ANNEX IISupplementary provisions
ANNEX II In the Annex to Regulation (EU) No 231/2012, the respective entries for E 160b Annatto, Bixin, Norbixin: (i) Solvent-extracted bixin and norbixin, (ii) alkali extracted annatto and (iii) oil extracted annatto are replaced by the following: ‘E 160 b (i) ANNATTO BIXIN (I) SOLVENT-EXTRACTED BIXIN Synonyms Annatto B, Orlean, Terre orellana, L. Orange, CI Natural Orange 4 Definition Solvent-extracted bixin is obtained by the extraction of the outer coating of the seeds of the annatto tree ( Bixa orellana L.) with one or more of the following food grade solvents: acetone, methanol, hexane, ethanol, isopropyl alcohol, ethyl acetate, alkaline alcohol or supercritical carbon dioxide. The resulting preparation may be acidified, followed by the removal of the solvent, drying and milling. Solvent-extracted bixin contains several coloured components; the major colouring principle is cis-bixin, a minor colouring principle is trans-bixin; thermal degradation products of bixin may also be present as a result of processing. Colour Index No 75120 Einecs 230-248-7 Chemical name cis-Bixin: Methyl (9-cis)-hydrogen-6,6'-diapo-Ψ,Ψ-carotenedioate Chemical formula cis-Bixin: C 25 H 30 O 4 Molecular weight 394,5 Assay Not less than 85 % colouring matter (expressed as bixin) E 1 % 1cm 3090 at ca. 487 nm in tetrahydrofuran and acetone Description Dark red-brown to red-purple powder Identification Solubility Insoluble in water, slightly soluble in ethanol Spectrometry The sample in acetone shows absorbance maxima at about 425, 457 and 487 nm Purity Norbixin Not more than 5 % of total colouring matters 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 (II) AQUEOUS-PROCESSED BIXIN Synonyms Annatto E, Orlean, Terre orellana, L. Orange, CI Natural Orange 4 Definition Aqueous-processed bixin is prepared by the extraction of the outer coating of the seeds of the annatto tree ( Bixa orellana L.) by abrading the seeds in the presence of cold, mildly-alkaline water. The resultant preparation is acidified to precipitate bixin which is then filtered, dried and milled. Aqueous-processed bixin contains several coloured components; the major colouring principle is cis-bixin, a minor colouring principle is trans-bixin; thermal degradation products of bixin may also be present as a result of processing. Colour Index No 75120 Einecs 230-248-7 Chemical name cis-Bixin: Methyl (9-cis)-hydrogen-6,6'-diapo-Ψ,Ψ-carotenedioate Chemical formula cis-Bixin: C 25 H 30 O 4 Molecular weight 394,5 Assay Not less than 25 % colouring matter (expressed as bixin) E 1 % 1cm 3090 at ca. 487 nm in tetrahydrofuran and acetone Description Dark red-brown to red-purple powder Identification Solubility Insoluble in water, slightly soluble in ethanol Spectrometry The sample in acetone shows absorbance maxima at about 425, 457 and 487 nm Purity Norbixin Not more than 7 % of total colouring matters 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 E 160 b (ii) ANNATTO NORBIXIN (I) SOLVENT-EXTRACTED NORBIXIN 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: C 24 H 28 O 4 cis -Norbixin dipotassium salt: C 24 H 26 K 2 O 4 cis -Norbixin disodium salt: C 24 H 26 Na 2 O 4 Molecular weight 380,5 (acid), 456,7 (dipotassium salt), 424,5 (disodium salt) Assay Not less than 85 % colouring matter (expressed as norbixin) E 1 % 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 (II) ALKALI-PROCESSED NORBIXIN, ACID-PRECIPITATED 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: C 24 H 28 O 4 cis -Norbixin dipotassium salt: C 24 H 26 K 2 O 4 cis -Norbixin disodium salt: C 24 H 26 Na 2 O 4 Molecular weight 380,5 (acid), 456,7 (dipotassium salt), 424,5 (disodium salt) Assay Not less than 35 % colouring matter (expressed as norbixin) E 1 % 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 (III) ALKALI-PROCESSED NORBIXIN, NOT ACID-PRECIPITATED 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: C 24 H 28 O 4 cis -Norbixin dipotassium salt: C 24 H 26 K 2 O 4 cis -Norbixin disodium salt: C 24 H 26 Na 2 O 4 Molecular weight 380,5 (acid), 456,7 (dipotassium salt), 424,5 (disodium salt) Assay Not less than 15 % colouring matter (expressed as norbixin) E 1 % 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’