Sodium hydrogen N-(1-oxotetradecyl)-L-glutamate

Genetic toxicity: in vitro

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 487 (In vitro Mammalian Cell Micronucleus Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

other: in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

L-Glutamic acid, N-coco acyl derivs., disodium salts

EC Number:

269-085-1

EC Name:

L-Glutamic acid, N-coco acyl derivs., disodium salts

Cas Number:

68187-30-4

IUPAC Name:

L-glutamic acid, N-coco acyl derivs., disodium salts

Test material form:

solid

Details on test material:

Organic

Method

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

From 0 to 1600 µg/mL

Vehicle / solvent:

Water

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

ethylmethanesulphonate

Details on test system and experimental conditions:

- Exposure of 4 or 24 hours
- Concentrations from 0 to 1600 µg/mL
- With or without metabolic activation (S9 mix)
- Negative (vehicle) and positive controls included

Rationale for test conditions:

- Guideline
- Cytotoxicity

Evaluation criteria:

- Chromosome damages
- Number of chromosomal aberrations

Statistics:

According to guideline

Results and discussion

Test results

Key result

Species / strain:

Chinese hamster lung fibroblasts (V79)

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

Any other information on results incl. tables

Table 1. Summary of results – experimental parts without S9 mix

Exp.	Exposure	Test groups	S9	Prec.*	Genotoxicity	Cytotoxicity
	period		mix		Micronucleated cells**	Proliferation index (PI)	RICC***
					[%]	absolute	[%]
1	4 hrs	Negative control	-	n.d.	0.7	2.77	100.0
		10.94 µg/mL	-	-	n.d.	n.d.	941.5
		21.88 µg/mL	-	-	n.d.	n.d.	91.3
		43.75 µg/mL	-	-	n.d.	n.d.	91.1
		87.50 µg/mL	-	-	n.d.	n.d.	89.1
		175.00 µg/mL	-	-	0.6	2.73	88.9
		350.00 µg/mL	-	-	0.8	2.58	86.1
		700.00 µg/mL^+	-	-	4.3^S	2.28	41.4
		Positive control^2	-	n.d.	2.7^S	2.52	n.t.
3	4 hrs	Negative control	-	n.d.	0.8	2.32	100.0
		200.00 µg/mL	-	-	n.d.	n.d.	107.0
		400.00 µg/mL	-	-	0.5	2.16	94.5
		600.00 µg/mL	-	-	0.7	2.38	85.4
		700.00 µg/mL^+	-	-	1.0	2.23	78.1
		800.00 µg/mL	-	-	n.s.	n.s.	-0.5
		Positive control^1	-	n.d.	2.4^S	2.30	n.t.
2	24 hrs	Negative control	-	n.d.	1.2	2.35	100.0
		200.00 µg/mL	-	-	n.d.	n.d.	93.3
		400.00 µg/mL	-	-	n.d.	n.d.	90.6
		600.00 µg/mL	-	-	1.4	2.08	84.0
		700.00 µg/mL	-	-	0.3	2.23	88.4
		800.00 µg/mL	-	-	0.6	2.09	48.1
		Positive control^1	-	n.d.	9.1^S	2.32	n.t.

*      Precipitation in culture medium at the end of exposure period

**     Relative number of micronucleated cells per 1000 cells scored per test group

***   Relative increase in cell count (RICC)

⁺      To confirm the results an increased sample of 4 000 cells was scorted.

^S      Frequency statistically significant higher than corresponding control values

n.d. Not determined     n.t.     Not tested

n.s. Not scorable due to strong cytotoxicity

¹      EMS 500 µg/mL            ²        EMS 600 µg/mL

Table 2. Summary of results – experimental parts with S9 mix

Exp.	Exposure	Test groups	S9	Prec.*	Genotoxicity	Cytotoxicity
	period		mix		Micronucleated cells**	Proliferation index (PI)	RICC***
					[%]	absolute	[%]
1	4 hrs	Negative control	+	n.d.	1.5	2.70	100.0
		5.47 µg/mL	+	-	n.d.	n.d.	98.3
		10.94 µg/mL	+	-	n.d.	n.d.	96.1
		21.88 µg/mL	+	-	n.d.	n.d.	125.4
		43.75 µg/mL	+	-	n.d.	n.d.	114.8
		87.50 µg/mL	+	-	1.7	2.56	117.6
		175.00 µg/mL	+	-	0.5	2.47	97.2
		350.00 µg/mL	+	-	0.9	2.43	93.6
		Positive control^1	+	n.d.	8.5^S	2.32	n.t.
2	4 hrs	Negative control	+	n.d.	0.8	2.31	100.0
		100.00 µg/mL	+	-	n.d.	n.d.	81.7
		200.00 µg/mL	+	-	1.0	2.27	70.5
		400.00 µg/mL	+	-	1.1	2.07	54.1
		800.00 µg/mL	+	-	1.3	2.05	39.4
		1 600.00 µg/mL	+	-	n.s.	n.s.	32.7
		Positive control^1	+	n.d.	15.2^S	2.09	n.t.
3	4 hrs	Negative control	+	n.d.	1.5	2.03	100.0
		75.00 µg/mL	+	-	n.d.	n.d.	107.0
		150.00 µg/mL	+	-	n.d.	n.d.	109.3
		300.00 µg/mL	+	-	n.d.	n.d.	99.5
		600.00 µg/mL	+	-	0.8	1.98	89.2
		1 200.00 µg/mL	+	-	1.3	2.23	91.5
		1 600.00 µg/mL^+	+	-	1.6	2.03	76.4
		Positive control^1	+	n.d.	12.5^S	1.80	n.t.

*      Precipitation in culture medium at the end of exposure period

**     Relative number of micronucleated cells per 1000 cells scored per test group

***   Relative increase in cell count (RICC)

⁺      To confirm the results an increased sample of 4 000 cells was scorted.

^S      Frequency statistically significant higher than corresponding control values

n.d. Not determined              n.t.     Not tested

n.s. Not scorable due to strong cytotoxicity

¹      CPP 2.5 µg/mL

Applicant's summary and conclusion

Conclusions:

Under the study conditions, the substance was not clastogenic in Chinese hamster lung fibroblast V79 cells.

Executive summary:

A study was conducted to determine the in vitro genetic toxicity of the substance according to OECD Guideline 487, in compliance with GLP. Two independent experiments were performed in Chinese hamster lung fibroblasts (V79 cells). Cells were exposed to the test substance (diluted in water) at concentrations from 0 to 1600 µg/mL for either 4 or 24 h with and without metabolic activation (S9 mix). Cytotoxicity, indicated by a clearly reduced proliferation index, cell numbers or a low quality of the slides was observed in the absence of S9 mix at 700 µg/mL (1st experiment) and at 800 µg/mL (2nd and 3rd experiment). In the presence of S9 mix, cytotoxic effects were obtained 800 µg/mL onwards but only in the 2nd experiment. In the 1st experiment a single statistically significant increase in the number of micronucleated cells was observed at 700 µg/mL after 4 h exposure in the absence of metabolic activation. However, this finding was neither confirmed in the repeat experiment under similar conditions (3rd experiment) nor in any additional experiment under modified test conditions (24 h exposure in the absence of metabolic activation or 4 h exposure in the presence of metabolic activation). The finding was considered artifactual due to the strong cytotoxicity in this test group (RICC 41.4%). Under the study conditions, the substance was not clastogenic in Chinese hamster lung fibroblast V79 cells (BASF SE, 2013).