-rich extract production using low-pressure solvent extraction method employing ethanol, meanwhile tocotrienols-rich extract is obtained as a first product. Nevertheless, the economic evaluation showed that increasing tocotrienols-rich extract production has a more pronounced positive impact on the economic viability of the concept.

Antibacterial activity of Bixa orellana L. (achiote) against Streptococcus mutans and Streptococcus sanguinis

Objective: To evaluate the cytotoxic and antibacterial effect of Bixa orellana L. (B. orellana) (achiote) methanol extract against Streptococcus mutans (ATCC 25175) (S. mutans) and Streptococcus sanguinis (ATCC 10556) (S. sanguinis). Methods: Two methanol extracts of B. orellana were prepared in vitro, from the seeds and leaves. The antibacterial activity of extracts against S. mutans and S. sanguinis was evaluated using the cup-plate agar diffusion method. The minimum inhibitory concentration (MIC) was determined using the microdilution method and the cytotoxic activity was determinated by using the cell line MDCK. Results: A stronger antibacterial effect was observed with the leaves methanolic extract with an inhibition zone of (19.97 ± 1.31) mm against S. mutans and (19.97 ± 1.26) mm against S. sanguinis. The methanolic extract of the seeds had an activity of (15.11 ± 1.03) mm and (16.15 ± 2.15) mm against S. mutans and S. sanguinis, respectively. The MIC of the leaf and the seed extracts against S. sanguinis was 62.5 and 125 μg/mL, respectively, and the MIC of the leaf extract against S. mutans was 62.5 μg/mL, and for the seed extract it was 31.25 μg/mL. The 50% cytotoxic concentration was 366.45 and 325.05 μg/mL for the leaves and seeds extracts, respectively. Conclusions: The experimental findings demonstrated the antibacterial effect of the methanolic extract of B. orellana (achiote) on S. mutans and S. sanguinis. The extract of this plant is cytotoxic at high concentrations.

Influence of the degree of inulin polymerization on the ultrasound-assisted encapsulation of annatto seed oil.

The effect of the degree of polymerization (DP) of inulin was evaluated on its encapsulant characteristics. We assessed the influence of the average inulin DP (DP ≥10 and DP ≥23) in the ultrasound-assisted encapsulation of annatto seed oil using the freeze-drying technique for particle formation. The intensification of the homogenization process with ultrasound did not improve the characteristics of the emulsions due to the physicochemical limitations of the inulin molecular chain (molecules do not exhibit surface activity). The particle morphology, oil entrapment efficiency, encapsulation efficiency, X-ray diffraction, thermogravimetric analysis and Rancimat analyses proved the effectiveness of inulin as a wall material. The properties influenced by the DP were the surface oil, encapsulation efficiency, water activity, particle size and oxidative stability of the encapsulated oil because the highest DP promoted the formation of microparticles with lower surface oil content, greater encapsulation efficiency, low water activity, larger size and greater protection against oil oxidation.

Análise preliminar da citotoxicidade dos aditivos alimentares urucum e cúrcuma / Cytotoxicity of the food additive annatto and turmeric

Este trabalho teve por objetivo avaliar a aþÒo dos aditivos alimentares urucum e cúrcuma em células meristemáticas de pontas de raízes de Allium cepa nas doses de 5 e 10 g, nos tempos de exposiþÒo de 24 e 48 horas. Utilizou-se para cada dose um grupo de cinco bulbos de cebolas, que primeiramente foram enraizados em água destilada, e em seguida transferidos para as suas respectivas doses. Analisaram-se células em todo ciclo celular, totalizando 5.000 para cada controle e tempo de exposiþÒo. Os dados obtidos foram submetidos O análise estatística do Qui-quadrado (p<0,05). A partir dos resultados observou-se que as doses do urucum e do cúrcuma tiveram efeito antiproliferativo significativo sobre o ciclo celular deste sistema-teste. Também foi verificado número significativo de aberraþ§es celulares nos dois tempos de exposiþÒo avaliados de todas as doses estudadas. Portanto, nas condiþ§es analisadas, o urucum e o cúrcuma mostraram-se citotóxicos e genotóxicos.(AU)

This study aimed to evaluate the effect of food additives annatto and turmeric tip cells of Allium cepa roots in doses of 5 or 10 g in exposure times of 24 and 48 hours. A group of five onion bulbs was used for each dose. Each dose was first embedded in distilled water and then transferred to their respective doses. Cells were analyzed throughout the cell cycle, totaling 5000 for each control and exposure time. The obtained data were subjected to statistical analysis Chi-square (p <.05). From the results it was observed that the doses of turmeric and annatto had significant antiproliferative effect on the cell cycle of this test system. They also found a significant number of cellular aberrations in the two exposure times evaluated all doses studied. Therefore, under the conditions studied, annatto and turmeric proved cytotoxic and genotoxic.(AU)

Análise preliminar da citotoxicidade dos aditivos alimentares urucum e cúrcuma / Cytotoxicity of the food additive annatto and turmeric

Este trabalho teve por objetivo avaliar a ação dos aditivos alimentares urucum e cúrcuma em células meristemáticas de pontas de raízes de Allium cepa nas doses de 5 e 10 g, nos tempos de exposição de 24 e 48 horas. Utilizou-se para cada dose um grupo de cinco bulbos de cebolas, que primeiramente foram enraizados em água destilada, e em seguida transferidos para as suas respectivas doses. Analisaram-se células em todo ciclo celular, totalizando 5.000 para cada controle e tempo de exposição. Os dados obtidos foram submetidos à análise estatística do Qui-quadrado (p<0,05). A partir dos resultados observou-se que as doses do urucum e do cúrcuma tiveram efeito antiproliferativo significativo sobre o ciclo celular deste sistema-teste. Também foi verificado número significativo de aberrações celulares nos dois tempos de exposição avaliados de todas as doses estudadas. Portanto, nas condições analisadas, o urucum e o cúrcuma mostraram-se citotóxicos e genotóxicos.

This study aimed to evaluate the effect of food additives annatto and turmeric tip cells of Allium cepa roots in doses of 5 or 10 g in exposure times of 24 and 48 hours. A group of five onion bulbs was used for each dose. Each dose was first embedded in distilled water and then transferred to their respective doses. Cells were analyzed throughout the cell cycle, totaling 5000 for each control and exposure time. The obtained data were subjected to statistical analysis Chi-square (p <.05). From the results it was observed that the doses of turmeric and annatto had significant antiproliferative effect on the cell cycle of this test system. They also found a significant number of cellular aberrations in the two exposure times evaluated all doses studied. Therefore, under the conditions studied, annatto and turmeric proved cytotoxic and genotoxic.

Secagem solar de grãos residuais e farinha de grãos residuais de urucum / Solar drying of annatto grains and waste grains flour of annatto

O subproduto constituído pelos grãos residuais de urucum é resultante do processo de extração da bixina, principalmente na indústria alimentícia, utilizando-se como meio extrator óleo vegetal. Atualmente, a maioria desses grãos vem sendo descartada, no entanto vê-se a possibilidade de incorporá-los na dieta humana, uma vez que, como todos os grãos, são ricos em carboidratos, proteínas e fibras alimentares. Neste trabalho objetivou-se secar, em um secador solar, grãos residuais e farinha de grãos residuais de urucum, com e sem a camada de óleo proveniente do processo de extração industrial da bixina, ajustar diferentes modelos matemáticos as curvas de secagem e determinar a difusividade efetiva das amostras. Durante as secagens solares foi verificado, no interior do secador, temperaturas médias superiores a 50 °C. Observou-se que as amostras de grãos residuais de urucum com e sem a camada de óleo demoram um maior intervalo de tempo para atingirem o teor de água de aproximadamente 5% b.u., enquanto que as amostras de farinhas de grãos residuais de urucum com e sem óleo tiveram as maiores taxas de secagens. Todos os modelos matemáticos avaliados representaram de forma satisfatória a cinética de secagem das amostras testadas, com valores de R2 superiores a 0,96 e valores de DQM inferiores a 1,0, sendo que o modelo de Midilli foi o que apresentou os melhores parâmetros de ajustes. A difusividade efetiva apresentou-se na ordem de 10-11 m2 s-1, para todas as amostras estudadas, sendo que o maior valor foi encontrado para a amostra de grão sem óleo (6,47 × 10-11 m2 s-1) e o menor para a farinha sem óleo (0,69 × 10-11 m2 s-1).

The byproduct consisting of annatto grains is waste resulting from the extraction of bixin, especially in the food industry, using as extractant vegetable oil. Currently, most of these grains has been ruled out, however sees the possibility of incorporating them in the human diet, since, like all grains are rich in carbohydrates, protein and dietary fiber. The objective of this study was to dry annatto grains and waste grains flour of annatto, with and without the oil layer from the industrial process of extraction of bixin, in a solar dryer and fit different mathematical models of the drying and determine the diffusivity effective samples. During the solar drying was observed inside the dryer, average temperatures exceeding 50 °C. It was observed that samples of grain annatto with and without oil layer takes a longer time to achieve moisture content of approximately 5% wb, whereas the samples of waste grains flour of annatto with and without oil had the highest rates of drying. All mathematical models evaluated represent satisfactorily the drying kinetics of the samples tested, with R2 values greater than 0.96 and DQM values below 1.0, and the Midilli model presented the best parameters adjustments. The effective diffusivity presented in the order of 10-11 m2 s-1 for all samples, while the highest value was found for the sample of grain without oil (6.47 × 10-11 m2 s-1) and lowest for flour without oil (0.69 × 10-11 m2 s-1).

Bacterial growth inhibition potential of annatto plant parts

Objective: To study the antibacterial efficacy of Bixa orellana leaves and deseeded fruit capsule extracts against both Gram positive and Gram negative bacteria. Methods: The antibacterial activity of the ethanolic, methanolic, acetone and dimethyl sulphoxide extracts of B. orellana were tested against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus subtilis,Bacillus cereus and Staphylococus aureus by disc diffusion method. Results: The antibacterial activity of leaf was more pronounced even at low concentrations and fruit extracts exhibited the same at relatively higher concentrations. Only DMSO extract of seeds showed growth inhibition of S. aureus, B. subtilis, B. cereus, and P. aeruginosa. Conclusions: The present study suggested that the leaves and deseeded capsule extracts of B. orellana possess significant antibacterial activity thereby providing substantial support for the ethanobotanical applications of this plant.

La Bixa orellana L en el tratamiento de afecciones estomatológicas, un tema aún por estudiar / The Bixa orellana L in treatment of stomatology affections: a subject that hasn't studied yet

De las semillas de la planta tropical Bixa orellana L (bija) se extrae un pigmento (el annatto) que està compuesto bßsicamente por carotenoides (bixina/nor-bixina). Por su resistencia a la acción de los agentes químicos, pero no a los efectos del sol, se emplea con preferencia para colorear alimentos y bebidas. La dosis diaria permitida recomendada por la OMS es de 0-0,065 mg/kg de peso corporal. El annato es un colorante con muy baja toxicidad para el humano, reportàndose escasos incidentes de eventos alérgicos. A los extractos de bija le han sido demostrados diversas propiedades farmacológicas, entre las cuales se encuentran: antitumoral, antiinflamatoria, astringente, emoliente, antiséptica, antibacteriana, antioxidante, cicatrizante entre otras y se ha indicado en el tratamiento de infecciones bacterianas de forma general, estomatitis, y en la curación de heridas y quemaduras. Teniendo en cuenta que en el arsenal terapéutico al alcance del estomatólogo son escasos los productos de acción local cicatrizante, antioxidante, antiinflamatoria y antimicrobiana y mucho menos un producto que reúna todas estas propiedades juntas, es que se procedió a realizar una actualización bibliogràfica del tema Bixa orellana L, referido a su composición química, toxicología y efectos farmacológicos de interés para el tratamiento potencial de desórdenes en el campo de la estomatología. Existen evidencias de la utilidad de estos extractos para el tratamiento de desórdenes estomatológicos, pero estas deben aún ser validadas desde el punto de vista clínico con ensayos realizados de acuerdo con las buenas pràcticas clínicas, antes de cualquier aplicación en los servicios estomatológicos cubanos.

From the Bixa orellana L (bija) tropical plant seeds a pigment (annatto) was extracted which is basically composed by carotenoids (bixine/nor-bixine). Due to its resistance to chemical agent's action but not to sun effects, it is used preferably to food and drinks coloring. Daily dose allowance recommended by HWO is of 0-0,065 mg/kg/bw. Annate is a coloring agent with a very low toxicity for humans, even when some low rates of allergic events have been reported. Annate extracts have different pharmacological properties including the antitumor, anti-inflammatory, astringent, emollient, antiseptic, antibacterial, antioxidant, cicatrizant among others and has been prescribed in wounds and burns cure. Considering that in the therapeutical array available for stomatologist the cicatrizant antioxidant, anti-inflammatory and antimicrobial local actions are limited and by no means a product fulfilling all these properties together, we made a bibliographic updating of Bixa orellana L subject related to its chemical, toxicology composition and interesting pharmacological effects for potential treatment of stomatology disorders. There are evidences on usefulness of these extracts for stomatology disorders, but these must to be still validated from the clinical point of view with trials performed according to the good clinical practices before any application in Cuban stomatology services.

La Bixa orellana L en el tratamiento de afecciones estomatológicas, un tema aún por estudiar / The Bixa orellana L in treatment of stomatology affections: a subject that hasn't studied yet

De las semillas de la planta tropical Bixa orellana L (bija) se extrae un pigmento (el annatto) que està compuesto bßsicamente por carotenoides (bixina/nor-bixina). Por su resistencia a la acción de los agentes químicos, pero no a los efectos del sol, se emplea con preferencia para colorear alimentos y bebidas. La dosis diaria permitida recomendada por la OMS es de 0-0,065 mg/kg de peso corporal. El annato es un colorante con muy baja toxicidad para el humano, reportàndose escasos incidentes de eventos alérgicos. A los extractos de bija le han sido demostrados diversas propiedades farmacológicas, entre las cuales se encuentran: antitumoral, antiinflamatoria, astringente, emoliente, antiséptica, antibacteriana, antioxidante, cicatrizante entre otras y se ha indicado en el tratamiento de infecciones bacterianas de forma general, estomatitis, y en la curación de heridas y quemaduras. Teniendo en cuenta que en el arsenal terapéutico al alcance del estomatólogo son escasos los productos de acción local cicatrizante, antioxidante, antiinflamatoria y antimicrobiana y mucho menos un producto que reúna todas estas propiedades juntas, es que se procedió a realizar una actualización bibliogràfica del tema Bixa orellana L, referido a su composición química, toxicología y efectos farmacológicos de interés para el tratamiento potencial de desórdenes en el campo de la estomatología. Existen evidencias de la utilidad de estos extractos para el tratamiento de desórdenes estomatológicos, pero estas deben aún ser validadas desde el punto de vista clínico con ensayos realizados de acuerdo con las buenas pràcticas clínicas, antes de cualquier aplicación en los servicios estomatológicos cubanos(AU)

From the Bixa orellana L (bija) tropical plant seeds a pigment (annatto) was extracted which is basically composed by carotenoids (bixine/nor-bixine). Due to its resistance to chemical agent's action but not to sun effects, it is used preferably to food and drinks coloring. Daily dose allowance recommended by HWO is of 0-0,065 mg/kg/bw. Annate is a coloring agent with a very low toxicity for humans, even when some low rates of allergic events have been reported. Annate extracts have different pharmacological properties including the antitumor, anti-inflammatory, astringent, emollient, antiseptic, antibacterial, antioxidant, cicatrizant among others and has been prescribed in wounds and burns cure. Considering that in the therapeutical array available for stomatologist the cicatrizant antioxidant, anti-inflammatory and antimicrobial local actions are limited and by no means a product fulfilling all these properties together, we made a bibliographic updating of Bixa orellana L subject related to its chemical, toxicology composition and interesting pharmacological effects for potential treatment of stomatology disorders. There are evidences on usefulness of these extracts for stomatology disorders, but these must to be still validated from the clinical point of view with trials performed according to the good clinical practices before any application in Cuban stomatology services(AU)