norbixin and bixin

Colored Cheddar cheeses are prepared by adding an aqueous annatto extract (norbixin) to cheese milk; however, a considerable proportion (∼20%) of such colorant is transferred to whey, which can limit the end use applications of whey products. Different geographical regions have adopted various strategies for handling whey derived from colored cheeses production. For example, in the United States, whey products are treated with oxidizing agents such as hydrogen peroxide and benzoyl peroxide to obtain white and colorless spray-dried products; however, chemical bleaching of whey is prohibited in Europe and China. Fundamental studies have focused on understanding the interactions between colorants molecules and various components of cheese. In addition, the selective delivery of colorants to the cheese curd through approaches such as encapsulated norbixin and microcapsules of bixin or use of alternative colorants, including fat-soluble/emulsified versions of annatto or beta-carotene, has been studied. This review provides a critical analysis of pertinent scientific and patent literature pertaining to colorant delivery in cheese and various types of colorant products on the market for cheese manufacture, and also considers interactions between colorant molecules and cheese components; various strategies for elimination of color transfer to whey during cheese manufacture are also discussed.

Topics: Bixaceae; Carotenoids; Cheese; Food Coloring Agents; Food Handling; Oxidants; Plant Extracts; Whey

Organic compounds have been employed in developing new green energy solutions with good cost-efficiency compromise, such as photovoltaics. The light-harvesting process in these applications is a crucial feature that still needs improvements. Here, we studied natural dyes to propose an alternative for enhancing the light-harvesting capability of photovoltaics. We performed density functional theory calculations to investigate the electronic and optical properties of the four natural dyes found in achiote seeds (

Topics: Bixaceae; Carotenoids; Coloring Agents; Electronics; Seeds

We have previously reported how the natural food colorant, bixin, was enzymatically modified by appending sorbitol to the bixin scaffold. The resulted product, sorbitol ester of norbixin (SEN) was expected to be more hydrophilic. The present study aimed to investigate the physical behaviour of SEN in aqueous media. The property of SEN was studied together with non-reacted bixin as separation of the two compounds was unsuccessful. The SEN molecules behaved as a bolaamphiphile in aqueous media, underwent self-association and develop a hydrophilic aggregate. SEN-aggregates could uptake the non-reacted bixin molecules inside its hydrophobic moiety and dispersed it in aqueous media. Aggregation of SEN molecules with incorporated bixin resulted in a hypsochromic shift of the absorption spectra indicting H-aggregation. Dynamic light scattering showed the formation of aggregates with an average hydrodynamic radius 38 ± 2 nm. The dispersibility of the aggregates was affected by pH and the ionic strength of the media.

Topics: Carotenoids; Dynamic Light Scattering; Esters; Food Coloring Agents; Furans; Hydrogen-Ion Concentration; Hydrophobic and Hydrophilic Interactions; Nanostructures; Osmolar Concentration; Pyridones; Sorbitol; Water

The aim of this study was to evaluate the effect of annatto carotenoids intake associated to a single high-calorie meal (high fat and high carbohydrate) in postprandial biochemical, inflammatory and oxidative stress markers. Twelve healthy subjects (6 men, 6 women) were included in this randomised, controlled crossover study. Baseline blood samples were collected from fasting subjects that immediately received high-calorie meal without carotenoid (placebo) or containing 1.2mg/kg bixin (BIX) or 0.06mg/kg norbixin (NBIX). Blood samples were taken 60, 120 and 240min after meal intake. NBIX intake did not affect biochemical blood markers but reduced the postprandial levels of inflammatory cytokines (IL-1, IL-6 and TNF-α) and lipid oxidation 60-120min after meal. BIX only partially prevented postprandial-induced lipid oxidation. Results indicate that the intake of NBIX may be an alternative to reduce the postprandial inflammatory and oxidative stress responses to high-calorie meals.

Topics: Adult; Bixaceae; Blood Glucose; Blood Pressure; Carotenoids; Cytokines; Diet; Female; Humans; Inflammation; Male; Oxidative Stress; Plant Extracts; Postprandial Period; Young Adult

In this present study, the inhibitory mechanism of three selected apocarotenoids (bixin, norbixin and crocin) on the diphenolase activity of tyrosinase has been investigated. The preliminary screening results indicated that apocarotenoids inhibited tyrosinase activity in a dose-dependent manner. Kinetic analysis revealed that apocarotenoids reversibly inhibited tyrosinase activity. Analysis of fluorescence spectra showed that apocarotenoids quenched the intrinsic fluorescence intensity of the tyrosinase. Further, molecular docking results implied that apocarotenoids were allosterically bound to tyrosinase through hydrophobic interactions. The results of the in vitro studies suggested that higher concentrations of bixin and norbixin inhibited tyrosinase activity in B16F0 melanoma cells. Our results suggested that apocarotenoids could form the basis for the design of novel tyrosinase inhibitors.

Topics: Allosteric Site; Animals; Carotenoids; Cell Line, Tumor; Dose-Response Relationship, Drug; Hydrophobic and Hydrophilic Interactions; Kinetics; Melanins; Melanoma; Mice; Molecular Docking Simulation; Monophenol Monooxygenase; Protein Binding; Spectrometry, Fluorescence

The Cheddar cheese colorant annatto is present in whey and must be removed by bleaching. Chemical bleaching negatively affects the flavor of dried whey ingredients, which has established a need for a better understanding of the primary colorant in annatto, norbixin, along with cheese color alternatives. The objective of this study was to determine norbixin partitioning in cheese and whey from full-fat and fat-free Cheddar cheese and to determine the viability of bixin, the nonpolar form of norbixin, as an alternative Cheddar cheese colorant. Full-fat and fat-free Cheddar cheeses and wheys were manufactured from colored pasteurized milk. Three norbixin (4% wt/vol) levels (7.5, 15, and 30 mL of annatto/454 kg of milk) were used for full-fat Cheddar cheese manufacture, and 1 norbixin level was evaluated in fat-free Cheddar cheese (15 mL of annatto/454 kg of milk). For bixin incorporation, pasteurized whole milk was cooled to 55 °C, and then 60 mL of bixin/454 kg of milk (3.8% wt/vol bixin) was added and the milk homogenized (single stage, 8 MPa). Milk with no colorant and milk with norbixin at 15 mL/454 kg of milk were processed analogously as controls. No difference was found between the norbixin partition levels of full-fat and fat-free cheese and whey (cheese mean: 79%, whey: 11.2%). In contrast to norbixin recovery (9.3% in whey, 80% in cheese), 1.3% of added bixin to cheese milk was recovered in the homogenized, unseparated cheese whey, concurrent with higher recoveries of bixin in cheese (94.5%). These results indicate that fat content has no effect on norbixin binding or entrapment in Cheddar cheese and that bixin may be a viable alternative colorant to norbixin in the dairy industry.

Topics: Animals; Bixaceae; Carotenoids; Cheese; Food Coloring Agents; Food Handling; Plant Extracts; Taste; Whey Proteins

Bixin and norbixin are the main components of annatto, which is extracted from Bixa orellana and largely used as natural colorant in the food and pharmaceutical industries. Annatto can enhance CYP1A and CYP2B activity in rats; however, the inducer effect has not been investigated in human cell lines. In this study, the ability of bixin and norbixin to induce the cytochrome P450 (CYP) enzymes was assessed in HepG2 human hepatoma cell line. HepG2 cells were treated with bixin and norbixin, and the expression of the CYP genes quantified by real-time reverse transcription polymerase chain reaction (RT-PCR). Expression of CYP1A1 and CYP1A2 was significantly increased by bixin treatment, while CYP2B6, 2C9, 2E1 and 3A4 were unaffected. Cells were treated with norbixin showed no inducer effect. The results suggest that the inducer potential of annatto is attributed to bixin, but not to norbixin, despite their similarities in molecular structure.

Topics: Carotenoids; Cell Survival; Cytochrome P-450 Enzyme System; Enzyme Induction; Gene Expression; Hep G2 Cells; Humans; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction

Populations in the Amazon are exposed to organic mercury via consumption of contaminated foods. These ethnic groups consume a specific plant seed "annatto" which contains certain carotenoids. The aim of this study was to find out if these compounds (bixin, BIX and norbixin, NOR), protect against DNA-damage caused by the metal. Therefore, rats were treated orally with methylmercury (MeHg) and with the carotenoids under conditions that are relevant to humans. The animals were treated either with MeHg (30 μg/kg/bw/day), BIX (0.1-10 mg/kg/bw/day), NOR (0.01-1.0 mg/kg/bw/day) or combinations of the metal compound and the carotenoids consecutively for 45 days. Subsequently, the glutathione levels (GSH) and the activity of catalase were determined, and DNA-damage was measured in hepatocytes and leukocytes using single cell gel electrophoresis assays. Treatment with the metal alone caused a decrease in the GSH levels (35%) and induced DNA damage, which resulted in increased DNA migration after electrophoresis in liver and blood cells, whereas no effects were seen with the carotenoids alone. When BIX or NOR were given in combination with organic mercury, the intermediate and the highest concentrations of the carotenoids (1.0 and 10.0 mg/kg/bw/day BIX and 0.1 and 1.0 mg/kg/bw/day NOR) protected against DNA-damage. Furthermore, we found with both carotenoids, a moderate increase in the GSH levels in both metal-treated and untreated animals, while the activities of catalase remained unchanged. Our results indicate that consumption of BIX and NOR may protect humans against the adverse health effects caused by exposure to organic mercury.

Topics: Analysis of Variance; Animals; Bixaceae; Carotenoids; Catalase; Comet Assay; DNA Damage; Environmental Pollutants; Glutathione; Methylmercury Compounds; Molecular Structure; Oxidation-Reduction; Plant Extracts; Rats; Rats, Wistar

Peroxisome proliferator-activated receptor α (PPARα) is a ligand-activated transcription factor that regulates the expression of the genes involved in fatty acid oxidation. PPARα activators induce fatty acid oxidation in the liver, thereby improving lipid and carbohydrate metabolism in obese mice. In this study, the dietary cis-carotenoids bixin and norbixin, which are commonly used in the food coloring industry, were found to activate PPARα by luciferase reporter assays using GAL4/PPARα chimeric and full-length PPARα systems. Treatment with bixin and norbixin induced the mRNA expression of PPARα target genes involved in fatty acid oxidation in PPARα-expressing HepG2 hepatocytes. In obese KK-Ay mice, bixin treatment suppressed the development of hyperlipidemia and hepatic lipid accumulation. In the livers of bixin-treated mice, the mRNA levels of PPARα target genes related to fatty acid oxidation were up-regulated. Moreover, bixin treatment also improved obesity-induced dysfunctions of carbohydrate metabolism, such as hyperglycemia, hyperinsulinemia, and hypoadiponectinemia. Glucose tolerance test and insulin tolerance test revealed that glucose intolerance and insulin resistance in KK-Ay obese mice were attenuated by the treatment with bixin. These results indicate that bixin acts as a food-derived agonist of PPARα, and bixin treatment is useful for the management of obesity-induced metabolic dysfunctions in mice.

Topics: Adiponectin; Animals; Carbohydrate Metabolism; Carotenoids; Diet, High-Fat; Fatty Liver; Gene Expression Regulation; Glucose Tolerance Test; Hepatocytes; Hyperlipidemias; Lipid Metabolism; Male; Mice; Mice, Obese; Obesity; PPAR alpha

The present studies were performed to examine the contact allergenic effects of an annatto extract (ANT) in female BALB/c mice. ANT at 5-10% induced a greater than threefold increase in lymph node cell proliferation when compared to the control in the LLNA. Moreover, a significant increase in the percent ear swelling at 24h after ANT challenge was observed in the MEST. A significant increase in the percentage of B cells was also observed. To determine which of the two predominant coloring components (norbixin and bixin) in ANT was responsible for the sensitizing effects of ANT, norbixin was subsequently examined, with negative results being observed in both the LLNA and MEST following treatment with norbixin (1-20%). These findings suggested that perhaps bixin was responsible for the positive responses in both the LLNA and MEST following exposure to ANT. Therefore, further studies using a partially purified cis-bixin extract were conducted. Positive responses in both the LLNA and MEST were observed in mice treated with cis-bixin at the concentrations as low as 0.1-0.5%. These results have demonstrated that cis-bixin, but not norbixin, is likely a contact sensitizer and contributes to the contact hypersensitivity effects observed following dermal exposure to ANT in mice.

Topics: Animals; Bixaceae; Carotenoids; Dermatitis, Contact; Female; Flow Cytometry; Food Coloring Agents; Local Lymph Node Assay; Mice; Mice, Inbred BALB C; Plant Extracts; Random Allocation

A supramolecular solvent (SUPRA) made up of octanoic acid aggregates is proposed for the microextraction of bixin and norbixin, the two major components of the natural food colouring annatto, in food. The procedure involved the extraction of sub-gram quantities (200mg) of homogenized food with 0.8mL of the supramolecular solvent. The overall sample treatment took about 20 min, and several samples could be simultaneously treated using conventional lab equipment. No clean-up or solvent evaporation were required. Extraction efficiencies mainly depended on the major components making up the SUPRAS (i.e. octanoic acid and tetrahydrofuran) and were not affected by the pH or the temperature in the ranges studied (1-4 and 10-80°C, respectively). Bixin and norbixin in the extracts were quantified by liquid chromatography (LC) and diode array detection (DAD). They were separated in a Hypersil C18 column using a mobile phase consisting of 5% acetic acid and methanol (15:85, v/v). The retention times for norbixin and bixin standards were 5.1 and 8.6 min, respectively. Recoveries in samples ranged between about 78% and 113%. The precision of the method, expressed as relative standard deviation, was about 1.5% and the quantitation limits for bixin and norbixin were 0.19 and 0.23 mg kg(-1), respectively, which were far below the maximum limits permitted by the European Union for the level of addition to food. Concentration of norbixin in samples belonging to the five major groups of food commodities defined in the literature, ranged between 3.75 and 43.8 mg kg(-1) whereas bixin was only found in one snack sample (6.6 mg kg(-1)). The method is simple and rapid, while delivering accurate and precise results, and can be used for the routine determination of annatto in food for the control of the compliance of current legislation.

Topics: Bixaceae; Caprylates; Carotenoids; Chromatography, Liquid; Food Analysis; Food Coloring Agents; Furans; Hydrogen-Ion Concentration; Plant Extracts; Reproducibility of Results; Sensitivity and Specificity; Temperature

In pursuit of the anticancer effects of seeds of the rain forest plant Bixa orellana (annatto), we found that its constituent cis-bixin induced cytotoxicity in a wide variety of tumor cell lines (IC(50) values from 10 to 50 microM, 24-h exposures) and, importantly, also selectively killed freshly collected patient multiple myeloma cells and highly drug-resistant multiple myeloma cell lines. Mechanistic studies indicated that cis-bixin-induced cytotoxicity was greatly attenuated by co-treatment with glutathione or N-acetylcysteine (NAC); whereas fluorescence-activated cell sorting (FACS) assays using the cell-permeable dyes 5-(and-6) chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H(2)DCFDA), or dihydroethidium demonstrated that cis-bixin rapidly induced cellular reactive oxygen species (ROS) in dose- and time-dependent fashions, collectively implicating ROS as contributory to cis-bixin-induced cytotoxicity. In pursuit of potential contributors to ROS imposition by cis-bixin, we found that cis-bixin inhibited both thioredoxin (Trx) and thioredoxin reductase (TrxR1) activities at concentrations comparable to those required for cytotoxicity, implicating the inhibition of these redox enzymes as potentially contributing to its ability to impose cellular ROS and to kill cancer cells. Collectively, our studies indicate that the annatto constituent cis-bixin has intriguing selective antimyeloma activity that appears to be mediated through effects on redox signaling.

Topics: Acetylcysteine; Antineoplastic Agents; Apoptosis; Bixaceae; Carotenoids; Cell Line, Tumor; Cell Survival; Drug Screening Assays, Antitumor; Glutathione; Humans; Multiple Myeloma; Oxidation-Reduction; Oxidative Stress; Phytotherapy; Plant Extracts; Reactive Oxygen Species; Seeds; Thioredoxin-Disulfide Reductase; Thioredoxins

The development of an analytical method that enables routine analysis of annatto dye, specifically bixin and norbixin, in meat tissue is described. Liquid-solid extraction was carried out using acetonitrile. Analysis was by HPLC with photodiode array detection using two fixed wavelengths (458 and 486 nm). The possibilities of ion trap mass spectrometry (MS) were also assessed. Method performance characteristics, according to Commission Decision 2002/657/EC, were determined, with recoveries between 99 and 102% and calibration curves being linear in the 0.5-10 mg kg(-1) range. The limit of quantification was 0.5 mg kg(-1).

Topics: Animals; Bixaceae; Carotenoids; Chromatography, Liquid; Food Analysis; Food Coloring Agents; Mass Spectrometry; Meat; Plant Extracts; Reproducibility of Results

Insulin resistance is partly due to suppression of insulin-induced glucose uptake into adipocytes. The uptake is dependent on adipocyte differentiation, which is controlled at mRNA transcription level. The peroxisome proliferator-activated receptor (PPAR), a ligand-regulated nuclear receptor, is involved in the differentiation. Many food-derived compounds serve as ligands to activate or inactivate PPAR. In this study, we demonstrated that bixin and norbixin (annatto extracts) activate PPARgamma by luciferase reporter assay using GAL4-PPAR chimera proteins. To examine the effects of bixin on adipocytes, 3T3-L1 adipocytes were treated with bixin or norbixin. The treatment induced mRNA expression of PPARgamma target genes such as adipocyte-specific fatty acid-binding protein (aP2), lipoprotein lipase (LPL), and adiponectin in differentiated 3T3-L1 adipocytes and enhanced insulin-dependent glucose uptake. The observations indicate that bixin acts as an agonist of PPARgamma and enhances insulin sensitivity in 3T3-L1 adipocytes, suggesting that bixin is a valuable food-derived compound as a PPAR ligand to regulate lipid metabolism and to ameliorate metabolic syndrome.

Topics: 3T3-L1 Cells; Adipocytes; Adipogenesis; Animals; Carotenoids; Genes, Reporter; Glucose; Insulin; Insulin Resistance; Lipid Metabolism; Luciferases; Mice; PPAR gamma; RNA, Messenger

An increasing trend in the food and pharmaceutical industries is toward replacing synthetic additives with natural products. However, in this regard, difficulties may be encountered due to the instability of such compounds. Encapsulation has become an important process to protect natural pigments. This paper reports on the encapsulation of the natural urucum pigment with chitosan using different techniques and its release under different pH conditions. The material loaded with pigment was evaluated by infrared spectroscopy, scanning electron microscopy and thermal analysis. Chitosan was found to be an effective encapsulating agent for urucum pigment. No investigations have previously been reported on the relation of chitosan to the stability of encapsulated natural pigments.

Topics: Antioxidants; Bixaceae; Calorimetry, Differential Scanning; Carotenoids; Chitosan; Delayed-Action Preparations; Drug Compounding; Hydrogen-Ion Concentration; Microscopy, Electron, Scanning; Microspheres; Particle Size; Spectroscopy, Fourier Transform Infrared; Surface Properties; Thermogravimetry

Bixin, also known as annatto, is a seed-specific pigment widely used in foods and cosmetics since pre-Columbian times. We show that three genes from Bixa orellana, native to tropical America, govern bixin biosynthesis. These genes code for lycopene cleavage dioxygenase, bixin aldehyde dehydrogenase, and norbixin carboxyl methyltransferase, which catalyze the sequential conversion of lycopene into bixin. Introduction of these three genes in Escherichia coli engineered to produce lycopene induced bixin synthesis, thus expanding the supply of this economically important plant product.

Topics: Aldehyde Dehydrogenase; Amino Acid Sequence; Bixaceae; Carotenoids; Catalysis; Cloning, Molecular; DNA, Complementary; Escherichia coli; Gene Library; Genes, Plant; Lycopene; Methyltransferases; Molecular Sequence Data; Oxygenases; Recombinant Proteins; Seeds; Transformation, Bacterial

Analytical methods for the determination of the permitted food colouring annatto (E160b) have been developed or refined to encompass the wide range of food commodity types permitted to contain it. Specific solvent extraction regimens have been used depending upon the food commodity analysed and HPLC analysis techniques coupled with spectral confirmation have been used for the determination of the major colouring components. Qualitative and quantitative data on the annatto content of 165 composite and two single retail food samples covering a wide range of foods at levels above the limit of quantification (0.1 mg kg(-1)) is reported. Quantitative results are given for the major colour principals 9'-cis-bixin, 9'-cis-norbixin and trans-bixin. Semi-quantitative results are given for the minor bixin and norbixin isomers monocis- (not 9'-), di-cis- and trans-norbixin, for which authentic reference standards were not available. Repeat analyses (n = 4-9) of 12 different types of food commodity (covering the permitted range) spiked with annatto at levels between 1.7 and 27.7 mg kg(-1) gave mean recoveries between 61 and 96%. The corresponding relative SDs (RSD) were between 2.1 and 7.9%.

Topics: Animals; Bixaceae; Carotenoids; Chromatography, High Pressure Liquid; Fishes; Food Analysis; Food Coloring Agents; Humans; Plant Extracts

A procedure was developed for the detection and determination of bixin and norbixin in human plasma by reversed-phase HPLC with a sensitivity limit of 5 micrograms l-1. A group of seven volunteers ingested a single dose of 1 ml of a commercial Annatto Food Color (16 mg of cis-bixin in soybean oil). The presence of bixin (cis and trans) and norbixin (cis and trans) was demonstrated in the plasma at average levels of 11.6, 10.1, 2.8 and 0 micrograms l-1 of bixin and 48, 58, 53 and 29 micrograms l-1 of norbixin after 2, 4, 6 and 8 h, respectively. Considerable individual variations were observed. Complete plasma clearance generally occurred for bixin by 8 h and for norbixin by 24 h after ingestion of cis-bixin.

Topics: Carotenoids; Food Coloring Agents; Humans; Intestinal Absorption; Plants, Edible; Stereoisomerism

During a study of natural food colours, a simple and reliable high-performance liquid chromatography system was developed for use with cochineal and annato. An isocratic mobile phase, consisting of methanol and 6% aqueous acetic acid, resolved bixin and norbixin, while a gradient system was used to separate carminic acid and the annato compounds. The carminic acid contents of cochineal extract, carmine and carmine hydrosoluble were determined using an isocratic mobile phase (40:60, v/v). The detection limit for carminic acid in the various products was approximately 100 ng/g. Carminic acid was determined quantitatively in fruit beverages, yogurt and candies. It was demonstrated that, because of decomposition, carminic acid was not suitable for use in candies when manufacturing temperatures above 100 degrees C were required. Most membrane filters are not suitable for use with cochineal solutions, but a cellulose membrane filter did not adsorb carminic acid and was used successfully to remove impurities from water-based cochineal products and food extracts containing carminic acid.

Topics: Carmine; Carotenoids; Chromatography, High Pressure Liquid; Food Additives; Food Analysis; Isomerism