The safety of annatto extracts (E 160b) as a food additive
2016; Wiley; Volume: 14; Issue: 8 Linguagem: Inglês
10.2903/j.efsa.2016.4544
ISSN1831-4732
Tópico(s)Pesticide Residue Analysis and Safety
ResumoEFSA JournalVolume 14, Issue 8 e04544 Scientific OpinionOpen Access The safety of annatto extracts (E 160b) as a food additive EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS), EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS)Search for more papers by this author EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS), EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS)Search for more papers by this author First published: 24 August 2016 https://doi.org/10.2903/j.efsa.2016.4544Citations: 5 Correspondence: fip@efsa.europa.eu Requestor: European Commission Question numbers: EFSA-Q-2008-395, EFSA-Q-2011-00362 Panel members: Fernando Aguilar, Riccardo Crebelli, Alessandro Di Domenico, Birgit Dusemund, Maria Jose Frutos, Pierre Galtier, David Gott, Ursula Gundert-Remy, Claude Lambré, Jean-Charles Leblanc, Oliver Lindtner, Peter Moldeus, Alicja Mortensen, Pasquale Mosesso, Dominique Parent-Massin, Agneta Oskarsson, Ivan Stankovic, Ine Waalkens-Berendsen, Rudolf Antonius Woutersen, Matthew Wright and Maged Younes Acknowledgements: The Panel wishes to thank the members of the Standing Working Group on the re-evaluation of food colours: Fernando Aguilar, Riccardo Crebelli, Alessandro Di Domenico, Maria Jose Frutos, Pierre Galtier, David Gott, Claude Lambré, Jean-Charles Leblanc, Agneta Oskarsson, Jeanne Stadler, Paul Tobback, Ine Waalkens-Berendsen and Rudolf Antonius Woutersen for the preparatory work on this scientific opinion and EFSA staff members: Andrea Altieri, Ana Rincon, Alexandra Tard and Stavroula Tasiopoulou for the support provided to this scientific opinion. The ANS Panel wishes to acknowledge all European competent institutions, Member State bodies and other organisations that provided data for this scientific opinion. Adopted: 29 June 2016 AboutSectionsPDF ToolsExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Abstract Following a request from the European Commission to EFSA, the EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS) was asked to re-evaluate the safety of annatto extracts, bixin-based and norbixin-based, when used as a food additive and to evaluate the safety of aqueous-processed bixin (Annatto E); solvent-extracted bixin (Annatto B); alkali-processed norbixin, acid-precipitated (Annatto F); alkali-processed norbixin, not acid-precipitated (Annatto G) and solvent-extracted norbixin (Annatto C) with the view to replace the currently authorised annatto extracts (E 160b). Given: (a) that read-across among the five bixin- and norbixin-based annatto extracts was feasible; (b) the availability of adequate 90-day toxicity studies with Annatto B, C, E and F; (c) the absence of concern for mutagenicity, carcinogenicity, reproductive and developmental toxicity of Annatto B, C, F and G, whereas the mutagenicity of Annatto E is equivocal, the Panel concluded that 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; solvent-extracted bixin (Annatto B), solvent-extracted norbixin (Annatto C), alkali-processed, acid-precipitated norbixin (Annatto F) and alkali-processed, not acid-precipitated norbixin (Annatto G and its norbixin salts) should comply with the specifications as recommended by the Panel. The toxicological database is sufficient to derive an acceptable daily intake (ADI) of 6 mg bixin/kg body weight (bw) per day and an ADI of 0.3 mg norbixin/kg bw per day. Exposure estimates for bixin were below the ADI for all population groups and for all refined exposure scenarios, including for the extension of use. For norbixin, exceedance was observed for the extension of use at the 95th percentile for some population groups. Summary Following a request from the European Commission to the European Food Safety Authority (EFSA), the EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS) was asked to deliver a scientific opinion re-evaluating the safety of annatto extracts (E 160b(i), (ii), (iii)) when used as a food additive. Furthermore, the ANS Panel was requested to evaluate the safety of five other annatto extracts, following the request of the Annatto Interest Group of the Natural Food Colours Association (NATCOL) for authorisation of these extracts, with the view to replace the currently authorised food additive (E 160b). These include the two bixin-based annatto extracts: aqueous-processed bixin (Annatto E) (≥ 25% colouring matter) and solvent-extracted bixin (Annatto B) (≥ 85% colouring matter), and the three norbixin-based annatto extracts: alkali-processed, acid-precipitated norbixin (Annatto F) (≥ 35% colouring matter), alkali-processed, not acid-precipitated norbixin (Annatto G) (≥ 15% colouring matter) and solvent-extracted norbixin (Annatto C) (≥ 85% colouring matter). The qualitative and quantitative composition of the non-pigment fraction of the annatto extracts proposed by the applicant revealed that the non-pigment fraction contains several well-known plant constituents, including proteins (≤ 6%), lignocelluloses (< 16%), fatty acids (≤ 4%; probably as oil), polyphenols (≤ 4%) and ash (0.1–12%). Unidentified terpenoids (up to 13.4%) were only present in Annatto E. The applicant indicated that results for the solvent-extracted bixin (Annatto B) and the alkali-processed (acid-precipitated) norbixin (Annatto F) would be applicable also to the solvent-extracted norbixin (Annatto C). Based on studies in rats and humans, the Panel concluded that bixin and norbixin are absorbed. In all studies, norbixin was the major component present in the plasma and urine, even following administration of bixin-based extracts, most likely derived from both the norbixin present in the extract, as well as metabolism of bixin to norbixin. Four of the annatto extracts (Annatto B, C, E and F) evaluated in the present opinion were tested in 90-day studies in rats. The administration of all the four test materials to rats for 13 weeks did not result in any treatment-related deaths, and the general condition and behaviour of the animals were not affected by treatment. Some toxicological effects were observed. The effects observed related generally to increases in liver and kidney weights, with some indications of impaired function at high-dose levels. No evidence of histopathological damage was observed in any tissue, except for hepatocellular necrosis at the two highest dose levels of solvent-extracted norbixin (Annatto C). With the alkali-processed, acid-precipitated norbixin (Annatto F), haematological changes were also observed. The studies with the solvent-extracted bixin (Annatto B) revealed an effect on the kidney, with raised protein concentrations noted in urine samples obtained from males receiving 50,000 mg/kg diet. Blood plasma phosphorus concentrations were also increased in these animals, indicating a possible reduction in the glomerular filtration rate. Studies with the aqueous-processed bixin (Annatto E) revealed increased thyroid and kidney weights. No-observed-adverse-effect level (NOAEL) values were identified by the Panel from these 90-day studies. These NOAEL values were in accordance with the NOEL values identified in these studies by the Joint Food and Agriculture Organization (FAO)/World Health Organization (WHO) Expert Committee on Food Additives (JECFA) in 2006. No conclusion can be drawn from published genotoxicity studies with water-soluble annatto extracts, which used not validated test methods and/or limited protocols. However, a set of unpublished properly performed in vitro and in vivo genotoxicity studies on Annatto B, E and F were also available to the Panel for evaluation. In these studies, all the three annatto extracts produced weakly positive or equivocal results in vitro, especially at high doses, with no consistent pattern of activity. As an in vivo follow-up, a micronucleus test in mouse bone marrow and a comet assay in rat stomach and liver, the main target tissues following oral exposure, were performed. In these studies, the oral administration of Annatto B, E and F up to the maximum recommended dose did not induce any detectable genetic damage in mouse bone marrow, in which, however, no evidence of exposure was obtained; clearly negative results were, however, also obtained with comet assays in rat stomach and, for Annatto B and F, in rat liver. An unexplained heterogenicity in response was instead observed in the liver of animals treated with Annatto E. Negative results were obtained for Annatto C (solvent-extracted norbixin) in a separate bacterial mutation assay. Overall, based on the available experimental results, and according to the EFSA Scientific Committee Recommendations on Genotoxicity Testing Strategy (2011), Annatto B and F (and following read-across, also Annatto C and G) were evaluated by the Panel as not genotoxic, whereas more information on the identity of the components of the extract and further testing were considered necessary to clarify the equivocal response elicited by Annatto E in rat liver. Annatto extracts (E 160b) of lower purity than those evaluated in the present opinion were without significant toxicity when administered in long-term studies to mice and rats. The applicant considered it appropriate to extrapolate the conclusions from these old carcinogenicity studies to the annatto extracts Annatto B, C, E, F and G. The Panel agreed with this assumption. Based on the absence of evidence for carcinogenic response, the Panel concluded that the annatto extracts were of no concern with respect to carcinogenicity. A prenatal developmental toxicity study revealed that oral gavage administration of the alkali-processed, acid-precipitated norbixin (Annatto F) during gestation from the time of implantation until just before delivery resulted in no treatment-related effects on the progress or outcome of pregnancy for female CD (Sprague–Dawley) rats. No adverse effects on development were observed after treatment of rats with the aqueous-processed bixin (Annatto E). Moreover, in a long-term study with the annatto extracts of lower purity, no adverse effects on reproduction and no teratogenic effects were seen in rats. 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. 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). For the refined exposure scenarios, the Panel calculated exposure estimates separately for bixin and norbixin, based on the information provided by industry. From the refined estimated exposure scenarios, mean exposure to bixin ranged from 0.001 mg/kg body weight (bw) per day in adults to 0.10 mg/kg bw per day in toddlers. The 95th percentile exposure to bixin ranged from 0.01 mg/kg bw per day in infants, children, adolescents, adults and the elderly to 0.32 mg/kg bw per day in toddlers. For norbixin, mean exposure ranged from 0.002 mg/kg bw per day in infants and the elderly to 0.11 mg/kg bw per day in toddlers. The 95th percentile exposure to norbixin ranged from 0.01 mg/kg bw per day in infants, adults and the elderly to 0.24 mg/kg bw per day in toddlers. 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. For bixin, from the extension of use scenario considering additional exposure from food categories and levels proposed by the applicant, mean exposure ranged from 0.004 mg/kg bw per day for infants to 0.33 mg/kg bw per day for toddlers. The 95th percentile ranged from 0.01 mg/kg bw per day in the elderly to 0.65 mg/kg bw per day in toddlers. For norbixin, from the extension of use scenario considering additional exposure from food categories and levels proposed by the applicant, mean exposure ranged from 0.003 mg/kg bw per day for infants to 0.24 mg/kg bw per day for toddlers. The 95th percentile ranged from 0.02 mg/kg bw per day for infants, adults and the elderly to 0.46 mg/kg bw per day in infants and toddlers. The Panel noted that raising the acceptable level for norbixin in the bixin-based annatto extract Annatto B from 2.5% to 5%, as proposed by the applicant, would result in an additional exposure to norbixin of up to 0.017 mg/kg bw per day (considering the extension of use scenario, 95th percentile in toddlers). Given: that read-across among the five bixin- and norbixin-based annatto extracts was feasible; the availability of adequate 90-day toxicity studies with annatto extracts B, C, E and F; the absence of concern for mutagenicity, carcinogenicity, reproductive and developmental toxicity of annatto extracts B, C, F and G, whereas the mutagenicity of Annatto E is equivocal, the Panel concluded that: 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 the new annatto extracts: solvent-extracted bixin (Annatto B), solvent-extracted norbixin (Annatto C), alkali-processed, acid-precipitated norbixin (Annatto F) and alkali-processed, not acid-precipitated norbixin (Annatto G) and its salts: – they should comply with the specifications as recommended by the Panel; – the toxicological database is sufficient to derive an acceptable daily intake (ADI) of 6 mg bixin/kg bw per day and an ADI of 0.3 mg norbixin/kg bw per day, applying an uncertainty factor of 200 to the NOAEL values derived from the 90-day studies (1,206 mg/kg bw and 63 mg/kg bw, respectively). Based on the reported current use levels provided by the industry, the Panel concluded that exposure estimates were below the ADI of 6 mg/kg bw per day for bixin and below the ADI of 0.3 mg/kg bw per day for norbixin for all population groups and for all refined exposure scenarios. Considering the extension of use for the additional 16 food categories, all refined exposure estimates for bixin were below the ADI of 6 mg/kg bw per day for all populations. For norbixin, the ADI of 0.3 mg/kg bw per day was not exceeded in the non-brand-loyal scenario and in the brand-loyal scenario at the mean. The only exceedance observed for norbixin was in the brand-loyal scenario at the 95th percentile for infants (in one country), toddlers (in three countries) and children (in one country). However, the Panel noted that this exceedance results from the overestimation of the contribution from at least one food category (i.e. unripened cheese). as regards Annatto E, due to the equivocal results obtained with the in vivo comet assay, the Panel could not conclude on its safety. The Panel recommended that the alkali-extracted annatto (E 160b(ii)), the oil-extracted annatto (E 160b(iii)) and the solvent-extracted bixin and norbixin (E 160b(i)), currently authorised in the EU, should be replaced by the solvent-extracted bixin (Annatto B); solvent-extracted norbixin (Annatto C); alkali-processed, acid-precipitated norbixin (Annatto F) and alkali-processed, not acid-precipitated norbixin (Annatto G). In addition, the Panel recommended that the specifications for bixin- and norbixin-based annatto extracts (E 160b) according to the Commission Regulation (EU) No 231/2012 should be replaced by the specifications for the annatto extracts (Annatto B, C, F and G) as given by JECFA (2007, 2015). However, the maximum limits for the impurities of toxic elements (arsenic, lead, mercury) should be revised in order to ascertain that the annatto extracts as food additives will not be a significant source of exposure to these toxic elements in foods. Moreover, the Panel recommended that a maximum limit for cadmium should also be included in the specifications. Background as provided by the European Commission Regulation (EC) No 1333/20081 of the European Parliament and of the Council on food additives requires that food additives are subject to a safety evaluation by the European Food Safety Authority (EFSA) before they are permitted for use in the European Union (EU). In addition, it is foreseen that food additives must be kept under continuous observation and must be re-evaluated by EFSA. For this purpose, a programme for the re-evaluation of food additives that were already permitted in the EU before 20 January 2009 has been set up under Regulation (EU) No 257/20102. This Regulation also foresees that food additives are re-evaluated whenever necessary in the light of changing conditions of use and new scientific information. For efficiency and practical purposes, the re-evaluation should, as far as possible, be conducted by group of food additives according to the main functional class to which they belong. 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 EU3 of 2001. The report 'Food additives in Europe 2000' submitted by the Nordic Council of Ministers to the Commission, provides additional information for the prioritisation of additives for re-evaluation. As colours were among the first additives to be evaluated, these food additives should be re-evaluated with the highest priority. In 2003, the Commission already requested EFSA to start a systematic re-evaluation of authorised food additives. However, as a result of adoption of Regulation (EU) 257/2010, the 2003 Terms of References are replaced by those below. Annex II to Regulation (EC) No 1333/2008 authorises the use of annatto, bixin, norbixin (E 160b) in certain food categories only. Commission Regulation (EU) No 231/2012 laying down specifications for food additives specifies three annatto products (i) solvent-extracted bixin and norbixin, (ii) alkali-extracted annatto and (iii) oil-extracted annatto. The SCF has previously evaluated the annatto extracts, the last time, in 1979 when the SCF allocated an acceptable daily intake (ADI) of 2.5 mg/kg body weight (bw) for an annatto extract (containing 2.6% carotenoid expressed as bixin) equivalent to 0–0.065 mg/kg bw for carotenoids of annatto expressed as bixin. The Joint Food and Agriculture Organization (FAO)/World Health Organization (WHO) Expert Committee on Food Additives (JECFA) re-evaluated six annatto extracts on the basis of new data in 2003 and 2006. An ADI for bixin of 0–12 mg/kg bw and a group ADI for norbixin and its disodium and dipotassium salts of 0–0.6 mg/kg bw (expressed as norbixin) were established at the 67th JECFA (2006). Five specifications have been established for the annatto extracts covered by the established ADIs, but only tentative specifications were established for the oil-processed bixin for which no ADI has been established. JECFA requested data on the non-colouring fraction of the oil-processed bixin extract by the end of 2008. The Natural Food Colours Association (NATCOL)/Annatto Interest Group requested the authorisation of five annatto extracts categorised as bixin- or norbixin-based, with the view to replacing the currently authorised annatto extracts (E 160b). NATCOL's application covers the five annatto extracts for which JECFA has established full ADIs: (i) aqueous-processed annatto extracts, bixin-based, (ii) solvent-processed annatto extracts, bixin-based, (iii) alkali-processed, acid-precipitated annatto extracts, norbixin-based, (iv) alkali-processed, not acid-precipitated annatto extracts, norbixin-based and (v) solvent-processed annatto extracts, norbixin-based. Terms of Reference as provided by the European Commission The Commission asks EFSA to re-evaluate the safety of food additives already permitted in the Union before 2009 and to issue scientific opinions on these additives, taking especially into account the priorities, procedures and deadlines that are enshrined in the Regulation (EU) No 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. In accordance with Article 29 (1) (a) of Regulation (EC) No 178/2002, the European Commission asks EFSA to provide a scientific opinion on the safety of the annatto extracts covered by this application as a food colour for the uses proposed by the applicant. Interpretation of Terms of Reference The Panel noted that the Terms of Reference include raising the acceptable level for norbixin in the bixin-based annatto extract Annatto B from 2.5% to 5%, and the extension of use in 16 additional food categories. Assessment 1 Introduction 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. Furthermore, at the request of the European Commission, the present opinion evaluated the safety of five annatto extracts categorised as bixin-based (Annatto E and B) or norbixin-based (Annatto C, F and G), with the view to replace the currently authorised annatto extracts (E 160b) by the latter five. In 1974, JECFA established a temporary ADI of 1.25 mg/kg bw for the sum of bixin and norbixin (expressed as bixin) (JECFA, 1975), which was endorsed by the EU SCF in 1975 (SCF, 1975). In 1978, the SCF assessed the results of pharmacokinetic and metabolism studies of annatto extracts in rats following short- and long-time exposure and reviewed the results of acute metabolic studies in man. The Committee allocated an ADI of 0–2.5 mg/kg bw per day for annatto extracts containing 2.6% carotenoids expressed as bixin, equivalent to 0–0.065 mg/kg bw per day of carotenoids, expressed as bixin (SCF, 1979). At its 26th meeting in 1982, JECFA allocated an ADI of 0–0.065 mg/kg bw 'in terms of the carotenoid content expressed as bixin' (JECFA, 1982). The ADIs of the SCF (1979) and JECFA (1982) were both based on a long-term rat study performed with well-defined annatto extracts, which varied in total bixin content from 0.2% to 2.6%. Within the food colour legislation of the EU (Commission Regulation (EU) No 231/20124 laying down specifications for food additives), three different annatto extracts are described: (i) solvent-extracted bixin and norbixin (E 160b(i)); (ii) alkali-extracted annatto (E 160b(ii)); and (iii) oil-extracted annatto (E 160b(iii)). In 2006, JECFA agreed upon two separate ADIs for bixin- and norbixin-containing annatto extracts: an ADI of 12 mg/kg bw for bixin (92% bixin pure) and a group ADI of 0.6 mg/kg bw for norbixin (91.6% norbixin pure) and its sodium and potassium salts (JECFA, 2007). The request of the NATCOL/Annatto Interest Group to evaluate five annatto extracts categorised as bixin- or norbixin-based, with the view to replacing the currently authorised annatto extracts (E 160b) is included in the present opinion. The application by NATCOL covers the five annatto extracts for which JECFA has established full ADIs (JECFA, 2007): (i) aqueous-processed bixin (Annatto E); (ii) solvent-extracted bixin (Annatto B); (iii) alkali-processed norbixin, acid-precipitated (Annatto F); (iv) alkali-processed norbixin, not acid-precipitated (Annatto G); and (v) solvent-extracted norbixin (Annatto C). The Panel based its evaluation on the dossier submitted by the applicant, previous evaluations and reviews, additional literature that came available since then and the data available following public calls for data.5–7 The Panel noted that not all original studies on which previous evaluations were based were available for re-evaluation by the Panel. 2 Technical data 2.1 Identity of the substance The food colour annatto extracts (E 160b) is obtained from the outer layer (pericarp) – specifically, the outer coating – of the seeds of the tropical tree Bixa orellana. The tree is indigenous to Central and South America, where its seeds are used as a spice in traditional cooking. 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). In Brazil, Mexico and other Latin American countries, as well as in Asia (the Philippines), substantial quantities of processed annatto seeds are sold in retail outlets, often blended with other ingredients for addition to soups and meat dishes similar to the use of paprika seasonings in Europe. These condiments are known locally under various names; like paprika preparations, they impart not only colour, but also a distinct flavour. The principle pigment in the seeds is the carotenoid cis-bixin; processing may involve aqueous alkaline hydrolysis with simultaneous production of cis-norbixin (Figure 1). Annatto seeds contain cis-bixin (> 80% of the total carotenoid content), mainly in the 9′-cis configuration, with small quantities of trans-bixin and norbixin. In annatto seeds, the amount of norbixin is typically below 5% of the total colouring matter (Doc. provided to EFSA n. 3). Under specific conditions of temperature and pH, bixin can be hydrolysed into norbixin – the dicarboxylic acid – which can readily be turned into its sodium or potassium salt (Figure 1). Figure 1Open in figure viewerPowerPoint Structural formulas of cis-bixin and cis-norbixin, the latter also being used as the disodium or dipotassium salt. Position C-9′, eventually referred to in the text and coming from a different carbon atom numbering of the apocarotenoid structure, corresponds to position C-17 in the illustration above A common chemical name for cis-bixin is methyl (9′-cis)-hydrogen-6,6′-diapo-ψ,ψ-carotenedioate; its molecular formula and molecular weight are C25H30O4 and 394.50 g/mol. 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). Their molecular formulas and molecular weights (g/mol) are: C24H28O4 and 380.48 (acid); C24H26K2O4 and 456.66 (dipotassium salt); C24H26Na2O4 and 424.44 (disodium salt). 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. Table 1. CAS Registry and EINECS list numbers identifying the annatto extract principles of interest in the present opinion Substance CAS registry no. EINECS list no. cis-Bixin 6983-79-5 230-248-7 trans-Bixin 39937-23-0 – cis-Norbixin (free acid) 626-76-6 – cis-Norbixin, dipotassium salt – – cis-Norbixin, disodium salt – – trans-Norbixin (free acid) 542-40-5 208-810-8 trans-Norbixin, dipotassium salt 33261-80-2 251-431-8 trans-Norbixin, disodium salt 33261-81-3 251-432-3 CAS: Chemical Abstracts Service; EINECS: European Inventory of Existing Commercial chemical Substances. The CAS Registry and EINECS Nos for annatto are 1393-63-1 and 215-735-4, respectively. Annatto seed extracts are identified by CAS Registry and EINECS Nos 89957-43-7 and 289-561-2, respectively. A few synonyms, trade names and abbreviations for annatto extracts are: Achiote; Annatto; Annatto colour; Annatto extract; Annatto pigment; C.I. 75120; C.I. Natural Orange 4; FEMA No 2103; FEMA No 2103 (annatto extract); FEMA No 2104 (annatto seed); Orlean; Rocou or Roucou; Terre orellana; INS 160b; Urucum (JECFA, 2007, 2015; Doc. provided to EFSA n. 3; Smith and Wallin, 2006). The structural difference between bixin and norbixin – namely, an esterified carboxyl group – leads to appreciable differences in the physicochemical properties of the two molecules. Bixin- and norbixin-based annatto extracts are substantially insoluble in pure water and slightly soluble in ethanol; norbixin-based annatto extracts are soluble in alkaline water, but norbixin will precipitate in
Referência(s)