Alcohols, C10-16, ethoxylated, sulfates, mono(hydroxyethyl)ammonium salts

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The assessment is based on the data currently available. New studies, based on the category review and the final decisions issued for some of the category substances, which are also relevant for this assessment, are currently being conducted. The hazard assessment with respect to genetic toxicity will be updated once all ongoing studies have been finalised.

In vitro gene mutation in bacteria has been tested using the target substance.

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

No data on genotoxicity in mammalian cells are available for the target substance Alcohols, C10-16, ethoxylated (1-2.5 EO), sulfates, monoeth. salts (CAS 157627-92-4). Therefore, read-across from structural analogue substances has been applied.

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

Read-across from structural analogue source substance Alcohols, C12-14, ethoxylated, sulfates, sodium salts (CAS 68891-38-3).

Link to relevant study records

Reference

Reference 1

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:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

No E.coli strains tested (nor the TA102 strain)

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Target gene:

His operon

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Additional strain / cell type characteristics:

not applicable

Species / strain / cell type:

S. typhimurium TA 1538

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

0, 10, 100, 1000, 10000, 100000 µg/plate

Vehicle / solvent:

distilled water

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

distilled water

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

Evaluation criteria:

According to Guideline.

Statistics:

yes

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Table 1: Results of the Plate-Incorporation Test

Without S9-Mix	Test substance concentration (µg/plate)	Mean number of revertant colonies per plate (average of 3 plate ± SD)
		Base-pair substitution type		Frameshift type
		TA 100	TA 1535	TA 1538	TA 98	TA 1537
	0	222 ± 20	12 ± 3	12 ± 2	10 ± 3	11 ± 2
	10	187 ± 31	13 ± 1	18 ± 3	11 ± 1	5 ± 2
	10^2	231 ± 8	9 ± 2	12 ± 1	13 ± 4	6 ± 0
	10^3	102 ± 13	18 ± 4	32 ± 1	16 ± 4	7± 3
	10^4	/	/	/	/	/
	10^5	/	/	/	/	/
Positive controls (-S9)	Name	Na azide	Na azide	2-NF	2-NF	9-AA
	Concentration (µg/plate)	5	5	10	10	40
	Mean No. of colonies/plate (average of ± SD)	/	/	50 ± 4	47 ± 1	37 ± 2
With S9-Mix	0	155 ± 15	8 ± 2	15 ± 5	15 ± 2	12 ± 1
	10	199 ± 46	12 ± 3	8 ± 3	14 ± 4	9 ± 1
	10^2	177 ± 7	9 ± 3	11 ± 4	13 ± 2	9 ± 3
	10^3	151 ± 14	10 ± 1	7 ± 4	12 ± 3	11 ± 2
	10^4	135 ± 7	/	/	/	/
	10^5	123 ± 4	/	/	/	/
	Name	2AA	2AA	2AA	2AA	2AA
	Concentration (µg/plate)	1	1	1	1	1
	Mean No. of colonies/plate (average of ± SD)	/	81 ± 9	/	/	88 ± 18

Conclusions:

In the present in vitro bacterial reverse mutation assay (Ames test), the test substance did not show any genotoxic potential in presence or absence of metabolic activation (S9-Mix) when tested in the bacterial strains TA1535, TA1537, TA98, TA100 or TA1538.

Reference 2

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Deviations:

no

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Target gene:

TK operon

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: 2.44, 4.88, 9.76, 19.5, 39.1, 58.6 µg/mL
+S9: 2.44, 4.88, 9.76, 19.5, 39.1, 78.1, 117 µg/mL

Vehicle / solvent:

n.a.

Untreated negative controls:

yes

Remarks:

untreated cells

Negative solvent / vehicle controls:

no

Remarks:

no vehicle used

Positive controls:

yes

Positive control substance:

other: ethylmethanesulphonate, 7,12-dimethylbenzanthracene

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:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Conclusions:

Interpretation of results: negative

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

No data on in vivo genotoxicity are available for the target substance Alcohols, C10-16, ethoxylated (1-2.5 EO), sulfates, monoeth. salts (CAS 157627-92-4). Therefore, read-across from structural analogue substances has been applied.

Mammalian Bone Marrow Chromosome Aberration Test (CA / OECD 475): negative

Read-across from structural analogue source substance Alcohols, C12-14, ethoxylated, sulfates, sodium salts (CAS 68891-38-3).

Link to relevant study records

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)

Deviations:

no

GLP compliance:

yes

Type of assay:

chromosome aberration assay

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

According to Guideline.

Route of administration:

oral: gavage

Vehicle:

Distilled water

Duration of treatment / exposure:

n.a.

Frequency of treatment:

Once

Post exposure period:

test group: 10, 24 and 48 h
positive control: 26 h

Remarks:

Doses / Concentrations:
0, 1000, 2000 mg/kg bw
Basis:
nominal conc.

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Positive control(s):

Cyclophosphamide (50 mg/kg bw)

Tissues and cell types examined:

Femoral bone marrow cells

Details of tissue and slide preparation:

SAMPLING TIMES
test groups: 10, 24 and 48 h after treatment
positive control group: 26 h after treatment

Evaluation criteria:

According to Guideline.

Statistics:

Yes

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Conclusions:

Interpretation of results: negative

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

The assessment is based on the data currently available. New studies, based on the category review and the final decisions issued for some of the category substances, which are also relevant for this assessment, are currently being conducted. The hazard assessment with respect to genetic toxicity will be updated once all ongoing studies have been finalised.

Only data regarding mutation in bacteria have been obtained using the target substance. No data are available for other genotoxicity endpoints, i.e. mutation in mammalian cells and clastogenicity. Therefore, the latter endpoints are covered by read-across from structurally related AES. The AES reported within the category show similar structural, physico-chemical, environmental and toxicological properties. The approach of grouping different AES for the evaluation of their effects on human health and the environment was also made by the Danish EPA (2001) and HERA (2003), supporting the read-across approach between structurally related AES.

Genetic toxicity

There is one key study available addressing genetic toxicity for AES (C12-14; < 2.5 EO) C2H7NO (CAS No. 157627-92-4). Mutagenicity in bacteria is assessed in this key study performed similar to OECD Guideline 471 using Salmonelly typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100; the tester strain E.coli was not considered in this study (Z&S, 1996). The tester strains were treated using the plate incorporation method with and without the addition of a rat liver S9-mix. The dose range for the plate incorporation test was 0, 10, 100, 1000, 10000 and 100000 µg/plate. Results achieved with vehicle (distilled water) and positive controls were valid. Cytotoxicity was seen in presence and absence of metabolic activation while no genotoxicity was observed under both circumstances for AES (C12-14; < 2.5 EO) monoeth salt (CAS No. 157627-92-4).

There are two additional studies available addressing in vitro genotoxicity for the analogue substance AES (C12-14; 1-2.5EO) Na (CAS 68891-38-3).

Mutagenicity in bacteria was assessed in a key study performed similar to OECD Guideline 471. Tester strain TA 102 or E.coli were not used during the conduct of the study (Sasol, 1994). In this study, 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. The dose range for the plate incorporation test was 11, 56, 280, 1400 and 7000 µg/plate. Results achieved with vehicle (distilled water) and positive controls were valid. Cytotoxicity was seen in the presence and absence of metabolic activation while no genotoxicity was observed under both circumstances for AES (C12-14; 1-2.5 EO) Na (CAS 68891-38-3).

The mutagenicity of AES (C12-14) Na (CAS 68891-38-3, analytical purity 27.6% a.i., no data on grade of ethoxylation) in a mammalian cell line was investigated according to OECD guideline 476 using the mouse lymphoma L5178Y cells with and without metabolic activation (BASF, 1995a). The test concentrations were 2.44, 4.88, 9.76, 19.5, 39.1 and 58.6 µg/mL without metabolic activation as well as 2.44, 4.88, 9.76, 19.5, 39.1, 78.1 and 117 µg/mL with metabolic activation. Results achieved with the negative (distilled water) and positive controls were valid. Cytotoxicity was seen in presence and absence of metabolic activation while no genotoxicity was observed under both circumstances for Na AES (C12-14, 1-2.5 EO) (CAS 68891-38-3).

Referring to in vivo genotoxicity, the in vivo clastogenic potential of the analogue compound AES(C12-14) Na (CAS 68891-38-3, analytical purity 27-29% a.i., no data on grade of ethoxylation) was assessed in a mammalian bone marrow chromosomal aberration test with CD-1 mouse according to OECD Guideline 475 (BASF, 1995b). The test substance was administered via gavage at doses of 1000 and 2000 mg/kg bw to five animals per sex per dose. Distilled water was used as vehicle. The post exposure period were 10, 24 and 48 h for the test group including the vehicle control and 26 h for the positive control group. Results achieved with the negative (distilled water) and positive controls were valid. No signs of toxicity and no increased number of chromosome aberration were seen at 1000 and 2000 mg/kg bw. Thus the test substance did not show clastogenicity at 1000 and 2000 mg/kg bw based on the test material and 270 to 290 and 540 to 580 mg/kg bw based on the active ingredient.

In conclusion, the test substance did not show any genotoxic potential. This is supported by the conclusions of the HERA (2003) report for AES were it is stated that: “In all available in vitro and in vivo genotoxicity assays, there is no indication of genetic toxicity of AES.”

Influence of counter ions on genotoxicity

Since monoethanolamine (MEA) in its protonated form (monoethanolammonium) is the counter-ion present in Alcohols C10-16, ethoxylated (1-2,5 EO) sulphated, monoeth salts (CAS 157627-92-4), MEA might have an impact on the toxicological properties of the registered substance. Therefore, the toxicological profile of MEA is considered. MEA is listed in Annex VI of Regulation 1272/2008. It is classified as Acute Tox. 4; H302, H312 and H332 as well as Skin Corr. 1B; H314. Additionally a specific concentration limit is established for STOT SE3, H335 at concentrations ≥ 5% in Annex VI of Regulation 1272/2008.The effects of MEA on human health were assessed by the OECD in the SIDS initial assessment Report (2013). MEA was found to be not genotoxic in vitro and in vivo. Thus, MEA will not confer adverse effects regarding this endpoint.

References:

Danish EPA - Environmental and Health Assessment of Substances in Household Detergents and Cosmetic Detergent Products (2001). Environmental Project No. 615, pp. 24-28

HERA (2003). Human & Environmental Risk Assessment on ingredients of European household cleaning products Alcohol Ethoxysulphates, Human Health Risk Assessment Draft, 2003. http: //www. heraproject. com.

SIDS initial assessment report, (2013); http://webnet.oecd.org/HPV/UI/SIDS_Details.aspx?key=8aefe41b-8499-4052-943f-f6dd6f8c5997&idx=0

Justification for classification or non-classification

The available data on genetic toxicity do not meet the criteria for classification according to Regulation (EC) No. 1272/2008 (CLP) and are therefore conclusive but not sufficient for classification.