Amines, C12-18-alkyl, reaction products with acrylic acid, sodium salts

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 Dec 1989 - 06 Aug 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Acceptable, well-documented study which meets basic scientific principles, with acceptable restrictions. The study was performed on an analogue substance (for justification of read-across, please refer to the corresponding assessment report in Section 13).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Deviations:

yes

Remarks:

lack of study details, purity of test substance not specified

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

Sodium N-(2-carboxyethyl)-N-dodecyl-β-alaninate

EC Number:

239-032-7

EC Name:

Sodium N-(2-carboxyethyl)-N-dodecyl-β-alaninate

Cas Number:

14960-06-6

IUPAC Name:

Sodium Lauriminodipropionate

Test material form:

other: liquid

Details on test material:

- Common name : Sodium lauriminodipropionate
- For more details, see below the Confidential details for test material

Specific details on test material used for the study:

The study was performed on a commercial product (aqueous solution) as the test item is manufactured and used in a liquid form.

Method

Target gene:

not applicable

Species / strain

Species / strain / cell type:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254

Test concentrations with justification for top dose:

With S9:
. 8 hours harvest - 0.21, 0.28, 2.38 µl/mL
. 12 hour harvest - 0.28, 0.38, 0.50 µl/mL

Without S9:
. 8 hour harvest - 0.48, 0.63, 0.84 µl/mL
. 12 hour harvest - 0.63, 0.84, 1.13 µl/mL

Vehicle / solvent:

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

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Triethylenemelamine (TEM, 0.25 µg/mL, -S9); Cyclophosphamide (CP, 20 µg/mL, +S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 8 and 12 hours

NUMBER OF REPLICATIONS: 2

NUMBER OF CELLS EVALUATED: 100 per culture

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index

Statistics:

The cytotoxic effects of treatment are expressed relative to the solvent control (relative cloning efficiency). The number and types of aberrations found are presented for each treatment flask. Duplicate treatment flasks were compared using the Fisher's exact test and, if not statistically significant, combined for treatment comparison. The percentage of damaged cells (numerical and structural) in the total population of cells examined was calculated for each group. A statistical analysis of the percent aberrant cells per dose was made using the Fisher's exact test. The average number of aberrations per cell was reported but no statistical analysis was applied. The Cochran-Armitage trend test was performed between the solvent and treatment groups for each treatment condition and harvest time to test for evidence of a dose response.

Results and discussion

Test results

Species / strain:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Relative cloning efficiency at the highest dose level tested of 34% and 1% in the 8 hour and 12 hour non-activated studies, respectively, and 70% and 38% in the 8 hour and 12 hour S9 activated studies, respectively.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: stock concentrations were adjusted to pH7 in order to ensure neutrality of the treatment medium
- Effects of osmolality: osmolality was measured before the main experiments and concentrations chosen where no difference in osmolality was observed

RANGE-FINDING/SCREENING STUDIES:
Dose levels were chosen according to a range-finding toxicity study which was based on cloning efficiency relative to the solvent control. Based upon the findings of the toxicity study dose levels 0.5 and 1.5 µl/mL were the high doses selected for further study in the non-activated and S9 activated studies, respectively.
The average cell generation time (AGT) was calculated in the non-activated and S9 activated studies for the solvent and the two highest test concentrations yielding metaphase cells. At the absence of cell cycle delay harvest times were set at 8 and 12 hours after initiation of treatment for non-activated and S9 activated studies.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: summary of cytogenetic assay results

Treatment^1	Harvest time (hour)	Mean mitotic index^2	Cells scored	Cels with structural aberrations (%)^3,4	Structural aberrations per cell (mean ± SD)^5
Non-activated study
untreated cells	8	3.4	100	0	0 .000 ± 0.000
water	8	4.1	100	1	0.010 ± 0.100
0.21 µL/mL	8	3.9	100	0	0.000 ± 0.000
0.28 µL/mL	8	2.2	100	1	0.010 ± 0.100
0.38 µL/mL	8	0.2	67	0	0.000 ± 0.000
untreated cells	12	4.2	100	0	0.000 ± 0.000
water	12	3.5	100	0	0.000 ± 0.000
0.28 µL/mL	12	2.5	100	1	0.010 ± 0.100
0.38 µL/mL	12	0.8	100	2	0.020 ± 0.141
0.50 µL/mL	12	0.2	64	1	0.016 ± 0.125
TEM, 0,25 µg/mL	12	0.9	100	11	0.130 ± 0.393
S9 activated study
untreated cells	8	2.5	100	1	0.010 ± 0.100
water	8	2.3	100	3	0.030 ± 0.171
0.48 µL/mL	8	1.9	100	1	0.010 ± 0.100
0.63 µL/mL	8	2.1	100	1	0.010 ± 0.100
0.84 µL/mL	8	0.8	100	1	0.010 ± 0.100
untreated cells	12	3.7	100	1	0.010 ± 0.100
water	12	3.3	100	1	0.010 ± 0.100
0.63 µL/mL	12	3.7	100	3	0.030 ± 0.171
0.84 µL/mL	12	4.1	100	2	0.020 ± 0.141
1.13 µL/mL	12	0.9	55	0	0.000 ± 0.000
CP, 20µg/mL	12	1.8	100	12	0.130 ± 0.367

¹ CHO cells were treated at 37°C for 4 hours

² Mitotic Index = cells in mitosis per 500 cells counted, expressed as a percentage

³ Structural aberrations include unanalyzable cells and cells with one ore more aberrations, excluding gaps

⁴ significantly increased above the control using Fisher's exact test; *p<0.025

⁵ Gaps and unanalyzable cells are not included; SD = standard deviation

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Under the conditions of the assay described in this report, sodium lauriminodipropionate did not induce a significant increase in chromosome aberration in the CHO cytogenicity study.

Executive summary:

Sodium lauriminodipropionate was tested in a chromosome aberration assay using Chinese hamster ovary cells. The assay was conducted both in the absence and presence of an Aroclor induced rat liver S9 activation system. Dose levels of 0.21, 0.28 and 0.38 µl/mL in the 8 hour non-activated study, 0.28, 0.38 and 0.5 µl/mL in the 12 hour non-activated study, 0.48, 0.63 and 0.84 µl/mL in the 8 hour S9-activated study and dose levels 0.63, 0.84 and 1.13 µl/mL in the 12 hour S9-activated study were selected as the highest, non-precipitating concentrations with or without S9 which could be evaluated for chromosome aberrations.

Survival (relative cloning efficiency) at the highest dose level scored was 34% and 1% in the 8 and 12 hour non-activated studies, respectively, and 70% and 38% in the 8 and 12 hour S9 activated studies, respectively.

The test article did not induce a significant increase in chromosome aberrations at either harvest time, 8 or 12 hours, in the absence of presence of S9 activaton. The test substance was concluded to be negative in the CHO cytogenicity study.