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Diss Factsheets

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Information is available from three bacterial mutagenicity studies on N-[3-(dimethoxymethylsilyl)propyl]ethylenediamine (CAS No. 3069-29-2, EC No. 221-336-6). No further information is available for the registered substance, however, reliable data are available for the closely related substance, N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS No. 1760-24-3, EC No. 217-164-6).

Gene mutation (Bacterial reverse mutation assay / Ames test): negative with and without metabolic activation in Salmonella typhimurium strains TA 98, 100, 102, 1535, 1537 (OECD Test Guideline 471) (LPT, 2002).
Mutagenicity in mammalian cells: read-across from closely related substance N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS 1760-24-3): negative with and without metabolic activation in Chinese hamster ovary cells (similar to OECD Test Guideline 476) (BRRC, 1988a).

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2002-04-10 to 2002-06-13
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay
Target gene:
Histidine operon
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 102
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254 induced rat liver S9
Test concentrations with justification for top dose:
31.6, 100, 316, 1000 and 3160 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Ethylene glycol dimethylether

- Justification for choice of solvent/vehicle: Solubility properties and relative non-toxicity to bacteria
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA 1535, TA 100 (without metabolic activation): 10 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
TA 98 (without metabolic activation): 10 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
TA 1537 (without metabolic activation): 100 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
TA 102 (without metabolic activation): 1300 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-anthracene amide 2 µg/plate
Remarks:
TA 98, TA 102, TA 1537 (with metabolic activation)
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
TA 100, TA 1535 (with metabolic activation): 1500 µg/plate
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION
- Preincubation period: 60 minutes
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 3 plates for each test concentration x 2

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth


METHOD OF APPLICATION in agar (plate incorporation)

ACTIVATION: S9 mix contained glucose-6-phosphate and NADP as co-factors. 0.5 ml S9 mix was added to 2 ml top agar, 0.1 ml test solution, 0.1 ml salmonella cell suspension giving a final concentration of 0.9% S9.

DURATION
- Preincubation period: 1 hour (second experiment)
- Exposure duration: 48 hours



NUMBER OF REPLICATIONS: 3 replicates per concentration, experiment repeated


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


Evaluation criteria:
A result is positive if the number of revertants is significantly increased compared with the solvent control to at least 2-fold of the solvent control for TA 98, TA 100 and TA 102 and 3-fold of the solvent control for TA 1535 and TA 1537 in both experiments.

Positive results have to be reproducible and the histidine independence of the revertants has to be confirmed by streaking on histidine-free agar plates.

Cytotoxicity is defined as a reduction in the number of colonies by >50% compared with the solvent control and/or a sparse background lawn.
Statistics:
MANN and WHITNEY and Spearman’s rank.
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
background lawn reduced in preincubation experiment at 100 µg/plate without metabolic activation and at 316 µg/plate with metabolic activation. Top dose limited to 3160 µg/plate based on cytotoxicity observed in preliminary test test.
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
True 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:
cytotoxicity
Remarks:
background lawn reduced in preincubation experiment at 316 µg/plate without metabolic activation and at 1000 µg/plate with metabolic activation. Top dose limited to 3160 µg/plate based on cytotoxicity observed in preliminary test.
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
True 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:
cytotoxicity
Remarks:
background lawn reduced in preincubation experiment at 3160 µg/plate without metabolic activation and at 316 µg/plate with metabolic activation. Top dose limited to 3160 µg/plate based on cytoxicity observed in preliminary test.
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
True 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:
cytotoxicity
Remarks:
background lawn reduced at 3160 µg/plate in plate incorporation assay and at 316 µg/plate in preincubation experiment
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
True negative controls validity:
not applicable
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
background lawn reduced in preincubation experiment at 316 µg/plate without metabolic activation and at 31.6 µg/plate with metabolic activation. Top dose limited to 3160 µg/plate based on cytotoxicity observed in preliminary test.
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
True negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
COMPARISON WITH HISTORICAL CONTROL DATA: Controls were within range of historical controls.

Cytotoxicity: In the plate incorporation test slight cytotoxicity was noted in the test substance in TA 100, with and without metabolic activation.

In the preincubation test without metabolic activation pronounced cytotoxicity was noted at 3160 µg/plate in TA98, from 316 µg/plate to 3160 µg/plate in TA 100, TA 102 and TA 1537, and from 100 µg/plate up to 3160 µg/plate in TA 1535.

With metabolic activation, pronounced cytotoxicity was noted at concentrations 316 µg/plate – 3160 µg/plate in TA 98 and TA 1535, at 1000 µg/plate and 3160 µg/plate in TA 100 and TA 1537 and from 100 µg/plate – 3160 µg/plate in TA 102.


Table 2: Dose range-finding study Number of revertants per plate (2 plates)

 

TA100

Conc.
(µg/plate)

Plate 1

Plate 2

Cytotoxic
(yes/no)

0*

145

138

No

0.316

145

141

No

1

128

148

No

3.16

145

135

No

10

174

139

No

31.6

137

141

No

100

148

146

No

316

163

140

No

1000

156

150

No

3160

0

0

Yes

5000

0

0

Yes

*solvent control with ethylene glycol dimethyl ether

Table 3: Experiment 1 Plate incorporation Number of revertants per plate (mean of 3 plates)

 

TA98

TA100

TA102

Conc.
(
µg/plate)

— MA

+ MA

Cytotoxic
(yes/no)

— MA

+ MA

Cytotoxic
(yes/no)

— MA

+ MA

Cytotoxic
(yes/no)

0*

22.7

27.7

No

133.3

138.7

No

264.7

268

No

31.6

25

21.7

No

117

156

No

296.3

285.7

No

100

23.3

30.7

No

123.7

136.7

No

268

269.7

No

316

28.7

33.3

No

110.3

141.7

No

267

274.7

No

1000

26.3

29

No

125

124.3

No

275

264

No

3160

25

27.3

No

114.7

150.3

Yes**

249

244

No

Positive control

1026.3

1066

No

1161

1175.3

No

1191

1170.7

No

*solvent control with ethylene glycol dimethyl ether

** scarce background bacterial lawn

Table 3: Experiment 1 Plate incorporation Number of revertants per plate (mean of 3 plates)

 

TA1535

TA1537

Conc.
(
µg/plate)

— MA

+ MA

Cytotoxic
(yes/no)

— MA

+ MA

Cytotoxic
(yes/no)

0*

16.3

411.7

No

11.3

13.3

No

31.6

14

18

No

12

14.3

No

100

15.3

21.7

No

13.7

17.3

No

316

17.3

22.7

No

12.7

16.3

No

1000

17.3

19.7

No

14.7

12.3

No

3160

17.7

18.7

No

14.7

11.7

No

Positive control

401

411.7

No

974.3

945.7

No

*solvent control with ethylene glycol dimethyl ether

Table 4: Experiment 2 Preincubation Number of revertants per plate (mean of 3 plates)

 

TA98

TA100

TA102

Conc.
(
µg/plate)

— MA

+ MA

Cytotoxic
(yes/no)

— MA

+ MA

Cytotoxic
(yes/no)

— MA

+ MA

Cytotoxic
(yes/no)

0*

42.7

38.7

No

128.3

155.7

No

291.7

287.3

No

3.16

27.3

32

No

119

147.3

No

254.7

300

No

10

29

34.3

No

128.7

151

No

258.3

277.3

No

31.6

34

30.3

No

120.7

140.7

No

252.7

290.3

No

100

22.7

27

No

126

153.7

No

257

0

Yes

316

24.7

0

Yes

0

170.7

Yes

0

0

Yes

1000

24.3

0

Yes

0

0

Yes

0

0

Yes

3160

0

0

Yes

0

0

Yes

0

0

Yes

Positive control

983.3

1085.3

No

1471

1443.3

No

1488.3

1445.3

No

*solvent control with ethylene glycol dimethyl ether

Table 4: Experiment 2 Preincubation Number of revertants per plate (mean of 3 plates)

 

TA1535

TA1537

Conc.
(
µg/plate)

— MA

+ MA

Cytotoxic
(yes/no)

— MA

+ MA

Cytotoxic
(yes/no)

0*

15.3

15.3

No

9.7

14

No

3.16

13

17

No

8.3

9

No

10

13.3

15

No

8

11.7

No

31.6

13

16.3

No

8.7

10.7

No

100

0

13

Yes

9

10.7

No

316

0

0

Yes

0

10.3

Yes

1000

0

0

Yes

0

0

Yes

3160

0

0

Yes

0

0

Yes

Positive control

684.3

680.3

No

529.3

540.3

No

*solvent control with ethylene glycol dimethyl ether

Conclusions:
N-[3-(dimethoxymethylsilyl)propyl]ethylenediamine has been tested in a reliable bacterial mutagenicity study conducted according to OECD 471, and in compliance with GLP conditions. No increase in revertant frequency was observed with or without metabolic activation when the substance was tested up to a concentration of 3160 µg/plate in Salmonella typhimurium strains TA 98, TA 100, TA 102, TA 1535 and TA 1537 in either the initial plate incorporation assay or the repeat experiment using the pre-incubation method. Cytotoxicity was observed at the highest concentration tested in strain TA 100 in the plate incorporation test, and in all strains in the pre-incubation assay. Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.
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:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
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 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

(Total of 2 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

(Total of 2 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 in compliance with 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:
in vitro gene mutation study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
See attachment to Section 13 for justification of read across
Reason / purpose for cross-reference:
read-across source
Species / strain:
Chinese hamster Ovary (CHO)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
4 mg/ml with metabolic activation
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

Micronucleus assay in mouse (ip administration): read-across from closely related substance N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS 1760-24-3): negative in peripheral lymphocytes (similar to OECD Test Guideline 474) (BRRC, 1988)

Link to relevant study records

Referenceopen allclose all

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:
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
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
- Acclimation period: 5 days


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)
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 in compliance with 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:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
See attachment to Section 13 for justification of read-across.
Reason / purpose for cross-reference:
read-across source
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
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Information is available from three bacterial mutagenicity studies on N-[3-(dimethoxymethylsilyl)propyl]ethylenediamine. No further information is available for the registered substance, however, reliable data are available for the closely related substance, N-[3-(trimethoxysilyl)propyl]ethylenediamine.

N-[3-(dimethoxymethylsilyl)propyl]ethylenediamine has been tested in a reliable bacterial mutagenicity study conducted according to OECD Test Guideline 471 and in compliance with GLP. No increase in revertant frequency was observed with or without metabolic activation when the substance was tested up to a concentration of 3160 µg/plate in Salmonella typhimurium strains TA 98, TA 100, TA 102, TA 1535 and TA 1537 in either the initial plate incorporation assay or the repeat experiment using the pre-incubation method. Cytotoxicity was observed at the highest concentration tested in strain TA 100 (plate incorporation) and in all strains in the pre-incubation assay. Appropriate solvent and positive controls were included and gave expected results (LPT, 2002).

Two additional bacterial mutagenicity studies also gave negative results (Microtest Research Ltd., 1988 and Huntingdon Life Sciences, 1999). The most reliable study was chosen as key study.

In vitro cytogenicity testing is not required as there is an in vivo cytogenicity study is available for the analogue substance.

The analogue substance, 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 Test Guideline 476 and in compliance with 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 or without metabolic 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 (BRRC, 1988a).

The analogue substance, 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 Test Guideline 474, and in compliance with 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 (BRRC, 1988b).

Justification for classification or non-classification

Based on the available in vitro and in vivo genotoxicity data, N-[3-(dimethoxymethylsilyl)propyl]ethylenediamine is not classified for mutagenicity according to Regulation (EC) No. 1272/2008.