Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

No mutagenicity in bacterial cells can be expected for Calcium glucoheptonate since all in vitro tests (reliable and less reliable) in bacterial cells conducted with the structural analogues gluconates, organic and inorganic calcium compounds were overwhelmingly negative.

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: other: summry of available in vitro and in vivo genetic toxicity data
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable well documented peer-reviewed report.
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
The study was made on one yeast strain : Saccharomyces cerevisiae, strain D4 and 3 bacteria strains: S. typhimuriumTA1535, TA1537 and TA 1538. Positive controls are different from those in the OECD 471; only 3 concentration tested.
Principles of method if other than guideline:
Summary of genetic toxicity data on glucono-delta-lactone, sodium or calcium gluconate
GLP compliance:
not specified
Type of assay:
other: summary of a variety of data
Target gene:
his-
Species / strain / cell type:
other: S.typhimurium TA 1535, TA 1537, TA 1538
Species / strain / cell type:
Saccharomyces cerevisiae
Details on mammalian cell type (if applicable):
strain D4
Metabolic activation:
with and without
Metabolic activation system:
The tissue homogenates and supernatants (9000 g) were prepared from tissues of mouse (ICR random bred adult males); rat (Sprague-Dawnley adult males) and monkey (Macaca mulatta adult males).
Test concentrations with justification for top dose:
Sodium gluconate: 0.06, 0.012, 0.024 µg/mL (Salmonella typhimurium); 12.5, 25 and 50 µg/mL (yeast);
Glucono-delta-lactone: 2.5, 5 (5 µg/mL plate test; Salmonella typhimurium); 12.5 and 25 µg/mL (yeast);
Calcium gluconate: 12.5, 25 and 50 µg/mL (Salmonella typhimurium); 7.5, 15 and 30 µg/mL (yeast).
Vehicle / solvent:
- Solvent used: 0.067 M phosphate buffer, pH 7.4
Untreated negative controls:
yes
Remarks:
solvent control
Negative solvent / vehicle controls:
yes
Remarks:
vehicle control
True negative controls:
no
Positive controls:
yes
Remarks:
without S9
Positive control substance:
2-nitrofluorene
ethylmethanesulphonate
other: Quinacrine or quinacrinemustard (QM)
Untreated negative controls:
yes
Remarks:
solvent control
Negative solvent / vehicle controls:
yes
Remarks:
vehicle control
True negative controls:
no
Positive controls:
yes
Remarks:
with S9
Positive control substance:
2-acetylaminofluorene
N-dimethylnitrosamine
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation) and in suspension

DURATION
- Exposure duration:
- Glucono-delta-lactone: 4 days: bacteria and yeasts (plate test); 4 hours (yeasts) and 1 hour (bacteria) in suspension test.
- Sodium gluconate: 48 to 72 hours bacteria and yeasts (plate test); 4 hours (yeasts) and 1 hour (bacteria) in suspension test.
- Calcium gluconate: 4 days: bacteria and yeasts (plate test); 4 hours (yeasts) and 1 hour (bacteria) in suspension test.

DETERMINATION OF CYTOTOXICITY
- Glucono-delta-lactone: 50% survival in bacteria calculated was at 1% (10 μg/mL) test substance and 5% (50 μg/mL) for yeast;
- Sodium gluconate: 50% survival in bacteria calculated was at 0.0024 % test substance and 5% for yeast;
- Calcium gluconate: 50% survival in bacteria calculated was at 5.00 % test substance and 3.00% for yeast.

Tests in suspension without S9 mix: Bacterial plates were scored after incubation for 48 hours at 37°C. The yeast plates were incubated at 30°C for 3-5days before scoring.
Species / strain:
S. typhimurium, other: TA 1535; TA 1537 and TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
valid for glucono-delta-lactone, sodium and calcium gluconate
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Cytotoxic concentration (50% survival) (μg/ml):

Sodium gluconate: 0.024 (bacteria), 50 (yeast);

Glucono-delta-lactone: 10 (bacteria), 50 (yeast);

Caclium gluconate: 50 (bacteria), 30 (yeast).

Conclusions:
Interpretation of results (migrated information):
negative

The available in vitro mutagenicity data with glucono-delta-lactone, sodium or calcium gluconate were negative.
Executive summary:

Sodium gluconate, glucono-delta-lactone and calcium gluconate were tested on Saccharomyces cerevisiae and Salmonella typhimurium with and without metabolic activation. OECD Guideline 471 was deviated for the number of strains tested and the choice of positive controls. The substances were tested on Saccharomyces cerevisiae (strain D4) and Salmonella typhimurium (3 strains) with and without metabolic activation. Only 3 concentrations were tested where OECD guideline recommends at least 5 concentrations. None of the test substances showed mutagenicity on the strains tested.

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable well documented publication which meets basic scientific principles.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Mutagenicity of Calcium Citrate, Calcium Lactate, Calcium Sulphate and Calcium Phosphate Tribasic, dietary supplements, 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 chemical was tested with and without S9 mix.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Target gene:
his-
Species / strain / cell type:
S. typhimurium TA 97
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:
- Calcium Citrate and Calcium Lactate: 0. 0.1, 0.5, 1.0, 5.0 and 10.0 mg/plate;
- Calcium sulphate and Calcium Phosphate Tribasic: 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).
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)
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:
Statisticaly significant defference by Kruskal-Wallis test ( p <0.05) and dose-related incleasing by regression analysis (p < 0.01).
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
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
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Table 1. Results of Mutation Test with calcium dietary supplements

Dose (mg/plate)

No of Revertants /plate

Calcium Citrate

 

TA97

TA102

 

-S9

+S9

-S9

+S9

10

109

185

199

348

5

136

189

253

419

1

130

204

292

491

0.5

158

196

302

476

0.1

144

200

313

460

0

154

210

303

450

Positive control

215

3.118

4,524

1,233

Calcium Lactate

 

-S9

+S9

-S9

+S9

10

138

171

252

314

5

157

183

299

323

1

150

205

285

415

0.5

134

193

300

475

0.1

143

187

321

484

0

148

197

308

475

Positive control

219

3,393

4.060

1,189

Calcium Phosphate Tribasic

 

-S9

+S9

-S9

+S9

1

126

172

197

395

0.5

122

165

230

420

0.1

114

179

287

429

0.05

119

179

265

396

0.01

124

175

270

372

0

130

178

305

377

Positive control

208

2,455

3,546

1,363

Calcium sulphate

 

-S9

+S9

-S9

+S9

1

122

174

207

356

0.5

109

195

197

320

0.1

153

191

226

360

0.05

119

184

210

339

0.01

130

199

226

384

0

128

192

212

375

Positive control

237

3,085

4,095

1,673

Solvent: DW (not specified)

Conclusions:
Interpretation of results (migrated information):
negative

Calcium citrate, Calcium Lactate, Calcium sulphate and Calcium phosphate tribasic were 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. Calcium dietary supplements: Calcium citrate, Calcium Lactate, Calcium sulphate and Calcium sulphate Tribasic were negative in two tester strains.

Endpoint:
genetic toxicity in vitro
Remarks:
Type of genotoxicity: other: summary of mutagenicity data
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable well-documented peer-reviewed report.
Qualifier:
according to
Principles of method if other than guideline:
Summary of genetic toxicity study results conducted with monosaccharides, disaccharides, and related ingredients as used in cosmetics.
GLP compliance:
not specified
Species / strain:
other: The genotoxicity of a number of the mono- and disaccharides has been evaluated in in vitro and in vivo studies. The results of these studies are overwhelmingly negative.
Metabolic activation:
with and without
Genotoxicity:
negative
Vehicle controls validity:
other: not applicable (summary result)
Untreated negative controls validity:
other: not applicable (summary result)
Positive controls validity:
other: not applicable (summary result)
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Conclusions:
Interpretation of results (migrated information):
negative

The genotoxicity of a number of the mono- and disaccharides has been evaluated in in vitro and in vivo studies. The results of these studies are overwhelmingly negative.
Endpoint:
in vitro DNA damage and/or repair study
Remarks:
Type of genotoxicity: DNA damage and/or repair
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
1980
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Documentation insufficient for assessment.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Study results from the old IUCLID attached to OECD SIDS report
GLP compliance:
no
Type of assay:
Bacillus subtilis recombination assay
Target gene:
Bacillus subtilis: H17 (rec+), H45 (rec-)
Species / strain / cell type:
bacteria, other: Bacillus subtilis H17 (rec+), H45 (rec-)
Metabolic activation:
without
Metabolic activation system:
no data
Test concentrations with justification for top dose:
0.005-0.5 M (Bacillus subtilis)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
Untreated negative controls:
yes
Remarks:
solvent control
Negative solvent / vehicle controls:
yes
Remarks:
negative control
True negative controls:
no
Positive controls:
not specified
Details on test system and experimental conditions:
CaCl2 was dissolved in distilled water. On the day of the experiments, each bacterial stock was melted and streaked radially from small pipettes onto B-2 agar. A 0.05-ml portion of each metal solution (0.005-0.5 M) was dropped on to a filter paper disk (diameter 10 mm), and the disk was
placed on the starting point of the streak. The plates were kept at 4°C for 24h and then incubated at 37°C overnight;
Evaluation criteria:
When a chemical inhibits the growth of recombination-deficient cells (H45 cells) much stronger than that of wild-type cells (H17 cells), the chemical is considered positive for mutagenicity.
Species / strain:
bacteria, other: Bacillus subtilis H17 (rec+), H45 (rec-)
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Conclusions:
Interpretation of results (migrated information):
negative without metabolic activation

Calcium chloride was negative in Bacillus subtilis H17 (rec+), H45 (rec-) recombination assay without metabolic activation.
Executive summary:

In a Bacillus subtilis mutagenicity assay (no OECD guideline study) the potential of calcium chloride to damage cellular DNA was examined at concentrations up to 0.5 M (Tokuyama Corporation). The result of the test was negative.

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
1987
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Documentation insufficient for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
The study result originates from the OECD SIDS report (attached IUCLID)
GLP compliance:
no
Type of assay:
other: SOS Chromotest
Target gene:
sfiA
Species / strain / cell type:
E. coli, other: PQ37
Metabolic activation:
without
Metabolic activation system:
no data
Test concentrations with justification for top dose:
1 - 1000 μmol/L (0.1-111 mg/L)
Species / strain:
E. coli, other: PQ37
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Conclusions:
Interpretation of results (migrated information):
negative without metabolic activation

Calcium chloride was negative in a SOS Chromotest in E. coli PQ37 without metabolic activation.
Executive summary:

In an Escherichia coli test (no OECD guideline study) the potential to induce an SOS response was tested at doses up to 1 mM, also giving a negative result (Tokuyama Crporation).

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
1984 (S. typhimurium TA92, TA1535, TA100, TA1537, TA94, TA98
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable well-documented OECD SIDS Report (Comparable to guideline study with acceptable restrictions; Critical study for SIDS endpoint).
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Principles of method if other than guideline:
First study: The method was in principle equivalent to OECD Guideline 471, except that the test was carried out only with metabolic activation (S. typhimurium TA92, TA1535, TA100, TA1537, TA94, TA98);
second study: The method was in principle equivalent to OECD Guideline 471, except that some of the recommended test strains were not included (used only S.typhimurium TA97 and TA102).
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Species / strain / cell type:
S. typhimurium, other: TA92, TA1535, TA100, TA1537, TA94, TA98
Species / strain / cell type:
S. typhimurium, other: TA97, TA102
Metabolic activation:
with
Metabolic activation system:
The liver microsome fraction (S9) was prepared from the liver of Fischer rats pretreated 5 days before with polychlorinated biphenyls (500 mg/kg body weight of Kanechlor KC-400 in olive oil, ip).
Test concentrations with justification for top dose:
- First study: Duplicate plates were used for each of six different concentrations of the sample (max 5.0 mg/plate): in Salmonella typhimurium TA92, TA1535, TA100, TA1537, TA94, TA98;
- Second study: Triplicate plates were used for each of five different concentrations of the sample (dose range9: 0.1 - 10 mg/plate (Salmonella typhimurium TA97, TA102)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: phosphate buffer (S.typhimurium TA92, TA1535, TA100, TA1537, TA94, TA98);
water (S.typhimurium TA97 and TA102)
Untreated negative controls:
yes
Remarks:
solvent control
Negative solvent / vehicle controls:
yes
Remarks:
negative control
True negative controls:
no
Positive controls:
no
Remarks:
in test with S. typhimurium TA92, TA1535, TA100, TA1537, TA94, TA98
Untreated negative controls:
yes
Remarks:
solvent control
Negative solvent / vehicle controls:
yes
Remarks:
negative control
True negative controls:
no
Positive controls:
yes
Remarks:
in test with S. typhimurium TA97 and TA102
Positive control substance:
9-aminoacridine
mitomycin C
other: 2-aminoanthracene
Remarks:
9-aminoacridine (20 μg/plate) for TA97 without S9; 2-aminoanthracene (5 μg/plate) for TA97 with S9; mytomycin C (0.5 μg/plate) for TA102 without S9; and 2-aminoanthracene (5 μg/plate) for TA102 with S9.
Details on test system and experimental conditions:
CaCl2 was dissolved in water and 0.1 ml solution containing the indicated amount of CaCl2 was used for each plate (in the test with S.typhimurium TA97 and TA102).
Evaluation criteria:
The result was considered positive if the number of colonies found was twice the number of colonies of the control, which was exposed to phosphate buffer, the solvent for CaCl2 (S. typhimurium TA92, TA1535, TA100, TA1537, TA94, TA98).
Species / strain:
S. typhimurium, other: TA92, TA1535, TA100, TA1537, TA94, TA98
Metabolic activation:
with
Genotoxicity:
negative
Remarks:
A negative result indicates that no significant increases in the number of revertant colonies were detected in any S. Typhimurium strains at the maximum dose.
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
not examined
Species / strain:
S. typhimurium, other: TA97 and TA102
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
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Table 1. Mutation test with CaCl2

Dose (mg/plate)

No. of revertants

 

TA97

TA102

 

-S9

+S9

-S9

+S9

10

140

173

168

287

5

141

170

230

340

1

132

206

291

376

0.5

140

213

286

424

0.1

149

225

313

437

0

146

212

325

447

PC

205

2292

4930

1428

PC: positive control

Conclusions:
Interpretation of results (migrated information):
negative

Ca chloride was negative in two in vitro reverse mutation studies.
Executive summary:

"Two studies were carried out by the method similar to OECD Test Guideline 471. In a Salmonella mutation test, using TA92, TA94, TA98, TA100, TA1535 and TA1537, doses of calcium chloride up to 5 mg/plate were examined with metabolic activation [39]. In another Salmonella mutation test, using TA97 and TA102, doses up to 10 mg/plate were examined with or without metabolic activation [40]. No significant increases in mutation frequencies were observed in either study".

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Target gene:
his- (S.typhimurium)
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
5 % v/v S-9 in the S-9 mix from Aroclor-1254 induced Rat (Sprague-Dawley strain), male, liverhomogenate
Test concentrations with justification for top dose:
12, 37, 111, 333, 1,000 and 3,000 μg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: sterile distilled water
Untreated negative controls:
yes
Remarks:
solvent control
Negative solvent / vehicle controls:
yes
Remarks:
negative control
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
sodium azide
other: 2-aminoanthracene (2-AA)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

NUMBER OF REPLICATIONS: 1


Statistics:
Not used.
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Table 1. Result of bacterial reverse mutation assay with calcium sulfate, dihydrate

Tester strain

Chemical treated

Doseg/plate)

Colonies/plate (mean)[Factor]

Without S-9 mix

With S-9 mx

TA100

 

 

 

 

 

Test item

0

132

114

12

139 [1.0]

115 [1.0]

37

143 [1.1]

122 [1.1]

111

125 [0.9]

115 [1.0]

333

129 [1.0]

125 [1.0]

1,000

124 [0.9]

106 [1.0]

3,000

124 [0.9]

115 [1.0]

TA1535

 

 

 

 

 

Test item

0

13

10

12

20 [1.5]

11 [1.1]

37

22 [1.7]

12 [1.2]

111

17 [1.3]

13 [1.3]

333

16 [1.2]

14 [1.4]

1,000

17 [1.3]

13 [1.3]

3,000

14 [1.1]

9 [0.9]

TA98

 

 

 

 

 

Test item

0

22

34

12

22 [1.0]

28 [0.8]

37

21 [0.9]

26 [0.8]

111

29 [1.3]

37 [1.1]

333

25 [1.1]

31 [0.9]

1,000

23 [1.0]

33 [1.0]

3,000

22 [1.0]

36 [1.0]

TA 1537

 

 

 

 

 

Test item

0

15

17

12

16 [1.0]

17 [1.0]

37

11 [0.7]

22 [1.3]

111

15 [1.0]

19 [1.1]

333

14 [0.9]

17 [1.0]

1,000

13 [0.9]

16 [0.9]

3,000

12 [0.8]

16 [0.9]

E.coli

WP2 uvrA

Test item

0

8

12

 

12

7 [0.9]

8 [0.7]

 

37

6 [0.8]

13 [1.1]

 

111

8 [1.0]

8 [0.7]

 

333

8 [1.0]

8 [0.7]

 

1,000

6 [0.8]

9 [0.8]

 

3,000

6 [0.8]

9 [0.8]

Positive controls

 

 

 

 

TA100

SA

0.5

493 [3.7]

 

TA1535

SA

0.5

371 [28.5]

 

TA98

4NQO

0.5

426 [19.4]

 

TA1537

9-AA

50

740 [49.3]

 

WP2 uvrA

4NQO

0.5

377 [47.1]

 

TA100

2-AA

0.4

 

491 [4.3]

TA1535

2-AA

2

 

394 [39.4]

TA98

2-AA

0.4

14 [0.6]

289 [8.5]

TA1537

2-AA

2

 

326 [19.2]

WP2 uvrA

2-AA

4

 

311 [25.9]

[Factor]: No. of colonies of treated plate/No. of colonies of negative control plate

SA: Sodium azide 9-AA: 9-Amino acridine

4NQQ: 4-nitroquinoline-1-oxide 2-AA: 2-aminoanthracene

Conclusions:
Interpretation of results (migrated information):
negative

The mutation in the Salmonella tryphimurium (strains TA 98, TA 100, TA 1535, and TA 1537) and in the Escherchia coli WP2 uvrA did not occur with calcium sulfate, dihydrate.
Executive summary:

"A preliminary test was carried out to decide the appropriate starting dose level of the main study at the concentrations of 1.6, 8, 40, 200, 1,000 and 3,000 mg/plate. In the main study, calcium sulfate, dihydrate was negative in the bacterial reverse mutation assay with Salmonella tryphimurium (strains TA98, TA100, TA1535, and TA1537) and Escherichia coli WP2 uvrA with and without metabolic activation tested at concentrations of 12, 37, 111, 333, 1,000 and 3,000 mg/ plate in accordance with OECD TG 471 and 472. The number of revertant colonies in the plate was counted after 2 day-incubation at 37 °C. In conclusion, no mutation occurred with calcium sulfate, dihydrate with and without metabolic activation".

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

In vivo chromosome aberrations studies conducted with glucono-delta-lactone and sodium gluconate were negative clear showing that no genetic toxicity can be attributed to gluconate and glucoheptonate ions.

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vivo mammalian germ cell study: cytogenicity / chromosome aberration
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable well-documented peer-reviewed report.
Qualifier:
no guideline available
Deviations:
not specified
Remarks:
on GLP, guidelines, conditions, mitotic index, but sufficient for initial assessment.
GLP compliance:
no
Remarks:
the study was conducted prior to adoption of guidelines (in 1974).
Type of assay:
chromosome aberration assay
Species:
mouse
Strain:
C57BL
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 12 or 13 weeks
Route of administration:
oral: feed
Vehicle:
- Vehicle(s)/solvent(s) used: physiol. saline;

- Concentration of test material in vehicle:
- Amount of vehicle (if gavage or dermal): 1 mL/mouse
Duration of treatment / exposure:
single dose and 4 days
Frequency of treatment:
not specified
Post exposure period:
The animals were sacrified at 24 hours (single dose) and 27 hours after last administration (4-days repeated dose).
Remarks:
Doses / Concentrations:
2, 4 and 8 g/kg
Basis:
other: single dose administration
Remarks:
Doses / Concentrations:
2 and 4 g/kg
Basis:
other: 4 day repeated dose
No. of animals per sex per dose:
Single dose administration: 3 (vehicle control and test groups); 2 (positive control);
4-day repeated dose administration: 2 (vehicle control); 3 (test group 1: 4 g/kg); 2 (test group 2: 2 g/kg); 2 (positive control).
Control animals:
yes, concurrent vehicle
Positive control(s):
MMC (mitomycin C) dissolved with 0.9% physiological saline solution and administered intraperitoneally at a dose of 0.5 mL/mouse (= 5mg/kg bw).
Tissues and cell types examined:
At least 200 metaphase cells per mouse were examined for the presence or absence of chromosomal aberrations (gaps, breaks, translocation, fragments, ring chromosomes and minutes chromosomes).
Details of tissue and slide preparation:
TREATMENT AND SAMPLING TIMES: After receiving the single dose and the repeated dose test substance, the animals were sacrified at 24 hours (single dose) and 27 hours after last administration (4-days repeated dose). 0.3 mL of 500 μg/mL colchicine was intraperitoneally injected to each mouse at one hour before sacrifice so that the metaphase cells could be observed.

DETAILS OF SLIDE PREPARATION: After the bone marrow cells were washed, treated and fixed with a fixing solution (1:3 acetic acid:ethanol solution), the cells were suspended and dripped on a slide glass and stained with Giemsa solution and examined.
Sex:
male
Genotoxicity:
negative
Remarks:
in both experiments: single administration and 4-d repeated dose administration.
Toxicity:
yes
Remarks:
At 8 g/kg, all mice died (single dose administration experiment)
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid

Single dose administration:

At 8 g/kg, all mice died.

MMC induced chromosomal aberrations in at least 20% of bone marrow cells.

GDL induced chromosomal aberrations in the cells at a frequency of about 0.5% comparable to the control.

4-day repeated dose administration:

MMC induced chromosomal aberrations at about 30% cells.

The frequency of cells with chromosomal aberrations was 1 % or less in the test groups which is comparable to the control group. Induction of chromosomal aberration by GDL was not detected after in vivo single and repeated dose treatment.

Conclusions:
Interpretation of results (migrated information): negative
The frequency of cells with chromosomal aberrations in the test groups was comparable to the control group in both experiments: single administartion and 4-d repeat administration.
Endpoint:
in vivo mammalian germ cell study: cytogenicity / chromosome aberration
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable well-documented peer-reviewed reports.
Qualifier:
no guideline available
Deviations:
not specified
Remarks:
on GLP, guidelines, conditions, mitotic index, but sufficient for initial assessment.
GLP compliance:
no
Remarks:
the study was conducted prior to adoption of guidelines (in 1974).
Type of assay:
chromosome aberration assay
Species:
mouse
Strain:
C57BL
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 12 or 13 weeks
Route of administration:
oral: feed
Vehicle:
- Vehicle(s)/solvent(s) used: physiol. saline;

- Concentration of test material in vehicle:
- Amount of vehicle (if gavage or dermal): 1 mL/mouse
Duration of treatment / exposure:
single dose and 4 days
Frequency of treatment:
not specified
Post exposure period:
The animals were sacrified at 24 hours (single dose) and 27 hours after last administration (4-days repeated dose).
Remarks:
Doses / Concentrations:
2.5, 5 and 10 g/kg
Basis:
other: single dose administration
Remarks:
Doses / Concentrations:
1.25 and 2.5 g/kg
Basis:
other: 4 day repeated dose
No. of animals per sex per dose:
Single dose administration: 3 (vehicle control and test groups); 2 (positive control);
4-day repeated dose administration: 2 (vehicle control); 3 (test group 1: 2.5 g/kg); 2 (test group 2: 1.25 g/kg); 2 (positive control).
Control animals:
yes, concurrent vehicle
Positive control(s):
MMC (mitomycin C) dissolved with 0.9% physiological saline solution and administered intraperitoneally at a dose of 0.5 mL/mouse (= 5mg/kg bw).
Tissues and cell types examined:
At least 200 metaphase cells per mouse were examined for the presence or absence of chromosomal aberrations (gaps, breaks, translocation, fragments, ring chromosomes and minutes chromosomes).
Details of tissue and slide preparation:
TREATMENT AND SAMPLING TIMES: After receiving the single dose and the repeated dose test substance, the animals were sacrified at 24 hours (single dose) and 27 hours after last administration (4-days repeated dose). 0.3 mL of 500 μg/mL colchicine was intraperitoneally injected to each mouse at one hour before sacrifice so that the metaphase cells could be observed.

DETAILS OF SLIDE PREPARATION: After the bone marrow cells were washed, treated and fixed with a fixing solution (1:3 acetic acid:ethanol solution), the cells were suspended and dripped on a slide glass and stained with Giemsa solution and examined.
Sex:
male
Genotoxicity:
negative
Remarks:
in both experiments: single dose administration and 4-d repeated dose administration.
Toxicity:
yes
Remarks:
At 10 and 5 g/kg, all mice died (single dose administration); at 1.25 and 2.5 g/kg, one mouse died in each group (4-day repeated dose administration).
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid

Single dose administration:

At 10 and 5 g/kg, all mice died.

At 2.5 g/kg, observation could be made only on 2 animals (preparation of the chromosome specimen failed).

MMC induced chromosomal aberrations in at least 20% of bone marrow cells. Sodium gluconate induced chromosomal aberrations in the cells at a frequency of about 0.5% is comparable to the control. (1 gap and 1 minute chromosome for 283 cells).

4-day repeated dose administration:

At 1.25 and 2.5 g/kg, one mouse died in each group.

MMC induced chromosomal aberrations at about 30% cells. The frequency of cells with chromosomal aberrations was 0.5% in the test groups which is comparable to the control group.

Conclusions:
Interpretation of results (migrated information): negative
Induction of chromosomal aberration by sodium gluconate was not detected after in vivo single and repeated dose treatment.

Additional information

There are no genetic toxicity in vitro studies available for Calcium glucoheptonate. Since calcium glucoheptonate is expected to be diccociated into calcium and glucoheptonate ions, data on structurally similar substances: gluconic acid, its derivatives and some inorganic calcium compounds have been taken into account to assess mutagenicity potential of calcium and glucoheptonate ions in bacterial cells.

In vitro genetic toxicity studies with Calcium gluconate, Sodium gluconate and glucono-delta-lactone

Sodium gluconate, glucono-delta-lactone and calcium gluconate were tested by plate incorporation method and in suspension on Saccharomyces cerevisiae and Salmonella typhimurium with and without metabolic activation (SIDS, 2004). OECD Guideline 471 was deviated for the number of strains tested and the choice of positive controls. The substances were tested on Saccharomyces cerevisiae (strain D4) and Salmonella typhimurium (3 strains: TA 1535, TA 1537, TA 1538) with and without metabolic activation. Only 3 concentrations were tested where OECD guideline recommends at least 5 concentrations. None of the test substances showed mutagenicity on the strains tested.

The genotoxicity of a number of sugar-like substances the mono- and disaccharides has been evaluated in in vitro and in vivo studies. The results of these studies are overwhelmingly negative (CIR, 2014).

In vivo genetic toxicity studies with glucono-delta-lactone and sodium gluconate.

Since glucono-delta-lactone, sodium and calcium gluconates were tested only in three strains of Salmonella typhimurium with three concentrations, the results of in vivo studies conducted with glucono-delta-lactone and sodium gluconate (SIDS, 2004) can be taken into account to assess genetic toxicity potential of glucoheptonate ion.

Glucono-delta-lactone and sodium gluconate did not induce chromosomal aberrations in mice. The frequency of cells with chromosomal aberrations in the test groups was comparable to the control group in both experiments: single administartion and 4-d repeat administration.

Based on these data, no genetic toxicity can be attributed to gluconate ions, its derivatives and other sugar-like compounds including glucoheptonate ion.

In vitro genetic toxicity studies with organic and inorganic calcium compounds

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 (Fujita et al., 1988). 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. The concentrations tested were:0. 0.01, 0.05, 0.1, 0.5 and 1.0 mg/plate. Calcium citrate, Calcium sulphate, Calcium lactate and Calcium Phosphate Tribasic were negative in two tester strains.

In OECD SIDS report (2002) on calcium chloride (CAS 10043 -52 -4) results of genetic toxicity studies are reported: "Two studies were carried out by the method similar to OECD Test Guideline 471. In a Salmonella mutation test, using TA92, TA94, TA98, TA100, TA1535 and TA1537, doses of calcium chloride up to 5 mg/plate were examined with metabolic activation (Ishidate et al., 1984). In another Salmonella mutation test, using TA97 and TA102, doses up to 10 mg/plate were examined with or without metabolic activation. No significant increases in mutation frequencies were observed in either study. Two genetic toxicity studies with bacteria have further been reported although the studies are not OECD guideline studies. In a Bacillus subtilis mutagenicity assay the potential of calcium chloride to damage cellular DNA was examined at concentrations up to 0.5 M. The result of the test was negative. In an Escherichia coli test the potential to induce an SOS response was tested at doses up to 1 mM, also giving a negative result".

Bacterial gene reverse mutation tests with calcium sulphate, dihydrate (CAS 10101 -41 -4) in Salmonella typhimurium (strains TA98, TA100, TA1535 and TA1537) and Escherichia coli WP2 uvrA with and without metabolic activation gave negative results (SIDS, 2003).

Based on these data, no genetic toxicity can be attributed to calcium ion.


Justification for classification or non-classification

All available genetic toxicity studies in vitro in bacterial cells and yeasts conducted with calcium gluconate, sodium gluconate, glucono-delta-lactone, sugar-like compounds and their derivatives as well as with inorganic calcium compounds were negative. Additionally, in vivo studies conducted with glucono-delta-lactone and sodium gluconate did not induce chromosomal aberrations in mice, clear showing that no genetic toxicity can be attributed to glucoheptonate ion. Based on these data, Calcium glucoheptonate can be considered as not mutagenic as well.

According to European Regulation (EC) No. 1272/2008, Calcium glucoheptonate does not need to be classified and labelled as mutagenic substance.