Poly(oxy-1,2-ethanediyl), α-(carboxymethyl)-ω-hydroxy-, C16-18 (even numbered) and C18-unsatd. alkyl ethers

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

No significant concern can be reliably derived for the registration substance with respect to the endpoint genotoxicity based on the experimental data on the registration substance and on the read-across supporting substances.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Neither E. coli WP2 nor S. typhimurium TA102 were tested as required in the current OECD TG 471.

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted May 26, 1983

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted December 29, 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL- Batch No.of test material: 137304133- Expiration date of the lot/batch: 2001, JanuarySTABILITY AND STORAGE CONDITIONS OF TEST MATERIAL- Storage condition of test material: romm temperature (20°C)- Stability under test conditions: Min . 24 h (0 .1% solution)- Solubility and stability of the test substance in the solvent/vehicle: soluble in waterTREATMENT OF TEST MATERIAL PRIOR TO TESTING- Treatment of test material prior to testing: stock solutions 1. study 5000 mg/L, 2. study 50000mg/L- Preliminary purification step (if any): no- Final dilution of a dissolved solid, stock liquid or gel: in water

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium, other: 97a

Species / strain / cell type:

S. typhimurium TA 98

Species / strain / cell type:

S. typhimurium TA 100

Species / strain / cell type:

S. typhimurium TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9 mix (male wistar rats induced with Phenobarbital and ß-Naphtoflavone)

Test concentrations with justification for top dose:

cytotoxcicty was evident in TA97a without S9 in concentrations ≥ 500 µg/plate and in TA97a with S9 at 5000 µg/plateall other strains were tested up to the limit dose (5000 µg/plate)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: ICR191 (TA97a - S9); 4-Nitro-o-phenylen-diamine (TA9 -S9); Nitrofurantoine (TA 100 - S9); 2-Aminoanthracene (TA 97a, TA 98, TA 100 + S9); Cumene Hydroperoxide (TA 102 - S9); Danthron (TA 102 + S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)DURATION- Exposure duration: 48 hNUMBER OF REPLICATIONS: 3DETERMINATION OF CYTOTOXICITY- Method: evaluation of the background lawn

Statistics:

Arithmetic mean values and standard deviations were calculated out of colonies per plate of three replicates .For evaluation of the results the induction rate of the mean values was calculated:revertant colonies of test substance/revertant colonies of the corresponding control = Induction rateThe test substance is to be interpretated mutagenic if there is a concentration effect relationstup and the induction rate is ≥ 2.

Key result

Species / strain:

S. typhimurium, other: 97a

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

without S9 ≥ 500 µg/plate, with S9 ≥ 5000 µg/plate

Vehicle controls validity:

valid

Positive controls validity:

valid

Key result

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

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 100

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

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 102

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

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:1st study all test strains 5, 50, 500 µg/plate2nd studyTA97a - S9 16, 50, 160, 500, 1600 µg/plateTA97a + S9 16, 50, 160, 500, 1600, 5000 µg/plateTA98, TA100, TA102 ± S9 50, 160, 500, 1600, 5000 µg/plate

Conclusions:

In this study the test substance was found to have no mutagenic effects on Salmonella typhimurium strains TA 97 a, TA 98, TA 100 and TA 102 with (+) and without (-) the metabolic activation system S9 at concentrations of 0.005 - 5.0 mg/plate.

Executive summary:

The genotoxicity of the registration substance was investigated according to the OECD Guideline 471.

The strains TA97a, TA98, TA100, and TA102 of S. Typhimurium were exposed to the test item in water at concentrations of 5 to 5000 µg/plate.

The test was performed with the pre-incubation method in all bacterial strains in both the presence and the absence of metabolic activation.

The test item was tested up to cytotoxic concentrations for S. typhimurium TA97a (500 µg/plate) or up to limit concentrations for S. typhimurium TA 98, TA100, TA102 (5000 µg/plate). Precipitation of the test substance was not detected in both the presence and the absence of metabolic activation. The positive controls induced the appropriate responses in the corresponding strains.  There was no evidence of induced mutant colonies over background.

Reference 2

Endpoint:

in vitro cytogenicity / chromosome aberration 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 473 (In Vitro Mammalian Chromosome Aberration Test)

Deviations:

no

GLP compliance:

yes

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 mix

Test concentrations with justification for top dose:

+S9: 313, 625, 1250, 2500, 5000 µg/mL
-S9: 1.25, 2.5, 5, 10, 20, 39, 78 µg/mL

Vehicle / solvent:

1% ethanol

Untreated negative controls:

yes

Remarks:

untreated cells

Negative solvent / vehicle controls:

yes

Remarks:

1% ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: methylmethanesulfonate (10-40 µg/mL), cyclophosphamide (20-60 µg/mL)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

Evaluation criteria:

According to Guideline.

Key result

Species / strain:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Not clastogenic in in-vitro chromosome aberration test.

Executive summary:

The genotoxicity of C16 -18AE1 (CAS 68439 -49 -6) was investigated according to the OECD Guideline 473. The test material exhibited no significant clastogenic property in this test.

Reference 3

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

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 (incl. QA statement)

Type of assay:

mammalian cell gene mutation assay

Target gene:

HPRT locus

Species / strain / cell type:

Chinese hamster Ovary (CHO)

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

dose finding test: 0, 1 - 100 µg/mL (solubility limit)
main test: 0, 1.8, 6, 18, 60, 100 µg/mL

Vehicle / solvent:

H0 medium with 1% ethanol

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

H0 medium with 1% ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: ethylmethanesulfonate (300 µg/mL), 3-methylcholanthrene (10 µg/mL)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

Evaluation criteria:

According to Guideline.

Key result

Species / strain:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Not mutagenic in in-vitro mammalian cell gene mutation test

Executive summary:

The genotoxicity of C9 -11AE6 (CAS 68439 -46 -3) was investigated according to the OECD Guideline 476. The test material exhibited no significant mutagenic property in this test.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

No significant concern can be reliably derived for the registration substance with respect to the endpoint genotoxicity based on the experimental data on the registration substance and on a read-across supporting substance.

Link to relevant study records

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

no

GLP compliance:

yes

Type of assay:

micronucleus assay

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Massachusetts, USA
- Age at study initiation: Pre-test: 9.5 weeks, Main study: 7 weeks
- Weight at study initiation: Pre-test: 37-43 and 30-33 g for males and females, respectively; Main study: 31-34 and 25-29 g for males and females, respectively
- Assigned to test groups randomly: yes, under following basis: body weight
- Housing: 5 per cage
- Diet (ad libitum): Wayne Lab Rodent Blox
- Water (ad libitum): Tap water
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3
- Humidity (%): 30 to 70
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

intraperitoneal

Vehicle:

- Vehicle used: corn oil
- Lot/batch no. : Lot# 19F-003 (Sigma Chemical Company)

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The test item was weighed and diluted with corn oil just prior to dosing.
Intraperitoneal administration was chosen to maximize bioavailability of the test item to the hematopoietic cells of the bone marrow.

Duration of treatment / exposure:

Single application

Frequency of treatment:

Single application

Post exposure period:

Mice were sacrificed 24, 48 and 72h after application of the test item.
Mice of the vehicle control were sacrificed 48 h after application.
Mice of the positive control were sacrificed 24 h after application.

Remarks:

Doses / Concentrations:
100 mg/kg bw
Basis:
nominal conc.

No. of animals per sex per dose:

Pre-test: 2
Main study: 5

Control animals:

yes, concurrent vehicle

Positive control(s):

triethylenemelamine (TEM) in 0.9% saline
- Dose: 0.5 mg/kg bw

Tissues and cell types examined:

Tissue: bone marrow
Cells: polychromatic erythrocytes and normochromatic erythrocytes

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION:
In a preliminary Dose-Range-Finding study, the test item was administered intraperitoneally to 5 groups of CD-1 mice at 50, 100, 250, 500 and 1000 mg/kg bw. Due to the mortality observed at the two highest doses evaluated and the severe pharmacotoxic signs observed at the dose of 250 mg/kg bw, 100 mg/kg bw was selected as the dose in the MNT, as an estimate of the maximum tolerated dose.

DETAILS OF SLIDE PREPARATION:
Bone marrow of the femur was collected in fetal bovine serum and centrifuged. The pelleted bone marrow cells were resuspended in FBS and smear slides were preapred. Slides were dried, dipped in absolute methanol and air dried. Dried slides were stained using polychrome methylene blue and eosin, and a staining machine. Slides were coverd by permaslip and randomly coded.

Evaluation criteria:

Criteria for Scoring Micronuclei:
Micronuclei are uniform, darkly stained, typically round bodies in the cytoplasm of erythrocytes. Occasionally, micronuclei appear almond or tear drop shaped, inclusions in PCEs which are reflective, improperly shaped or stained, or which are not in the focal plane of the cell were judged to be artifacts and were not scored. Cells containing more than one micronucleus are scored as one micronucleated erythrocyte.
Criteria for a Valid Test:
If the spontaneous rate of micronuclei in the PCE is less than 0.5% and the positivec ontrol is statistically significant higher( p<0.05) than the spontaneous rate and at least seven animals per group survived the treatment, the results are deemed acceptable.

Statistics:

One-tailed t-tests were used to make pairwise comparisons between each treatment group and the vehicle control for statistically significant increases in the number of micronucleated PCE. The ratio of PCE/NCE was calculated based on 1000 erythrocytes for eacha nimal.The proportion of PCE per 1000 erythrocytesper animal was evaluated by pairwise two-tailed t-tests after an arcsin-transformation was performed.

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Remarks:

clinical signs of toxicity were observed. However, the PCE/NCE ratio was not decreased but increased after 48 h treatment.

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

Immediately after dosing, 9/15 males and 8/15 females had decreased body tone and 6/15 males had piloerection. At 24 h, 5/15 males and 6/15 females had decreased body tone and 8/15 males and 4/15 females had piloerection. Body drop was seen in 1/15 males and 1/15 males had abnormal gait. At 48 h, 3/10 males and 2/10 females had decreased body tone and 3/10 males and 1/10 females had piloerection. At 72 h, 1/5 males had body drop, decreased body tone and piloerection.
No pharmacotoxic signs were observed in animals administered the positive or vehicle controls.
The vehicle and the positive control were within the historical control range.

Table 3: Results of the in vivo micronucleus assay

Treatment group	Dose [mg/kg bw]	Sampling time [h]	Mean frequency of PCE with MN	PCE/NCE ratio
Vehicle control	0	48	0.04	1.24
Test substance	100	24	0.05	0.98
		48	0.03	1.64 (p≤0.05)
		72	0.05	1.45
Positive control (TEM)	0.5	24	2.04 (p≤0.01)	0.84 (p≤0.05)

Conclusions:

No effect in in-vivo micronucleus assay in mouse

Executive summary:

The genotoxicity of C12 -14AE2 (CAS 68439 -50 -9) was investigated according to the OECD Guideline 474. Mice were treated at dose of 2000 mg/kg bw once per day for two days. 24 hours after the seconed application the animals were sacrificed and the bone marros cells were collected. The incidence of mucronucleated polychromatic erythrocytes was comparable for treated and control mice. The ratio of polychromatic erythrocytes to total erhythrocytes remained unaffected. No clastogenic effect was found.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

The genotoxicity of the registration substance is to be derived based on the available data on the registration substance and on the read-across supporting substances. The justification of the read-across is provided as attached document in the dossier.

 

Overview of Data Availability and Obtained Results Study Types		Results
In-vitro	Ames Test (OECD 471)	negative in three studies (registration substance and read-across supporting substance
	Chromosome Aberration Test (OECD 473)	negative (read-across supporting substance)
	HPRT	negative (read-across supporting substance)
In-vivo	Micronucleus Test in Mice (OECD 474)	negative in two studies (registration substance and read-across supporting substance

In conclusion, the registration is considered to be of no concern for the endpoint genotoxicity.

Justification for classification or non-classification

No significant concern could be reliably derived. No classificaiton is justified.