Bixa Orellana Seeds Research Articles

FORMULATION OF ANNATTO FEED CONCENTRATE FOR LAYERS AND THE EVALUATION OF EGG YOLK COLOR PREFERENCE OF CONSUMERS

ABSTRACT Nowadays, intensely farmed poultry eggs show almost no differences in egg yolk and albumin compared with domestic layers. Annatto shrub grows rapidly to produce seeds coated with bixin and norbixin dyes which are used to color foods requiring no limits according to codex standards because they are generally regarded as safe for food applications. In this research, annatto concentrate prepared at 1:1 feed to annatto seeds was applied at 1, 4, 7 and 10% over 8 months. The results show that such feeds had no effect on the egg size (P > 0.05) (mean = 62.25 g) as well as proximate composition of feed and flavor of the resulting egg yolks, while rate of application of concentrate show significant (P < 0.05) effect on overall preference of egg yolk of layers fed up to 7%. It is recommended that annatto concentrate can be applied at 1% to give generally acceptable color intensity. PRACTICAL APPLICATIONS The increasing demand of consumers for naturally colored egg products drive poultry farmers to seek for safer ways of achieving this preference. This research is opening a window of opportunity for these farmers to achieve this aim by using Bixa orellana seeds coated with annatto, which is abound in the forest zones of the tropics. Advantage can therefore be taken of this resource which has very little or no application in the poultry sector.

Protective effect of Bixa orellana L. against radiation induced chromosomal aberration in Swiss Albino Mice.

Radioprotective effect of hydroalcoholic extract of seeds of Bixa orellana have been studied by examining chromosome aberration in cells of bone marrow in irradiated mice. Healthy adult Swiss mice were injected intraperitoneally (ip) with 500 mg kg–1 body weight, 1000 mg kg–1 of or double distilled water (DDW). They were exposed to whole body irradiation of 2.0 Gy gamma radiation 30 min later. After 24 h, chromosomal aberrations were studied in the bone marrow of the femur by routine metaphase preparation after colchicine treatment. Radiation (4.0 Gy) increased the number of aberrant cells from less than 1% in controls to almost 20%. Pre-treatment with the extract compounds resulted in a significant reduction in the percentage of aberrant metaphases as well as in the different types of aberration scored. The extract was not toxic at 1500 mg kg–1 body weight. Being non toxic and easily available natural source Bixa orellana extract may be use for as radioprotective for human beings. Keywords: – radioprotective, Bixa orellana

Annatto Powder Production in a Spouted Bed: An Experimental and CFD Study

The mechanical extraction of the bixin from Bixa orellana seeds using a spouted bed was investigated in this work. The experimental program was divided into two main steps. In the first step, a two-level factorial experimental design was used to analyze the influence of the main process variables on the mechanical extraction responses. The second step of the experiment was carried out to evaluate the effect of the distance between the draft tube and the conical base (ht). Computational fluid dynamic technique was used to understand seed flow and the effect of ht on the mechanical extraction process. The results obtained showed that the presence of the draft tube was the variable that most strongly affected the powder extraction. The best condition for the bixin extraction from B. orellana seeds was the one when the draft tube was positioned at 4 cm from the air inlet.

Anatomical and biochemical changes in the composition of developing seed coats of annatto (Bixa orellana L.)

Seeds of Bixa orellana (L.) have a sclerified palisade cell layer, which constitutes a natural barrier to water uptake. In fact, newly fully developed B. orellana seeds are highly impermeable to water and thereby dormant. The purpose of this work is to investigate, from a developmental point of view, the histochemical and physical changes in the cell walls of the seed coat that are associated with the water impermeability. Seed coat samples were analyzed by histochemical and polarization microscopy techniques, as well as by fractionation/HPAEC-PAD. For histochemical analysis the tissue samples were fixed, dehydrated, embedded in paraffin and the slides were dewaxed and tested with appropriate stains for different cell wall components. Throughout the development of B. orellana seeds, there was a gradual thickening of the seed coat at the palisade region. This thickening was due to the deposition of cellulose and hemicelluloses in the palisade layer cell walls, which resulted in a highly water impermeable seed coat. The carbohydrate composition of the cell walls changed dramatically at the late developmental stages due to the intense deposition of hemicelluloses. Hemicelluloses were mainly deposited in the outer region of the palisade layer cell walls and altered the birefringent pattern of the walls. Xylans were by far the most abundant hemicellulosic component of the cell walls. Deposition of cellulose and hemicelluloses, especially xylans, could be responsible for the impermeability to water observed in fully developed B. orellana seeds.

Supercritical carbon dioxide extraction of pigments from Bixa orellana seeds (experiments and modeling)

Supercritical CO2 extraction of the pigments from Bixa orellana seeds was carried out in a flow apparatus at a pressure of 200 bar and a temperature of 40 oC at two fluid flow rates (0.67g/min and 1.12g/min). The efficiency of the extraction was low (only about 1% of the pigment was extracted). The increase in flow rate led to a decrease in pigment recovery. A large increase in recovery (from 1% to 45%) was achieved using supercritical carbon dioxide with 5 mol % ethanol as extraction fluid at pressures of 200 and 300 bar and temperatures of 40 and 60 oC. Although the increase in temperature and pressure led to an increase in recovery, the changes in flow rate did not seem to affect it. Furthermore, two plug flow models were applied to describe the supercritical extraction of the pigments from annatto seeds. Mass transfer coefficients were determined and compared well with those obtained by other researchers with similar models for the supercritical extraction of solutes from plant materials.

Evaluation of the clastogenicity and anticlastogenicity of the carotenoid bixin in human lymphocyte cultures

Carotenoids are regarded as effective antioxidants, antimutagenic and anticarcinogenic agents. Annatto, a red-yellow extract obtained from seeds of Bixa orellana L. is a mixture of several carotenoids and one of them bixin (BXN), is known as its major coloring compound. Studies on BXN clastogenicity and anticlastogenicity in cultured human lymphocytes have not been reported so far. Therefore, the present study was undertaken to investigate the ability of BXN to induce chromosomal aberrations in human lymphocytes in vitro and to examine the possible anticlastogenic effect of this carotenoid in chromosomal damage induced by the clastogen cisplatin (cDDP). Human blood samples were obtained from six healthy, non-smoking volunteers; two females and four males aged 18–35 years. The concentrations of BXN (1.0; 2.5; 5.0 or 10 μg/mL) tested in combination with cDDP were established on the basis of mitotic index (MI) measurements. The data showed that BXN was not cytotoxic or clastogenic, when compared to untreated control. A marked decrease in the MI values compared to the untreated control and an increased percentage of aberrant metaphases was seen in all cultures treated with cDDP. The carotenoid efficiency in reducing the inhibitory effect of cDDP on lymphocyte MI is concentration-dependent. Cultures simultaneously treated with BXN and cDDP showed a statistically significant reduction in total chromosomal aberrations and aberrant metaphases. In our experiments, BXN may have acted as an antioxidant by intercepting free radicals generated by cDDP. The data obtained in the present study suggest that dietary carotenoids may act as protective agents against clastogenic effects of antitumor agents. However, extensive studies are necessary to elucidate the mechanism of action of BXN before its therapeutic use.

The use of nonlinearity measures to discriminate the equilibrium moisture equations for Bixa orellana seeds

In this work, equilibrium moisture data for Bixa orellana seeds at different temperatures and relative humidities were determined using the Thermoconstanter Novasina instrument. Since the majority of the sorption equilibrium equations presented in the literature are nonlinear, care should be taken in estimating their parameters, because in some situations, the least squares estimators may not be appropriate. Therefore, some procedures are available in the literature to validate the statistical properties of the least squares estimators of nonlinear models. In this work, the nonlinear measures are used to discriminate between five model equations that represent the sorption equilibrium isotherms of Bixa orellana seeds. Results show that the Halsey modified equation is the best one to describe the experimental data because it has nonsignificance bias and nonlinear measurement parameters.

ToF-SIMS applied to probe bixin in Bixa orellana seeds

Due to the high daily intake of Annatto, a natural food colorant, it is important to determine its constituents in order to attain some information of nutritional significance. First, an Annatto extract was obtained. The ToF-SIMS spectra measured after depositing a drop of this extract on a silicon substrate exhibit strong molecular peaks clearly related to the bixin molecule (C 25H 30O 4), which has a molar mass M=394. The most intense characteristic ion appears at m/ z=396 ( M+2), but other molecular ions (e.g. at m/ z=337 obtained by the loss of C 2O 2H or even a dimer at m/ z=790, or 2 M+2) were assigned to the bixin. On a silver substrate, M+Ag cations are also observed, confirming the straightforward identification of the colorant. The spectra recorded on the crude seed were remarkably similar to those obtained on the extract confirming the technique’s ability to probe the seed’s chemistry without preliminary treatment. After a prolonged storage in ambient light, a drastic reduction of the bixin molecular ion is observed, along with the appearance of xylene fragments.

Time-of-flight secondary ion mass spectrometry and X-ray photoelectron spectroscopy analyses of Bixa orellana seeds.

Three different experiments were performed in order to obtain the major carotenoid composition of the natural colorant annatto (E160b) through ToF-SIMS (time-of-flight secondary ion mass spectrometry) and XPS (X-ray photoelectron spectroscopy) analyses. In the first experiment, Bixa orellana seeds aril as well as its interior part were analyzed. The analysis of the seeds aril by ToF-SIMS gives the colorant fingerprint without any sample treatment, showing the presence of bixin and its characteristic fragments. The analysis performed in the interior part of the seeds indicates the presence of Fe. The second set of measurements was conducted on the seeds organic extract right after extraction revealing the same components observed by in situ measurement. A third set of measurements was performed aiming to determine the reason for the organic extract color shift observed after 3 months of exposure to ambient light at room temperature. In this case, it was possible to evidence the degradation of bixin by the loss of xylene molecules through ToF-SIMS and the probable carotenoid oxidation based on the C1s XPS spectrum of the degraded extract.

Antimicrobial activity of the leaves and seeds of Bixa orellana

The ethanolic extracts of the leaves and seeds of Bixa orellana showed a broad spectrum of antimicrobial activity. The activity of the leaf extract was more pronounced.

Induction of liver monooxygenases by annatto and bixin in female rats

Annatto or urucum is an orange-yellow dye obtained from Bixa orellana seeds. It has been used as a natural dye in a variety of food products, drugs and cosmetics, and also in Brazilian cuisine as a condiment ('colorau'). Bixin, a carotenoid devoid of provitamin A activity, is the main pigment found in annatto. Some carotenoids (canthaxanthin, astaxanthin and beta-Apo-8'-carotenal) are known to be potent inducers of CYP1A1, a property not shared by others (beta-carotene, lycopene and lutein). Little is known, however, about the CYP1A1-inducing properties of bixin and annatto. The present study was performed to determine the effects of an annatto extract (28% bixin) and bixin (95% pure) on rat liver monooxygenases. Adult female Wistar rats were treated by gavage with daily doses of annatto (250 mg/kg body weight, which contains approximately 70 mg bixin/kg body weight), bixin (250 mg/kg body weight) or the vehicle only (corn oil, 3.75 g/kg body weight) for 5 consecutive days, or were not treated (untreated control). The activities of aniline-4-hydroxylase (A4H), ethoxycoumarin-O-deethylase (ECOD), ethoxy- (EROD), methoxy- (MROD), pentoxy- (PROD) and benzyloxy- (BROD) resorufin-O-dealkylases were measured in liver microsomes. Annatto (250 mg/kg containing 70 mg bixin/kg) induced EROD (3.8x), MROD (4.2x), BROD (3.3x) and PROD (2.4x). Bixin (250 mg/kg) was a weaker inducer of EROD (2.7x), MROD (2.3x) and BROD (1.9x) and did not alter PROD, A4H or ECOD activities. These results suggest that constituents of the extract other than bixin play an important role in the induction of CYP1A and CYP2B observed with annatto food colorings.

Evaluation of the developmental toxicity of annatto in the rat

Annatto, a dye extracted from Bixa orellana seeds, is used as a color additive in butter, cheese and in a variety of other foods as well as in drugs and cosmetics. Toxicological data on annatto and on its main carotenoid pigment bixin are still scarce. In this study we evaluated the developmental toxicity of annatto (28% of bixin). Annatto (0, 31.2, 62.5, 125, 250 and 500 mg/kg body weight/day) was given by gavage to Wistar rats on days 6–15 of pregnancy. Ceasarean sections were performed on day 21. Implantations, living and dead fetuses and resorptions were recorded. Fetuses were weighed and examined for externally-visible anomalies. One-third of fetuses from each litter was examined for visceral anomalies by a microsectioning technique. The remaining fetuses were cleared and stained with Alizarin Red S for skeleton evaluation. No adverse effect of annatto on the mothers was noted. No increase in embryolethality and no reduction of fetal body weight were observed among annatto-exposed rats. Annatto did not induce any increase in the incidence of externally-visible, visceral or skeletal anomalies in the exposed offspring. These findings suggest that annatto was neither maternally toxic nor embryotoxic in the rat. Therefore, the no-observed-adverse-effect level (NOAEL) for annatto-induced maternal and developmental toxicity was 500 mg/kg body weight/day or greater (or ⩾140 mg bixin/kg body weight/day) by the oral route.

Norbixin ingestion did not induce any detectable DNA breakage in liver and kidney but caused a considerable impairment in plasma glucose levels of rats and mice

From the seeds of Bixa orellana are extracted the carotenoids bixin and norbixin that have been widely used for coloring food. In this study, the toxicity of norbixin, purified or not (annatto extract containing 50% norbixin), was investigated in mice and rats after 21 days of ingestion through drinking water. Mice were exposed to doses of 56 and 351 mg/kg (annatto extract) and 0.8, 7.6, 66 and 274 mg/kg (norbixin). Rats were exposed to doses of 0.8, 7.5 and 68 mg/kg (annatto extract) and 0.8, 8.5 and 74 mg/kg (norbixin). In rats, no toxicity was detected by plasma chemistry. In mice, norbixin induced an increase in plasma alanine aminotransferase activity (ALT) while both norbixin and annatto extract induced a decrease in plasma total protein and globulins ( P < 0.05). However, no signs of toxicity were detected in liver by histopathological analysis. No enhancement in DNA breakage was detected in liver or kidney from mice treated with annatto pigments, as evaluated by the comet assay. Nevertheless, there was a remarkable effect of norbixin on the glycemia of both rodent species. In rats, norbixin induced hyperglycemia that ranged from 26.9% (8.5 mg/kg norbixin, to 52.6% (74 mg/kg norbixin, P < 0.01) above control levels. In mice, norbixin induced hypoglycemia that ranged from 14.4% (0.8 mg/kg norbixin, P < 0.05) to 21.5% (66 mg/kg norbixin, P < 0.001) below control levels. Rats and mice treated with annatto pigments showed hyperinsulinemia and hypoinsulinemia, respectively indicating that pancreatic β-cells were functional. More studies should be performed to fully understand of how species-related differences influences the biological fate of norbixin.

Germinação de sementes de urucum (Bixa orellana L.)

O urucuzeiro ou urucueiro é a única espécie do gênero Bixa, sendo nativa da América tropical. Entre os corantes naturais, o urucum é o segundo em importância econômica, sendo os maiores produtores o Peru, Brasil e Quênia. A propagação por sementes vêm apresentando problemas, pois a germinação é baixa devido à dormência imposta pelo tegumento. O objetivo do trabalho foi estudar o efeito, da separação por peso, das sementes de urucum, na qualidade fisiológica, assim como, tratamentos para promover a germinação. O experimento foi conduzido separando-se o lote original em quatro frações denominadas sementes leves, médias, pesadas e extra pesadas. A seguir as sementes foram avaliadas através de determinação do peso de 100 sementes, determinação do grau de umidade, teste de germinação e avaliação de tratamentos para superar a dormência sendo eles: água quente por um, três e cinco minutos, pré-embebição por 24 horas seguido de água quente por um, três e cinco minutos e escarificação mecânica. Pôde-se concluir que a separação de sementes de urucum, por peso, influi na qualidade fisiológica, sendo que sementes mais pesadas são de qualidade superior, porém apresentam maior incidência de sementes duras, assim, o tratamento de escarificação mecânica é recomendado, na superação da dormência de sementes de urucum.

Biochemical behaviour of norbixin during in vitro DNA damage induced by reactive oxygen species.

Naturally occurring antioxidants such as carotenoids are extensively studied for their potential in reducing the risk for cancer and other chronic diseases. In the present study, the radical-scavenger activity of the food additive norbixin, a water-soluble carotenoid extracted from Bixa orellana seeds and commercialized as annatto, was evaluated under conditions of DNA damage induced by reactive oxygen species, particularly by hydroxyl radicals. The cell-free scavenger activity of norbixin was evaluated using plasmid DNA as target molecule and Sn2+ or Fe2+ as oxidant. The addition of H2O2 enhanced DNA breakage induced by metal ions, particularly Fe2+. Under these conditions, norbixin started to protect plasmid DNA against single- and double-strand breakage at a metal:norbixin ratio of 1:1 (Sn2+) and 1:10 (Fe2+). However, at lower ratios to Sn2+, norbixin enhanced Sn2+-induced DNA breakage (P < 0.05). The ability of norbixin to protect genomic DNA against oxidative damage was assessed in murine fibroblasts submitted to H2O2-induced oxidative stress and the results were evaluated by the comet assay. Under low serum conditions (2 % fetal bovine serum (FBS)), a protective effect of norbixin against H2O2-induced DNA breakage was inversely related to its concentration, a protection ranging from 41 % (10 microm) to 21 % (50 microm). At higher concentrations of norbixin, however, oxidative DNA breakage was still enhanced, even in the presence of a high serum concentration (10 % FBS). Under normal conditions, norbixin per se has no detectable genotoxic or cytotoxic effects on murine fibroblasts. The antimutagenic potential of norbixin against oxidative mutagens was also evaluated by the Salmonella typhimurium assay, with a maximum inhibition of 87 % against the mutagenicity induced by H2O2. Although plasmid DNA and Ames data indicated that norbixin can protect DNA against oxidative damage, it seems to be a risky guardian of genomic DNA as it can also increase the extent of oxidative damage.

Germinação de sementes em desenvolvimento de Bixa orellana

Com este trabalho objetivou-se investigar a aquisição da capacidade germinativa e o estabelecimento da dormência em sementes em desenvolvimento de Bixa orellana L. (urucum). Foram caracterizados 7 estádios de desenvolvimento das sementes, baseados nas características morfológicas, massa fresca, massa seca e teor de umidade dos frutos e das sementes. Sementes imaturas de B. orellana não germinaram durante os estádios iniciais de seu desenvolvimento. As sementes adquiriram a capacidade germinativa máxima quando os níveis de umidade declinaram para valores em torno de 60%. Nessa fase, as sementes apresentaram acúmulo máximo de massa seca. A dormência foi estabelecida quando as sementes exibiam um conteúdo de água em torno de 10%.

BIXIN POWDER PRODUCTION IN CONICAL SPOUTED BED UNITS

ABSTRACT Recent work has demonstrated that the red carotenoid bixin is easily extracted from Bixa orellana seeds by particle attrition and impact rather than by solvent extraction. This technique may require a previous step for drying seeds at a safe temperature to preserve pigment quality. A spouted bed (SB) with a draft tube is the most appropriate equipment to process both seed drying and bixin extraction at low operational costs. Operational parameters of the SB unit are optimized as a function of the powder production rate. The maximum air inlet temperature for drying the seeds is also specified. A high performance liquid chromatography method has been developed for the semi-quantitative determination of bixin content in the powder produced. Results show that this SB unit can be more competitive than ball mill equipment.

Detection of Bixin, Lycopene, Canthaxanthin, and β-Apo-8′-Carotenal in Products Derived from Red Pepper

An analytical method using either thin layer or liquid chromatography is proposed for the detection of 4 pigments (bixin, lycopene, canthaxanthin and beta-apo-8'-carotenal) that can be used fraudulently to intensify the natural color of products derived from red pepper (oleoresins, paprika, paprika paste, etc.). Similarly, the addition of other colorant natural products containing some of these pigments as major pigments (such as tomato for lycopene and Bixa orellana seeds for bixin) can be detected. The method proposed can also be used to control the aforementioned pigments in their natural sources as well as in food products.

Production of annatto concentrates in spouted beds

AbstractAnnatto is composed of a natural reddish‐yellow pigment, which is commonly used in the food, cosmetic and pharmaceutical industries. This pigment is easily extracted from Bixa orellana seeds by particle impact and attrition rather than by solvent extraction. Seeds must be dried at a safe temperature to preserve the pigment quality. Experiments show that the annatto powder produced in a non‐conventional spouted bed has a high pigment content and is thus suitable for commercial use. A comparative analysis of the bixin extraction in different systems demonstrates that this spouted bed unit can be competitive for commercial applications. Hydrodynamic and drying parameters were studied to improve the powder production rate in this unit.

Terpenoids and an apocarotenoid from seeds of Bixa orellana

all- E-Geranylgeraniol is present in the oleoresin from Bixa orellana to the extent of 57%, or approximately 1% of dry seeds which makes B. orellana the richest known source of this important C 20-terpene alcohol. Other compounds isolated for the first time from seeds of Bixa were the terpenes farnesylacetone, geranylgeranyl octadecanoate and geranylgeranyl formate, δ-tocotrienol, and an apocarotenoid.