3. In the appropriate place, insert the following entry—
“E 960c 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 or the DNA of the yeasts K. phaffi UGT-a or K. phaffi UGT-b must 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 | ||
| Molecular formula | Trivial name | Formula | Conversion factor |
| Rebaudioside M | C56 H90 O33 | 0.25 | |
| Molecular weight and CAS number | Trivial name | CAS Number | Molecular weight (g/mol) |
| Rebaudioside M | 1220616-44-3 | 1291.29 | |
| Assay | 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) | ||
| Identification | |||
| Solubility | Freely soluble to slightly soluble in water | ||
| pH | Between 4.5 and 7.0 (1 in 100 solution) | ||
| Purity | |||
| Total ash | Not more than 1% | ||
| Loss on drying | Not more than 6% (105°C, 2h) | ||
| Residual solvent | Not more than 5000 mg/kg ethanol | ||
| Arsenic | Not more than 0.015 mg/kg | ||
| Lead | Not more than 0.2 mg/kg | ||
| Cadmium | Not more than 0.015 mg/kg | ||
| Mercury | Not more than 0.07 mg/kg | ||
| Residual protein | 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)” | ||