Alcohols, C12-14, ethoxylated, sulfates, sodium 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 genotoxicity:

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

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

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

26 Apr - 24 May 1994

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 a TA102 strain).

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

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:

Type and composition of metabolic activation system: phonobarbital-induced liver microsomes
- source of S9: Cytotest cell research GmbH & Co.KG (Lot-no: 140194)
- method of preparation of S9 mix: S9 mix contained 23.5 mL bidest. water, 35 mL Na-phosphate solution, 1.4 mL KCl-MgCl2-solution, 0.35 mL Glucose-6-phosphate solution, 2.8 mL NADP-solution and 7.0 mL S9 fraction.
- concentration or volume of S9 mix and S9 in the final culture medium: 7.0 mL
- quality controls of S9: enzymatic activity tested with aminoanthracene in all strains.

Test concentrations with justification for top dose:

0, 11, 56, 280, 1400, 7000 µg/plate
(corresponding to 0, 40, 200, 1001 and 5003 µg a.s./plate)

Vehicle / solvent:

water

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

destilled water

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

other: aminoanthracene

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration: triplicates
- Number of independent experiments: 2

METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding: Exp. I: 88 - 145 x 10^7; Exp. II: 123 - 145 x 10^7
- Test substance added in medium; in agar (plate incorporation, Exp. I) and preincubation (Exp. II)

METHODS FOR MEASUREMENTS OF CYTOTOXICITY
background growth inhibition

METHODS FOR MEASUREMENTS OF GENOTOXICIY
Number of revertant colonies

Evaluation criteria:

According to Guideline.

Statistics:

yes

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Exp. I: at 7000 µg/plate, ± S9 Exp. II: at ≥ 1400 µg/plate, ± S9

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Exp. I+II: at ≥ 1400 µg/plate, ± S9

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1538

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

True negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

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

True negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Exp. II: at 7000 µg/plate, ± S9

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation and time of the determination: no precipitation observed at any concentration

TEST RESULTS
For details on the test results for the two experiments, please refer to Tables 1 and 2 and Attachment 1.

Table 1: Results of the plate-incorporation test

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type		Frameshift type
		TA 100	TA1535	TA1538	TA98	TA1537
–	0	143 ± 24	12 ± 9	20 ± 2	29 ± 5	15 ± 4
–	11	144 ± 4	15 ± 2	18 ± 6	38 ± 3	14 ± 1
–	56	160 ± 13	12 ± 4	23 ± 4	37 ± 2	12 ± 2
–	280	142 ± 5	8 ± 3	23 ± 9	37 ± 3	11 ± 4
–	1400	101 ± 14	9 ± 3	18 ± 2	34 ± 3	6 ± 1
–	7000	124 ± 7	3 ± 1	18 ± 4	29 ± 2	0
Positive controls, –S9	Name	Na azide	Na azide	2-NF	2-NF	9-AA
	Concentrations (μg/plate)	2.5	2.5	2.5	2.5	25
	Mean No. of colonies/plate (average of 3 ± SD)	438 ± 34	358 ± 23	231 ± 13	224 ± 11	51 ± 12
+	0	152 ± 8	12 ± 6	26 ± 1	43 ± 9	20 ± 5
+	11	150 ± 14	15 ± 3	27 ± 4	40 ± 7	19 ± 5
+	56	168 ± 15	15 ± 5	31 ± 3	41 ± 3	13 ± 2
+	280	150 ± 3	12 ± 3	25 ± 3	41 ± 7	12 ± 2
+	1400	159 ± 4	9 ± 2	28 ± 3	44 ± 1	3 ± 2
+	7000	137 ± 13	5 ± 3	19 ± 3	36 ± 6	0
Positive controls, +S9	Name	2-AA	2-AA	2-AA	2-AA	2-AA
	Concentrations (μg/plate)	2.5	2.5	2.5	2.5	2.5
	Mean No. of colonies/plate (average of 3 ± SD)	1829 ± 160	182 ± 11	1247 ± 32	1457 ± 148	213 ± 4

 

Table 2: Results of the Pre-incubation Test

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type		Frameshift type
		TA 100	TA1535	TA1538	TA98	TA1537
–	0	190 ± 10	16 ± 7	29 ± 5	44 ± 3	23 ± 6
–	11	206 ± 9	19 ± 3	32 ± 6	48 ± 6	29 ± 5
–	56	191 ± 14	19 ± 3	31 ± 3	45  ± 8	23 ± 2
–	280	201 ± 5	10 ± 3	30 ± 1	42 ± 7	17 ± 9
–	1400	143 ± 13	0 ± 0	23 ± 2	52 ± 17	0 ± 0
–	7000	107 ± 2	0 ± 0	22 ± 2	36 ± 4	0 ± 0
Positive controls, –S9	Name	Na azide	Na azide	2-NF	2-NF	9-AA
	Concentrations (μg/plate)	2.5	2.5	2.5	2.5	25
	Mean No. of colonies/plate (average of 3 ± SD)	460 ± 41	387 ± 17	200 ± 17	222 ± 30	98 ± 14
+	0	158 ± 15	17 ± 8	38 ± 4	47 ± 8	20 ± 5
+	11	181 ± 4	14 ± 2	37 ± 6	48 ± 16	14 ± 5
+	56	172 ± 3	18 ± 5	50 ± 3	52 ± 3	19 ± 3
+	280	173 ± 5	19 ± 6	39 ± 4	49 ± 10	15 ± 6
+	1400	178 ± 10	7 ± 1	49 ± 7	34 ± 12	7 ± 3
+	7000	106 ± 6	0 ± 0	35 ± 7	35 ± 3	0 ± 0
Positive controls, +S9	Name	2-AA	2-AA	2-AA	2-AA	2-AA
	Concentrations (μg/plate)	2.5	2.5	2.5	2.5	2.5
	Mean No. of colonies/plate (average of 3 ± SD)	1402 ± 68	108 ± 8	704 ± 32	1033 ± 19	154 ± 9

Conclusions:

This study was conducted according to test guideline 92/62/EWG B.14 and similar to OECD 471 and was compliant with GLP. Both the plate-incorporation method (Exp. I) and the preincubation method (Exp. II) were applied in this experiment, examining the possible mutagenicity / genotoxicity of the test substance in the five Salmonella typhimurium tester strains TA1535, TA 1537, TA100, TA98, and TA1538. Under the conditions of the test, no indications of mutagenic effects of the test item could be found at assessable doses of up to 5000 µg/plate in any of the S. typhimurium strains used. This is true for both experiments with and without metabolic activation (S9).

Reference 2

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 Oct. 1994 - 25 Nov. 1994

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 (incl. QA statement)

Type of assay:

mammalian cell gene mutation assay

Target gene:

TK locus

Species / strain / cell type:

mouse lymphoma L5178Y cells

Details on mammalian cell type (if applicable):

CELLS USED
- Type and source of cells: L5178Y TK+/- mouse lymphoma cells; obtained form American Type Culture Collection, Rockville, MD
- Suitability of cells: recommended in TG 476
- Absence of Mycoplasma contamination: yes
- generation time, plating efficiency and mutation rates (spontaneous and induced) were checked

MEDIA USED
- Type and composition of media, CO2 concentration, humidity level, temperature, if applicable: RPMI 1640 minimal medium supplemented with 10% horse serum heat-inactivated at 56°C for 20 min before use (complete medium). All incubations at 37°C in a 5% CO2 atmosphere (100% humidity).

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Type and composition of metabolic activation system:
- source of S9: Co-factor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of five young male Sprague-Dawley rats treated with phenobarbitone and betanaphthoflavone.
- quality controls of S9: The efficacy was checked in an Ames test and produced acceptable responses with 2-aminoanthracene and benzo(a)pyrene in S. typhimurium TA100; protein content (28.4 ± 0.37 and 24.2 ± 4.19 mg/mL) and aminopyrine demethylase activity (2.59 ± 0.08 and 3.48 ± 0.13 µM/g liver/5 min formaldehyde production) checked.

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

The selection of the concentrations used in the main experiments was based on data from a preliminary cytotoxicity assay. Severe toxicity was observed at the seven highest dose-levels (with and without S9). Based on these results maximum concentrations of 58.6 and 117 µg/mL were selected for treatments in the absence and presence of S9 mix, respectively.

Vehicle / solvent:

Since no solvent vehicle was employed in this study the negative controls consisted of untreated cultures.

Untreated negative controls:

yes

Remarks:

untreated cells

Negative solvent / vehicle controls:

no

Remarks:

no vehicle used

True negative controls:

no

Positive controls:

yes

Positive control substance:

7,12-dimethylbenzanthracene

ethylmethanesulphonate

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration : single
- Number of independent experiments : two

METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable): 1 x 10E6 cells/mL
- Test substance added in medium

TREATMENT AND HARVEST SCHEDULE:
- Exposure duration/duration of treatment: 4 h at 37 °C

FOR GENE MUTATION:
- Expression time (cells in growth medium between treatment and selection): 72 h
- Selection time (if incubation with a selective agent): 12 - 16 days
- Method used: agar
- Selection agent: Trifluorothymidine (final concentration 4.0 µg/mL)
- Number of cells seeded and method to enumerate numbers of viable and mutants cells: 500,000 cells; after incubation, the plates are scored either manually or using a calibrated Artek Model 890 Automatic colony counter; the mutatioh frequency at each test point is calculated.

METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method: Percentage survival relative to the solvent controls is calculated for each treatment in a preliminary cytotoxicity experiment. Dose-levels giving a predicted 20% survival are estimated with and without S9. The estimated concentrations are chosen as the highest dose-levels for the mutation assays.

Evaluation criteria:

For a test substance to be considered mutagenic in this assay, it is required that:
(i) There is a two-fold (or more) increase in mutation frequency compared with the solvent control values, over two consecutive test substance treatment levels. If only the highest practicable dose-level (or the highest dose-level not to cause unacceptable toxicity) gives such an increase, then a single treatment-level will suffice.
(ii) The increases must be reproduced in an independent experiment.
(iii) There must be evidence for a dose-relation (i.e. statistically significant effect in the ANOVA analysis).

Statistics:

The results of the experiments were subjected to an Analysis of Variance, in which the contribution of experiment number and dose-level to the observed variation was examined.

Key result

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 58.6 µg/mL -S9 and 177 µg/mL + S9

Vehicle controls validity:

not examined

Remarks:

no vehicle used

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Data on pH: The addition of the test substance solution did not have any obvious effect on the pH of the treatment medium.
- Data on osmolality: The addition of the test substance solution did not have any obvious effect on the osmolality of the treatment medium.
- Possibility of evaporation from medium: The test substance is not volatile.
- Water solubility: The test substance was soluble in complete medium at a concentration of 100 mg/mL. On the basis of these results, a maximum concentration of 10,000 µg/mL was selected for the cytotoxicity test.

RANGE-FINDING/SCREENING STUDIES
The test substance was assayed at a maximum concentration of 10,000 µg/mL and 8 lower dose levels spaced at two-fold intervals. Following treatment both in the absence and presence of S9 mix, severe toxicity was observed at the 7 highest dose levels, reducing total suspension growth to below the limit of detection. In the absence of S9 mix, at the next lower dose level (78.1 µg/mL) the total suspension growth value was reduced to 2% of the negative control. In the presence of S9 mix, no toxicity was observed at the next lower dose level (78.1 µg/mL). Maximum concentrations of 58.6 and 117 µg/mL were selected for the main study in the absence and presence of S9 mix, respectively.

TEST RESULTS:
For details please refer to Tables 1-4 and the attachment.

Table 1: Results of Experiment I without metabolic activation

Concentration [µg/mL]	Relative Plating Efficiency [%]	Total number of mutant colonies	Mean of mutation plate counts	S.D.
0	100	217	21.7	2.8
2.44	94	78	15.6	1.1
4.88	102	92	18.4	2.7
9.76	92	92	18.4	3.2
19.5	103	107	21.4	3
39.1	93	118	23.6	4
58.6	104	117	23.4	3
EMS, 2.5	71	710	142	13.1
EMS, 5.0	37	1039	207.8	11.8

 

Table 2: Results of Experiment I with metabolic activation

Concentration [µg/mL]	Relative Plating Efficiency [%]	Total number of mutant colonies	Mean of mutation plate counts	S.D.
0	100	149	14.9	2.4
2.44	98	104	20.8	1.3
4.88	100	112	22.4	4
9.76	101	57	11.4	1.5
19.5	96	67	13.4	2.1
39.1	97	57	11.4	2.7
78.1	97	67	13.4	2.1
117	90	65	13	3.4
DMBA, 2.0	64	229	45.8	5.7
DMBA, 3.0	52	236	59	5

 

Table 3: Results of Experiment II without metabolic activation

Concentration [µg/mL]	Relative Plating Efficiency [%]	Total number of mutant colonies	Mean of mutation plate counts	S.D.
0	100	178	17.8	3.7
2.44	99	93	18.6	3
4.88	95	76	15.2	1.8
9.76	99	79	15.8	3.1
19.5	94	75	15	2.3
39.1	95	83	16.6	3
58.6	115	76	15.2	2.2
EMS, 2.5	62	604	120.8	5.8
EMS, 5.0	27	588	117.6	13.6

 

Table 4: Results of Experiment II with metabolic activation

 

Concentration [µg/mL]	Relative Plating Efficiency [%]	Total number of mutant colonies	Mean of mutation plate counts	S.D.
0	100	164	16.4	4.2
2.44	85	72	14.4	1.8
4.88	95	87	17.4	2.7
9.76	83	68	13.6	2.1
19.5	114	114	22.8	3.1
39.1	92	75	15.0	3.0
78.1	99	126	25.2	5.0
117	-	-	-	-
DMBA, 2.0	114	164	82.0	7.1
DMBA, 3.0	122	205	102.5	14.8

'-': Not plated

Conclusions:

In the present in vitro gene mutation study in mouse lymphoma (L5178Y) cells, the test substance did not induce gene mutations when tested at concentrations 117 and 58.6 µg/plate with and without metabolic activation, respectively. Therefore it is concluded that the test substance is not mutagenic in vitro in L5178Y mouse lymphoma cells, under the reported experimental conditions.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

In vivo genotoxicity:

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

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

Study period:

6 Oct. 1994 - 6 Feb. 1995

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 (incl. QA statement)

Type of assay:

mammalian bone marrow chromosome aberration test

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Italia S.p.A., Calco, Como, Italy
- Age at study initiation: 5 - 6 weeks
- Weight at study initiation: 25 - 32 g (males); 20 - 27 g (females)
- Assigned to test groups randomly: yes, immediately after arrival at test facility
- Fasting period before study: yes, overnight
- Housing: 5 animals/cage, separated by sexes, in clear polycarbonate cages (type 2b: Techniplast) with a stainless steel mesh lid and floor; each cage equiped with absorbent bedding which was inspected daily and changed as necessary.
- Diet: Altromin MT; ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATE: 26 Oct. 1994

Route of administration:

oral: gavage

Vehicle:

Vehicle(s)/solvent(s) used: injectable grade sterile distilled water
- Lot/batch no.: 22143-lB (Don Baxter S.p.A., Trieste, italy)

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Fresh solutions of the test substance were prepared for each day's work; solutions were prepared on a weight/volume basis. Solutions of the test substance, following corrections of active constituent were prepared in injectable grade sterile distilled water.

Duration of treatment / exposure:

not applicable

Frequency of treatment:

single treatment

Post exposure period:

Test groups: 10, 24 and 48 h
Positive control group: approx. 26 h

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

Remarks:

corrected for a.i. content of test material

Dose / conc.:

2 000 mg/kg bw/day (actual dose received)

Remarks:

corrected for a.i. content of test material; represents the recommended limit dose for administration periods < 14 days according to OECD TG 475

No. of animals per sex per dose:

Test groups: 5 for each sampling time (10, 24 and 48 h)
Positive control group: 5

Control animals:

yes, concurrent vehicle

Positive control(s):

Cyclophosphamide (50 mg/kg bw)

Tissues and cell types examined:

Tissue: femoral bone marrow
Cell type: 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

CRITERIA FOR DOSE SELECTION:
Range finding study performed to find the maximum tolerated dose.

TREATMENT AND SAMPLING TIMES:
Following treatment and prior to sample collection, animals are injected intraperitoneally with 4 mg/mL colchicine and samples are collected 3 h thereafter. Cells are harvested from the bone marrow, swollen, fixed and stained, and analysed for chromosomal aberrations.

DETAILS OF SLIDE PREPARATION:
Slides were fixed with a methanol:acetic acid solution, air-dried and stained with Giemsa.

METHOD OF ANALYSIS:
50 metaphase spreads per animal were examined microscopically at high magnification for the presence of chromosomal aberrations. Only metaphases containing 40 chromosomes are scored for aberrations. The number of chromosomes, the specific types and numbers of aberrations are recorded. The Vernier readings of aberrant or equivocal metaphases are recorded.

OTHER:
The mitotic index is calculated. This is based on the number of metaphases observed per 1000 cells and is expressed as a percentage.

Evaluation criteria:

A test substance is considered to have clastogenic properties if a statistically significant increase in the incidence of cells bearing aberrations is observed at any test point, excluding gaps. The lesions observed must be consistent with those expected at the sampling time at which the increases are observed. The internal consistency of the findings within groups are also taken into account.

Statistics:

The experimental unit investigated in the present study is the proportion of cells from an animal which exhibit chromosomal aberrations. It is therefore assumed that the data has a binomial error structure, each cell out of a total of 'n' for each animal either bearing or not-bearing aberrations.
A logistic-linear model is fitted to the data adding the factors 'Sex', 'Treatment' and 'Sex-Treatment Interaction' in turn. The change in deviance with each factor is compared with the Chi-squared distribution to determine the level of significance. The deviance associated with the residual term is compared with the Chi-squared distribution to determine the goodness-of-fit of the logistic-linear model. This allows the construction of a table similar to a classical analysis of variance.
Data from in vivo cytogenetic assays often contain extra-binomial variation, which may result in part from animal-to-animal variation, or from the responses of different populations of cells within animals. This situation will be highlighted by a significant lack of fit of the logistic-linear model. When this is the case, an allowance for the extra-binomial variation can be made using a generalised 'heterogeneity factor' to weight the logistic-linear model variables.
The statistical analysis is performed using the values for the total number of aberration-bearing cells observed for each animal, excluding gaps from consideration.

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- Dose range:
250, 500, 1000 and 2000 mg/kg bw
- Clinical signs of toxicity in test animals:
No clinical signs were observed at any dose level tested.
- Evidence of cytotoxicity in tissue analysed:
No reduction in the mitotic index was observed in any treatment group.

A summary of the test results can be found under "Any other information on results incl. tables", Table 1 - 2.

Table 1: Results of the 24 h exposure experiment

	Dose (mg/kg bw/d)
	0	1000	2000	CCP
Postexposure period [h]	24	24	24	24
Total no. of animals	10	10	10	10
No. of animals with aberrant metaphases	0	2	2	10
Analyzed metaphases	500	500	500	500
Aberrant metaphases	0	2	2	72
Including gaps	0	2	2	72
Excluding gaps	0	2	2	72
Exchanges	0	0	0	7
Polyploidy	0	2	1	1

 

Table 2: Summary table of the results

	Dose (mg/kg bw/d)
	0	1000	2000	CCP
Postexposure period [h]: 24
Total no. of animals	10	10	10	10
Analyzed metaphases	500	500	500	500
Percent abberrant cells (Mean ± SD)	0.0 ± 0.0	0.2  ±  0.6	0.4  ±  0.8	17.6*** ±  4.4
Mitotic index (Mean ± SD)	41.0 ± 12.7	52.6  ± 9.1	61.7  ±  13.6	41.6  ±  11.8
Postexposure period [h]: 10
Total no. of animals	10	10	10	10
Analyzed metaphases	500	500	500	500
Percent abberrant cells (Mean ± SD)	0.2 ± 0.6	0.2 ± 0.6	34.1 ± 9.3	-
Mitotic index (Mean ± SD)	30.2 ± 13.0	0.6 ± 10.0	34.1 ± 8.0	-
Postexposure period [h]: 48
Total no. of animals	10	10	10	10
Analyzed metaphases	500	500	500	500
Percent abberrant cells (Mean ± SD)	0.0 ± 0.0	0.2 ± 0.6	0.8 ± 1.4	-
Mitotic index (Mean ± SD)	68.8 ± 5.3	64.5 ± 5.3	67.5 ± 5.7	-

Conclusions:

In the present in vivo cytogenicity study in mice, compliant with GLP and according to OECD test guideline 475, the test substance did not induce chromosomal aberrations in bone marrow cells of mice after oral administration at 1000 and 2000 mg/kg bw to both male and female animals.

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.

The genetic toxicity of alcohol ethoxysulfates (AES) was evaluated in several in vitro and in vivo genotoxicity tests. In summary, all Ames tests conducted with the substance AES (C12-14, 1-2.5 EO) Na (CAS 68891-38-3) using several strains of S. typhimurium showed a negative result. Testing for mammalian genotoxicity in vitro revealed a negative outcome as well. The result of in vivo testing revealed that the substance AES (C12-14, 1-2.5 EO) Na (CAS 68891-38-3) has no clastogenic potential. According to Regulation (EC) No. 1907/2006 (REACH) VIII 8.4.2, column 2, an in vitro cytogenicity study in mammalian cells does not need to be conducted if adequate data from an in vivo cytogenicity test are available. Based on the outcome of these tests, AES (C12-14, 1-2.5 EO) Na (CAS 68891-38-3) can be considered as non-genotoxic.

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, and are therefore conclusive but not sufficient for classification.