| 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 D is a steviol glycoside composed predominantly of rebaudioside D with minor amounts of other steviol glycosides such as rebaudioside A, rebaudioside B, and rebaudioside M.
Rebaudioside M or D is obtained via enzymatic bioconversion of purified steviol glycoside leaf extracts (95% steviol glycosides) using UDP-glucosyltransferase and sucrose synthase enzymes produced by the genetically modified yeasts K. phaffi (formerly known as Pichia pastoris) UGT-a and/or K. phaffi UGT-b that facilitate the transfer of glucose from sucrose and UDP-glucose to steviol glycosides via glycosidic bonds. Rebaudioside M is produced using enzymes from both yeasts, whereas rebaudioside D is produced using enzymes only from K. phaffi UGT-a.
After removal of the enzymes by solid-liquid separation and heat treatment, the purification involves concentration of the rebaudioside M or D by resin adsorption, followed by recrystallisation of rebaudioside M or D resulting in a final product containing not less than 95 % of rebaudioside M, or not less than 95 % rebaudioside D. Viable cells of the yeasts K. phaffii UGT-a and/or K. phaffii UGT-b or 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 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; 13-[(O-b-D-Glucopyranosyl-(1-2)-O-[b-D-glucosylpyranosyl-(1-3)]-b-Dglucosylpyranosyl)oxy]kaur-16-en-18-oic acid (4-)-O-b-D-glucosylpyranosyl-(1-2)-O-[b-Dglucosylpyranosyl-(1-3)]-b-Dglycosylpyranosyl ester |
| Trivial name, molecular formula and conversion factors |
Rebaudioside D, C50H80O28, conversion factor 0.29.
Rebaudioside M, C56H90O33, conversion factor 0.25. |
| Trivial name, CAS number and molecular weight (g/mol) |
Rebaudioside D, 63279-13-0, 1 129.15.
Rebaudioside M, 1220616-44-3, 1 291.30. |
| Assay |
Not less than 95 % rebaudioside M, on the dried basis; or Not less than 95% rebaudioside D, 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. |
| Identification (pH) |
Between 4.5 and 7.0 (1 in 100 solution). |
| Purity (Total ash) |
Not more than 1 %. |
| Purity (Loss on drying) |
Not more than 6 % (105 °C, 2h). |
| Purity (Residual solvent) |
Not more than 5 000 mg/kg ethanol. |
| Purity (Arsenic) |
Not more than 0.5 mg/kg. |
| Purity (Lead) |
Not more than 0.25 mg/kg. |
| Purity (Cadmium) |
Not more than 0.25 mg/kg. |
| Purity (Mercury) |
Not more than 0.1 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]. |