Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Bacterial reverse mutation assay (Ames / OECD guideline 471): negative

In vitro mammalian chromosome aberration test (ChrAb / OECD guideline 473): negative

In vitro mammalian cell gene mutation assay (MLA / OECD guideline 476): negative

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 1538
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
1. test: 8, 40, 200, 1000, 5000 µg/plate2 test: 5, 10, 20, 40, 80 µg/plate (-S9 mix); 2.5, 10, 40, 160, 640 µg/plate (+S9 mix)
Vehicle / solvent:
Water
Untreated negative controls:
yes
Remarks:
untreated cells
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: sodium azide, 4-nitroquinoline-N-oxide, 9-aminoacridine, 2-aminoanthracene
Species / strain:
other: all strains tested
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 200 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Conclusions:
negative
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Species / strain / cell type:
other: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Test concentrations with justification for top dose:
1st and 2nd test: 8, 40, 200, 1000, and 5000 µg/plate
Species / strain:
other: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
5000 µg/plate
Vehicle controls validity:
valid
Positive controls validity:
valid
Conclusions:
negative
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Species / strain / cell type:
other: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Test concentrations with justification for top dose:
1st and 2nd test: 8, 40, 200, 1000, and 5000 µg/plate
Species / strain:
other: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
5000 µg/plate
Conclusions:
negative
Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Target gene:
n.a.
Species / strain / cell type:
Chinese hamster Ovary (CHO)
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
501, 1500, 5010 µg/mL
Vehicle / solvent:
water
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: mitomycin, 5 µg/mL (-S9); cyclophosphamide, 50µg/mL (+S9)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium
Evaluation criteria:
According to guideline.
Species / strain:
Chinese hamster Ovary (CHO)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Conclusions:
negativeThe test item was not considered to be clastogenic.
Executive summary:

No enhanced aberration rate in the presence or absence of metabolic activation was observed in this chromosomal aberration assay. Therefore, the test substance was not considered to be clastogenic.

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Target gene:
Thymidine kinase locus (tk)
Species / strain / cell type:
mouse lymphoma L5178Y cells
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
-S9: 3.125, 6.25, 10, 12.5, 20, 25, 30, 40, 50, 55, 60,65, 70, 80 and 100 µg/mL+S9: 50, 55, 60, 65, 70, 75, 80, 85, 90 and 95 µg/mL
Vehicle / solvent:
DMSO
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: methylmethanesulfonate, ethylmethanesulphonate
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium
Evaluation criteria:
According to guideline.
Statistics:
Yes.
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Conclusions:
negativeThe test item was not considered to be mutagenic.
Executive summary:

No enhanced mutation rate in the S9 treated or untreated cells was observed in this mouse lymphoma assay. Therefore, the test substance was not considered to be mutagenic.

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Target gene:
Thymidine kinase locus (tk)
Species / strain / cell type:
mouse lymphoma L5178Y cells
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
-S9: 156.25, 312.5, 625, 1250, 2500 µg/mL (Trial 1)200, 1000, 1800, 2600, 3400, 4200 µg/mL (Trial 2)1000, 1800, 2600, 3400, 4200 µg/mL (Trial 3)1000, 1800, 2600, 3400, 4200, 5000 µg/mL (Trial 4)+S9: 200, 1000, 1800, 2600, 3400, 4200 µg/mL (Trial 1)1000, 1800, 2600, 3400, 4200 µg/mL (Trial 2&3)2600, 3000, 3400, 3800, 4200 µg/mL (Trial 4)
Vehicle / solvent:
DMSO
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: methylmethanesulfonate, ethylmethanesulfonate(-S9); methylcholanthrene (+S9)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium
Evaluation criteria:
According to guideline.
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Conclusions:
negativeThe test item was not considered to be mutagenic.
Executive summary:

No significant mutagenic responses were observed in three of four trials without S9 mix. In trial 1 there was a single, statistically significant response at one dose level but this was not supported by similar responses at higher dose levels in either the same or two other trials. Thus trial 1 was inconclusive because higher dose levels could have been tested. In trial 3 without S9 mix, all doses tested and permitting survival were associated with significant responses. With S9, two experiments were negative and two were inconclusive because higher doses could have been tested. The weight of evidence strongly suggested that sodium (2-ethylhexyl) alcohol sulfate was not mutagenic in this assay, though the anomalous response in one trial without S9 mix has not been explained.

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Remarks:
No E.coli strains tested.
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 1538
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
1. test: 8, 40, 200, 1000, 5000 µg/plate2 test: 5, 10, 20, 40, 80 µg/plate (-S9 mix); 2.5, 10, 40, 160, 640 µg/plate (+S9 mix)
Vehicle / solvent:
Water
Untreated negative controls:
yes
Remarks:
untreated cells
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: sodium azide, 4-nitroquinoline-N-oxide, 9-aminoacridine, 2-aminoanthracene
Species / strain:
other: all strains tested
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 200 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Conclusions:
negative
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
Neither S. typhimurium TA102 or E. coli WP2 used
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Species / strain / cell type:
other: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Test concentrations with justification for top dose:
1st and 2nd test: 8, 40, 200, 1000, and 5000 µg/plate
Species / strain:
other: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
5000 µg/plate
Vehicle controls validity:
valid
Positive controls validity:
valid
Conclusions:
negative
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
no tester strain E. coli WP2 or S. typhimurium TA102 was used
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Species / strain / cell type:
other: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Test concentrations with justification for top dose:
1st and 2nd test: 8, 40, 200, 1000, and 5000 µg/plate
Species / strain:
other: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
5000 µg/plate
Conclusions:
negative
Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Deviations:
yes
Remarks:
Not all results displayed.
GLP compliance:
not specified
Type of assay:
in vitro mammalian chromosome aberration test
Target gene:
n.a.
Species / strain / cell type:
Chinese hamster Ovary (CHO)
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
501, 1500, 5010 µg/mL
Vehicle / solvent:
water
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: mitomycin, 5 µg/mL (-S9); cyclophosphamide, 50µg/mL (+S9)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium
Evaluation criteria:
According to guideline.
Species / strain:
Chinese hamster Ovary (CHO)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Conclusions:
negativeThe test item was not considered to be clastogenic.
Executive summary:

No enhanced aberration rate in the presence or absence of metabolic activation was observed in this chromosomal aberration assay. Therefore, the test substance was not considered to be clastogenic.

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
yes
Remarks:
lack of details on test substance
GLP compliance:
no
Type of assay:
mammalian cell gene mutation assay
Target gene:
Thymidine kinase locus (tk)
Species / strain / cell type:
mouse lymphoma L5178Y cells
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
-S9: 3.125, 6.25, 10, 12.5, 20, 25, 30, 40, 50, 55, 60,65, 70, 80 and 100 µg/mL+S9: 50, 55, 60, 65, 70, 75, 80, 85, 90 and 95 µg/mL
Vehicle / solvent:
DMSO
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: methylmethanesulfonate, ethylmethanesulphonate
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium
Evaluation criteria:
According to guideline.
Statistics:
Yes.
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Conclusions:
negativeThe test item was not considered to be mutagenic.
Executive summary:

No enhanced mutation rate in the S9 treated or untreated cells was observed in this mouse lymphoma assay. Therefore, the test substance was not considered to be mutagenic.

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
yes
Remarks:
lack of details on test substance
GLP compliance:
not specified
Type of assay:
mammalian cell gene mutation assay
Target gene:
Thymidine kinase locus (tk)
Species / strain / cell type:
mouse lymphoma L5178Y cells
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
-S9: 156.25, 312.5, 625, 1250, 2500 µg/mL (Trial 1)200, 1000, 1800, 2600, 3400, 4200 µg/mL (Trial 2)1000, 1800, 2600, 3400, 4200 µg/mL (Trial 3)1000, 1800, 2600, 3400, 4200, 5000 µg/mL (Trial 4)+S9: 200, 1000, 1800, 2600, 3400, 4200 µg/mL (Trial 1)1000, 1800, 2600, 3400, 4200 µg/mL (Trial 2&3)2600, 3000, 3400, 3800, 4200 µg/mL (Trial 4)
Vehicle / solvent:
DMSO
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: methylmethanesulfonate, ethylmethanesulfonate(-S9); methylcholanthrene (+S9)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium
Evaluation criteria:
According to guideline.
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Conclusions:
negativeThe test item was not considered to be mutagenic.
Executive summary:

No significant mutagenic responses were observed in three of four trials without S9 mix. In trial 1 there was a single, statistically significant response at one dose level but this was not supported by similar responses at higher dose levels in either the same or two other trials. Thus trial 1 was inconclusive because higher dose levels could have been tested. In trial 3 without S9 mix, all doses tested and permitting survival were associated with significant responses. With S9, two experiments were negative and two were inconclusive because higher doses could have been tested. The weight of evidence strongly suggested that sodium (2-ethylhexyl) alcohol sulfate was not mutagenic in this assay, though the anomalous response in one trial without S9 mix has not been explained.

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

Genetic toxicity in vivo

Description of key information

Rodent Dominant Lethal Test (DLA / OECD guideline 478): negative

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vivo mammalian germ cell study: cytogenicity / chromosome aberration
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Species:
mouse
Strain:
CD-1
Sex:
male/female
Route of administration:
oral: gavage
Duration of treatment / exposure:
single dose
Remarks:
Doses / Concentrations:0, 120, 380 or 1200 mg/kg bwBasis:
Sex:
male/female
Genotoxicity:
negative

no adverse effects on pregnancy frequency, number of implantations or 

frequency of early deaths

Conclusions:
negative
Endpoint:
in vivo mammalian germ cell study: cytogenicity / chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study well documented, meets generally accepted scientific principles, acceptable for assessment Adopted according to OECD SIDS or other official EU regulation procedures (public available peer reviewed source).
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 478 (Genetic Toxicology: Rodent Dominant Lethal Test)
Deviations:
yes
Remarks:
Only limited data on test material and methodology available
GLP compliance:
no
Type of assay:
rodent dominant lethal assay
Species:
mouse
Strain:
CD-1
Sex:
male/female
Route of administration:
oral: gavage
Duration of treatment / exposure:
single dose
Remarks:
Doses / Concentrations:0, 120, 380 or 1200 mg/kg bwBasis:
Sex:
male/female
Genotoxicity:
negative

no adverse effects on pregnancy frequency, number of implantations or 

frequency of early deaths

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

Additional information

No data on genetic toxicity is available for C9-12AS Na (CAS n.a.). To assess the mutagenic potential on mammalian cells in-vitro a read-across to structurally related alkyl sulfate (AS), i.e. C8AS Na (CAS 142-31-4), C10AS Na (CAS 142-87-0), C8 branched AS Na (CAS 126-92-1) and C12AS Na (CAS 151-21-3) was performed. The possibility of a read-across to other alkyl sulfates in accordance with Regulation (EC) No 1907/2006 Annex XI 1.5. Grouping of substances and read-across approach was assessed. In Annex XI 1.5 it is given that a read-across approach is possible for substances, whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity. The AS reported within the AS category show structural similarity. The most important common structural feature of the category members is the presence of a predominantly linear aliphatic hydrocarbon chain with a polar sulfate group, neutralized with a counter ion. This structural feature confers the surfactant properties of the alkyl sulfates. The surfactant property of the members of the AS category in turn represent the predominant attribute in mediating effects on mammalian health. Therefore, the AS of the AS category have similar physico-chemical, environmental and toxicological properties, validating the read across approach within the category. The approach of grouping different AS for the evaluation of their effects on human health and the environment was also made by the OECD in the SIDS initial assessment profile [1] and by a voluntary industry programme carrying out Human and Environmental Risk Assessments (HERA [2]) further supporting the read across approach between structurally related AS. During data processing of all alkyl sulfates within the category it turned out that alkyl sulfates with a carbon chain length of C12 exert the most prominent effects on human health. Alkyl sulfates with this chain length turned out to be harmful if swallowed and also to elicit the most prominent effects after application to the skin (assessed among others as induction of erythema in human volunteers and in vitro swelling of skin after application of alkyl sulfates with various carbon chain length; cf IUCLID chapter 7.12). Therefore C12AS Na (CAS 151-21-3) was additionally chosen as read-across substance as worst case assumption.

There are three studies available addressing mutagenicity in bacteria.

In one study, performed according to OECD Guideline 471, Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 1538 and TA 100 were treated with C10AS Na (CAS 142-87-0, analytical purity 29-30%) in presence and absence of metabolic activation. The tester strains TA 102 or E.coli WP2 were not used during the conduct of the study (Banduhn, 1992). The concentrations tested were 8, 40, 200, 1000 and 5000 µg/plate in both experiments. Results achieved with negative control (untreated cell suspension and medium), vehicle (water) and positive controls were valid. Cytotoxicity was observed at 5000 µg/plate. No genotoxicity was observed.

In the second study, performed according to OECD Guideline 471, Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 1538 and TA 100 were treated with C8AS Na (CAS 142-31-4, analytical purity 40%) in presence and absence of metabolic activation. The tester strains TA 102 or E.coli WP2 were not used during the conduct of the study (Banduhn, 1989). The concentrations tested were 8, 40, 200, 1000 and 5000 µg/plate in both experiments. Results achieved with negative control (untreated and medium), vehicle (water) and positive controls were valid. Cytotoxicity was observed at 5000 µg/plate. No genotoxicity was observed.

In the third study according to OECD Guideline 471 with C12AS Na (CAS 151-21-3) Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 1538 and TA 100 were treated using the plate incorporation method with and without the addition of a rat liver S9-mix (Banduhn, 1988). Tester strain TA 102 or E.coli were not used during the conduct of the study. The dose range was 8, 40, 200, 1000, 5000 µg/plate for the first experiment (with and without S9 mix) as well as 5, 10, 20, 40, 80 µg/plate (-S9 mix) and 2.5, 10, 40, 160, 640 µg/plate (+S9 mix) for the second experiment. Results achieved with vehicle (water) and positive controls were valid. Cytotoxicity was observed in presence and absence of metabolic activation occasionally at around 200 µg/plate while no genotoxicity was observed for C12AS Na (CAS 151-21-3) at all.

The clastogenicity of C8 branched AS Na (CAS 126-92-1, analytical purity 39.6%) in a mammalian cell line was investigated similar to OECD guideline 473 using Chinese hamster Ovary cells with and without metabolic activation (Loveday, 1990). The test concentrations were 501, 1500 and 5010 µg/mL. Results achieved with the vehicle (water) and positive controls were valid. No cytotoxicity was observed in presence and absence of metabolic activation however the test was conducted up to the limit concentration of 5000 µg/mL. No enhanced chromosome aberration was observed under any conditions of the study.

The potential of C8 branched AS Na (CAS 126-92-1, no data on analytical purity) to induce genotoxicity in mammalian cells in vitro was assessed in a study conducted similar to OECD guideline 476 using the mouse lymphoma L5178Y cells with and without metabolic activation (McGregor, 1991). The study comprised of 4 trials with and without metabolic activation. The concentrations tested in the absence of metabolic activation were 156.25, 312.5, 625, 1250, 2500 µg/mL (Trial 1); 200, 1000, 1800, 2600, 3400, 4200 µg/mL (Trial 2); 1000, 1800, 2600, 3400, 4200 µg/mL (Trial 3) and 1000, 1800, 2600, 3400, 4200, 5000 µg/mL (Trial 4). The concentrations tested in the presence of metabolic activation were 200, 1000, 1800, 2600, 3400, 4200 µg/mL (Trial 1), 1000, 1800, 2600, 3400, 4200 µg/mL (Trial 2&3) and 2600, 3000, 3400, 3800, 4200 µg/mL (Trial 4). No treatment related increased mutation frequencies were observed in three of four trials without S9 mix. However, in trial 3, all concentrations showing no marked cytotoxicity were associated with significantly increased mutation frequencies in the absence of metabolic activation. There was no reasonable explanation for this striking result of trial 3. In trial 1 without metabolic activation a statistically significant increased mutation frequency occurred at a single concentration level. At higher concentrations no increased mutation frequencies were observed within this trial. Taken together no increased mutation frequency was evidenced when L5178Y cells were exposed to the test substance without metabolic activation. In the presence of metabolic activation no increased mutation frequencies were observed. In two trials the top dose of 4200 µg/L produced marked cytotoxicity. Therefore also no increased mutation frequency was evidenced when L5178Y cells were exposed to the test substance with metabolic activation.

The mutagenicity of C12AS Na (CAS 151-21-3, no data on analytical purity) in a mammalian cell line was investigated similar to OECD guideline 476 using the mouse lymphoma L5178Y cells with and without metabolic activation (McGregor, 1988). The test concentrations were 3.125, 6.25, 10, 12.5, 20, 25, 30, 40, 50, 55, 60, 65, 70, 80 and 100 µg/mL without and 50, 55, 60, 65, 70, 75, 80, 85, 90 and 95 µg/mL with metabolic activation. Results achieved with the negative (untreated), vehicle (DMSO) and positive controls were valid. Cytotoxicity was observed in presence and absence of metabolic activation while no genotoxicity was observed under both circumstances for C12AS Na (CAS 151-21-3).

The potential of C12AS Na (CAS 151-21-3) to induce in-vivo chromosomal aberration was assessed in a study comparable to the dominant lethal test with CD-1 mouse (Unilever, 1976). The test substance was administered via gavage at doses of 120, 380 and 1200 mg/kg bw to a total of 225 males. Each male was caged with 2 virgin females for 7 days. Thereafter males were caged with another two virgin females for 7 days. This was repeated another 6 times. The males were not further examined. Females were sacrificed 13 days after the assumed date of fertilization, i.e.15 or 16 days after caging females with male and the frequency of early death, frequency of pregnancy and number of implantations was assessed. No adverse effects on and the frequency of early death, frequency of pregnancy and number of implantations occurred. Thus the test substance did not show clastogenicity at doses of 120, 380 and 1200.

 

In conclusion, the structurally related substances did not show any genotoxic potential. This is supported by the conclusions of the HERA Draft report “AS are not genotoxic, mutagenic or carcinogenic…” and the conclusions of the SIDS initial assessment profile “Alkyl sulfates of different chain length and with different counter ions were not mutagenic in standard bacterial and mammalian cell systems [...]. There was also no indication for a genotoxic potential of alkyl sulfates in various in vivo studies on mice […].”


Justification for classification or non-classification

According to the classification criteria of Directive 67/548/EEC and Regulation (EC) No 1272/2008 the substance does not need to be classified for genotoxicity.