Commission Regulation (EU) 2022/1922 of 10 October 2022 amending the Annex to Regulation (EU) No 231/2012 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 as regards specifications for rebaudiosides M, D and AM produced via enzymatic conversion of puriﬁed stevia leaf extracts and the specifications for rebaudioside M produced via enzyme modification of steviol glycosides from Stevia (E 960c(i)) (Text with EEA relevance)

Article 1

The Annex to Regulation (EU) No 231/2012 is amended in accordance with the Annex to this Regulation.

Article 2

This Regulation shall enter into force on the twentieth day following that of its publication in the Official Journal of the European Union .

ANNEX

ANNEX
The Annex to Regulation (EU) No 231/2012 is amended as follows:
(1)
in the entry for E 960c(i) Rebaudioside M produced via enzyme modification of Steviol glycosides from stevia, in the row ‘Definition’ the last sentence is replaced by the following:
‘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.’;
(2)
the following entries are inserted after the entry for E 960c(i):
‘
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.
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
C 56 H 90 O 33
0,25
Molecular weight and CAS No
Trivial name
CAS Number
Molecular weight (g/mol)
Rebaudioside M
1220616-44-3
1 291,29
Assay
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).
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 5 000 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]
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.
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
Molecular formula
Trivial name
Formula
Conversion factor
Rebaudioside D
C 50 H 80 O 28
0,29
Rebaudioside A
C 44 H 70 O 23
0,33
Molecular weight and CAS No
Trivial name
CAS Number
Molecular weight (g/mol)
Rebaudioside D
63279-13-0
1 291,15
Rebaudioside A
58543-16-1
967,01
Assay
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).
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 5 000 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]
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.
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.
Molecular formula
Trivial name
Formula
Conversion factor
Rebaudioside AM
C 50 H 80 O 28
0,29
Molecular weight and CAS No
Trivial name
CAS Number
Molecular weight (g/mol)
Rebaudioside AM
2222580-26-7
1 291,15
Assay
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).
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 5 000 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]’

Schedules & Appendices

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