Annatto

Ethanol-Based Extraction of Annatto (Bixa Orellana L.) and Characterization of the Bixin and Norbixin.[Pubmed:38680327]

ACS Omega. 2024 Apr 13;9(16):18273-18277.

Bixa orellana dye was extracted from the seeds by the method of Soxhlet extraction using ethanol where a percentage yield of 31.4 was obtained. Recrystallization of the dye was performed using chloroform and ethanol to increase the degree of purity. Thin layer chromatography was used to separate the dye into Bixin and Norbixin with R(f) values of 0.56 and 0.42, respectively. The extracted dye was pH sensitive, insoluble in water but soluble in some organic solvents like ethanol and acetic acid. Characterization by UV-vis spectroscopy showed maximum absorption for Bixin and Norbixin at 457 and 453 nm, respectively. Fourier transform infrared spectra of Bixin and Norbixin showed bands confirming similar functional groups for both Bixin and Norbixin except for -CO- stretching and methylene vibration which was absent in Norbixin. With the yield, purity level, and other properties of the extracted dye, the comparative advantages of ethanol-based Soxhlet extraction of the dye were well established.

First Report of Colletotrichum siamense Causing Anthracnose on Bixa orellana in China.[Pubmed:38679590]

Plant Dis. 2024 Apr 28.

Annatto (Bixa orellana L.) is widely cultivated in China. Its seed is used as medicine and as an astringent antipyretic. Since 2019, anthracnose-type lesions have been observed on the Annatto leaves in the field (about 30 hectares) in Zhanjiang (21 18'12''N, 110 17'22''E), Guangdong Province, China. Disease incidence was around 70% (n = 100 investigated plants from about 3 ha). The early symptoms were yellow spots on the edge or tip of leaves. The spots gradually expanded and became dark brown, eventually coalescing into large irregular or circular lesions (Supplemental Figure 1-A). Ten symptomatic leaves from 10 plants were sampled. The margins of the lesions were cut into 2 x 2 mm pieces and the surfaces were disinfected with 75% ethanol for 30 sec and 2% sodium hypochlorite for 60 sec. After that, pieces were rinsed thrice in sterile water, placed on potato dextrose agar(PDA) medium, and incubated at 28 ℃ for 3 days. Pure cultures were obtained by transferring hyphal tips to new PDA plates. Twenty isolates were obtained. Three representative single-spore isolates (BOC-1, BOC-2, and BOC-3) from the twenty isolates were confirmed to be identical based on morphological characteristics and ITS analysis and used for further study. Besides, the three isolates were deposited in the fungus collection at Aquatic Organisms Museum of Guangdong Ocean University. Colonies on PDA were white to gray with cottony mycelia after incubating in the dark for 6 days at 28 ℃. Conidia were one-celled, hyaline, cylindrical, clavate, and obtuse at both ends; they measured 9.6 to 18.5 microm x 3.5 to 5.5 microm (n = 50). Appressoria were oval to irregular in shape and dark brown, and they measured 6 to 9 microm x 4.5 to 8 microm (n = 30) (Supplemental Figure 1-D, E and F). These morphological characteristics matched the description of Colletotrichum siamense (Prihastuti et al. 2009; Sharma et al. 2013). For molecular identification, the internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), chitin synthase (CHS-1), and actin (ACT) loci of the isolates were amplified using primer pairs ITS1/ITS4, GDF1/GDR1, CHS-79F/CHS-354R, and ACT-512F/ACT-783R, respectively (Weir et al. 2012). Sequences were deposited in GenBank under nos. MZ047377-MZ047379 (ITS), MZ126934-MZ1269346 (GAPDH), MZ126904-MZ1269046 (CHS-1), and MZ126844-MZ1268446 (ACT). A phylogenetic tree was generated on the basis of the concatenated data from ITS, GAPDH, CHS-1, and ACT sequences that clustered the three isolates with C. siamense (the type strain MFLU 090230), (Supplemental Figure 2). The pathogenicity of the three isolates was tested respectively in a greenhouse maintained at 25 to 29℃ and 80% relative humidity. Annatto seeding ( n =5, 2-month-old) were inoculated with a spore solution (1 x 105 per mL) until it run-off. Whereas control plants were sprayed with sterile distilled water.. The experient was repeated three times. Anthracnose lesions were observed on the inoculated leaves after 10 days while the control plants remained healthy (Supplemental Figure 1-G, and H). The same pathogen was re-isolated from all the inoculated leaves based on morphology and ITS analysis. C. siamense has been reported to cause anthracnose in a broad range of hosts (Weir et al. 2012; Wang et al. 2017; Liu et al. 2017; Zhuo et al. 2017 ), but not in B. orellana. To our knowledge, this is the first report of C. siamense causing anthracnose on B. orellana in China. Our study provides important reference information for controlling this disease.

Sustainable appraisal of lipstick tree seeds (Bixa orellana)-based bixin natural orange colorant for green mordanted silk fabrics and wool yarns.[Pubmed:38614468]

Sci Prog. 2024 Apr-Jun;107(2):368504241242282.

This research aims to optimize the silk and wool dyeing process using natural dyes from Bixa orellana (Annatto) through response surface methodology. Central composite design experiments highlight the significant enhancement of color outcomes achieved through microwave treatment. For silk, the optimal conditions (80 degrees C for 40 min) with Annatto extract yield a color strength (K/S) of 17.8588, while wool achieves a K/S of 7.5329. Introducing eco-friendly bio-mordants, such as pomegranate peel and red sumac tannins, enhances color strength. Pre-dyeing treatments with 2% red sumac, 1.5% pomegranate peel, and weld flower extracts for silk produce high color strength, with K/S values of 16.4063, 16.3784, and 12.1658, respectively. Post-dyeing, the K/S values increase to 40.1178, 17.4779, and 21.6494. Wool yarn exhibits similar improvements, with pre-dyeing K/S values of 13.1353, 13.5060, and 16.3232, escalating to 10.5892, 15.3141, and 23.4850 post-dyeing. Furthermore, this research underscores improved colorfastness properties, including notable enhancements in light, wash, and rubbing fastness for both silk fabric and wool yarn. These findings underscore the efficacy of the proposed sustainable dyeing methods, offering valuable insights for eco-friendly textile production.

Consumer perception of Cheddar cheese color.[Pubmed:38608957]

J Dairy Sci. 2024 Apr 10:S0022-0302(24)00730-6.

The color of Cheddar cheese in the US is influenced by many factors, primarily the amount of Annatto added as a colorant. The US FDA is currently reviewing its definition of the term "natural" on food labels, which may result in the use of colorants being restricted in natural cheeses. The objective of this study was to evaluate how consumers perceive Cheddar cheese color to better understand how changes to legislation surrounding colorants in natural Cheddar cheese may affect consumption. We were also interested in determining if a relationship exists between color and other perceived characteristics of Cheddar cheese. Two online surveys on Cheddar cheese color and flavor attributes (n = 1226 and n = 1183, respectively) were conducted, followed by a consumer acceptance test on 6 commercially available Cheddar cheeses (n = 196). Overall, consumers preferred light orange color in Cheddar cheese over dark orange or white Cheddar cheese, but segmentation was observed for Cheddar color preference. Light orange Cheddar and white Cheddar were perceived as approximately equal in terms of "naturalness." White and light orange Cheddars were perceived as more natural than dark orange Cheddars conceptually and in consumer acceptance testing. White Cheddar was considered most natural by 50.3% of n = 1283 survey participants and 43.4% of n = 196 consumer acceptance test participants, while light orange Cheddar was perceived as most natural by 40.6% and 45.9% of these groups respectively. A bimodal distribution was observed in both the online survey and in consumer acceptance testing for "naturalness" of Cheddar cheese color, with a subset of consumers (31.4% of n = 1183 survey participants and 30.6% of n = 196 consumer testing participants) indicating that white Cheddar was the least natural option. Consumers associated orange color in Cheddar cheese with more "sharp" flavor both in an online survey format and consumer acceptance testing.

Annatto-Enriched Egg Improves the Perception of Satiety in Healthy Adults-Randomized Clinical Trial: EGGANT Study.[Pubmed:38472844]

Foods. 2024 Feb 28;13(5):731.

Diet is one of the factors that prevents the development and death from cardiovascular diseases (CVD). It has been proposed that diets high in protein, which increase satiety, and with a high content of antioxidants, help reduce cardiovascular risk factors. The egg is one of the foods that produces greater satiety and provides antioxidants. In addition, due to its lipophilic matrix, it could improve the bioavailability of other dietary antioxidants such as Annatto. OBJECTIVE: This study evaluated the effects of egg and Annatto-enriched egg consumption on satiety markers and CVD risk factors in healthy adults from Colombia. METHODS: A parallel randomized clinical trial was conducted, where one hundred and five (n = 105) men and women, divided into three groups, consumed daily for 8 weeks: (a) two eggs (egg group), or (b) two eggs with Annatto (egg + Annatto group), or (c) two egg whites (placebo group). RESULTS: The three groups were similar in gender distribution. No significant changes were found over time (before vs. after) in any of the groups nor between the groups in anthropometric variables, physical activity, eating profile, and ghrelin as an objective marker of satiety. In the egg + Annatto group, subjective satiety increased (effect size 0.431; p < 0.05) after consumption. CONCLUSIONS: In healthy adults, the intake of two eggs, or two eggs with Annatto daily for 8 weeks, did not result in significant changes in ghrelin; but eggs with Annatto tend to increase the perception of satiety.

Tocotrienol isoforms: The molecular mechanisms underlying their effects in cancer therapy and their implementation in clinical trials.[Pubmed:38336507]

J Integr Med. 2024 Jan;22(1):1-11.

Tocotrienols are found in a variety of natural sources, like rice bran, Annatto seeds and palm oil, and have been shown to have several health-promoting properties, particularly against chronic diseases such as cancer. The incidence of cancer is rapidly increasing around the world, not only a result of continued aging and population growth, but also due to the adoption of aspects of the Western lifestyle, such as high-fat diets and low-physical activity. The literature provides strong evidence that tocotrienols are able to inhibit the growth of various cancers, including breast, lung, ovarian, prostate, liver, brain, colon, myeloma and pancreatic cancers. These findings, along with the reported safety profile of tocotrienols in healthy human volunteers, encourage further research into these compounds' potential use in cancer prevention and treatment. The current review provided detailed information about the molecular mechanisms of action of different tocotrienol isoforms in various cancer models and evaluated the potential therapeutic effects of different vitamin E analogues on important cancer hallmarks, such as cellular proliferation, apoptosis, angiogenesis and metastasis. MEDLINE/PubMed and Scopus databases were used to identify recently published articles that investigated the anticancer effects of vitamin E derivatives in various types of cancer in vitro and in vivo along with clinical evidence of adjuvant chemopreventive benefits. Following an overview of pre-clinical studies, we describe several completed and ongoing clinical trials that are paving the way for the successful implementation of tocotrienols in cancer chemotherapy.

Chromatic and medicinal properties of six natural textile dyes: A review of eucalyptus, weld, madder, annatto, indigo and woad.[Pubmed:38034740]

Heliyon. 2023 Nov 8;9(11):e22013.

The environmental and health-related impacts of synthetic dyes have led to growing interest in natural dyes as sustainable and eco-friendly alternatives. However, natural dyes have been used to dye textiles with limited color shade and poor fastness properties, and little research has been conducted in this field. Additionally, natural dyes also have the potential to provide added functionalities to textiles, such as antibacterial and anti-UV properties. A systematic literature review of 38 studies was conducted to analyze the use of six natural dyes derived from eucalyptus (Eucalyptus globulus Labill.), weld (Reseda luteola L.), madder (Rubia tinctorum L.), Annatto (Bixa Orellana L.), true indigo (Indigofera tinctoria L.) and woad (Isatis tinctoria L.). These dyes were selected after a preliminary analysis of studies on plant-based natural dyes with primary colors, considering their chromatic and potential medicinal properties. This study explores the influence of different dyeing parameters and auxiliary products in these properties. The research discussed how the chromatic and medicinal properties of natural dyes can be affected by various factors and provides a summary table with the chromatic palette possibilities according to the different materials and processes relationships. Exploring the combination of natural dyes with environmentally friendly auxiliary products can be a promising development area for creating a wide range of color shades. Further research is also needed to optimize the dyeing processes with natural dyes, towards more sustainable textile dyeing possibilities.

Comprehensive Update on Carotenoid Colorants from Plants and Microalgae: Challenges and Advances from Research Laboratories to Industry.[Pubmed:38002140]

Foods. 2023 Nov 10;12(22):4080.

The substitution of synthetic food dyes with natural colorants continues to be assiduously pursued. The current list of natural carotenoid colorants consists of plant-derived Annatto (bixin and norbixin), paprika (capsanthin and capsorubin), saffron (crocin), tomato and gac fruit lycopene, marigold lutein, and red palm oil (alpha- and beta-carotene), along with microalgal Dunaliella beta-carotene and Haematococcus astaxanthin and fungal Blakeslea trispora beta-carotene and lycopene. Potential microalgal sources are being sought, especially in relation to lutein, for which commercial plant sources are lacking. Research efforts, manifested in numerous reviews and research papers published in the last decade, have been directed to green extraction, microencapsulation/nanoencapsulation, and valorization of processing by-products. Extraction is shifting from conventional extraction with organic solvents to supercritical CO(2) extraction and different types of assisted extraction. Initially intended for the stabilization of the highly degradable carotenoids, additional benefits of encapsulation have been demonstrated, especially the improvement of carotenoid solubility and bioavailability. Instead of searching for new higher plant sources, enormous effort has been directed to the utilization of by-products of the fruit and vegetable processing industry, with the application of biorefinery and circular economy concepts. Amidst enormous research activities, however, the gap between research and industrial implementation remains wide.

Plant age-dependent dynamics of annatto pigment (bixin) biosynthesis in Bixa orellana.[Pubmed:37975812]

J Exp Bot. 2024 Feb 28;75(5):1390-1406.

Age affects the production of secondary metabolites, but how developmental cues regulate secondary metabolism remains poorly understood. The achiote tree (Bixa orellana L.) is a source of bixin, an apocarotenoid used in diverse industries worldwide. Understanding how age-dependent mechanisms control bixin biosynthesis is of great interest for plant biology and for economic reasons. Here we overexpressed miRNA156 (miR156) in B. orellana to comprehensively study the effects of the miR156-SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) module on age-dependent bixin biosynthesis in leaves. Overexpression of miR156 in Annatto plants (miR156ox) reduced BoSPL transcript levels, impacted leaf ontogeny, lessened bixin production, and increased abscisic acid levels. Modulation of expression of BoCCD4-4 and BoCCD1, key genes in carotenoid biosynthesis, was associated with diverting the carbon flux from bixin to abscisic acid in miR156ox leaves. Proteomic analyses revealed an overall low accumulation of most secondary metabolite-related enzymes in miR156ox leaves, suggesting that miR156-targeted BoSPLs may be required to activate several secondary metabolic pathways. Our findings suggest that the conserved BomiR156-BoSPL module is deployed to regulate leaf dynamics of bixin biosynthesis, and may create novel opportunities to fine-tune bixin output in B. orellana breeding programs.

Validation, Measurement Uncertainty, and Determination of Bixin and Norbixin in Processed Foods of Animal Resources Distributed in Korea.[Pubmed:37969328]

Food Sci Anim Resour. 2023 Nov;43(6):949-960.

This research aimed to validate a high-performance liquid chromatography method for the quantitative determination of bixin and norbixin in various foods. The Diode Array Detector (495 nm) technique was used. Method was validated for specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, and accuracy, and the measurement uncertainty was assessed. The calibration curve showed excellent linearity (r(2)>/=0.9999) over the tested concentration range of 0.2-25 mg/L. The LOD and LOQ were 0.03-0.11 and 0.02-0.05 mg/L for bixin and norbixin, respectively. The intra- and inter-day accuracies and precisions were 88.0+/-1.3-97.0+/-0.5% and 0.2%-2.6% relative SD (RSD) for bixin and 88.2+/-0.8-105.8+/-0.8% and 0.3%-2.7% RSD for norbixin, respectively. Inter-laboratory validation for accuracy and precision was conducted in three laboratories, and these results all met the AOAC guidelines. In addition, the relative expanded uncertainty (<22%) satisfied the CODEX recommendation. Furthermore, products distributed in Korea were monitored for Annatto extracts using the proposed method to demonstrate its application. The developed analytical method is reliable for quantifying bixin and norbixin in various foods.

Application of different concentrations of the natural dye potassium norbixinate (annatto) in polyamide 6.6 fabrics.[Pubmed:37950732]

Nat Prod Res. 2023 Nov 11:1-8.

Polyamide fabrics were dyed with concentrations ranging from 4% to 0.25% (o.w.f.) of the natural dye, potassium norbixinate (Annatto). The exhaustion, chromatic coordinates, colouristic intensity (K/S), and fastness to washing and rubbing were evaluated. The natural dye was characterised, and its maximum absorption peaks were identified at 452 nm and 482 nm through UV-vis scanning. Its main chemical groups were identified by FTIR-ATR. All dyeings exhibited high exhaustion percentage, with a maximum of 98.4% for 1% dye concentration. The dyed samples displayed visually appealing orange hues, with a maximum K/S value of 6.9. Most of the fastness test results were rated between 5 and 4/5, remaining within the standards established by most textile industries. Potassium norbixinate exhibited a similar tinctorial behaviour to synthetic acid dyes for polyamide, suggesting ionic chemical reaction interaction between dye and polyamide, highlighting the potential use in the textile industry.

Spatial variability of chemical indicators of Amazon agricultural soils through geomultivariate statistics, Brazil.[Pubmed:37682342]

Environ Monit Assess. 2023 Sep 8;195(10):1167.

This work focuses on evaluating the spatial variability of chemical attributes of soils under different agricultural use and native forest, indicating which are the possible indicator attributes of changes in environmental, through the use and management of the soil. The study was carried out in the southern region of the Amazonas state, in an Argissolo Vermelho-Amarelo (Ultisol). Sampling grids were established measuring: 90 m x 70 m with regular soil collection spacing of 10 m for the guarana and forest areas; 90 m x 56 m spaced at 10 m x 8 m for Annatto area; and 54 m x 42 m with spacing between points of 6 m for the cupuacu area, totaling 80 sampling points in each area, with soil samples collected at depths of 0.0-0.05; 0.05-0.10 m and 0.10-0.20 m. The following attributes were determined: pH, Al(3+), K(+), Ca(2+), Mg(2+), P, H + Al, CEC, V% and m%. Descriptive, geostatistical and multivariate statistical analyzes were performed. The results show that it is possible to state that the descriptive, geostatistical and multivariate statistical techniques were able to identify the difference between the spatial variability of the attributes according to each specific use of individual soils. The multivariate analysis made it possible to select the attributes that most contribute to the variability of these soils, and with that, it was found that the forest showed less spatial variability in the surface layer, with higher reach values by scaled semivariograms.

A Review of the Chemistry and Biological Activities of Natural Colorants, Dyes, and Pigments: Challenges, and Opportunities for Food, Cosmetics, and Pharmaceutical Application.[Pubmed:37471105]

Chem Biodivers. 2023 Aug;20(8):e202300561.

Natural pigments are important sources for the screening of bioactive lead compounds. This article reviewed the chemistry and therapeutic potentials of over 570 colored molecules from plants, fungi, bacteria, insects, algae, and marine sources. Moreover, related biological activities, advanced extraction, and identification approaches were reviewed. A variety of biological activities, including cytotoxicity against cancer cells, antioxidant, anti-inflammatory, wound healing, anti-microbial, antiviral, and anti-protozoal activities, have been reported for different pigments. Considering their structural backbone, they were classified as naphthoquinones, carotenoids, flavonoids, xanthones, anthocyanins, benzotropolones, alkaloids, terpenoids, isoprenoids, and non-isoprenoids. Alkaloid pigments were mostly isolated from bacteria and marine sources, while flavonoids were mostly found in plants and mushrooms. Colored quinones and xanthones were mostly extracted from plants and fungi, while colored polyketides and terpenoids are often found in marine sources and fungi. Carotenoids are mostly distributed among bacteria, followed by fungi and plants. The pigments isolated from insects have different structures, but among them, carotenoids and quinone/xanthone are the most important. Considering good manufacturing practices, the current permitted natural colorants are: Carotenoids (canthaxanthin, beta-carotene, beta-apo-8'-carotenal, Annatto, astaxanthin) and their sources, lycopene, anthocyanins, betanin, chlorophyllins, spirulina extract, carmine and cochineal extract, henna, riboflavin, pyrogallol, logwood extract, guaiazulene, turmeric, and soy leghemoglobin.

Dyes Used in Processed Meat Products in the Polish Market, and Their Possible Risks and Benefits for Consumer Health.[Pubmed:37444348]

Foods. 2023 Jul 6;12(13):2610.

Manufacturers are obliged to label processed meat products with information concerning the additives used and nutritional values. The aim of the study was to identify the dyes most frequently used in processed meat, evaluate their influence on specific food qualities, assess whether their use was correct and review their effect on health. The analysis was based on information on the labels and images of processed meat, and used a generalised linear model with a binary dependent variable. The risks and benefits for human health were defined based on the available literature. Twelve dyes were found to be used in the manufacture of processed meat. Carmine was found in 183 of 273 (67.03%) evaluated assortments containing dyes. The occurrence of water, flavourings and high fat and carbohydrate contents increased the chances that a dye would be present in a particular product. Unauthorised use of food additives was found in 20 products, with smoked meat products demonstrating the highest number of non-compliances. In general, the dyes used with food are considered safe; however, reservations are associated with the use of E150C and E150D caramels due to their potential carcinogenic effect, and carmine and Annatto due to their allergic effects.