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Toxicological information

Genetic toxicity: in vitro

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Administrative data

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:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions (analytical purity of test substance not reported).

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report date:
2007

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
(analytical purity of test substance not reported, 2-aminoanthracene was the sole indicator of the efficacy of the S9-mix)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Sorbitan stearate
EC Number:
215-664-9
EC Name:
Sorbitan stearate
Cas Number:
1338-41-6
IUPAC Name:
1,4-anhydro-6-O-stearoyl-D-glucitol
Details on test material:
- Name of test material (as cited in study report): Sorbitan, monooctadecanoate (SMO)
- Physical state: yellow crystalline pellet
- Analytical purity: no data
- Lot/batch No.: #60
- Stability under test conditions: verified
- Storage condition of test material: sealed and kept at room temperature

Method

Target gene:
his operon
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
E. coli WP2 uvr A pKM 101
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9 mix
Test concentrations with justification for top dose:
Range-finder: 50, 150, 500, 1500, 5000 µg/plate
Main study: 313, 625, 1250, 2500, 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Dimethyl sulfoxide
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: -S9: TA100: 2-(2-Furyl)-3-(5-nitro-2-furyl)aerylamide; TA1535: Sodium azide; WP2 uvrA: 2-(2-Furyl)-3-(5-nitro-2-furyl)aerylamide; TA98: 2-(2-Furyl)-3-(5-nitro-2-furyl)aerylamide; TA1537: 9-Aminoacridine; +S9: 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 3
Two independent experiments were performed, each in triplicate.

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth


Evaluation criteria:
If the average number of colonies on plates containing the test substance increased more than twice the negative control, the test is considered positive.

Results and discussion

Test resultsopen allclose all
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, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A pKM 101
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: at concentration of 625 µg/plate (without S9) and at 1250 µg/plate (with S9) and higher concentrations in both experiments.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
Cytotoxicity was observed at precipitating concentrations
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Range finding test:

With or without S9-Mix Test substance concentration Mean number of revertant colonies per plate 
(μg/plate)
Base-pair substitution type Frameshift type
TA 100 TA1535 WP2 uvrA TA98 TA1537
0 116 ± 10 9 ± 3 17 ± 2 18 ± 3 7 ± 4
50 107 10 21 27 5
150 125 12 28 19 4
500* 133 9 16 16 8
1500* 118 10 16 18 7
5000* 108 7 8 12 4
Positive controls, –S9 Name  AF-2 SA AF-2 AF-2 9AA
Concentrations (μg/plate) 0.01 0.5 0.01 0.1 80
Mean No. of colonies/plate (average of 3 ± SD) 488 ± 39 588 ± 36 112 ± 9 433 ± 39 371 ± 47
+ 0 119 ± 9 7 ± 1 21 ± 3 19 ± 1 12 ± 3
+ 50 116 11 17 23 10
+ 150 114 13 15 22 10
+ 500* 129 8 23 31 10
+ 1500* 120 3 20 21 4
+ 5000* 108 4 23 17 6
Positive controls, –S9 Name  2AA 2AA 2AA 2AA 2AA
Concentrations (μg/plate) 1 2 10 0.5 2
Mean No. of colonies/plate (average of 3 ± SD) 862 ± 7 371 ± 36 863 ± 18 353 ± 18 163 ± 14

Experiment I:

With or without S9-Mix Test substance concentration Mean number of revertant colonies per plate 
(μg/plate) (average of 3 plates ± Standard deviation)
Base-pair substitution type Frameshift type
TA 100 TA1535 WP2 uvrA TA98 TA1537
0 123 ± 15 11 ± 3 21 ± 6 20 ± 3 6 ± 1
313 99 ± 12 14 ± 2 17 3 18 ± 2 7 ± 2
625* 116 ± 9 9 ± 5 20 ± 8 17 ± 4 5 ± 1
1250* 126 ± 9 10 ± 2 19 ± 3 17 ± 8 8 ± 3
2500* 109 ± 10 8 ± 2 13 ± 3 17 ± 3 6 ± 2
5000* 99 ± 7 6 ± 1 17 ± 2 18 ± 4 6 ± 1
Positive controls, –S9 Name  AF-2 SA AF-2 AF-2 9AA
Concentrations (μg/plate) 0.01 0.5 0.01 0.1 80
Mean No. of colonies/plate (average of 3 ± SD) 493 ± 18 584 ± 50 109 ± 4 423 ± 38 311 ± 10
+ 0 135 ± 23 11 ± 3 22 ± 3 30 ± 8 12 ± 6
+ 313 140 ± 23 7 ± 4 34 ± 2 30 ± 9 15 ± 1
+ 625 140 ± 17 9 ± 2 25 ± 8 35 ± 4 11 ± 2
+ 1250* 142 ± 10 8 ± 4 26 ± 5 32 ± 3 14 ± 7
+ 2500* 153 ± 11 8 ± 2 28 ± 2 29 ± 3 12 ± 2
+ 5000* 153 ± 10 7 ± 4 27 ± 9 24 ± 3 11 ± 3
Positive controls, –S9 Name  2AA 2AA 2AA 2AA 2AA
Concentrations (μg/plate) 1 2 10 0.5 2
Mean No. of colonies/plate (average of 3 ± SD) 796 ± 19 335 ± 45 906 ± 51 356 ± 22 150 ± 2

Experiment II:

With or without S9-Mix Test substance concentration Mean number of revertant colonies per plate 
(μg/plate) (average of 3 plates ± Standard deviation)
Base-pair substitution type Frameshift type
TA 100 TA1535 WP2 uvrA TA98 TA1537
0 103 ± 4 12 ± 3 40 ± 2 16 ± 4 8 ± 1
313 99 ± 7 10 ± 4 38 ± 3 15 ± 7 8 ± 2
625 94 ± 5 10 ± 4 39 ± 1 19 ± 4 11 ± 2
1250 90 ± 9 8 ± 5 42 ± 4 14 ± 7 8 ± 3
2500 84 ± 10 11 ± 2 40 ± 6 17 ± 3 6 ± 2
5000 102 ± 5 6 ± 1 42 ± 6 18 ± 4 7 v 4
Positive controls, –S9 Name  AF-2 SA AF-2 AF-2 9AA
Concentrations (μg/plate) 0.01 0.5 0.01 0.1 80
Mean No. of colonies/plate (average of 3 ± SD) 542 ± 6 481 ± 9 131 ± 13 456 ± 24 380 ± 33
+ 0 90 ± 8 12 ± 4 45 ± 2 26 ± 10 10 ± 2
+ 313 100 ± 16 7 ± 1 33 ± 2 20 ± 3 12 ± 7
+ 625* 96 ± 9 8 ± 2 41 ± 5 22 ± 6 10 ± 3
+ 1250* 100 ± 7 9 ± 2 39 ± 5 23 ± 3 10 ± 2
+ 2500* 94 ± 5 9 ± 2 40 ± 3 23 ± 4 10 ± 2
+ 5000* 106 ± 11 15 ± 1 40 ± 11 19 ± 2 13 ± 3
Positive controls, –S9 Name  2AA 2AA 2AA 2AA 2AA
Concentrations (μg/plate) 1 2 10 0.5 2
Mean No. of colonies/plate (average of 3 ± SD) 775 ± 29 347 ± 12 817 ± 51 391 ± 30 197 ± 13

* Precipitation was observed on the surface of agar plats

AF-2: 2 -(2 -Fuyl)-3 -(5 -nitro-2 -furyl)acrylamide

SA: Sodium azide

9AA: 9 -Aminoacridine

2AA: 2 -Aminoanthracene

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative