Reaction products of sodium glucoheptonate with copper sulfate and ammonium hydroxide

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable well documented publication which meets basic scientific principles.

Data source

Reference

Reference Type:

publication

Title:

Mutagenicity Test of Food Additives with Salmonella typhimuriutn TA97 and TA102 (III)

Author:

Fujita, H., Nakano, M., Sasaki, M.

Year:

1998

Bibliographic source:

Ann. Rep Tokyo Metr. Res Lab. PH. (The Tokyo Metropolitan Research Laboratory of Public Health), 39, 343-350, 1988

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Mutagenicity of copper gluconate, a dietary supplement, was examined in Ames' tester strains, Salmonella typhimurium TA97 and TA102. The mutation test was carried out by the preincubation procedure described by Ames et al. The test chemical was tested with and without S9 mix.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Copper gluconate

Cas Number:

527-09-3

IUPAC Name:

Copper gluconate

Constituent 2

Reference substance name:

Copper di-D-gluconate

EC Number:

208-408-2

EC Name:

Copper di-D-gluconate

Cas Number:

527-09-3

Molecular formula:

C12H22CuO14

IUPAC Name:

copper(2+) bis(2,3,4,5,6-pentahydroxyhexanoate) (non-preferred name)

Test material form:

not specified

Method

Target gene:

his-

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 97

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 102

Metabolic activation:

with and without

Metabolic activation system:

no data

Test concentrations with justification for top dose:

0. 0.01, 0.05, 0.1, 0.5 and 1.0 mg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: phosphate buffer (pH 7.4).

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

Phosphate buffer (pH 7.4)

Negative solvent / vehicle controls:

yes

Remarks:

negative control

True negative controls:

no

Positive controls:

yes

Remarks:

in TA97

Positive control substance:

9-aminoacridine

other: 2-AA

Remarks:

-S9 mix: 9-AA (30 µg); +S9 mix: 2-AA (5 µg)

Controls 2

Untreated negative controls:

yes

Remarks:

Phosphate buffer (pH 7.4)

Negative solvent / vehicle controls:

yes

Remarks:

negative control

True negative controls:

no

Positive controls:

yes

Remarks:

in TA102

Positive control substance:

mitomycin C

other: 2-AA

Remarks:

-S9 mix: MMC (0.5 µg); +S9 mix: 2-AA (5 µg)

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 min

Statistics:

Statistically significant defference by Kruskal-Wallis test ( p <0.05) and dose-related increasing by regression analysis (p < 0.01).

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Table 1. Results of Mutation Test with copper (II) gluconate

Dose (mg/plate)	No of Revertants /plate
	TA97		TA102
	-S9	+S9	-S9	+S9
1	105	106	82	205
0.5	108	127	114	317
0.1	120	121	220	396
0.05	116	133	266	394
0.01	135	139	253	389
0	124	173	238	432
Positive control	179	2523	4105	1668
Solvent: DW (not specified)

Applicant's summary and conclusion

Conclusions:

Copper gluconate was negative in two tester strains Salmonella typhimurium TA97 and TA102.

Executive summary:

Mutagenicity of 28 food additives including 7 dietary supplements, 7 free flowing agents, 5 antioxidants, 3 thickening agents, 3 food colors, 2 color fixatives, and an anticaking agent were examined in Ames' tester strains, Salmonella typhimurium TA97 and TA102. The mutation test was carried out by the preincubation procedure described by Ames et al. The test chemicals were preincubated with S9 mix or phosphate buffer (pH7.4) for 20 min. Copper gluconate was negative in two tester strains.