N-(3-(trimethoxysilyl)propyl)ethylenediamine

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation (Bacterial reverse mutation assay / Ames test): negative with and without metabolic activation in Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and Escherichia coli WP2 CM 881 and CM891 (EPA OPPTS 870.5100, similar to OECD TG 471) (DCC (1999)).

Cytogenicity in mammalian cells: testing not required as a reliable in vivo micronucleus is available. Mutagenicity in mammalian cells: negative with and without metabolic activation in CHO cells (EPA Health Effects Test Guidelines HG-Gene Muta-Somatic Cells, similar to OECD TG 476) (Slesinski and Guzzie (1988)).

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

Study period:

1998-11-10 - 1998-11-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

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

Species / strain / cell type:

E. coli, other: E coli WP2 trp pKM101 (CM881)

Species / strain / cell type:

E. coli, other: E coli WP2 trp uvrA pKM101 (CM891)

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver S9

Test concentrations with justification for top dose:

50, 150, 500, 1500 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: Sponsors choice as completely miscible in water

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Remarks:

without metabolic activation TA1535 5 µg/plate, TA100 3 µg/plate, CM881 and CM891 2 µg/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA1537 without metabolic activation 30 µg/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA98 without metabolic activation 1 µg/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene 2 µg/plate TA1535, 10 µg/plate CM881 and CM891

Remarks:

with metabolic activation

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

TA1537, TA98 and TA100 with metabolic activation 5 µg/plate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation

ACTIVATION: S9 mix contained S9 fraction 10%, MgCl, KCl, sodium orthophosphate buffer pH 7.4, glucose-6-phosphate and NADP. 0.5 ml S9 mix and 0.1ml of test solution were added to 2ml agar.

DURATION

- Preincubation period: 30 minutes
- Exposure duration: 3 days

SELECTION AGENT (mutation assays): histidine deficient agar

NUMBER OF REPLICATIONS: triplicate plates, experiment repeated

DETERMINATION OF CYTOTOXICITY
- Method: other: condition of background lawn

ACTIVATION
- Aroclor induced rat liver S9 was present at 10% in the S9 mix, which included NADP and glucolse-6-phosphate as cofactors. 0.5 ml S9 mix was added to 2 ml top agar, 0.1 ml test solution or control and 0.1 ml bacterial culture giving a final concentration of approximately 2%.

Evaluation criteria:

A result is considered positive if there is a three-fold increase in the number of revertants in TA1535, TA1537 and TA98 over the solvent control and a two-fold increase for other strains. This should occur in both experiments and show some evidence of a dose-relationship. The positive controls must cause at least a doubling of mean revertant colony numbers over the mean of the solvent control.

Statistics:

None in report

Key result

Species / strain:

S. typhimurium TA 1535

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 applicable

Positive controls validity:

valid

Key result

Species / strain:

E. coli WP2 uvr A pKM 101

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 applicable

Positive controls validity:

valid

Key result

Species / strain:

E. coli, other: E. coli WP2 pKM101 (CM881)

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 applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1537

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 applicable

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

Untreated negative controls validity:

not applicable

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

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Average number of revertants per plate (mean of three plates)

Treatment µg/plate	TA 1535		TA 1537		TA 98		TA 100		CM 881		CM 891
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
Solvent control	14	16	9	10	19	23	108	127	46	53	179	180
50	14	19	7	6	22	27	110	133	56	67	201	190
150	14	20	7	7	19	24	111	132	63	77	189	203
500	14	16	7	8	24	24	109	113	46	56	193	203
1500	13	16	6	7	25	23	111	123	61	73	199	195
5000	10	11	9	9	15	23	87	114	61	63	188	197
Positive control	370	179	316	46	115	164	456	718	817	392	1703	1799
Average Revertants per plate with and without metabolic activation – Exp 2 pre-incubation
Treatment µg/plate	TA 1535		TA 1537		TA 98		TA 100		CM 881		CM 891
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9		+S9
Solvent control	18	15	9	9	29	27	134	143	61	69	247		242
50	20	16	8	11	27	31	115	134	55	73	238		253
150	18	18	9	10	21	30	110	135	47	66	216		237
500	13	21	10	9	26	26	106	129	58	62	261		253
1500	18	18	7	8	21	28	113	114	54	80	230		235
5000	15	18	8	6	25	28	105	97	48	60	215		268
Positive control	244	115	120	144	130	270	456	569	792	438	1569		758

Average Revertants per plate with and without metabolic activation – Exp 1

Conclusions:

N-(3-(trimethoxysilyl)propyl)ethylenediamine has been tested for mutagenicity to bacteria, in a study that was conducted according to the OECD TG 471 and in compliance with GLP. No evidence of test substance related increase in the number of revertants was observed with or without activation in the initial experiment using the plate incorporation method or the repeat pre-incubation assay in Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537, E coli WP2 trp uvrA pKM101 (CM891) and E coli WP2 trp pKM101 (CM881). The solvent used was water, and hydrolysis was likely to have occurred under test conditions, so it is assumed that exposure was to the hydrolysis product as well as the parent substance. It is not considered to invalidate the study, as it is considered that hydrolysis may occur in vivo. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.

Reference 2

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1988-01-13 to 1988-03-21

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Remarks:

Insufficient analysable concentrations to meet current guidelines.

Qualifier:

equivalent or similar to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

other: EPA Health Effects Test Guidelines HG-Gene Muta-Somatic Cells (1983)

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Target gene:

HGPRT (hypoxanthine-guanine phosphoribosyl transferase)

Species / strain / cell type:

Chinese hamster Ovary (CHO)

Details on mammalian cell type (if applicable):

- Type and identity of media: Ham's Modified F12 Medium
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: no
- Periodically "cleansed" against high spontaneous background: no

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Aroclor-1254 induced rat liver S9

Test concentrations with justification for top dose:

without metabolic activation: 2.5, 3.0, 3.25, 3.5 and 4.0 mg/ml, with metabolic activation: 2.0, 2.5, 3.0, 3.5, 4.0 and 4.5 mg/ml

Vehicle / solvent:

DMSO

Untreated negative controls:

yes

Remarks:

cell culture medium

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

Remarks:

without metabolic activation

Untreated negative controls:

yes

Remarks:

cell culture medium

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

N-dimethylnitrosamine

Remarks:

with metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 5 hours (with and without metabolic activation)
- Expression time (cells in growth medium): 9-12 days


SELECTION AGENT (mutation assays): TG selective medium

NUMBER OF REPLICATIONS: Duplicate cultures. Surviving fraction was determined at 18 and 24 hours after removal of chemical using 4 plates per culture and 100 cells per plate; mutant frequency was determined using 5 plates per culture. The test was repeated.



DETERMINATION OF CYTOTOXICITY
- Method: plating efficiency

ACTIVATION: S9 mix contained glucose-6-phosphate and NADP as co-factors, and 5% of S9. 1.0 ml of S9 mix was added to 4 ml of culture medium, giving a final concentration of 1% S9.

Evaluation criteria:

The criteria for interpretation of the test results as a positive or negative response depend upon both the level of statistical significance from the concurrent control and the evidence of a dose-response effect following treatment. When a definite dose-response relationship is not evident, but one or more marginally significant values are obtained, a careful examination of the data from the concurrent positive and negative controls and comparisons to historical control data may be used to evaluate the probable biological significance of the responses.

Statistics:

Box-Cox transformation

Key result

Species / strain:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

4.0 mg/ml with metabolic activation

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

CHO Mutation Assay: with and without metabolic activation

Results on evaluation of plating efficiency and mutant colonies - Experiment 1

	Mutant colonies		Plating efficiency
Test chemical	(Mean of 5 plates)		% of Combined Solvent Controls
	- S9	+S9	-S9	+S9
Negative control	0	18	106.3	105.9
Solvent control A	20	2	93.7	102.3
Solvent control B	0	0	106.3	97.8
2.0A	-	6	-	120.5
2.0B	-	13	-	104.4
2.5A	6	35	103.4	107.5
2.5B	1	15	110.0	100.5
3.0A	5	4	104.6	91.0
3.0B	2	0	124.4	109.2
3.25A	5	-	101.1	-
3.25B	5	-	96.0	-
3.5A	0	0	89.4	116.0
3.5B	1	0	103.7	107.7
4.0A	0	-**	114.7	-**
4.0B	5	-**	94.0	-**
Positive control	143	55	115.7	113.2

 

**= Cytotoxic, cultures did not grow

Results on evaluation of plating efficiency and mutant colonies - Experiment 2

Test chemical	Mutant colonies (Mean of 5 plates)	Plating efficiency
	+S9
Negative control	13	88.2
Solvent control A	0	104.1
Solvent control B	1	95.8
2.25A	12	118.7
2.25B	0	99.7
2.5A	0	106.8
2.5B	0	111.1
2.75A	0	101.9
2.75B	0	96.6
Positive control	63	92.6

Conclusions:

N-(3-trimethoxysilyl)propyl)ethylenediamine has been tested in a reliable study according to an appropriate US EPA test guideline that is equivalent to OECD 476 and under GLP. An increase in mutant frequency was observed in one of two duplicate cultures with metabolic activation at one concentration in the initial experiment. No other increase in mutant frequency was observed at any concentration with and without activation (5 hours treatment). The experiment was repeated (with metabolic activation): no increase in mutant frequency was observed. The appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for the induction of mutations in CHO cells under the conditions of the test.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

Micronucleus assay in mouse (ip administration): negative (EPA 560/6-83-001, similar to OECD 474) (BRRC (1988)). 

Link to relevant study records

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1988-03-15 to 1988-04-08

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Only 1000 cells/animal were scored for micronuclei.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

number of micronucleated PCEs scored per 1000 cells, not 2000

Qualifier:

according to guideline

Guideline:

other: EPA Health Effect T G, EPA Report 560/6-83-001

GLP compliance:

yes

Type of assay:

micronucleus assay

Species:

mouse

Strain:

Swiss Webster

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Portage, MI
- Age at study initiation: 5 weeks
- Weight at study initiation: male - 22.5 g - 28.0 g. female - 19.8 g - 21.7 g
- Assigned to test groups randomly: randomised separately by sex
- Fasting period before study: no
- Housing: 5 mice/sex/cage in shoe-box type plastic cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature (°C): controlled but not specified
- Humidity (%): controlled but not specified
- Air changes (per hr): not known
- Photoperiod (hrs dark / hrs light): 12 hr light/dark cycle

Route of administration:

intraperitoneal

Vehicle:

- Vehicle(s)/solvent(s) used: corn oil
- Justification for choice of solvent/vehicle: incompatible with water thus corn oil was used
- Concentration of test material in vehicle: 200 mg/kg

Duration of treatment / exposure:

single i p dose

Frequency of treatment:

One treatment only

Post exposure period:

30, 48 and 72 hours

Dose / conc.:

87.5 mg/kg bw/day

Dose / conc.:

175 mg/kg bw/day

Dose / conc.:

280 mg/kg bw/day

No. of animals per sex per dose:

5

Control animals:

yes

Positive control(s):

Triethylenemelamine
- Justification for choice of positive control(s): known to demonstrate the sensitivity and responsiveness of the animals in the definitive test.
- Route of administration: i p
- Doses / concentrations: 0.3 mg/kg bw

Tissues and cell types examined:

Blood was collected by nicking the tail of each animal with a scalpel and slides prepared for each animal per sampling time. Polychromatic erythrocytes (PCE's) were examined.

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: Determination of the PCE/NCE ratio for the groups of animals with partial mortality was used to evaluate the possibility of bone marrow cytotoxicity from the test chemical. Three dose levels of approximately 80%, 50% and 25% of the LD50 value were evaluated for effects upon the incidence of micronuclei.

TREATMENT AND SAMPLING TIMES (in addition to information in specific fields): i.p. injection followed by sampling times of 30, 48 and 72 hours

DETAILS OF SLIDE PREPARATION: Slides were stained with Gurr's R-66 Giemsa diluted in phosphate buffer. Slides were coded by animal number only and read blindly.

METHOD OF ANALYSIS: A minimum of 1000 PCE's were examined microscopically for each animal per sample time. The PCE/NCE ratio for approximately 1000 total cells was calculated and recorded. The number of micronucleated PCE/1000 NCE was recorded.

Evaluation criteria:

A test result is considered to be negative if no statistically significant or dose related increases are apparent between the vehicle control and groups of animals treated with Organofunctional Silane A-1120.

Statistics:

Fisher's Exact Test (Sokal and Rohlf, 1981)

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Remarks:

LD50 354 mg/kg bw and above. PCE/NCE ratio was reduced at highest concentration, 48 h exposure and at all doses, 72 h exposure.

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- Dose range: 125 mg/kg bw - 2000 mg/kg b
- Solubility: Incompatible with water thus corn oil was used
- Clinical signs of toxicity in test animals: All mice dosed at 1000 mg/kg bw and 2000 mg/kg bw died. 500 mg/kg bw was lethal to 4 out of 5 females and all the male mice. One female tested at 250 mg/kg bw died.
- Evidence of cytotoxicity in tissue analyzed: a decrease in the PCE/NCE ratio to 80% of the control value was observed at the 48 hour treatment interval. At 72 hours the PCE/NCE ratios had increased.
- Harvest times: 48 hours


RESULTS OF DEFINITIVE STUDY

- Induction of micronuclei (for Micronucleus assay): negative.
- Ratio of PCE/NCE (for Micronucleus assay): PCE/NCE ratio was slightly decreased at 72 hour treatment interval.
- Appropriateness of dose levels and route: Both dose levels and route were appropriate.
- Statistical evaluation: Analysis of variance indicated no sex-related differences in the incidence of micronuclei. No statistically significant (p ≤ 0.01) or treatment related increases in the numbers of micronuclei were observed at any dose or treatment interval.

Table 3: Summary of micronucleus results

	30 hours				48 hours				72 hours
Treatment groups mg/kg bw	Number of PCE’s with micronuclei per 1000 PCE’s		Mean PCE’s/1000 NCE (SD)		Number of PCE’s with micronuclei per 1000 PCE’s		Mean PCE’s/1000 NCE (SD)		Number of PCE’s with micronuclei per 1000 PCE’s		Mean PCE’s/1000 NCE (SD)
	Male	Female	Group mean	SD	Male	Female	Group mean	SD	Male	Female	Group mean	SD
Vehicle control	4.0 (2.00)	1.6 (0.89)	33.7	5.52	2.4 (2.70)	3.6 (1.14)	36.4	4.24	3.8 (2.05)	3.8 (2.68)	42.9	3.25
87.5	5.0 (2.55)	3.6 (1.52)	31.8	6.79	3.0 (1.73)	2.2 (0.84)	36.0	0.28	6.4 (2.19)	2.8 (1.64)	33.9	2.12
175	2.6 (1.52)	4.0 (1.22)	34.9	8.06	3.2 (1.10)	3.8 (1.64)	37.2	7.92	2.3 (0.96)	5.2 (3.96)	35.5	6.68
280	3.4 (2.19)	3.6 (2.19)	35.4	4.81	3.4 (3.29)	3.0 (1.22)	30.4	3.39	3.8 (1.92)	1.2 (0.84)	27.9	1.27
Positive control	36.2 (16.84)	28.6 (10.31)	27.4	9.05	Not evaluated		Not evaluated		Not evaluated		Not evaluated

Table 4 Summary of micronucleus frequency and statistical differences from controls (Fisher's exact test, one-tailed)

Treatment/dose mg/kg bw	No. PCE observed	PCE with micronuclei	Statistical significance
	30 hour sample
Vehicle control	10 000	28	NS
87.5	10 000	43	0.05>p>0.01*
175	10 000	33	NS
280	10 000	35	NS
Positive control	10 000	324	p<0.001
	48 hour sample
Vehicle control	10 000	30	NS
87.5	10 000	26	NS
175	10 000	35	NS
280	10 000	32	NS
	72 hour sample
Vehicle control	10 000	38	NS
87.5	10 000	46	NS
175	10 000	35	NS
280	10 000	25	NS

Results from sexes are combined as no sex-related differences were shown.

* not considered of biological significance

Conclusions:

N-(3-(trimethoxysilyl)propyl)ethylenediamine has been tested in a reliable study according to EPA Health Effect T G, Report 560/6-83-001, which is similar to OECD 474, and under GLP conditions. No treatment related increases in numbers of micronuclei in PCE's of Swiss-Webster mice were observed. Relatively high dose levels of the test compound were tested up to 80% of the LD50 with no indication of significant induction of micronuclei. The PCE/NCE ratio was reduced at the highest concentration, 48 h exposure and at all doses, 72 h exposure, indicating exposure of the target tissue to the test substance. The test compound is considered to be negative for the induction of micronuclei under the conditions of this test.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Information is available for all the required endpoints for N-(3 -(trimethoxysilyl)propyl)ethylenediamine.

N-(3-(trimethoxysilyl)propyl)ethylenediamine has been tested for mutagenicity to bacteria, in a study that was conducted according to the OECD TG 471 and in compliance with GLP (DCC (1999)). No evidence of test substance related increase in the number of revertants was observed with or without activation in the initial or the repeat experiments using the plate incorporation method in Salmonella typhimurium strains TA 98, TA 100, TA 153, TA 1537, E coli WP2 trp uvrA pKM101 (CM891) and E coli WP2 trp pKM101 (CM881). The solvent used was water, and hydrolysis was likely to have occurred under test conditions, so it is assumed that exposure was to the hydrolysis product as well as the parent substance. It is not considered to invalidate the study, as it is considered that hydrolysis may occur in vivo. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test. The result of this study is supported by four older studies, where non-aqueous solvents were used: the results of all the studies on bacterial mutagenicity are in agreement. The most reliable study was selected as key.

 

N-(3-trimethoxysilyl)propyl)ethylenediamine has been tested in a reliable study according to an appropriate US EPA test guideline that is equivalent to OECD 476 and under GLP (Slesinski and Guzzie (1988)). An increase in mutant frequency was observed in one of two duplicate cultures with metabolic activation at one concentration in the initial experiment. No other increase in mutant frequency was observed at any concentration with and without activation (5 hours treatment). The experiment was repeated (with metabolic activation): no increase in mutant frequency was observed. The appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for the induction of mutations in CHO cells under the conditions of the test.

 

No in vitro cytogenicity study is available; testing is not required as a reliable in vivo micronucleus assay is available.

 

A reliable in vitro sister chromatid assay is available, conducted according to a US EPA protocol that is similar to OECD TG 479 and under GLP (Slesinski and Guzzie (1988)). N-(3-(trimethoxysilyl)propyl)ethylenediamine did not induce statistically and biologically significant increases in the sister chromatid exchange frequency of CHO cells. A slight statistically significant increase in the sister chromatid exchange frequency of CHO cells was not considered of biological significance as the level of increase was very low, and was not dose related. Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for the induction of sister chromatid exchange in vitro under the conditions of the test. This study is not considered key as it does not correspond to a standard REACH endpoint.

 

N-(3-(trimethoxysilyl)propyl)ethylenediamine has been tested in a reliable in vivo micronucleus study according to EPA Health Effect T G, Report 560/6-83-001, which is similar to OECD 474, and under GLP conditions (BRRC (1988)). No treatment related increases in numbers of micronuclei in PCE's of Swiss-Webster mice were observed after intraperitoneal administration. Relatively high dose levels of the test compound were tested up to 80% of the LD50 with no indication of significant induction of micronuclei. The PCE/NCE ratio was reduced at the highest concentration, 48 h exposure and at all doses, 72 h exposure, indicating exposure of the target tissue to the test substance. The test compound is considered to be negative for the induction of micronuclei under the conditions of this test.

 

No evidence of mutagenicity or clastogenicity was observed in any of the available in vitro and in vivo studies.

Justification for classification or non-classification

Based on the available in vitro and in vivo genotoxicity data N-(3-(trimethoxysilyl)propyl)ethylenediamine is not classified for mutagenicity according to Regulation 1272/2008/EC.