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The Food Additives and Novel Foods (Authorisations and Miscellaneous Amendments) and Food Flavourings (Removal of Authorisations) (Scotland) Regulations 2024
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“E 960C(ii) REBAUDIOSIDE M, AM AND D PRODUCED VIA ENZYMATIC CONVERSION OF HIGHLY PURIFIED STEVIOL GLYCOSIDES FROM STEVIA LEAF EXTRACTS
| Synonyms | |||
|---|---|---|---|
| Definition | Steviol glycosides produced via enzymatic conversion of highly purified steviol glycosides (rebaudioside A or stevioside) from Stevia leaf extracts are composed predominantly of rebaudioside M, rebaudioside D, and rebaudioside AM. Rebaudiosides D, M and AM are produced via enzymatic conversion of highly purified steviol glycoside (rebaudioside A or stevioside) extracts (95% steviol glycosides) obtained from Stevia rebaudiana Bertoni plant using UDP-glucosyltransferase and sucrose synthase enzymes produced by genetically modified strains of Escherichia coli (pPM294, pFAH170, and pSK041) 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 steviol glycosides by resin adsorption, followed by recrystallisation of the steviol glycosides resulting in a final product containing not less than 95 % of total steviol glycosides, including one or more of rebaudiosides D, M and AM. Viable cells or DNA of Escherichia coli (pPM294, pFAH170, and pSK041) 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 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 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 |
| Rebaudioside M | C56 H90 O33 | 0.25 | |
| Rebaudioside D | C50 H80 O28 | 0.29 | |
| Rebaudioside AM | C50 H80 O28 | 0.29 | |
| Molecular weight and CAS No | Trivial name | CAS Number | Molecular weight (g/mol) |
| Rebaudioside M | 1220616-44-3 | 1291.29 | |
| Rebaudioside D | 63279-13-0 | 1129.15 | |
| Rebaudioside AM | 2222580-26-7 | 1129.15 | |
| Assay | Not less than 95 % of steviol glycosides on the dried basis, including one or more of rebaudiosides D, M and AM | ||
| 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”. | ||
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