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EC number: 239-415-9 | CAS number: 15396-00-6
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Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Gene mutation (Bacterial reverse mutation assay / Ames test): positive without activation in Salmonella typhimurium strain TA 98, negative with activation in strain TA 98; negative with and without activation in strains TA 100, TA 1535, TA 1537, TA 1538 (similar to OECD Test Guideline 471 and in compliance with GLP) (Bushy Run Research Center, 1994).
Cytogenicity in mammalian cells: data for the the hydrolysis product, 3-(trimethoxysilyl)propylamine (CAS 13822-56-5); negative in peripheral human lymphocytes with and without metabolic activation (according to OECD Test Guideline 443 and in compliance with GLP) (Charles River Laboratories, 2018).
Mutagenicity in mammalian cells: positive with and without metabolic activation in mouse lymphoma L5178Y cells (OECD Test Guideline 476 and in compliance with GLP) (BSL Bioservice, 2012).
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1994-07-06 - 1994-10-08
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- range of strains does not comply with current guideline
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- Histidine
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Species / strain / cell type:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor induced rat liver S9
- Test concentrations with justification for top dose:
- Test 1: 0.003 - 0.30 µg/plate. Test 2: 0.03 - 3.0 µg/plate
- Vehicle / solvent:
- DMSO
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 4-Nitro-o-Phenylenediamine 0.01 µg/plate
- Remarks:
- TA98, TA1538 without metabolic activation
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- TA100, TA1535 without metabolic activation; 0.01 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 0.06 µg/plate
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-Aminoanthracene 2.5 µg/plate
- Remarks:
- TA 98, TA100, TA1535, TA1537 and TA1538 with metabolic activation
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation
DURATION
- Preincubation period: ?
- Exposure duration: 48 - 72 hours
SELECTION AGENT (mutation assays):
NUMBER OF REPLICATIONS:
3 plates per dose, experiment repeated
DETERMINATION OF CYTOTOXICITY
- Method: Revertant colonies were counted
OTHER: - Evaluation criteria:
- A chemical is considered positive if it shows a statistically significant dose dependent and reproducible increase in the number of revertants relative to the solvent control.
- Statistics:
- None used
- 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
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Remarks:
- Produced a weak but consistent mutagenic effect
- 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
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with
- 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
- 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
- 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
- Species / strain:
- S. typhimurium TA 1538
- 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
- Conclusions:
- 3-(trimethoxysilyl)propyl isocyanate has been tested according to a protocol that is similar to OECD 471, and under GLP in Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538. A slight increase in the number of revertants was observed in strain TA 98 in the absence of metabolic activation, in the initial experiment at a single dose. This result was reproducible in the second experiment, in which the response appeared to be weakly dose-dependent. This was considered to be evidence of a weak mutagenic potential. It is concluded that the test substance is positive for mutagenicity to Salmonella typhimurium TA 98 in the absence of metabolic activation under the conditions of the test.
- 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)
- Version / remarks:
- 2016
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- other: in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- lymphocytes: human
- Details on mammalian cell type (if applicable):
- CELLS USED
- Source of cells: Blood was collected from healthy adult, non-smoking volunteers (approximately 18 to 35 years of age).
- Suitability of cells: recommended in OECD TG 473
- Cell cycle length, doubling time or proliferation index: Average generation time (AGT):
Dose-range finding study: age 26, AGT = 13.5 h and age 27, AGT = 15.2 h
First cytogenetic assay: age 28, AGT = 13.4 h (absence of S9-mix)
age 26, AGT = 14.4 h (presence of S9-mix)
Second cytogenetic assay: age 32, AGT = 14.6 h
- Sex, age and number of blood donors if applicable: see above
- Whether whole blood or separated lymphocytes were used if applicable: lymphocyte cultures: Whole blood (0.4 mL) treated with heparin was added to 5 mL or 4.8 mL culture medium (in the absence and presence of S9-mix, respectively). Per culture 0.1 mL (9 mg/mL) phytohaemagglutinin (Remel, Europe Ltd., Dartford, United Kingdom) was added
MEDIA USED
- Type and identity of media including CO2 concentration if applicable:
Culture medium consisted of RPMI 1640 medium (Life technologies), supplemented with 20% (v/v) heat-inactivated (56°C; 30 min) fetal calf serum (Life technologies), L-glutamine (2 mM) (Life technologies), penicillin/streptomycin (50 U/mL and 50 µg/mL respectively) (Life technologies) and 30 U/mL heparin (Sigma, Zwijndrecht, The Netherlands).
All incubations were carried out in a controlled environment, in which optimal conditions were a humid atmosphere of 80 - 100% (actual range 40 - 89%), containing 5.0 ± 0.5% CO2 in air in the dark at 37.0 ± 1.0°C (actual range 34.6 - 37.1°C) - Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital (80 mg/kg body weight) and ß-naphthoflavone (100 mg/kg) activated rat liver S9.
- Test concentrations with justification for top dose:
- 500, 1000 and 2000 µg/ml. No cytotoxicity was observed at the limit dose, which was the lowest dose at which precipitation was observed.
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used:DMSO
- Justification for choice of solvent/vehicle: none given in study report - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- mitomycin C
- Details on test system and experimental conditions:
- ACTIVATION: Phenobarbital (80 mg/kg body weight) and ß-naphthoflavone (100 mg/kg) activated rat liver S9.
S9-mix components contained per mL physiological saline: 1.63 mg MgCl2.6H2O; 2.46 mg KCl; 1.7 mg glucose-6-phosphate; 3.4 mg NADP ; 4 µmol HEPES. S9-mix contained 50% (v/v) S9-fraction.
0.2 mL S9-mix was added to 5.3 mL cell culture. The concentration of the S9-fraction in the exposure medium was 1.8% (v/v).
METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: 48 +/- 2 hours
- Exposure duration: Experiment 1: 3 hours with and without metabolic activation; Experiment 2: 24 and 48 hours without metabolic activation
- Fixation time (start of exposure up to fixation or harvest of cells): Experiment 1: 24 hours Experiment 2: 24 and 48 hours
SPINDLE INHIBITOR (cytogenetic assays): colchicine (0.5 µg/mL medium)
STAIN (for cytogenetic assays): 5% (v/v) Giemsa (Merck)
NUMBER OF REPLICATIONS: Duplicate cultures
METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED: During the last 2.5 - 3 h of the culture period, cell division was arrested by the addition of the spindle inhibitor colchicine (0.5 µg/mL medium). Thereafter the cell cultures were centrifuged for 5 min at 365 g and the supernatant was removed. Cells in the remaining cell pellet were swollen by a 5 min treatment with hypotonic 0.56% (w/v) potassium chloride solution at 37°C. After hypotonic treatment, cells were fixed with 3 changes of methanol: acetic acid fixative (3:1 v/v).
Fixed cells were dropped onto cleaned slides, which were immersed in a 1:1 mixture of 96% (v/v) ethanol/ether and cleaned with a tissue. The slides were marked with the Charles River Den Bosch study identification number and group number. At least two slides were prepared per culture. Slides were allowed to dry and thereafter stained for 10 - 30 min with 5% (v/v) Giemsa solution in Sörensen buffer pH 6.8. Thereafter slides were rinsed in water and allowed to dry. The dry slides were automatically embedded and mounted with a coverslip in an automated cover slipper
NUMBER OF CELLS EVALUATED: at least 1000 cells
NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells):
CRITERIA FOR MICRONUCLEUS IDENTIFICATION: Not applicable
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index;
- Any supplementary information relevant to cytotoxicity: The mitotic index of each culture was determined by counting the number of metaphases from at least 1000 cells (with a maximum deviation of 5%).
OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreplication: yes
- Methods, such as kinetochore antibody binding, to characterize whether micronuclei contain whole or fragmented chromosomes (if applicable): not applicable - Rationale for test conditions:
- According to protocol
- Evaluation criteria:
- A test item is considered positive (clastogenic) in the chromosome aberration test if:
a) At least one of the test concentrations exhibits a statistically significant (Fisher’s exact test, one-sided, p < 0.05) increase compared with the concurrent negative control.
b) The increase is dose related when evaluated with a trend test.
c) Any of the results are outside the 95% control limits of the historical control data range.
A test item is considered negative (not clastogenic) in the chromosome aberration test if:
a) None of the test concentrations exhibits a statistically significant (Fisher’s exact test, one-sided, p < 0.05) increase compared with the concurrent negative control.
b) There is no concentration-related increase when evaluated with a trend test.
c) All results are inside the 95% control limits of the negative historical control data range. - Statistics:
- Fisher’s exact test, one-sided
- Key result
- Species / strain:
- lymphocytes: peripheral human lymphocytes
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Remarks:
- . The test item precipitated in the culture medium at the limit dose.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH and osmolality: The pH and osmolarity of a concentration of 2000 µg/mL were 9.3 and 0.433 mOsm/kg, respectively, (compared to 8.2 and 0.433 mOsm/kg in the solvent control) in the solubility test. Concentrations of 1000 µg/ml and 500 µg/ml had a pH of 8.9 and 8.7, respectively.
- Precipitation: precipitation was observed only at the highest (limit) dose.
- Definition of acceptable cells for analysis: Only metaphases containing 46 ± 2 centromeres (chromosomes) were analysed..
- Other confounding effects: none
RANGE-FINDING/SCREENING STUDIES: the mitotic index of the cultures tested in the dose range-finding study was as follows (% of control):
Without metabolic activation (-S9-mix)
3 h exposure time, 24 h fixation time
Control 100
500 97
1000 96
2000 72
MMC-C; 0.5 µg/mL 87
MMC-C; 0.75 µg/mL 53
With metabolic activation (+S9-mix)
3 h exposure time, 24 h fixation time
Control 100
500 89
1000 90
2000 86
CP; 10 µg/mL 72
HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%)
- Positive historical control data: The positive control chemicals (MMC-C and CP) both produced statistically significant increases in the frequency of aberrant cells. In addition, the number of cells with chromosome aberrations found in the positive control cultures was within the 95% control limits of the distribution of the historical positive control database.
- Negative (solvent/vehicle) historical control data: The number of cells with chromosome aberrations found in the solvent control cultures was within the 95% control limits of the distribution of the historical negative control database . The number of polyploid cells and cells with endoreduplicated chromosomes in the solvent control cultures was within the 95% control limits of the distribution of the historical negative control database .
ADDITIONAL INFORMATION ON CYTOTOXICITY:
- Measurement of cytotoxicity used: mitotic index - Conclusions:
- 3-(Trimethoxysilyl)propylamine has been tested in a reliable in vitro cytogenetic assay according to OECD TG 473 and under GLP (CRL, 2018). The test substance did not induce statistically and biologically significant increases in the chromosomal aberration frequency of cultivated peripheral human lymphocytes in either the initial 3-hour exposure assay, with and without metabolic activation, or in the repeat experiment in which cells were exposed for 24 and 48 hours without metabolic activation. Appropriate positive and solvent controls were included and gave expected results. It is concluded that the test substance is negative for the induction of chromosome aberrations (not clastogenic) in vitro 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:
- 2011-11-03 to 2012-02-20
- 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)
- Version / remarks:
- 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5300 - In vitro Mammalian Cell Gene Mutation Test
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: IWGT Recommendations
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- (Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Germany)
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- Thymidine kinase
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: RPMI
- Properly maintained: Yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically "cleansed" against high spontaneous background: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital and ß-Naphthoflavone-induced rat liver S9 mix
- Test concentrations with justification for top dose:
- Pre-experiment
with and without metabolic activation:
0.2, 0.5, 2.5, 5.0, 7.5 and 10.0 mM
Main Experiment:
with metabolic activation:
0.01, 0.02, 0.05, 0.10, 0.20, 0.50, 1.00 and 2.00 mM
and without metabolic activation:
0.05, 0.10, 0.20, 0.50, 1.00, 1.50, 2.00 and 2.50 mM - Vehicle / solvent:
- The test item was dissolved in DMSO (final concentration 1% DMSO v/v).
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- Remarks:
- with metabolic activation 2.5 µg/mL
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- without metabolic activation 300 µg/mL
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Remarks:
- without metabolic activation M 10 µg/mL
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: dissolved in DMSO
DURATION: 4 h (short-term exposure)
Expression time (cells in growth medium): 48 h
Selection time (if incubation with selection agent): about 14 days
SELECTION AGENT ( mutation assay) 5 µg/mL trifluorothymidine
NUMBER OF REPLICATIONS: one experiment with single exposure; cells were seeded in 4 plates and evaluated
NUMBER OF CELLS SEEDED: 2000 cells per well
DETERMINATION OF CYTOTOXICITY: relative total growth (RTG)
METABOLIC ACTIVATION: S9 mix contained glucose-- phosphate and NADP as co-factors. Protein content of S9 mix was adjusted to give a final protein concentration of 0.75 mg/ml in the cultures. - Evaluation criteria:
- The test item is considered mutagenic if following criteria are met:
-The induced mutant frequency meets or exceeds the Global Evaluation factor (GEF) of 126 per 10^6 cells
- A dose-dependent increase in mutant frequency is detected.
Besides, combined with a positive effect in the mutant frequency, an increased occurrence of small colonies (=40% of total colonies) is an indication for potential clastogenic effects and/or chromosomal aberrations.
According to the OECD guideline, the biological relevance is considered first for the interpretation of results. Statistical methods might be used as an aid in evaluation the test result.
A test item is considered to be negative if the induced mutant frequency is below the GEF and the trend test is negative. - Statistics:
- The non-parametric Mann-Whitney test is applied to the mutation data to prove the dose groups for any significant difference in mutant frequency compared to the negative /solvent controls.
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Remarks:
- However, due to precipitation of the test item at all concentrations showing an increase in induced mutant frequency above the GEF, results should be evaluated with care.
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- RTG 12.3% (2 mM, +MA), 6.8% (2.5 mM, -MA)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Information on mutagenicity to mammalian cells is available for 3-(trimethoxysilyl)propyl isocyanate from a reliable study conducted according to OECD 476 and in compliance with GLP, using mouse lymphoma L5178Y cells. A statistically significant, biologically-relevant increase in mutant frequency was observed with and without metabolic activation. The Global Evaluation Factor (GEF) was exceeded at two highest evaluated doses without metabolic activation (2.0 and 2.5 mM, relative total growth (RTG) 21.7 and 6.8% respectively), and at the highest dose with metabolic activation (2.0 mM, RTG 12.3%). A dose-response relationship was observed in the main experiment without metabolic activation. In addition, slight evidence for clastogenicity was observed. Appropriate solvent, negative and positive controls were included and gave expected results. It is concluded that the test substance is positive for mutagenicity to mammalian cells, with some evidence for a clastogenic effect. Precipitation of the test item occurred at all concentrations showing an increase in induced mutant frequency above the GEF, so these results should be evaluated with care.
- Executive summary:
The test item 3-(trimethoxysilyl)propyl isocyanate was assessed for its potential to induce mutations at the mouse lymphoma thymidine kinase locus using the cell line L5178Y.
The selection of the concentrations used in the main experiment was based on data from the pre-experiment. In the main experiment with metabolic activation 2.00 mM and without metabolic activation2.50 mM were selected as the highest concentrations. The experiment with and without metabolic activation was performed as a 4 h short-term exposure assay.
The test item was dissolved in DMSO.
The test item was investigated at the following concentrations:
with metabolic activation:
0.01, 0.02, 0.05, 0.10, 0.20, 0.50, 1.00 and 2.00 mM
and without metabolic activation:
0.05, 0.10, 0.20, 0.50, 1.00, 1.50, 2.00 and 2.50 mM
Precipitation of the test item was noted in the pre-experimentwith and without metabolic activationand in the main experiment with metabolic activation.
Growth inhibition was observed in the main experiment with and without metabolic activation.
In the main experiment with metabolic activation the relative total growth (RTG) was 12.3% for the highest concentration (2.00 mM) evaluated. The highest concentration evaluated without metabolic activation was 2.50 mM with a RTG of 6.8%. Due to high cytotoxicity the highest concentration was not considered for evaluation of mutagenicity.
In the main experiment a biologically relevant increase of mutants was found after treatment with the test item (with and without metabolic activation).The Global Evaluation Factor (GEF; defined as the mean of the negative/vehicle mutant frequency plus one standard deviation; data gathered from ten laboratories) was exceeded by the induced mutant frequency in the higher dose ranges with and without metabolic activation.
In addition, in the main experiment without metabolic activation a dose-response relationship was observed.
Additionally, in the main experiment with and without metabolic activation colony sizing showed clastogenic effects at the highest evaluated concentrations induced by the test item under the experimental conditions .
EMS, MMS and B[a]P were used as positive controls and showed distinct and biologically relevant effects in mutation frequency. Additionally, MMS and B[a]P significantly increased the number of small colonies, thus proving the efficiency of the test system to indicate potential clastogenic effects.
This study is classified as acceptable. This study satisfies the requirements for Test Guidelines OPPTS 870.5300, OECD 476 for in vitro mutagenicity (mammalian forward mutation) data.
Referenceopen allclose all
Table 1a Experiment 1 Number of revertants per plate (mean of 3 plates)
Concentration µg/plate |
TA98 |
TA100 |
TA1535 |
|||
-MA |
+MA |
-MA |
+MA |
-MA |
+MA |
|
Solvent control |
7 |
18 |
90 |
88 |
25 |
8 |
Positive control |
294 |
1500 |
645 |
1310 |
596 |
90 |
3 |
10 |
- |
1 |
- |
3* |
- |
10 |
10 |
14 |
64 |
80 |
134 |
5 |
30 |
12 |
15 |
69 |
72 |
3* |
6 |
100 |
17 |
14 |
60 |
71 |
9* |
6 |
300 |
12 |
9 |
69 |
50 |
4* |
2* |
3000 |
- |
3* |
- |
8* |
- |
0* |
* Toxic, absence of background lawn, or mean number of colonies < ½ solvent control values.
Table 1b Experiment 1 Number of revertants per plate (mean of 3 plates)
Concentration |
TA1537 |
TA1538 |
||
µg/plate |
-MA |
+MA |
-MA |
+MA |
Solvent control |
4 |
3 |
9 |
14 |
Positive control |
76 |
150 |
451 |
1285 |
3 |
4 |
- |
5 |
3 |
10 |
4 |
4 |
11 |
11 |
30 |
4 |
5 |
7 |
9 |
100 |
3 |
5 |
12 |
12 |
300 |
2 |
3 |
2 |
6* |
3000 |
- |
* |
2* | 0* |
* Toxic: absence of background lawn, or mean number of colonies < ½ solvent control values.
Table 2a Experiment 2 Number of revertants per plate (mean of 3 plates)
Concentration |
TA98 |
TA100 |
TA1535 |
|||
µg/plate |
-MA |
+MA |
-MA |
+MA |
-MA |
+MA |
Solvent control |
11 |
17 |
59 |
83 |
3 |
5 |
Positive control |
341 |
1508 |
688 |
1572 |
664 |
101 |
3 |
12 |
- |
59 |
- |
4 |
- |
10 |
13 |
20 |
53 |
65 |
4 |
7 |
30 |
15 |
17 |
66 |
74 |
5 |
4 |
100 |
22 |
18 |
65 |
76 |
2 |
1* |
300 |
6 |
15 |
57 |
40* |
1* |
4 |
3000 |
- |
0* |
- |
14* |
- |
3* |
* Toxic, absence of background lawn, or mean number of colonies < ½ solvent control values.
Table 2b Experiment 2 Number of revertants per plate (mean of 3 plates)
Concentration |
TA1537 |
TA1538 |
||
µg/plate |
-MA |
+MA |
-MA |
+MA |
Solvent control |
3 |
6 |
10 |
9 |
Positive control |
100 |
175 |
338 |
1340 |
3 |
2 |
- |
9 |
- |
10 |
4 |
5 |
9 |
9 |
30 |
5 |
3 |
11 |
11 |
100 |
5 |
5 |
10 |
12 |
300 |
2 |
3 |
4* |
6 |
3000 |
- |
2* |
- |
2* |
* Toxic, absence of background lawn, or mean number of colonies < ½ solvent control values.
Table 1 Chromosome Aberrations in Human Lymphocyte Cultures
Treated with
3-(trimethoxysilyl)propylaminein the Absence of S9-Mix in the First
Cytogenetic Assay (3 H Exposure Time, 24 H Fixation Time)
Conc |
DMSO (1.0% v/v) |
500 µg/mL |
1000 µg/mL |
2000 µg/mL |
MMC-C 0.5 µg/mL |
MMC-C 0.75 µg/mL |
||||||||||||
Culture |
A |
B |
A+B |
A |
B |
A+B |
A |
B |
A+B |
A |
B |
A+B |
A |
B |
A+B |
|
|
B |
Mitotic Index (%) |
|
100 |
|
|
97 |
|
|
96 |
|
|
72 |
|
|
87 |
|
|
53 |
|
No. of Cells scored |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
143 |
293 |
No. of Cells with aberrations (+ gaps) a) |
3 |
1 |
4 |
0 |
1 |
1 |
0 |
0 |
0 |
1 |
1 |
2 |
16 |
8 |
24***) |
17 |
18 |
35***) |
No. of Cells with aberrations (- gaps) |
2 |
0 |
2 |
0 |
1 |
1 |
0 |
0 |
0 |
1 |
1 |
2 |
16 |
8 |
24***) |
17 |
17 |
34***) |
g’ |
1 |
1 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
1 |
|
g” |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
b’ |
1 |
|
|
|
|
|
|
|
|
|
|
|
8 |
7 |
|
10 |
7 |
|
b” |
1 |
|
|
|
|
|
|
|
|
|
|
|
3 |
2 |
|
4 |
3 |
|
m’ |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
m” |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
exch. |
|
|
|
|
1 |
|
|
|
|
1 |
1 |
|
6 |
1 |
|
6 |
8 |
|
dic |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
d’ |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
misc. |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
total aberr (+ gaps) |
3 |
1 |
|
0 |
1 |
|
0 |
0 |
|
1 |
1 |
|
17 |
9 |
|
20 |
19 |
|
total aberr (- gaps) |
2 |
0 |
|
0 |
1 |
|
0 |
0 |
|
1 |
1 |
|
17 |
9 |
|
20 |
18 |
|
a) Abbreviations
used for various types of aberrations are listed below the tables
misc. =
(miscellaneous) aberrations not belonging to the ones mentioned above.
*) Significantly
different from control group (Fisher’s exact test), * P < 0.05, ** P <
0.01 or
*** P < 0.001.
Table 2 Chromosome Aberrations in Human Lymphocyte Cultures
Treated with the test item in the Presence of S9-Mix in the First
Cytogenetic Assay (3 H Exposure Time, 24 H Fixation Time)
Conc |
DMSO (1.0% v/v) |
500 µg/mL |
1000 µg/mL |
2000 µg/mL |
CP 10 µg/mL |
||||||||||
Culture |
A |
B |
A+B |
A |
B |
A+B |
A |
B |
A+B |
A |
B |
A+B |
A |
B |
A+B |
Mitotic Index (%) |
|
100 |
|
|
86 |
|
|
84 |
|
|
81 |
|
|
58 |
|
No. of Cells scored |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
150 |
300 |
No. of Cells with aberrations (+ gaps) a) |
0 |
1 |
1 |
1 |
0 |
1 |
0 |
1 |
1 |
1 |
0 |
1 |
23 |
21 |
44***) |
No. of Cells with aberrations (- gaps) |
0 |
1 |
1 |
1 |
0 |
1 |
0 |
1 |
1 |
1 |
0 |
1 |
23 |
18 |
41***) |
g’ |
|
|
|
|
|
|
|
|
|
|
|
|
|
2 |
|
g” |
|
|
|
|
|
|
|
|
|
|
|
|
|
1 |
|
b’ |
|
|
|
|
|
|
|
|
|
1 |
|
|
14 |
10 |
|
b” |
|
|
|
|
|
|
|
|
|
|
|
|
5 |
5 |
|
m’ |
|
1 |
|
1 |
|
|
|
1 |
|
|
|
|
2 |
2 |
|
m” |
|
|
|
|
|
|
|
|
|
|
|
|
2 |
2 |
|
exch. |
|
|
|
|
|
|
|
|
|
|
|
|
3 |
1 |
|
dic |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
d’ |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Misc. |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
total aberr (+ gaps) |
0 |
1 |
|
1 |
0 |
|
0 |
1 |
|
1 |
0 |
|
26 |
23 |
|
total aberr (- gaps) |
0 |
1 |
|
1 |
0 |
|
0 |
1 |
|
1 |
0 |
|
26 |
20 |
|
a) Abbreviations
used for various types of aberrations are listed in below the tables.
misc. =
(miscellaneous) aberrations not belonging to the ones mentioned above.
*) Significantly
different from control group (Fisher’s exact test), * P < 0.05, ** P <
0.01 or
*** P < 0.001.
Table 3 Chromosome Aberrations in Human Lymphocyte Cultures
Treated with the test item in the Absence of S9-Mix in the Second
Cytogenetic Assay (24 H Exposure Time, 24 H Fixation Time)
Conc |
DMSO (1.0% v/v) |
500 µg/mL |
1000 µg/mL |
2000 µg/mL |
MMC-C 0.2 µg/mL |
||||||||||
Culture |
A B A+B |
A B A+B |
A B A+B |
A B A+B |
A B A+B |
||||||||||
Mitotic Index (%) |
100 |
99 |
101 |
99 |
47 |
||||||||||
No. of Cells scored |
150 150300 |
150 150300 |
150 150300 |
150 150300 |
150 150300 |
||||||||||
Culture |
A |
B |
A+B |
A |
B |
A+B |
A |
B |
A+B |
A |
B |
A+B |
A |
B |
A+B |
Mitotic Index (%) |
|
100 |
|
|
99 |
|
|
1201 |
|
|
99 |
|
|
47 |
|
No. of Cells scored |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
150 |
300 |
No. of Cells with aberrations (+ gaps) a) |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 |
0 |
0 |
0 |
37 |
41 |
78***) |
No. of Cells with aberrations (- gaps) |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 |
0 |
0 |
0 |
34 |
37 |
71***) |
g’ |
|
|
|
|
|
|
|
|
|
|
|
|
4 |
5 |
|
g” |
|
|
|
|
|
|
|
|
|
|
|
|
1 |
|
|
b’ |
|
|
|
|
|
|
|
|
|
|
|
|
7 |
21 |
|
b” |
|
|
|
|
|
|
1 |
|
|
|
|
|
7 |
7 |
|
m’ |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
m” |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
exch. |
|
|
|
|
|
|
|
|
|
|
|
|
22 |
18 |
|
dic |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
d’ |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
misc. |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
total aberr (+ gaps) |
0 |
0 |
|
0 |
0 |
|
1 |
0 |
|
0 |
0 |
|
41 |
51 |
|
total aberr (- gaps) |
0 |
0 |
|
0 |
0 |
|
1 |
0 |
|
0 |
0 |
|
36 |
46 |
|
a) Abbreviations
used for various types of aberrations are listed below the tables.
misc. =
(miscellaneous) aberrations not belonging to the ones mentioned above.
*) Significantly
different from control group (Fisher’s exact test), * P < 0.05, ** P <
0.01 or
*** P < 0.001.
Table 4 Chromosome Aberrations in Human Lymphocyte Cultures
Treated with the test item in the Absence of S9-Mix in the Second
Cytogenetic Assay (48 H Exposure Time, 48 H Fixation Time)
Conc |
DMSO (1.0% v/v) |
500 µg/mL |
1000 µg/mL |
2000 µg/mL |
MMC-C 0.1 µg/mL |
||||||||||
Culture |
A |
B |
A+B |
A |
B |
A+B |
A |
B |
A+B |
A |
B |
A+B |
A |
B |
A+B |
Mitotic Index (%) |
|
100 |
|
|
98 |
|
|
90 |
|
|
95 |
|
|
86 |
|
No. of Cells scored |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
150 |
300 |
150 |
150 |
300 |
No. of Cells with aberrations (+ gaps) a) |
0 |
0 |
0 |
2 |
0 |
2 |
0 |
1 |
1 |
1 |
0 |
1 |
38 |
29 |
67***) |
No. of Cells with aberrations (- gaps) |
0 |
0 |
0 |
1 |
0 |
1 |
0 |
0 |
0 |
1 |
0 |
1 |
33 |
27 |
60***) |
g’ |
|
|
|
1 |
|
|
|
1 |
|
|
|
|
4 |
3 |
|
g” |
|
|
|
|
|
|
|
|
|
|
|
|
1 |
|
|
b’ |
|
|
|
|
|
|
|
|
|
1 |
|
|
7 |
13 |
|
b” |
|
|
|
|
|
|
|
|
|
|
|
|
11 |
10 |
|
m’ |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
m” |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
exch. |
|
|
|
1 |
|
|
|
|
|
|
|
|
18 |
11 |
|
dic |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
d’ |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
misc. |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
total aberr (+ gaps) |
0 |
0 |
|
2 |
0 |
|
0 |
1 |
|
1 |
0 |
|
41 |
37 |
|
total aberr (- gaps) |
0 |
0 |
|
1 |
0 |
|
0 |
0 |
|
1 |
0 |
|
36 |
34 |
|
a) Abbreviations
used for various types of aberrations are listed below the tables.
misc. =
(miscellaneous) aberrations not belonging to the ones mentioned above.
*) Significantly
different from control group (Fisher’s exact test), * P < 0.05, ** P <
0.01 or
*** P < 0.001.
Aberration Abbreviation Description
Chromatid gap g' An achromatic lesion which appears as an unstained region in the chromatid arm, the size of which is equal to or smaller than the width of the chromatid and the apparently "broken" segments of the chromatid arm are in alignment.
Chromosome gap g" An achromatic lesion which appears as an unstained region in both chromatids at the same position, the size of which is equal to or smaller than the width of the chromatid and the apparently "broken" segments of the chromatids are in alignment.
Chromatid break b' An achromatic lesion in a chromatid arm, the size of which is larger than the width of the chromatid. The broken segments of the chromatid arm are aligned or unaligned.
Chromosome break b" An achromatic lesion in both chromatids at the same position, the size of which is larger than the width of the chromatid. The broken segments of the chromatids are aligned or unaligned.
Chromatid deletion d' Deleted material at the end of a chromatid arm.
Minute m' A single, usually circular, part of a chromatid lacking a centromere.
Double minutes m" Two, usually circular, parts of a chromatid lacking a centromere.
Dicentric chromosome dic A chromosome containing two centromeres.
Tricentric tric A chromosome containing three
chromosome centromeres.
Ring chromosome r A ring structure with a distinct lumen.
Exchange figure exch. An exchange(s) between two or more chromosomes resulting in the formation of a tri- or more-armed configuration.
Chromosome intra A chromosome intrachange is scored
intrachange after rejoining of a lesion within one
chromosome.
Pulverized p A fragmented or pulverized chromosome
chromosomes
Multiple ma A metaphase spread containing ten or
aberrations more of the above mentioned aberrations (chromatid and chromosome gaps not included). ma is counted as 10 aberrations.
Polyploidy poly A chromosome number that is a multiple of the normal diploid number.
Endoreduplication endo A form of polyploidy in which each centromere connects two or four pairs of chromatids instead of the normal one pair.
Pre-experiment for toxicity with and without metabolic activation
Concentration (mM) |
Number of cells 4 h after treatment |
Number of cells 24 h after treatment |
Number of cells 48 h after treatment |
SGa |
RSGb (%) |
|||||
|
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
Negative control |
31900 |
277000 |
1080000 |
771000 |
1590000 |
1490000 |
17.2 |
11.5 |
118.1 |
115.0 |
Negative control |
351000 |
324000 |
1140000 |
889000 |
1540000 |
1470000 |
17.6 |
13.1 |
120.8 |
130.8 |
Solvent control |
331000 |
258000 |
920000 |
644000 |
1610000 |
1480000 |
14.8 |
9.5 |
100.0 |
100.0 |
Solvent control |
331000 |
293000 |
926000 |
721000 |
1540000 |
1450000 |
14.3 |
10.5 |
||
0.2 |
338000 |
281000 |
916000 |
677000 |
1530000 |
1410000 |
14.0 |
9.5 |
96.4 |
95.5 |
0.5 |
343000 |
295000 |
941000 |
652000 |
1530000 |
1450000 |
14.4 |
9.5 |
99.0 |
94.6 |
2.5 |
323000 |
184000 |
779000 |
119000 |
1550000 |
1360000 |
12.1 |
4.1 |
83.1 |
40.8 |
5.0 (P) |
276000 |
73000 |
325000 |
51700 |
862000 |
45000 |
2.8 |
0.1 |
19.3 |
1.4 |
7.5 (P) |
175000 |
92300 |
158000 |
79400 |
152000 |
69900 |
0.5 |
0.2 |
3.1 |
2.1 |
10.0(P) |
172000 |
93700 |
128000 |
77500 |
104000 |
61000 |
0.3 |
0.2 |
2.1 |
1.8 |
P = precipitation RSG = relative suspension growth SG = suspension growth
Summary tables with and without metabolic activation
Treatment (mM) |
RTG (%) |
MF (mutants/106cells |
IMF (mutants/106cells) |
Precipitate |
|
-S9 |
-S9 |
-S9 |
S9 |
Negative control |
101.2 |
84.9 |
/ |
- |
Negative control |
97.9 |
/ |
- |
|
Solvent control |
100.0 |
73.7 |
/ |
- |
Solvent control |
/ |
- |
||
0.05 |
101.9 |
67.1 |
-6.7 |
- |
0.10 |
91.6 |
75.5 |
1.7 |
- |
0.20 |
93.2 |
99.0 |
25.3 |
- |
0.50 |
91.8 |
69.5 |
-4.2 |
- |
1.00 |
86.1 |
75.6 |
1.8 |
- |
1.50 |
43.6 |
158.1 |
84.3 |
- |
2.00 |
21.7 |
208.3 |
134.5 |
+ |
2.50 |
6.8 |
299.2 |
225.4 |
+ |
Positive control 1 |
80.7 |
455.2 |
381.5 |
- |
Positive control 2 |
63.7 |
381.5 |
307.7 |
- |
Treatment (mM) |
RTG (%) |
MF (mutants/106cells |
IMF (mutants/106cells) |
Precipitate |
|
+S9 |
+S9 |
+S9 |
+S9 |
Negative control |
90.8 |
87.0 |
/ |
- |
Negative control |
102.2 |
/ |
- |
|
Solvent control |
100.0 |
68.9 |
/ |
- |
Solvent control |
/ |
- |
||
0.01 |
88.2 |
41.9 |
-27.0 |
- |
0.02 |
92.5 |
98.8 |
30.0 |
- |
0.05 |
90.4 |
72.3 |
3.4 |
- |
0.10 |
103.2 |
57.6 |
-11.3 |
- |
0.20 |
102.4 |
55.1 |
-13.7 |
- |
0.50 |
82.2 |
153.9 |
85.0 |
- |
1.00 |
78.8 |
80.3 |
11.4 |
- |
2.00 |
12.3 |
218.1 |
149.3 |
+ |
Positive control |
53.9 |
518.2 |
449.3 |
- |
None of the test substance exceed GEF (global evaluation factor).
Positive control results exceed GEF and were statistically significant: test substance results did not exceed GEF and ere not statistically significant.
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (positive)
Genetic toxicity in vivo
Description of key information
Micronucleus assay in mouse (oral administration): negative at doses up to the maximum tolerated dose (MTD) of 1000 mg/kg bw (OECD Test Guideline 474 and in compliance with GLP) (Harlan Laboratories, 2011).
Link to relevant study records
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2011-02-16 - 2011-03-10
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- The study was compliant with the 1997 guideline, which was current at the time of the study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- Version / remarks:
- 2008
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- micronucleus assay
- Species:
- mouse
- Strain:
- other: Albino HSD
- Sex:
- male
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Harlan Laboratories UK Ltd
- Age at study initiation: 6 - 10 weeks old
- Weight at study initiation: 20 - 30 g
- Assigned to test groups randomly: yes
- Fasting period before study: no information
- Housing: Groups of up to 7 in solid floor polypropylene cages with wood flake bedding
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25°C
- Humidity (%): 30 - 70%
- Air changes (per hr): Approx 15 per hour
- Photoperiod (hrs dark / hrs light): 12 dark /12 light
IN-LIFE DATES: From: 2011-02-22 To: 2011-03-10 . - Route of administration:
- oral: gavage
- Vehicle:
- - Vehicle(s)/solvent(s) used: arachis oil
- Justification for choice of solvent/vehicle: None given
- Amount of vehicle (if gavage or dermal): 10 ml/kg
- Lot/batch no. (if required): V-4855
. - Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: Arachis oil was used as vehicle. Dosing volume was 10 ml/kg.
All animals were dosed once by gavage using a metal cannula attached to a graduated syringe. - Duration of treatment / exposure:
- 24 and 48 hours
- Frequency of treatment:
- Once
- Post exposure period:
- 24 and 48 hours
- Remarks:
- Doses / Concentrations:
250 -1000 mg/kg bw
Basis:
other: gavage - No. of animals per sex per dose:
- 7 male mice
- Control animals:
- yes
- Positive control(s):
- - cyclophosphamide;
- Justification for choice of positive control(s): Cyclophosphamide is known to produce micronuclei under the conditions of the test
- Route of administration: oral - gavage
- Doses / concentrations: 50 mg/kg - 5 mg/ml - Tissues and cell types examined:
- Polychromatic erythrocytes (PCE) were scored for micronuclei and normochromatic erythrocytes (NCE) were counted from bone marrow smears
- Details of tissue and slide preparation:
- CRITERIA FOR DOSE SELECTION: The maximum tolerated dose level or that which produces some evidence of toxicity up to a maximum recommended dose of 2000 mg/kg.
TREATMENT AND SAMPLING TIMES (in addition to information in specific fields): Animals were dosed once and sampled at 24 and 48 hours.
DETAILS OF SLIDE PREPARATION: Immediately following termination both femurs were dissected from each animal, prepared and stained in May-Grunwald/Giemsa, air dried and mounted with a cover slip.
METHOD OF ANALYSIS: Slides were coded and examined under a microscope x1000 magnification. - Evaluation criteria:
- A positive mutagenic response would be demonstrated when a statistically significant, dose-responsive, toxicologically relevant increase in the number of micronucleated polychromatic erythrocytes was observed for either the 24 or 48-hour kill times when compared to the vehicle control group. If these criteria were not fulfilled, then the test item would be considered non-genotoxic under the conditions of the test.
- Statistics:
- The Student's t-test (two tailed) was used and any significant results were confirmed using the one way analysis of variance.
- Sex:
- male
- Genotoxicity:
- negative
- Toxicity:
- yes
- Remarks:
- reduction in PCE/NCE ratio was observed in the 1000 mg/kg test group relative to the vehicle group.
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- RESULTS OF RANGE-FINDING STUDY
- Dose range: 1000 - 2000 mg/kg bw
- Solubility: The test item was formulated 2 hours before application and was assumed stable for this duration.
- Clinical signs of toxicity in test animals: Animals above 1000 mg/kg bw showed clinical signs of excessive toxicity which included hunched posture, pilo-erection, splayed gait, ataxia, ptosis, dehydration, lethargy, decreased respiratory rate and laboured respiration.
- Evidence of cytotoxicity in tissue analyzed: Not examined
- Rationale for exposure: "It was considered unnecessary to investigate the intraperitoneal route of administration as there was evidence of marked toxicity via the oral route. No sex-related differences were observed, so only male mice were treated in the micronucleus study."
- Harvest times: 24 and 48 hours
- High dose with and without activation: 1000 mg/kg bw, no metabolic activation was used
RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): negative
- Ratio of PCE/NCE (for Micronucleus assay): Modest decreases in PCE/NCE ratio were observed in both 24 and 48-hour 1000mg/kg dose groups when compared to the vehicle control group.
- Appropriateness of dose levels and route: Dose levels and route were appropriate.
- Statistical evaluation: no statistically significant effects were observed.
- Other: Two premature deaths were observed in the 48-hour 1000 mg/kg dose group and one premature death was observed in the 24-hour 500 mg/kg dose group. The deaths were considered to be due to variable sensitivity of the mice to the test item. - Conclusions:
- 3-(Trimethoxysilyl)propyl isocyanate has been tested in a reliable micronucleus assay according to OECD TG 474 and in compliance with GLP. No statistically significant increase in the frequency of micronucleated polychromatic erythrocytes was observed in peripheral erythrocytes of mice treated with the test substance by oral gavage. Appropriate vehicle and positive controls were included and gave expected results. It is concluded that 3-(trimethoxysilyl)propyl isocyanate is negative for the induction of micronuclei under the conditions of this test.
Reference
Table 3 Results of micronucleus assay
Treatment group mg/kg bw and sample time |
Number of PCE with micronuclei per 2000 PCE |
PCE/NCE Ratio |
||
Group Mean |
SD |
Group Mean |
SD |
|
Control 24 hour |
2.4 |
2.5 |
0.87 |
0.33 |
1000 48 hour |
1.6 |
2.3 |
0.67 |
0.16 |
1000 24 hour |
2.7 |
3.0 |
0.57 |
0.17 |
500 24 hour |
1.5 |
1.0 |
0.78 |
0.13 |
250 24 hour |
0.4 |
0.8 |
0.80 |
0.15 |
Positive control 24 hour |
31.2** |
14.4 |
0.71 |
0.09 |
** = P<0.001
Endpoint conclusion
- Endpoint conclusion:
- no study available (further information necessary)
Additional information
Data for the registered substance 3-(trimethoxysilyl)propyl isocyanate (CAS 15396-00-6) are available from reliable in vitro studies for mutagenicity to bacterial and mammalian cells, and an in vivo micronucleus assay. In vitro cytogenicity data are available for the hydrolysis product, 3-(trimethoxysilyl)propylamine, (CAS 13822-56-5).
3-(Trimethoxysilyl)propyl isocyanate has been tested according to a protocol similar to OECD Test Guideline 471 and in compliance with GLP in Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538. A slight increase in the number of revertants was observed in strain TA 98 in the absence of metabolic activation, in the initial experiment at a single dose. This result was reproducible in the second experiment, in which the response appeared to be weakly dose-dependent. This was considered to be evidence of a weak mutagenic potential. It is concluded that the test substance is positive for mutagenicity to Salmonella typhimurium TA 98 in the absence of metabolic activation under the conditions of the test (Bushy Run Research Center, 1994).
Bacterial mutagenicity data for the hydrolysis product, 3-(trimethoxysilyl)propylamine (CAS 13822-56-5), have been added to the dataset as suppoting information for completeness.
The hydrolysis product, 3-(trimethoxysilyl)propylamine has been tested in a reliable in vitro cytogenetic assay according to OECD Test Guideline 473 and in compliance with GLP (Charles River Laboratories, 2018). The test substance did not induce statistically and biologically significant increases in the chromosomal aberration frequency of cultivated peripheral human lymphocytes in either the initial 3-hour exposure assay, with and without metabolic activation, or in the repeat experiment in which cells were exposed for 24 and 48 hours without metabolic activation. Appropriate positive and solvent controls were included and gave expected results. It is concluded that the test substance is negative for the induction of chromosome aberrations (not clastogenic) in vitro under the conditions of the test.
Information on mutagenicity to mammalian cells is available for 3-(trimethoxysilyl)propyl isocyanate from a reliable study conducted according to OECD Test Guideline 476 (1997) and in compliance with GLP, using mouse lymphoma L5178Y cells (BSL Bioservice, 2012). A statistically significant, biologically-relevant increase in mutant frequency was observed with and without metabolic activation. The Global Evaluation Factor (GEF) was exceeded at two highest evaluated doses without metabolic activation (2.0 and 2.5 mM, relative total growth (RTG) 21.7 and 6.8% respectively), and at the highest dose with metabolic activation (2.0 mM, RTG 12.3%). A dose-response relationship was observed in the main experiment without metabolic activation. In addition, slight evidence for clastogenicity was observed. Appropriate solvent, negative and positive controls were included and gave expected results. It is concluded that the test substance is positive for mutagenicity to mammalian cells, with some evidence for a clastogenic effect. Precipitation of the test item occurred at all concentrations showing an increase in induced mutant frequency above the GEF, so these results should be evaluated with care.
3-(Trimethoxysilyl)propyl isocyanate has been tested in a reliable in vivo micronucleus assay according to OECD Test Guideline 474 (2008) and in compliance with GLP (Harlan Laboratories, 2011). No statistically significance increase in the frequency of micronucleated polychromatic erythrocytes was observed in peripheral erythrocytes of mice treated with the test substance by oral gavage. Appropriate vehicle and positive controls were included and gave expected results. It is concluded that 3-(trimethoxysilyl)propyl isocyanate is negative for the induction of micronuclei under the conditions of this test.
In vitro results for 3-(trimethoxysilyl)propyl isocyanate indicate that the substance has potential for mutagenicity. The in vivo micronucleus assay indicates that the substance is not clastogenic. Further information is required on the potential for mutagenicity, so an in vivo Comet assay is proposed to investigate the genetic toxicity potential further.
Justification for use of data for the hydrolysis product, 3-(trimethoxysilyl)propylamine, (CAS 13822-56-5)
The registered substance, 3-(trimethoxysilyl)propyl isocyanate (CAS 15396-00-6) is a trimethoxysilane with a propyl side-chain that has an isocyanate functional group at the opposite end from the trimethoxysilane group.
The isocyanate group in 3-(trimethoxysilyl)propyl isocyanate (CAS 15396-00-6) hydrolyses very rapidly under physiological conditions (half-life 1 minute at 37°C and pH 7), forming an amine. Therefore, the substance hydrolyses very rapidly to give the hydrolysis product, 3-(trimethoxysilyl)propylamine (CAS 13822-56-5). 3-(Trimethoxysilyl)propylamine (CAS 13822-56-5) has been predicted to hydrolyse rapidly under conditions relevant for oral exposure. The estimated hydrolysis half-life at pH 2 (relevant for oral exposure) and 37.5°C is approximately 5 seconds. Therefore, the test organism is expected to be mainly exposed to the hydrolysis products, 3-aminopropylsilanetriol and methanol.
Justification for classification or non-classification
Insufficient information is available to conclude on the classification for mutagenicity of 3-(trimethoxysilyl)propyl isocyanate according to Regulation (EC) No.1272/2008.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
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