# Commission Regulation (EU) 2020/771 of 11 June 2020 amending Annexes II and III to Regulation (EC) No 1333/2008 of the European Parliament and of the Council and the Annex to Commission Regulation (EU) No 231/2012 as regards the use of Annatto, Bixin, Norbixin (E 160b) (Text with EEA relevance), Division: E 160 b (ii) ANNATTO NORBIXIN Extent: E+W+S+N.I. Synonyms Annatto C, Orlean, Terre orellana, L. Orange, CI Natural Orange 4 Definition Solvent-extracted norbixin is obtained from the outer coating of the seeds of the annatto tree (Bixa orellana L.) by washing with one or more of the following food grade solvents: acetone, methanol, hexane, ethanol, isopropyl alcohol, ethyl acetate, alkaline alcohol or supercritical carbon dioxide followed by solvent removal, crystallization and drying. Aqueous alkali is added to the resultant powder, which is then heated to hydrolyse the colouring matter and cooled. The aqueous solution is filtered, and acidified to precipitate the norbixin. The precipitate is filtered, washed, dried and milled, to give a granular powder. Solvent-extracted norbixin contains several coloured components; the major colouring principle is cis-norbixin, a minor colouring principle is trans-norbixin; thermal degradation products of norbixin may also be present as a result of processing. Colour Index No 75120 Einecs 208-810-8 Chemical name cis-Norbixin: 6,6'-Diapo-Ψ,Ψ-carotenedioic acid cis-Norbixin dipotassium salt: Dipotassium 6,6'-diapo-Ψ,Ψ-carotenedioate cis-Norbixin disodium salt: Disodium 6,6'-diapo-Ψ,Ψ-carotenedioate Chemical formula cis-Norbixin: C24H28O4 cis-Norbixin dipotassium salt: C24H26K2O4 cis-Norbixin disodium salt: C24H26Na2O4 Molecular weight 380,5 (acid), 456,7 (dipotassium salt), 424,5 (disodium salt) Assay Not less than 85 % colouring matter (expressed as norbixin) E1 % 1cm 2870 at ca. 482 nm in 0,5 % potassium hydroxide solution Description Dark red-brown to red-purple powder Identification Solubility Soluble in alkaline water, slightly soluble in ethanol Spectrometry The sample in 0,5 % potassium hydroxide solution shows absorbance maxima at about 453 nm and 482 nm Purity Residual Solvents Acetone: Not more than 30 mg/kg Methanol: Not more than 50 mg/kg Hexane: Not more than 25 mg/kg Ethanol: Isopropyl alcohol: Ethyl acetate: not more than 50 mg/kg, singly or in combination Arsenic Not more than 2 mg/kg Lead Not more than 1 mg/kg Mercury Not more than 1 mg/kg Cadmium Not more than 0,5 mg/kg Synonyms Annatto F, Orlean, Terre orellana, L. Orange, CI Natural Orange 4 Definition Alkali-processed norbixin (acid-precipitated) is prepared by the extraction of the outer coating of the seeds of the annatto tree (Bixa orellana L.) with aqueous alkali. The bixin is hydrolysed to norbixin in hot alkaline solution and is acidified to precipitate the norbixin. The precipitate is filtered, dried and milled to give a granular powder. Alkali-processed norbixin contains several coloured components; the major colouring principle is cis-norbixin, a minor colouring principle is trans-norbixin; thermal degradation products of norbixin may also be present as a result of processing. Colour Index No 75120 Einecs 208-810-8 Chemical name cis-Norbixin: 6,6'-Diapo-Ψ,Ψ-carotenedioic acid cis-Norbixin dipotassium salt: Dipotassium 6,6'-diapo-Ψ,Ψ-carotenedioate cis-Norbixin disodium salt: Disodium 6,6'-diapo-Ψ,Ψ-carotenedioate Chemical formula cis-Norbixin: C24H28O4 cis-Norbixin dipotassium salt: C24H26K2O4 cis-Norbixin disodium salt: C24H26Na2O4 Molecular weight 380,5 (acid), 456,7 (dipotassium salt), 424,5 (disodium salt) Assay Not less than 35 % colouring matter (expressed as norbixin) E1 % 1cm 2870 at ca. 482 nm in 0,5 % potassium hydroxide solution Description Dark red-brown to red-purple powder Identification Solubility Soluble in alkaline water, slightly soluble in ethanol Spectrometry The sample in 0,5 % potassium hydroxide solution shows absorbance maxima at about 453 nm and 482 nm Purity Arsenic Not more than 2 mg/kg Lead Not more than 1 mg/kg Mercury Not more than 1 mg/kg Cadmium Not more than 0,5 mg/kg Synonyms Annatto G, Orlean, Terre orellana, L. Orange, CI Natural Orange 4 Definition Alkali-processed norbixin (not acid-precipitated) is prepared by the extraction of the outer coating of the seeds of the annatto tree (Bixa orellana L.) with aqueous alkali. The bixin is hydrolysed to norbixin in hot alkaline solution. The precipitate is filtered, dried and milled to give a granular powder. Extracts contain mainly the potassium or sodium salt of norbixin as the major colouring matter. Alkali-processed norbixin (not acid-precipitated) contains several coloured components; the major colouring principle is cis-norbixin, a minor colouring principle is trans-norbixin; thermal degradation products of norbixin may also be present as a result of processing. Colour Index No 75120 Einecs 208-810-8 Chemical name cis-Norbixin: 6,6'-Diapo-Ψ,Ψ-carotenedioic acid cis-Norbixin dipotassium salt: Dipotassium 6,6'-diapo-Ψ,Ψ-carotenedioate cis-Norbixin disodium salt: Disodium 6,6'-diapo-Ψ,Ψ-carotenedioate Chemical formula cis-Norbixin: C24H28O4 cis-Norbixin dipotassium salt: C24H26K2O4 cis-Norbixin disodium salt: C24H26Na2O4 Molecular weight 380,5 (acid), 456,7 (dipotassium salt), 424,5 (disodium salt) Assay Not less than 15 % colouring matter (expressed as norbixin) E1 % 1cm 2870 at ca. 482 nm in 0,5 % potassium hydroxide solution Description Dark red-brown to red-purple powder Identification Solubility Soluble in alkaline water, slightly soluble in ethanol Spectrometry The sample in 0,5 % potassium hydroxide solution shows absorbance maxima at about 453 nm and 482 nm Purity Arsenic Not more than 2 mg/kg Lead Not more than 1 mg/kg Mercury Not more than 1 mg/kg Cadmium Not more than 0,5 mg/kg’ --- Source: https://www.legislation.gov.uk/eur/2020/771/annex/II/division/2 (the official version) Page: https://openlegislation.uk/eu-regulation-2020-771/annex-II/division-2 Contains public sector information licensed under the Open Government Licence v3.0. Source: legislation.gov.uk, The National Archives.