LegalizeCommission Regulation (EU) No 231/2012 of 9 March 2012 laying down specifications for food additives listed in Annexes II and III to Regulation (EC) No 1333/2008 of the European Parliament and of the Council Text with EEA relevance
| Synonyms | 4,1’,6’-Trichlorogalactosucrose |
| Definition | Definition |
| Einecs | 259-952-2 |
| Chemical name | 1,6-Dichloro-1,6-dideoxy-β-D-fructofuranosyl-4-chloro-4-deoxy-α-D-galactopyranoside |
| Chemical formula | C12H19Cl3O8 |
| Molecular weight | 397,64 |
| Assay | Content not less than 98 % and not more than 102 % C12H19Cl3O8 calculated on an anhydrous basis. |
| Description | White to off-white, practically odourless, crystalline powder. |
| Identification | Identification |
| Solubility | Freely soluble in water, methanol and ethanol Slightly soluble in ethyl acetate |
| Infrared absorption spectrum | The infrared spectrum of a potassium bromide dispersion of the sample exhibits relative maxima at similar wave numbers as those shown in the reference spectrum obtained using a sucralose reference standard. |
| Thin layer chromatography | The main spot in the test solution has the same Rf value as that of the main spot of standard solution A referred to in the test for other chlorinated disaccharides. This standard solution is obtained by dissolving 1,0g of sucralose reference standard in 10 ml of methanol. |
| Specific rotation | [α]D 20 + 84,0° to + 87,5° calculated on the anhydrous basis (10 % w/v solution) |
| Purity | Purity |
| Water content | Not more than 2,0 % (Karl Fischer method) |
| Sulphated ash | Not more than 0,7 % |
| Other chlorinated disaccharides | Not more than 0,5 % |
| Chlorinated monosaccharides | Not more than 0,1 % |
| Triphenylphosphine oxide | Not more than 150 mg/kg |
| Methanol | Not more than 0,1 % |
| Lead | Not more than 1 mg/kg |
E 957 THAUMATIN
| Synonyms | |
| Definition | Definition |
| Einecs | 258-822-2 |
| Chemical name | Thaumatin is obtained by aqueous extraction (pH 2,5 to 4) of the arils of the fruit of strains of Thaumatococcus daniellii (Benth) and consists essentially of the proteins thaumatin I and thaumatin II together with minor amounts of plant constituents derived from the source material |
| Chemical formula | Polypeptide of 207 amino acids |
| Molecular weight | Thaumatin I 22209 Thaumatin II 22293 |
| Assay | Not less than 15,1 % nitrogen on the dried basis equivalent to not less than 93 % proteins (N × 6,2) |
| Description | Odourless, cream-coloured powder. Approximately 2 000 to 3 000 times as sweet as sucrose |
| Identification | Identification |
| Solubility | Very soluble in water, insoluble in acetone |
| Purity | Purity |
| Loss on drying | Not more than 9 % (105 °C to constant weight) |
| Carbohydrates | Not more than 3 % (expressed on dry weight basis) |
| Sulphated ash | Not more than 2 % (expressed on dry weight basis) |
| Aluminium | Not more than 100 mg/kg (expressed on dry weight basis) |
| Arsenic | Not more than 3 mg/kg (expressed on dry weight basis) |
| Lead | Not more than 3 mg/kg (expressed on dry weight basis) |
| Microbiological criteria | Microbiological criteria |
| Total aerobic microbial count | Not more than 1 000 colonies per gram |
| Escherichia coli | Absent in 1 g |
E 959 NEOHESPERIDINE DIHYDROCHALCONE
| Synonyms | Neohesperidin dihydrochalcone; NHDC; Hesperetin dihydrochalcone-4′-β-neohesperidoside; Neohesperidin DC |
| Definition | It is obtained by catalytic hydrogenation of neohesperidin |
| Einecs | 243-978-6 |
| Chemical name | 2-O-α-L-rhamnopyranosyl-4′-β-D-glucopyranosyl hesperetin dihydrochalcone. |
| Chemical formula | C28H36O15 |
| Molecular weight | 612,6 |
| Assay | Content not less than 96 % on the dried basis |
| Description | Off-white, odourless, crystalline powder. Approximately between 1 000 and 1 800 times as sweet as sucrose |
| Identification | Identification |
| Solubility | Freely soluble in hot water, very slightly soluble in cold water, practically insoluble in ether and benzene |
| Ultraviolet absorption maximum | 282 to 283 nm for a solution of 2 mg in 100 ml methanol |
| Neu's test | Dissolve about 10 mg of neohesperidine DC in 1 ml methanol, add 1 ml of a 1 % 2-aminoethyl diphenyl borate methanolic solution. A bright yellow colour is produced |
| Purity | Purity |
| Loss on drying | Not more than 11 % (105 °C, 3 hours) |
| Sulphated ash | Not more than 0,2 % (expressed on dry weight basis) |
| Arsenic | Not more than 3 mg/kg expressed on dry weight basis |
| Lead | Not more than 2 mg/kg (expressed on dry weight basis) |
E 960a STEVIOL GLYCOSIDES FROM STEVIA
| Synonyms | |||
| Definition | The manufacturing process comprises two main phases: the first involving water extraction of the leaves of the Stevia rebaudiana Bertoni plant and preliminary purification of the extract by employing ion exchange chromatography to yield a steviol glycoside primary extract, and the second involving recrystallisation of the steviol glycosides from methanol or aqueous ethanol resulting in a final product containing not less than 95 % of the below identified 11 related steviol glycosides, in any combination and ratio. The additive may contain residues of ion-exchange resins used in the manufacturing process. Several other related steviol glycosides that may be generated as a result of the production process, but do not occur naturally in the Stevia rebaudiana plant have been identified in small amounts (0,10 to 0,37 % w/w). | The manufacturing process comprises two main phases: the first involving water extraction of the leaves of the Stevia rebaudiana Bertoni plant and preliminary purification of the extract by employing ion exchange chromatography to yield a steviol glycoside primary extract, and the second involving recrystallisation of the steviol glycosides from methanol or aqueous ethanol resulting in a final product containing not less than 95 % of the below identified 11 related steviol glycosides, in any combination and ratio. The additive may contain residues of ion-exchange resins used in the manufacturing process. Several other related steviol glycosides that may be generated as a result of the production process, but do not occur naturally in the Stevia rebaudiana plant have been identified in small amounts (0,10 to 0,37 % w/w). | The manufacturing process comprises two main phases: the first involving water extraction of the leaves of the Stevia rebaudiana Bertoni plant and preliminary purification of the extract by employing ion exchange chromatography to yield a steviol glycoside primary extract, and the second involving recrystallisation of the steviol glycosides from methanol or aqueous ethanol resulting in a final product containing not less than 95 % of the below identified 11 related steviol glycosides, in any combination and ratio. The additive may contain residues of ion-exchange resins used in the manufacturing process. Several other related steviol glycosides that may be generated as a result of the production process, but do not occur naturally in the Stevia rebaudiana plant have been identified in small amounts (0,10 to 0,37 % w/w). |
| Chemical name | Steviolbioside: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rubusoside: 13-β-D-glucopyranosyloxykaur-16-en-18-oic acid, β-D-glucopyranosyl ester Dulcoside A: 13-[(2-O-α–L-rhamnopyranosyl-β–D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Stevioside: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside A: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D- glucopyranosyl ester Rebaudioside B: 13-[(2-O-β–D-glucopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rebaudioside C: 13-[(2-O-α–L-rhamnopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside D: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside E: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside F: 13[(2-O-β-D-xylofurananosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester | Steviolbioside: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rubusoside: 13-β-D-glucopyranosyloxykaur-16-en-18-oic acid, β-D-glucopyranosyl ester Dulcoside A: 13-[(2-O-α–L-rhamnopyranosyl-β–D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Stevioside: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside A: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D- glucopyranosyl ester Rebaudioside B: 13-[(2-O-β–D-glucopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rebaudioside C: 13-[(2-O-α–L-rhamnopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside D: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside E: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside F: 13[(2-O-β-D-xylofurananosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester | Steviolbioside: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rubusoside: 13-β-D-glucopyranosyloxykaur-16-en-18-oic acid, β-D-glucopyranosyl ester Dulcoside A: 13-[(2-O-α–L-rhamnopyranosyl-β–D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Stevioside: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside A: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D- glucopyranosyl ester Rebaudioside B: 13-[(2-O-β–D-glucopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rebaudioside C: 13-[(2-O-α–L-rhamnopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside D: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside E: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside F: 13[(2-O-β-D-xylofurananosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester |
| Molecular formula | Trivial name | Formula | Conversion factor |
| Molecular formula | Steviol | C20 H30 O3 | 1,00 |
| Molecular formula | Steviolbioside | C32 H50 O13 | 0,50 |
| Molecular formula | Rubusoside | C32 H50 O13 | 0,50 |
| Molecular formula | Dulcoside A | C38 H60 O17 | 0,40 |
| Molecular formula | Stevioside | C38 H60 O18 | 0,40 |
| Molecular formula | Rebaudioside A | C44 H70 O23 | 0,33 |
| Molecular formula | Rebaudioside B | C38 H60 O18 | 0,40 |
| Molecular formula | Rebaudioside C | C44 H70 O22 | 0,34 |
| Molecular formula | Rebaudioside D | C50 H80 O28 | 0,29 |
| Molecular formula | Rebaudioside E | C44 H70 O23 | 0,33 |
| Molecular formula | Rebaudioside F | C43 H68 O22 | 0,34 |
| Molecular formula | Rebaudioside M | C56 H90 O33 | 0,25 |
| Molecular weight and CAS No | Trivial name | CAS Number | *Molecular weight* (g/mol) |
| Molecular weight and CAS No | Steviol | 318,46 | |
| Molecular weight and CAS No | Steviolbioside | 41093-60-1 | 642,73 |
| Molecular weight and CAS No | Rubusoside | 64849-39-4 | 642,73 |
| Molecular weight and CAS No | Dulcoside A | 64432-06-0 | 788,87 |
| Molecular weight and CAS No | Stevioside | 57817-89-7 | 804,88 |
| Molecular weight and CAS No | Rebaudioside A | 58543-16-1 | 967,01 |
| Molecular weight and CAS No | Rebaudioside B | 58543-17-2 | 804,88 |
| Molecular weight and CAS No | Rebaudioside C | 63550-99-2 | 951,02 |
| Molecular weight and CAS No | Rebaudioside D | 63279-13-0 | 1 129,15 |
| Molecular weight and CAS No | Rebaudioside E | 63279-14-1 | 967,01 |
| Molecular weight and CAS No | Rebaudioside F | 438045-89-7 | 936,99 |
| Molecular weight and CAS No | Rebaudioside M | 1220616-44-3 | 1 291,30 |
| Assay | Not less than 95 % steviolbioside, rubusoside, dulcoside A, stevioside, rebaudiosides A, B, C, D, E, F and M on the dried basis, in any combination and ratio. | Not less than 95 % steviolbioside, rubusoside, dulcoside A, stevioside, rebaudiosides A, B, C, D, E, F and M on the dried basis, in any combination and ratio. | Not less than 95 % steviolbioside, rubusoside, dulcoside A, stevioside, rebaudiosides A, B, C, D, E, F and M on the dried basis, in any combination and ratio. |
| Description | White to light yellow powder, approximately between 200 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 200 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 200 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). |
| Identification | Identification | Identification | Identification |
| Solubility | Freely soluble to slightly soluble in water | Freely soluble to slightly soluble in water | Freely soluble to slightly soluble in water |
| pH | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) |
| Purity | Purity | Purity | Purity |
| Total ash | Not more than 1 % | Not more than 1 % | Not more than 1 % |
| Loss on drying | Not more than 6 % (105 °C, 2h) | Not more than 6 % (105 °C, 2h) | Not more than 6 % (105 °C, 2h) |
| Residual solvents | Not more than 200 mg/kg methanol Not more than 5 000 mg/kg ethanol | Not more than 200 mg/kg methanol Not more than 5 000 mg/kg ethanol | Not more than 200 mg/kg methanol Not more than 5 000 mg/kg ethanol |
| Arsenic | Not more than 1 mg/kg | Not more than 1 mg/kg | Not more than 1 mg/kg |
| Lead | Not more than 1 mg/kg | Not more than 1 mg/kg | Not more than 1 mg/kg |
E 960B(I) REBAUDIOSIDE M FROM FERMENTATION PRODUCED BY YARROWIA LIPOLYTICA
| Synonyms | |||
| Definition | Rebaudioside M from fermentation produced by Yarrowia lipolytica consist of a mixture of steviol glycosides composed of rebaudioside M as the main componant, with some rebaudioside D, and smaller amounts of rebaudioside A and rebaudioside B. The manufacturing process comprises two main phases. The first phase involves fermentation of a simple sugar source by a non-toxigenic non-pathogenic strain of Yarrowia lipolytica that has been genetically modified with heterologous genes to overexpress genes which are involved in the synthesis of steviol glycosides to result in the strain VRM (CBS 147477). Removal of biomass by solid-liquid separation and heat treatment is followed by concentration of the steviol glycosides. The second phase involves purification by employing ion exchange chromatography, followed by cristallisation of the steviol glycosides from ethanol, resulting in a final product containing not less than 95 % of rebaudiosides M, D, A and B. Viable cells and DNA of Yarrowia lipolytica VRM shall not be detected in the food additive. | Rebaudioside M from fermentation produced by Yarrowia lipolytica consist of a mixture of steviol glycosides composed of rebaudioside M as the main componant, with some rebaudioside D, and smaller amounts of rebaudioside A and rebaudioside B. The manufacturing process comprises two main phases. The first phase involves fermentation of a simple sugar source by a non-toxigenic non-pathogenic strain of Yarrowia lipolytica that has been genetically modified with heterologous genes to overexpress genes which are involved in the synthesis of steviol glycosides to result in the strain VRM (CBS 147477). Removal of biomass by solid-liquid separation and heat treatment is followed by concentration of the steviol glycosides. The second phase involves purification by employing ion exchange chromatography, followed by cristallisation of the steviol glycosides from ethanol, resulting in a final product containing not less than 95 % of rebaudiosides M, D, A and B. Viable cells and DNA of Yarrowia lipolytica VRM shall not be detected in the food additive. | Rebaudioside M from fermentation produced by Yarrowia lipolytica consist of a mixture of steviol glycosides composed of rebaudioside M as the main componant, with some rebaudioside D, and smaller amounts of rebaudioside A and rebaudioside B. The manufacturing process comprises two main phases. The first phase involves fermentation of a simple sugar source by a non-toxigenic non-pathogenic strain of Yarrowia lipolytica that has been genetically modified with heterologous genes to overexpress genes which are involved in the synthesis of steviol glycosides to result in the strain VRM (CBS 147477). Removal of biomass by solid-liquid separation and heat treatment is followed by concentration of the steviol glycosides. The second phase involves purification by employing ion exchange chromatography, followed by cristallisation of the steviol glycosides from ethanol, resulting in a final product containing not less than 95 % of rebaudiosides M, D, A and B. Viable cells and DNA of Yarrowia lipolytica VRM shall not be detected in the food additive. |
| Chemical name | Rebaudioside A: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D- glucopyranosyl ester Rebaudioside B: 13-[(2-O-β–D-glucopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rebaudioside D: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester | Rebaudioside A: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D- glucopyranosyl ester Rebaudioside B: 13-[(2-O-β–D-glucopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rebaudioside D: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester | Rebaudioside A: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D- glucopyranosyl ester Rebaudioside B: 13-[(2-O-β–D-glucopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rebaudioside D: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester |
| Molecular formula | Trivial name | Formula | Conversion factor |
| Rebaudioside A | C44 H70 O23 | 0,33 | |
| Rebaudioside B | C38 H60 O18 | 0,40 | |
| Rebaudioside D | C50H80O28 | 0,29 | |
| Rebaudioside M | C56H90O33 | 0,25 | |
| Molecular weight and CAS No | Trivial name | CAS Number | Molecular weight (g/mol) |
| Molecular weight and CAS No | Rebaudioside A | 58543-16-1 | 967,01 |
| Molecular weight and CAS No | Rebaudioside B | 58543-17-2 | 804,88 |
| Molecular weight and CAS No | Rebaudioside D | 63279-13-0 | 1 129,15 |
| Molecular weight and CAS No | Rebaudioside M | 1220616-44-3 | 1 291,30 |
| Assay | Not less than 95 % of rebaudioside M, rebaudioside D, rebaudioside A and rebaudioside B, on the dried basis. | Not less than 95 % of rebaudioside M, rebaudioside D, rebaudioside A and rebaudioside B, on the dried basis. | Not less than 95 % of rebaudioside M, rebaudioside D, rebaudioside A and rebaudioside B, on the dried basis. |
| Particle size | Not more than 0,5 % of particles of less than 300 nm (by STEM method) | Not more than 0,5 % of particles of less than 300 nm (by STEM method) | Not more than 0,5 % of particles of less than 300 nm (by STEM method) |
| Description | White to light yellow powder, approximately between 200 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 200 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 200 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). |
| Identification | Identification | Identification | Identification |
| Solubility | Slightly soluble in water | Slightly soluble in water | Slightly soluble in water |
| pH | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) |
| Purity | Purity | Purity | Purity |
| Total ash | Not more than 1 % | Not more than 1 % | Not more than 1 % |
| Loss on drying | Not more than 6 % (105 °C, 2h) | Not more than 6 % (105 °C, 2h) | Not more than 6 % (105 °C, 2h) |
| Residual solvent | Not more than 5 000 mg/kg ethanol | Not more than 5 000 mg/kg ethanol | Not more than 5 000 mg/kg ethanol |
| Kaurenoic acid | Not more than 0,3 mg/kg (measured by liquid chromatography-mass spectrometry detection) | Not more than 0,3 mg/kg (measured by liquid chromatography-mass spectrometry detection) | Not more than 0,3 mg/kg (measured by liquid chromatography-mass spectrometry detection) |
| Arsenic | Not more than 0,01 mg/kg | Not more than 0,01 mg/kg | Not more than 0,01 mg/kg |
| Lead | Not more than 0,01 mg/kg | Not more than 0,01 mg/kg | Not more than 0,01 mg/kg |
| Cadmium | Not more than 0,01 mg/kg | Not more than 0,01 mg/kg | Not more than 0,01 mg/kg |
| Mercury | Not more than 0,05 mg/kg | Not more than 0,05 mg/kg | Not more than 0,05 mg/kg |
| Microbiological criteria | Microbiological criteria | Microbiological criteria | Microbiological criteria |
| Total (aerobic) plate count | Not more than 1 000 CFU/g | Not more than 1 000 CFU/g | Not more than 1 000 CFU/g |
| Yeast and moulds | Not more than 100 CFU/g | Not more than 100 CFU/g | Not more than 100 CFU/g |
| Escherichia coli | Negative in 1 g | Negative in 1 g | Negative in 1 g |
| Salmonella spp. | Negative in 25 g | Negative in 25 g | Negative in 25 g |
| Residual protein | Not more than 20 mg/kg | Not more than 20 mg/kg | Not more than 20 mg/kg |
E 960c(i) REBAUDIOSIDE M PRODUCED VIA ENZYME MODIFICATION OF STEVIOL GLYCOSIDES FROM STEVIA
| Synonyms | |||
| Definition | Rebaudioside M is a steviol glycoside composed predominantly of rebaudioside M with minor amounts of other steviol glycosides such as rebaudioside A, rebaudioside B, rebaudioside D, rebaudioside I, and stevioside. Rebaudioside M is obtained via enzymatic bioconversion of purified steviol glycoside leaf extracts (95% steviol glycosides) of the Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified yeasts K. phaffi (formerly known as Pichia pastoris) UGT-a and K. phaffi UGT-b that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside M by resin adsorption, followed by recrystallisation of rebaudioside M resulting in a final product containing not less than 95 % of rebaudioside M. Viable cells of the yeasts K. phaffii UGT-a and K. phaffii UGT-b and their DNA shall not be detected in the food additive. | Rebaudioside M is a steviol glycoside composed predominantly of rebaudioside M with minor amounts of other steviol glycosides such as rebaudioside A, rebaudioside B, rebaudioside D, rebaudioside I, and stevioside. Rebaudioside M is obtained via enzymatic bioconversion of purified steviol glycoside leaf extracts (95% steviol glycosides) of the Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified yeasts K. phaffi (formerly known as Pichia pastoris) UGT-a and K. phaffi UGT-b that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside M by resin adsorption, followed by recrystallisation of rebaudioside M resulting in a final product containing not less than 95 % of rebaudioside M. Viable cells of the yeasts K. phaffii UGT-a and K. phaffii UGT-b and their DNA shall not be detected in the food additive. | Rebaudioside M is a steviol glycoside composed predominantly of rebaudioside M with minor amounts of other steviol glycosides such as rebaudioside A, rebaudioside B, rebaudioside D, rebaudioside I, and stevioside. Rebaudioside M is obtained via enzymatic bioconversion of purified steviol glycoside leaf extracts (95% steviol glycosides) of the Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified yeasts K. phaffi (formerly known as Pichia pastoris) UGT-a and K. phaffi UGT-b that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside M by resin adsorption, followed by recrystallisation of rebaudioside M resulting in a final product containing not less than 95 % of rebaudioside M. Viable cells of the yeasts K. phaffii UGT-a and K. phaffii UGT-b and their DNA shall not be detected in the food additive. |
| Chemical name | Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester | Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester | Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester |
| Molecular formula | Trivial name | Formula | Conversion factor |
| Molecular formula | Rebaudioside M | C56 H90 O33 | 0,25 |
| Molecular weight and CAS No | Trivial name | CAS Number | Molecular weight (g/mol) |
| Molecular weight and CAS No | |||
| Molecular weight and CAS No | Rebaudioside M | 1220616-44-3 | 1,291.29 |
| Assay | Not less than 95 % rebaudioside M on the dried basis. | Not less than 95 % rebaudioside M on the dried basis. | Not less than 95 % rebaudioside M on the dried basis. |
| Description | White to light yellow powder, approximately between 200 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 200 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 200 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). |
| Identification | Identification | Identification | Identification |
| Solubility | Freely soluble to slightly soluble in water | Freely soluble to slightly soluble in water | Freely soluble to slightly soluble in water |
| pH | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) |
| Purity | Purity | Purity | Purity |
| Total ash | Not more than 1 % | Not more than 1 % | Not more than 1 % |
| Loss on drying | Not more than 6 % (105 °C, 2h) | Not more than 6 % (105 °C, 2h) | Not more than 6 % (105 °C, 2h) |
| Residual solvent | Not more than 5 000 mg/kg ethanol | Not more than 5 000 mg/kg ethanol | Not more than 5 000 mg/kg ethanol |
| Arsenic | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg |
| Lead | Not more than 0,2 mg/kg | Not more than 0,2 mg/kg | Not more than 0,2 mg/kg |
| Cadmium | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg |
| Mercury | Not more than 0,07 mg/kg | Not more than 0,07 mg/kg | Not more than 0,07 mg/kg |
| Residual protein | Not more than 5 mg/kg | Not more than 5 mg/kg | Not more than 5 mg/kg |
| Particle size | Not less than 74 μm [using a mesh #200 sieve with a particle size limit of 74 μm] | Not less than 74 μm [using a mesh #200 sieve with a particle size limit of 74 μm] | Not less than 74 μm [using a mesh #200 sieve with a particle size limit of 74 μm] |
E 960c(ii) REBAUDIOSIDE M PRODUCED VIA ENZYMATIC CONVERSION OF HIGHLY PURIFIED REBAUDIOSIDE A STEVIA LEAF EXTRACTS
| Synonyms | |||
| Definition | Rebaudioside M produced via enzymatic conversion of highly purified rebaudioside A stevia leaf extracts is a steviol glycoside composed predominantly of rebaudioside M with minor amounts of other steviol glycosides such as rebaudioside A and rebaudioside D. Rebaudioside M is produced via enzymatic conversion of highly purified steviol glycoside rebaudioside A extracts (95 % steviol glycosides) obtained from Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified strains of E. coli (pPM294, pFAF170 and pSK401) that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside M by resin adsorption, followed by recrystallisation of the steviol glycosides resulting in a final product containing not less than 95 % of rebaudioside M. Viable cells of E. coli (pPM294, pFAF170 and pSK401) and their DNA shall not be detected in the food additive. | Rebaudioside M produced via enzymatic conversion of highly purified rebaudioside A stevia leaf extracts is a steviol glycoside composed predominantly of rebaudioside M with minor amounts of other steviol glycosides such as rebaudioside A and rebaudioside D. Rebaudioside M is produced via enzymatic conversion of highly purified steviol glycoside rebaudioside A extracts (95 % steviol glycosides) obtained from Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified strains of E. coli (pPM294, pFAF170 and pSK401) that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside M by resin adsorption, followed by recrystallisation of the steviol glycosides resulting in a final product containing not less than 95 % of rebaudioside M. Viable cells of E. coli (pPM294, pFAF170 and pSK401) and their DNA shall not be detected in the food additive. | Rebaudioside M produced via enzymatic conversion of highly purified rebaudioside A stevia leaf extracts is a steviol glycoside composed predominantly of rebaudioside M with minor amounts of other steviol glycosides such as rebaudioside A and rebaudioside D. Rebaudioside M is produced via enzymatic conversion of highly purified steviol glycoside rebaudioside A extracts (95 % steviol glycosides) obtained from Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified strains of E. coli (pPM294, pFAF170 and pSK401) that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside M by resin adsorption, followed by recrystallisation of the steviol glycosides resulting in a final product containing not less than 95 % of rebaudioside M. Viable cells of E. coli (pPM294, pFAF170 and pSK401) and their DNA shall not be detected in the food additive. |
| Chemical name | Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester | Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester | Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester |
| Molecular formula | Trivial name | Formula | Conversion factor |
| Molecular formula | Rebaudioside M | C56H90O33 | 0,25 |
| Molecular weight and CAS No | Trivial name | CAS Number | Molecular weight (g/mol) |
| Molecular weight and CAS No | Rebaudioside M | 1220616-44-3 | 1 291,29 |
| Assay | Not less than 95 % rebaudioside M on the dried basis. | Not less than 95 % rebaudioside M on the dried basis. | Not less than 95 % rebaudioside M on the dried basis. |
| Description | White to light yellow powder, approximately between 150 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 150 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 150 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). |
| Identification | Identification | Identification | Identification |
| Solubility | Freely soluble to slightly soluble in water | Freely soluble to slightly soluble in water | Freely soluble to slightly soluble in water |
| pH | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) |
| Purity | Purity | Purity | Purity |
| Total ash | Not more than 1 % | Not more than 1 % | Not more than 1 % |
| Loss on drying | Not more than 6 % (105 °C, 2h) | Not more than 6 % (105 °C, 2h) | Not more than 6 % (105 °C, 2h) |
| Residual solvent | Not more than 5 000 mg/kg ethanol | Not more than 5 000 mg/kg ethanol | Not more than 5 000 mg/kg ethanol |
| Arsenic | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg |
| Lead | Not more than 0,2 mg/kg | Not more than 0,2 mg/kg | Not more than 0,2 mg/kg |
| Cadmium | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg |
| Mercury | Not more than 0,07 mg/kg | Not more than 0,07 mg/kg | Not more than 0,07 mg/kg |
| Residual protein | Not more than 5 mg/kg | Not more than 5 mg/kg | Not more than 5 mg/kg |
| Particle size | Not less than 74 μm [using a mesh #200 sieve with a particle size limit of 74 μm] | Not less than 74 μm [using a mesh #200 sieve with a particle size limit of 74 μm] | Not less than 74 μm [using a mesh #200 sieve with a particle size limit of 74 μm] |
E 960c(iii) REBAUDIOSIDE D PRODUCED VIA ENZYMATIC CONVERSION OF HIGHLY PURIFIED REBAUDIOSIDE A STEVIA LEAF EXTRACTS
| Synonyms | |||
| Definition | Rebaudioside D produced via enzymatic conversion of highly purified rebaudioside A stevia leaf extracts is a steviol glycoside composed predominantly of rebaudioside D with minor amounts of other steviol glycosides such as rebaudioside A and rebaudioside M. Rebaudioside D is produced via enzymatic conversion of highly purified steviol glycoside rebaudioside A extracts (95 % steviol glycosides) obtained from Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified strains of E. coli (pPM294, pFAF170 and pSK401) that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside D by resin adsorption, followed by recrystallisation of the steviol glycosides resulting in a final product containing not less than 95 % of rebaudioside D and rebaudioside A. Viable cells of E. coli (pPM294, pFAF170 and pSK401) and their DNA shall not be detected in the food additive. | Rebaudioside D produced via enzymatic conversion of highly purified rebaudioside A stevia leaf extracts is a steviol glycoside composed predominantly of rebaudioside D with minor amounts of other steviol glycosides such as rebaudioside A and rebaudioside M. Rebaudioside D is produced via enzymatic conversion of highly purified steviol glycoside rebaudioside A extracts (95 % steviol glycosides) obtained from Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified strains of E. coli (pPM294, pFAF170 and pSK401) that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside D by resin adsorption, followed by recrystallisation of the steviol glycosides resulting in a final product containing not less than 95 % of rebaudioside D and rebaudioside A. Viable cells of E. coli (pPM294, pFAF170 and pSK401) and their DNA shall not be detected in the food additive. | Rebaudioside D produced via enzymatic conversion of highly purified rebaudioside A stevia leaf extracts is a steviol glycoside composed predominantly of rebaudioside D with minor amounts of other steviol glycosides such as rebaudioside A and rebaudioside M. Rebaudioside D is produced via enzymatic conversion of highly purified steviol glycoside rebaudioside A extracts (95 % steviol glycosides) obtained from Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified strains of E. coli (pPM294, pFAF170 and pSK401) that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside D by resin adsorption, followed by recrystallisation of the steviol glycosides resulting in a final product containing not less than 95 % of rebaudioside D and rebaudioside A. Viable cells of E. coli (pPM294, pFAF170 and pSK401) and their DNA shall not be detected in the food additive. |
| Chemical name | Rebaudioside D: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester. Rebaudioside A: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester | Rebaudioside D: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester. Rebaudioside A: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester | Rebaudioside D: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester. Rebaudioside A: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester |
| Molecular formula | Trivial name | Formula | Conversion factor |
| Molecular formula | Rebaudioside D | C50H80O28 | 0,29 |
| Molecular formula | Rebaudioside A | C44H70O23 | 0,33 |
| Molecular weight and CAS No | Trivial name | CAS Number | Molecular weight (g/mol) |
| Molecular weight and CAS No | Rebaudioside D | 63279-13-0 | 1 291,15 |
| Molecular weight and CAS No | Rebaudioside A | 58543-16-1 | 967,01 |
| Assay | Not less than 95 % rebaudiosides D and A on the dried basis. | Not less than 95 % rebaudiosides D and A on the dried basis. | Not less than 95 % rebaudiosides D and A on the dried basis. |
| Description | White to light yellow powder, approximately between 150 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 150 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 150 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). |
| Identification | Identification | Identification | Identification |
| Solubility | Freely soluble to slightly soluble in water | Freely soluble to slightly soluble in water | Freely soluble to slightly soluble in water |
| pH | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) |
| Purity | Purity | Purity | Purity |
| Total ash | Not more than 1 % | Not more than 1 % | Not more than 1 % |
| Loss on drying | Not more than 6 % (105 °C, 2h) | Not more than 6 % (105 °C, 2h) | Not more than 6 % (105 °C, 2h) |
| Residual solvent | Not more than 5 000 mg/kg ethanol | Not more than 5 000 mg/kg ethanol | Not more than 5 000 mg/kg ethanol |
| Arsenic | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg |
| Lead | Not more than 0,2 mg/kg | Not more than 0,2 mg/kg | Not more than 0,2 mg/kg |
| Cadmium | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg |
| Mercury | Not more than 0,07 mg/kg | Not more than 0,07 mg/kg | Not more than 0,07 mg/kg |
| Residual protein | Not more than 5 mg/kg | Not more than 5 mg/kg | Not more than 5 mg/kg |
| Particle size | Not less than 74 μm [using a mesh #200 sieve with a particle size limit of 74 μm] | Not less than 74 μm [using a mesh #200 sieve with a particle size limit of 74 μm] | Not less than 74 μm [using a mesh #200 sieve with a particle size limit of 74 μm] |
E 960c(iv) REBAUDIOSIDE AM PRODUCED VIA ENZYMATIC CONVERSION OF HIGHLY PURIFIED STEVIOSIDE STEVIA LEAF EXTRACTS
| Synonyms | |||
| Definition | Rebaudioside AM produced via enzymatic convertion of highly purified stevioside stevia leaf extracts is a steviol glycoside composed predominantly of rebaudioside AM with minor amounts of other steviol glycosides such as stevioside and rebaudioside E. Rebaudioside AM is produced via enzymatic conversion of highly purified steviol glycoside stevioside extracts (95 % steviol glycosides) obtained from Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified strains of E. coli (pPM294, pFAF170 and pSK401) that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside AM by resin adsorption, followed by recrystallisation of the steviol glycosides resulting in a final product containing not less than 95 % of rebaudioside AM. Viable cells of E. coli (pPM294, pFAF170 and pSK401) and their DNA shall not be detected in the food additive. | Rebaudioside AM produced via enzymatic convertion of highly purified stevioside stevia leaf extracts is a steviol glycoside composed predominantly of rebaudioside AM with minor amounts of other steviol glycosides such as stevioside and rebaudioside E. Rebaudioside AM is produced via enzymatic conversion of highly purified steviol glycoside stevioside extracts (95 % steviol glycosides) obtained from Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified strains of E. coli (pPM294, pFAF170 and pSK401) that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside AM by resin adsorption, followed by recrystallisation of the steviol glycosides resulting in a final product containing not less than 95 % of rebaudioside AM. Viable cells of E. coli (pPM294, pFAF170 and pSK401) and their DNA shall not be detected in the food additive. | Rebaudioside AM produced via enzymatic convertion of highly purified stevioside stevia leaf extracts is a steviol glycoside composed predominantly of rebaudioside AM with minor amounts of other steviol glycosides such as stevioside and rebaudioside E. Rebaudioside AM is produced via enzymatic conversion of highly purified steviol glycoside stevioside extracts (95 % steviol glycosides) obtained from Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified strains of E. coli (pPM294, pFAF170 and pSK401) that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside AM by resin adsorption, followed by recrystallisation of the steviol glycosides resulting in a final product containing not less than 95 % of rebaudioside AM. Viable cells of E. coli (pPM294, pFAF170 and pSK401) and their DNA shall not be detected in the food additive. |
| Chemical name | Rebaudioside AM: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2- O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester. | Rebaudioside AM: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2- O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester. | Rebaudioside AM: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2- O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester. |
| Molecular formula | Trivial name | Formula | Conversion factor |
| Molecular formula | Rebaudioside AM | C50 H80 O28 | 0,29 |
| Molecular weight and CAS No | Trivial name | CAS Number | Molecular weight (g/mol) |
| Molecular weight and CAS No | Rebaudioside AM | 2222580-26-7 | 1 291,15 |
| Assay | Not less than 95 % rebaudioside AM on the dried basis. | Not less than 95 % rebaudioside AM on the dried basis. | Not less than 95 % rebaudioside AM on the dried basis. |
| Description | White to light yellow powder, approximately between 150 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 150 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 150 and 350 times sweeter than sucrose (at 5 % sucrose equivalency). |
| Identification | Identification | Identification | Identification |
| Solubility | Freely soluble to slightly soluble in water | Freely soluble to slightly soluble in water | Freely soluble to slightly soluble in water |
| pH | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) |
| Purity | Purity | Purity | Purity |
| Total ash | Not more than 1 % | Not more than 1 % | Not more than 1 % |
| Loss on drying | Not more than 6 % (105 °C, 2h) | Not more than 6 % (105 °C, 2h) | Not more than 6 % (105 °C, 2h) |
| Residual solvent | Not more than 5 000 mg/kg ethanol | Not more than 5 000 mg/kg ethanol | Not more than 5 000 mg/kg ethanol |
| Arsenic | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg |
| Lead | Not more than 0,2 mg/kg | Not more than 0,2 mg/kg | Not more than 0,2 mg/kg |
| Cadmium | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg |
| Mercury | Not more than 0,07 mg/kg | Not more than 0,07 mg/kg | Not more than 0,07 mg/kg |
| Residual protein | Not more than 5 mg/kg | Not more than 5 mg/kg | Not more than 5 mg/kg |
| Particle size | Not less than 74 μm [using a mesh #200 sieve with a particle size limit of 74 μm] | Not less than 74 μm [using a mesh #200 sieve with a particle size limit of 74 μm] | Not less than 74 μm [using a mesh #200 sieve with a particle size limit of 74 μm] |
E 960d GLUCOSYLATED STEVIOL GLYCOSIDES
| Synonyms | |||
| Definition | Mixture of larger glycosides of steviol derived by glucosylation of steviol glycosides extracted from leaves of Stevia rebaudiana Bertoni plant. The mixture is composed of glucosylated steviol glycosides and residual parent steviol glycosides from Stevia leaf. Glucosylated steviol glycosides are produced by treating the steviol glycosides, extracted from Stevia leaf, and starch suitable for human consumption with Cyclomaltodextrin glucanotransferase (EC 2.4.1.19) derived from a non-GMO strain of Anoxybacillus caldiproteolyticus St-88. The enzyme transfers glucose units from the starch to the steviol glycosides. The resulting material is heated and treated with activated carbon to remove the enzyme, then passed through adsorption/desorption resin to remove residual hydrolysed starch (dextrin), followed by purification and preparation of the final product using processes that may include decolourisation, concentration and spray drying. | Mixture of larger glycosides of steviol derived by glucosylation of steviol glycosides extracted from leaves of Stevia rebaudiana Bertoni plant. The mixture is composed of glucosylated steviol glycosides and residual parent steviol glycosides from Stevia leaf. Glucosylated steviol glycosides are produced by treating the steviol glycosides, extracted from Stevia leaf, and starch suitable for human consumption with Cyclomaltodextrin glucanotransferase (EC 2.4.1.19) derived from a non-GMO strain of Anoxybacillus caldiproteolyticus St-88. The enzyme transfers glucose units from the starch to the steviol glycosides. The resulting material is heated and treated with activated carbon to remove the enzyme, then passed through adsorption/desorption resin to remove residual hydrolysed starch (dextrin), followed by purification and preparation of the final product using processes that may include decolourisation, concentration and spray drying. | Mixture of larger glycosides of steviol derived by glucosylation of steviol glycosides extracted from leaves of Stevia rebaudiana Bertoni plant. The mixture is composed of glucosylated steviol glycosides and residual parent steviol glycosides from Stevia leaf. Glucosylated steviol glycosides are produced by treating the steviol glycosides, extracted from Stevia leaf, and starch suitable for human consumption with Cyclomaltodextrin glucanotransferase (EC 2.4.1.19) derived from a non-GMO strain of Anoxybacillus caldiproteolyticus St-88. The enzyme transfers glucose units from the starch to the steviol glycosides. The resulting material is heated and treated with activated carbon to remove the enzyme, then passed through adsorption/desorption resin to remove residual hydrolysed starch (dextrin), followed by purification and preparation of the final product using processes that may include decolourisation, concentration and spray drying. |
| Chemical name | Steviolbioside: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rubusoside: 13-β-D-glucopyranosyloxykaur-16-en-18-oic acid, β-D-glucopyranosyl ester Dulcoside A: 13-[(2-O-α–L-rhamnopyranosyl-β–D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Stevioside: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside A: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D- glucopyranosyl ester Rebaudioside B: 13-[(2-O-β–D-glucopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rebaudioside C: 13-[(2-O-α–L-rhamnopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside D: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside E: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside F: 13-[(2-O-β-D-xylofurananosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester And their glucosylated derivatives (1-20 added glucose units) | Steviolbioside: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rubusoside: 13-β-D-glucopyranosyloxykaur-16-en-18-oic acid, β-D-glucopyranosyl ester Dulcoside A: 13-[(2-O-α–L-rhamnopyranosyl-β–D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Stevioside: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside A: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D- glucopyranosyl ester Rebaudioside B: 13-[(2-O-β–D-glucopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rebaudioside C: 13-[(2-O-α–L-rhamnopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside D: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside E: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside F: 13-[(2-O-β-D-xylofurananosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester And their glucosylated derivatives (1-20 added glucose units) | Steviolbioside: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rubusoside: 13-β-D-glucopyranosyloxykaur-16-en-18-oic acid, β-D-glucopyranosyl ester Dulcoside A: 13-[(2-O-α–L-rhamnopyranosyl-β–D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Stevioside: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside A: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D- glucopyranosyl ester Rebaudioside B: 13-[(2-O-β–D-glucopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid Rebaudioside C: 13-[(2-O-α–L-rhamnopyranosyl-3-O-β–D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside D: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside E: 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester Rebaudioside F: 13-[(2-O-β-D-xylofurananosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, β-D-glucopyranosyl ester Rebaudioside M: 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid, 2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl ester And their glucosylated derivatives (1-20 added glucose units) |
| Molecular formula | Trivial name | Formula | Conversion factor |
| n-Glucosylated Steviolbioside | C(32+n6)H(50+n10)O(13+n*5) | ||
| n-Glucosylated Rubusoside | C(32+n6)H(50+n10)O(13+n*5) | ||
| n-Glucosylated Dulcoside A | C(38+n6)H(60+n10)O(17+n*5) | ||
| n-Glucosylated Stevioside | C(38+n6)H(60+n10)O(18+n*5) | ||
| n-Glucosylated Rebaudioside A | C(44+n6)H(70+n10)O(23+n*5) | ||
| n-Glucosylated Rebaudioside B | C(38+n6)H(60+n10)O(18+n*5) | ||
| n-Glucosylated Rebaudioside C | C(44+n6)H(70+n10)O(22+n*5) | ||
| n-Glucosylated Rebaudioside D | C(50+n6)H(80+n10)O(28+n*5) | ||
| n-Glucosylated Rebaudioside E | C(44+n6)H(70+n10)O(23+n*5) | ||
| n-Glucosylated Rebaudioside F | C(43+n6)H(68+n10)O(22+n*5) | ||
| n-Glucosylated Rebaudioside M | C(56+n6)H(90+n10)O(33+n*5) | ||
| n: number of glucose units enzymatically added to the parent steviol glycoside (n = 1-20) Typical conversion factor for glucosylated steviol glycosides mixtures = 0,20 (on the dried, dextrin-free, basis) | n: number of glucose units enzymatically added to the parent steviol glycoside (n = 1-20) Typical conversion factor for glucosylated steviol glycosides mixtures = 0,20 (on the dried, dextrin-free, basis) | n: number of glucose units enzymatically added to the parent steviol glycoside (n = 1-20) Typical conversion factor for glucosylated steviol glycosides mixtures = 0,20 (on the dried, dextrin-free, basis) | |
| Steviol | C20H30O3 | 1,00 | |
| Steviolbioside | C32H50O13 | 0,50 | |
| Rubusoside | C32H50O13 | 0,50 | |
| Dulcoside A | C38H60O17 | 0,40 | |
| Stevioside | C38H60O18 | 0,40 | |
| Rebaudioside A | C44H70O23 | 0,33 | |
| Rebaudioside B | C38H60O18 | 0,40 | |
| Rebaudioside C | C44H70O22 | 0,34 | |
| Rebaudioside D | C50H80O28 | 0,29 | |
| Rebaudioside E | C44H70O23 | 0,33 | |
| Rebaudioside F | C43H68O22 | 0,34 | |
| Rebaudioside M | C56H90O33 | 0,25 | |
| Molecular weight and CAS No | Trivial name | CAS Number | Molecular weight (g/mol) |
| Molecular weight and CAS No | n-Glucosylated Steviolbioside | Not available | 642,73+n*162,15 |
| Molecular weight and CAS No | n-Glucosylated Rubusoside | Not available | 642,73+n*162,15 |
| Molecular weight and CAS No | n-Glucosylated Dulcoside A | Not available | 788,87+n*162,15 |
| Molecular weight and CAS No | n-Glucosylated Stevioside | Not available | 804,88+n*162,15 |
| Molecular weight and CAS No | n-Glucosylated Rebaudioside A | Not available | 967,01+n*162,15 |
| Molecular weight and CAS No | n-Glucosylated Rebaudioside B | Not available | 804,88+n*162,15 |
| Molecular weight and CAS No | n-Glucosylated Rebaudioside C | Not available | 951,02+n*162,15 |
| Molecular weight and CAS No | n-Glucosylated Rebaudioside D | Not available | 1129,15+n*162,15 |
| Molecular weight and CAS No | n-Glucosylated Rebaudioside E | Not available | 967,01+n*162,15 |
| Molecular weight and CAS No | n-Glucosylated Rebaudioside F | Not available | 936,99+n*162,15 |
| Molecular weight and CAS No | n-Glucosylated Rebaudioside M | Not available | 1291,30+n*162,15 |
| Molecular weight and CAS No | Steviol | 318,46 | |
| Molecular weight and CAS No | Steviolbioside | 41093-60-1 | 642,73 |
| Molecular weight and CAS No | Rubusoside | 64849-39-4 | 642,73 |
| Molecular weight and CAS No | Dulcoside A | 64432-06-0 | 788,87 |
| Molecular weight and CAS No | Stevioside | 57817-89-7 | 804,88 |
| Molecular weight and CAS No | Rebaudioside A | 58543-16-1 | 967,01 |
| Molecular weight and CAS No | Rebaudioside B | 58543-17-2 | 804,88 |
| Molecular weight and CAS No | Rebaudioside C | 63550-99-2 | 951,02 |
| Molecular weight and CAS No | Rebaudioside D | 63279-13-0 | 1 129,15 |
| Molecular weight and CAS No | Rebaudioside E | 63279-14-1 | 967,01 |
| Molecular weight and CAS No | Rebaudioside F | 438045-89-7 | 936,99 |
| Molecular weight and CAS No | Rebaudioside M | 1220616-44-3 | 1 291,30 |
| Assay | Not less than 95 % of total steviol glycosides, comprised of above mentioned steviol glycosides along with their glucosylated derivatives (1-20 added glucose units), on the dried, dextrin-free, basis. | Not less than 95 % of total steviol glycosides, comprised of above mentioned steviol glycosides along with their glucosylated derivatives (1-20 added glucose units), on the dried, dextrin-free, basis. | Not less than 95 % of total steviol glycosides, comprised of above mentioned steviol glycosides along with their glucosylated derivatives (1-20 added glucose units), on the dried, dextrin-free, basis. |
| Description | White to light yellow powder, approximately between 100 and 200 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 100 and 200 times sweeter than sucrose (at 5 % sucrose equivalency). | White to light yellow powder, approximately between 100 and 200 times sweeter than sucrose (at 5 % sucrose equivalency). |
| Identification | Identification | Identification | Identification |
| Solubility | Soluble in water | Soluble in water | Soluble in water |
| pH | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) | Between 4,5 and 7,0 (1 in 100 solution) |
| Purity | Purity | Purity | Purity |
| Total ash | Not more than 1 % | Not more than 1 % | Not more than 1 % |
| Loss on drying | Not more than 6 % (105 °C, 2 h) | Not more than 6 % (105 °C, 2 h) | Not more than 6 % (105 °C, 2 h) |
| Residual solvent | Not more than 200 mg/kg methanol Not more than 3 000 mg/kg ethanol | Not more than 200 mg/kg methanol Not more than 3 000 mg/kg ethanol | Not more than 200 mg/kg methanol Not more than 3 000 mg/kg ethanol |
| Arsenic | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg | Not more than 0,015 mg/kg |
| Lead | Not more than 0,1 mg/kg | Not more than 0,1 mg/kg | Not more than 0,1 mg/kg |
| Cadmium | Not more than 0,1 mg/kg | Not more than 0,1 mg/kg | Not more than 0,1 mg/kg |
| Mercury | Not more than 0,1 mg/kg | Not more than 0,1 mg/kg | Not more than 0,1 mg/kg |
| Microbiological criteria | Microbiological criteria | Microbiological criteria | Microbiological criteria |
| Total (aerobic) plate count | Not more than 1 000 CFU/g | Not more than 1 000 CFU/g | Not more than 1 000 CFU/g |
| Yeast and moulds | Not more than 200 CFU/g | Not more than 200 CFU/g | Not more than 200 CFU/g |
| E. coli | Negative in 1 g | Negative in 1 g | Negative in 1 g |
| Salmonella | Negative in 25 g | Negative in 25 g | Negative in 25 g |
E 961 NEOTAME
| Synonyms | N-[N-(3,3-dimethylbutyl)-L-α-aspartyl]-L-phenylalanine 1-methyl ester; N(3,3-dimethylbutyl)-L-aspartyl-L-phenylalanine methyl ester. |
| Definition | Neotame is manufactured by reaction under hydrogen pressure of aspartame with 3,3,-dimethylbutyraldehyde in methanol in presence of a palladium/carbon catalyst. It is isolated and purified by filtration, where diatomaceous earth may be used. After solvent removal via distillation, neotame is washed with water, isolated by centrifugation and finally vacuum dried. |
| CAS Nr. | 165450-17-9 |
| Chemical name | N-[N-(3,3-dimethylbutyl)-L-α-aspartyl]-L-phenylalanine 1-methyl ester |
| Chemical formula | C20H30N2O5 |
| Molecular weight | 378,47 |
| Description | white to off-white powder |
| Assay | Not less than 97,0 % on the dried basis |
| Identification | Identification |
| Solubility | 4,75 % (w/w) at 60 °C in water, soluble in ethanol and ethyl acetate |
| Purity | Purity |
| Water content | Not more than 5 % (Karl Fischer, sample size 25 ± 5mg) |
| pH | 5,0-7,0 (0,5 % aqueous solution) |
| Melting range | 81 °C to 84 °C |
| N-[(3,3-dimethylbutyl)-L-α-aspartyl]-L-phenylalanine | Not more than 1,5 % |
| Lead | Not more than 1 mg/kg |
E 962 SALT OF ASPARTAME-ACESULFAME
| Synonyms | Aspartame-acesulfame; Aspartame-acesulfame salt |
| Definition | The salt is prepared by heating an approximately 2:1 ratio (w/w) of aspartame and acesulfame K in solution at acidic pH and allowing crystallisation to occur. The potassium and moisture are eliminated. The product is more stable than aspartame alone. |
| Einecs | |
| Chemical name | 6-Methyl-1,2,3-oxathiazine-4(3H)-one-2,2-dioxide salt of L-phenylalanyl-2-methyl-L-α-aspartic acid |
| Chemical formula | C18H23O9N3S |
| Molecular weight | 457,46 |
| Assay | 63,0 % to 66,0 % aspartame (dry basis) and 34,0 % to 37,0 % acesulfame (acid form on a dry basis) |
| Description | A white, odourless, crystalline powder |
| Identification | Identification |
| Solubility | Sparingly soluble water; slightly soluble in ethanol |
| Transmittance | The transmittance of a 1 % solution in water determined in a 1 cm cell at 430 nm with a suitable spectrophotometer using water as a reference, is not less than 0,95, equivalent to an absorbance of not more than approximately 0,022. |
| Specific rotation | [α]D 20 + 14,5° to + 16,5° Determine at a concentration of 6,2 g in 100 ml formic acid (15N) within 30 min of preparation of the solution. Divide the calculated specific rotation by 0,646 to correct for the aspartame content of the salt of aspartame-acesulfame |
| Purity | Purity |
| Loss on drying | Not more than 0,5 % (105 °C, 4 hours) |
| 5-Benzyl-3,6-dioxo-2-piperazineacetic acid | Not more than 0,5 % |
| Lead | Not more than 1 mg/kg |
E 964 POLYGLYCITOL SYRUP
| Synonyms | Hydrogenated starch hydrolysate, hydrogenated glucose syrup and polyglucitol |
| Definition | A mixture consisting mainly of maltitol and sorbitol and lesser amounts of hydrogenated oligo- and polysaccharides and maltrotriitol. It is manufactured by the catalytic hydrogenation of a mixture of starch hydrolysates consisting of glucose, maltose and higher glucose polymers, similar to the catalytic hydrogenation process used for the manufacture of maltitol syrup. The resulting syrup is desalted by ion exchange and concentrated to the desired level. |
| Einecs | |
| Chemical name | Sorbitol: D-glucitol |
| Chemical name | Maltitol: (α)-D-Glucopyranosyl-1,4-D-glucitol |
| Chemical formula | Sorbitol: C6H14O6 |
| Chemical formula | Maltitol: C12H24O11 |
| Molecular weight | Sorbitol: 182,2 |
| Molecular weight | Maltitol: 344,3 |
| Assay | Content not less than 99 % of total hydrogenated saccharides on the anhydrous basis, not less than 50 % higher molecular weight polyols, not more than 50 % of maltitol and not more than 20 % of sorbitol on the anhydrous basis. |
| Description | Colourless and odourless clear viscous liquid |
| Identification | |
| Solubility | Very soluble in water and slightly soluble in ethanol |
| Test for maltitol | Passes test |
| Test for sorbitol | To 5 g of the sample add 7 ml of methanol, 1 ml of benzaldehyde and 1 ml of hydrochloric acid. Mix and shake in a mechanical shaker until crystals appear. Filter the crystals and dissolve in 20 ml of boiling water containing 1 g of sodium bicarbonate. Filter the crystals, wash with 5 ml of a water-methanol mixture (1 in 2) and dry in the air. The crystals of the monobenzylidine derivative of sorbitol so obtained melt between 173 and 179 °C. |
| Purity | |
| Water content | Not more than 31 % (Karl Fischer method) |
| Chlorides | Not more than 50 mg/kg |
| Sulphates | Not more than 100 mg/kg |
| Reducing sugars | Not more than 0,3 % |
| Nickel | Not more than 2 mg/kg |
| Lead | Not more than 1 mg/kg |
E 965 (i) MALTITOL
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