Both products are soluble in ethyl ether and oils. E160b Annatto, bixin, norbixin. Available online: http://www.efsa.europa.eu/en/dataclosed/call/140324 The Panel noted that not all original studies on which previous evaluations were based were available for re‐evaluation by the Panel. Published 23 March 2014. In the absence of any macroscopic or histopathological changes noted in the kidneys, the Panel considered these findings to be of only minor, if any toxicological importance. In the review by Scotter (2009), several other methods are also mentioned. The Panel concluded that the changes in renal function observed at the two highest dose levels, point to a NOAEL of 1,000 mg Annatto F/kg diet, equivalent to 79 mg Annatto F/kg bw per day in males and 86 mg Annatto F/kg bw per day in females, corresponding to 30 mg norbixin/kg bw per day in males and 33 mg norbixin/kg bw per day in females. Blood plasma phosphorus concentrations were also increased in these animals, indicating a possible reduction in the glomerular filtration rate. Based on the manufacturing processes described for bixin‐based and norbixin‐based annatto extracts (Figure 2), the following read‐across approaches were proposed by the applicant (NATCOL, 2015). (2002). The Panel noted that according to the EU specifications for the food additive annatto, bixin, norbixin, (E 160b), impurities of the toxic elements arsenic, lead, mercury and cadmium are accepted up to a concentration of 3, 2, 1 and 1 mg/kg, respectively. The mutagenicity was decreased or even abolished in presence of S9, and was observed only at high doses, which were associated with visible precipitation of the dye. The authors noted that a minimum of 0.06 g pure trans‐bixin/kg bw was required to cause toxicity (a diabetogenic effect) in the dog, which was equivalent to 4.2 g of pure trans‐bixin (or 63 g of the crude extract) in a 70‐kg (possibly undernourished) man. 01.7.2 Ripened cheese, only ripened orange, yellow and broken‐white cheese, and red and green pesto cheese; 02.1 Fats and oils essentially free from water (excluding anhydrous milkfat), only fats; 15.1 Potato‐, cereal‐, flour‐ or starch‐based snacks, only extruded or expanded savoury snack products/excluding extruded or expanded savoury snack products; 15.2 Processed nuts, only savoury‐coated nuts. Overall, based on the read‐across from Annatto F to Annatto G and from Annatto E to Annatto B and Annatto C, and considering the results of older studies with annatto preparations of low purity, the Panel concluded that no adverse effects on reproduction or development were to be expected from the five annatto extracts described in the present opinion.  They are commonly found on food labels throughout the European Union. Up to 2.5 mg/kg body weight for annatto, and 0.065 mg/kg body weight for bixin. To assess the dietary exposure to annatto (E 160b) from its use as a food additive, the exposure was calculated based on (1) maximum permitted levels (MPL) set out in the European Union (EU) legislation (defined as the regulatory maximum level exposure assessment scenario); (2) the reported use levels (defined as the refined exposure assessment scenario); and (3) use levels proposed by the applicant for the extension of use (defined as extension of use scenario). The presence of several carotenoids, fatty acids esters and resins with a bitter taste was reported in the 1970s by Hager (Hager, 1972). The controls (Group 1) received untreated diet for 4 weeks. E160d. Commission Regulation (EU) 257/2010 of 25 March 2010 setting up a programme for the re‐evaluation of approved food additives in accordance with Regulation (EC) No 1333/2008 of the European Parliament and of the Council on food additives. For Annatto C, only a negative bacterial mutation assay was available, whereas no data were available for Annatto G. However, the Panel considered that, as Annatto C is derived from Annatto B, and Annatto G has a similar composition (on a dry weight basis) to Annatto F, read‐across from Annatto B and Annatto F to Annatto C and Annatto G, respectively, was possible, and that the latter could be considered not to raise a genotoxic concern either. Its scent is described as "slightly peppery with a hint of nutmeg" and flavor as "slightly nutty, sweet and peppery". There were also other manufacturing methods used previously (e.g. The Panel noted that no evidence of bone marrow exposure, highlighted by the altered proportion of young to mature erythrocytes, was achieved in these studies. Haematological and biochemical examination and necropsy were performed on 10 rats of each sex on day 29, and on the remaining five rats of each sex after recovery at day 43. Annatto extracts. The alkali‐processed, acid‐precipitated norbixin (Annatto F) induced a marginal increase in chromosomal aberrations, just above the 95th percentile of the historical control range, only in the presence of S9 at the two highest tested doses (8 and 12 μg/mL). the safety of the currently authorised solvent‐extracted bixin and norbixin (E 160b(i)), alkali‐extracted annatto (E 160b(ii)) and oil‐extracted annatto (E 160b(iii)), with the specifications defined in Commission Regulation (EU) No 231/2012, could not be assessed due to the lack of data, both in terms of identification and toxicological studies; as regards Annatto E, due to the equivocal results obtained with the. provided to EFSA n. 4). Dose levels were chosen based on a preliminary 2‐week range‐finding study (Aoki, 2001). NATCOL (Natural Food Colours Association), 2008. Studies with the aqueous‐processed bixin (Annatto E) revealed increased thyroid and kidney weights. Specifications for annatto extracts (E 160b) have been defined in Commission Regulation (EU) No 231/2012 laying down specifications for food additives (Table 3). The applicant has requested the extension of use of bixin‐ and norbixin‐based annatto extracts in 16 additional food categories, of which 15 uses for bixin‐based and seven for norbixin‐based annatto extracts. In 2003, the Commission already requested EFSA to start a systematic re‐evaluation of authorised food additives. However, as the annatto extracts were given in combination with other food colours, it was not always possible to know whether the observed effects were due to the annatto extracts or to one of the other colours present in the challenging material (Mikkelsen et al., 1978; Juhlin, 1981; Vien et al., 1987; Young et al., 1987; Fuglsang et al., 1993, 1994). CAS: Chemical Abstracts Service; EINECS: European Inventory of Existing Commercial chemical Substances. The present opinion deals with the re‐evaluation of the safety of annatto extracts (E 160b), when used as a food additive and permitted in the EU before 2009. For the analysis of several samples of extruded corn snack food, De Oliveira and Mercadante (2004) described a method involving enzymatic digestion with α‐amylase at room temperature of the ground sample (pretreatment), followed by multiple extractions of the aqueous phase with ethyl acetate. Based on the information made available by industry, separate exposure estimates for bixin‐ and norbixin‐based extracts were calculated (except for the regulatory maximum level exposure assessment scenario based on the MPLs). Aqueous‐processed bixin (Annatto E) is prepared by removal of the outer coating of the seeds of the annatto tree by abrading them in the presence of cold, mildly alkaline water (alkalised with potassium or sodium hydroxide). Yearly world production of annatto seeds was estimated to be 14,500 metric tonnes (dry weight) in the late 1990s: important producers of the seeds are Brazil, Guatemala, India, Peru, the Philippines and a few African countries (Smith and Wallin, 2006; Raghavan, 2007; Giridhar and Parimalan, 2010). According to the study authors, treatment with CE produced a significant increase in the proportion of micronucleated polychromatic erythrocytes. The pertinent Chemical Abstracts Service (CAS) and European Inventory of Existing Commercial chemical Substances (EINECS) (EC) number identifiers for the cis and trans configurations of bixin and norbixin are presented in Table 1. In addition, slightly reduced urinary output was seen in males receiving 3,000 or 9,000 mg/kg diet, and slightly low pH values were recorded for males receiving 9,000 mg/kg diet. Preliminary toxicity study by dietary administration to CD rats for 4 weeks, Annatto E. Preliminary Toxicity Study by Dietary Administration to CD Rats for 4 Weeks, Annatto F Preliminary toxicity study by dietary administration to CD rats for 4 weeks, Annatto G. Preliminary toxicity study by dietary administration to CD rats for 4 weeks, Annatto B. What are food additive E numbers? Similarly, a common chemical name for cis‐norbixin is (9′‐cis)‐6,6′‐diapo‐ψ,ψ‐carotenedioic acid; cis‐norbixin also occurs as the dipotassium or disodium salt (dipotassium or disodium 6,6′‐diapo‐ψ,ψ‐carotenedioate). Mechanical abrasion using food grade vegetable oil … The order of priorities for the re‐evaluation of the currently approved food additives should be set on the basis of the following criteria: the time since the last evaluation of a food additive by the Scientific Committee on Food (SCF) or by EFSA, the availability of new scientific evidence, the extent of use of a food additive in food and the human exposure to the food additive taking also into account the outcome of the Report from the Commission on Dietary Food Additive Intake in the EU33 Origin: Natural colour isolated from the seeds of the annatto tree (Bixa orellana ).Annatto is the name of the crude extract, whereas bixin is the fat-soluble colour and norbixin the water-soluble colour No treatment‐related histopathological lesions were seen in any of the organs examined. Mean recoveries obtained from spiked food samples of 12 different food matrices spiked with annatto at levels up to 27.7 mg/kg and ranged from 61% to 96%; the LOQ was reported to be 0.1 mg/kg (LOD, 0.01 mg/kg). Mitochondrial damage in the liver and pancreas was produced with the purified material. curry powder, tandoori), pickles, relishes, chutney, piccalilli, From 12 weeks up to and including 11 months of age, Bulgaria, Denmark, Finland, Germany, Italy, United Kingdom, From 12 up to and including 35 months of age, Belgium, Bulgaria, Denmark, Germany, Spain, Finland, United Kingdom, Italy, Netherlands, From 36 months up to and including 9 years of age, Austria, Belgium, Bulgaria, Czech Republic, Germany, Denmark, Spain, Finland, France, United Kingdom, Greece, Italy, Latvia, Netherlands, Sweden, From 10 up to and including 17 years of age, Austria, Belgium, Cyprus, Czech Republic, Germany, Denmark, Spain, Finland, France, United Kingdom, Italy, Latvia, Netherlands, Sweden, From 18 up to and including 64 years of age, Austria, Belgium, Czech Republic, Germany, Denmark, Spain, Finland, France, United Kingdom, Hungary, Ireland, Italy, Latvia, Netherlands, Romania, Sweden, Austria, Belgium, Germany, Denmark, Finland, France, United Kingdom, Hungary, Ireland, Italy, Netherlands, Romania, Sweden, Consumption data: different methodologies/representativeness/underreporting/misreporting/no portion size standard, Use of data from food consumption survey of a few days to estimate long‐term (chronic) exposure for high percentiles (95th percentile), Correspondence of reported use levels to the food items in the EFSA Comprehensive Database: uncertainties to which types of food the levels refer to, Food categories selected for the exposure assessment: exclusion of food categories due to missing FoodEx linkage (n = 6), Food categories selected for the exposure assessment: inclusion of food categories without considering the restrictions/exceptions (n = 4 for currently authorised uses and n =2 for the extension of use), Concentration data: levels considered applicable to all food items within the entire food category, Uncertainty in possible national differences in use levels of food categories, Alkali‐processed, acid‐precipitated norbixin (Annatto F), 0.1% solution of annatto extract in vegetable oil, 0.02% suspension of annatto extract in vegetable oil, 0.1% water soluble preparation of annatto extract, Alkali‐processed norbixin (acid‐precipitated) (Annatto F), Decorations, coatings and fillings, except fruit‐Based fillings covered by category 4.2.4, Desserts excluding products covered in category 1, 3 and 4, Butter and concentrated butter and butter oil and anhydrous milkfat, Salads and savoury‐based sandwich spreads, Dietary foods for special medical purposes defined in Directive 1999/21/EC (excluding products from food category 13.1.5), Processed foods not covered by categories 1–17, excluding foods for infants and young children, Non‐individually wrapped chocolate pieces, Other fat and oil emulsions including spreads as defined by Regulation (EC) No 1234/2007 and liquid emulsions, Fruit and vegetable preparations excluding compote (fruit preparations for use in desserts/yogurts), Meat preparations as defined by Regulation (EC) No 853/2004 (breakfast sausages with a minimum cereal content of 6%; burger meat with a minimum vegetable and/or cereal content of 4%), Heat‐treated processed meat (sausages, pâtés and terrines), Processed fish and fishery products including molluscs and crustaceans − only surimi and similar products and salmon substitutes, Seasonings and condiments (seasonings (e.g. In line with the extracts tested toxicologically, a minimum assay value of 0.2% total carotenoids was set. In rats dosed with OSB, the blood level was 62 mg/100 mL after 3 h and 19 mg/100 mL after 24 h. Thus, OSB was metabolised less rapidly than WSA, presumably because the OSB pigments would need to be metabolised to water‐soluble pigments, similar to those of WSA, before being metabolised further. The annatto pigments in rat's blood were determined 3 h and 24 h after treatment. Overall, 16 out of 22 food categories were considered in the present exposure assessment for the current permitted uses of the annatto extracts (E 160b): six were not taken into account in the exposure assessment because these and/or their relative restrictions/exceptions are not referenced in the EFSA Comprehensive Database; four food categories were included in the exposure assessment without considering their restrictions/exceptions. The chemical properties of the colouring principles bixin and norbixin, lead to a solubility profile that distinguishes annatto extracts from other carotenoids (Freund et al., 1988; Collins, 1992; Levy and Rivadeneira, 2000). Therefore, the Panel concluded that the NOAEL in this study for both males and females is 10,000 mg Annatto E/kg diet. The Mintel's Global New Products Database (GNPD) is an online database which monitors product introductions in consumer packaged goods markets worldwide. 184.108.40.206 Food categories considered for the exposure assessment of E 160b. For norbixin, from the extension of use scenario considering additional exposure from food categories and levels proposed by the applicant, in the brand‐loyal scenario, mean exposure ranged from 0.005 mg/kg bw per day for infants to 0.24 mg/kg bw per day for toddlers. 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