hydrolysis products of 3-(triethoxysilyl)propan-1-amine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 Jan - 23 Feb 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP - Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Behörde für Soziales, Familie, Gesundheit und Verbraucherschutz, Hamburg, Germany

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

hydrolysis products of 3-(triethoxysilyl)propan-1-amine

EC Number:

939-125-9

Molecular formula:

not applicable for UVCB substance

IUPAC Name:

hydrolysis products of 3-(triethoxysilyl)propan-1-amine

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

co-factor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Arochlor 1254

Test concentrations with justification for top dose:

Preliminary cytotoxicity test: 0.316, 1.0, 3.16, 10.0, 31.6, 100, 316, 1000, 3160 and 5000 µg/plate with and without metabolic activation
Experiment I and II: 10, 31.6, 100, 316, 1000 and 3160 μg/plate with and without metabolic activation

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

-S9: 10 µg/plate SA (TA 1535, TA 100), 10 µg/plate 2-NF (TA 98), 100 μg/plate 9-AA (TA 1537), 1300 μg/plate MMS (TA 102); + S9: 2 μg/plate 2-AA (TA 98, TA 102, TA 1537), 1500 μg/plate CA (TA 100, TA 1535)

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

cyclophosphamide

methylmethanesulfonate

other: 2-amino-anthracene

Remarks:

SA: sodium azide; 2-NF: 2-nitro-fluorene; 9-AA: 9-amino-acridine; MMS: methylmethanesulfonate; 2-AA: 2-amino-anthracene; CA: cyclophosphamide

Details on test system and experimental conditions:

METHOD OF APPLICATION 1: preliminary cytotoxicity test and experiment I: in agar (plate incorporation)

DURATION
- Exposure duration: 48 to 72 h

METHOD OF APPLICATION 2: experiment II: preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 to 72 h

NUMBER OF REPLICATIONS: triplicates each in two independent experiments for the main assay

DETERMINATION OF CYTOTOXICITY
- Method: inspection of background lawn and reduction on the number of revertants

Evaluation criteria:

The test item was considered to show a positive response if:
- the number of revertants was significantly increased (p ≤ 0.05, U-test according to MANN and WHITNEY) 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 independent experiments;
- in addition, a significant (p ≤ 0.05) concentration (log value)-related effect (Spearman’s rank correlation coefficient) was observed;
- positive results were reproducible and the histidine independence of the revertants was confirmed by streaking random samples on histidine-free agar plates.

Statistics:

Mean values and standard deviations of revertant colonies were calculated. Statistical analysis between treated and solvent control samples was performed using the U-test according to MANN and WHITNEY and statistical significance was indicated at p ≤ 0.05. Concentration (log value)-related effects were analysed using the Spearman’s rank correlation coefficient (statistical significance indicated at p ≤ 0.05).

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

≥ 3160 µg/plate (±S9)

Vehicle controls validity:

not valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: in two preliminary cytotoxicity tests, the Salmonella typhimurium strain TA 100 was tested at concentrations ranging from 0.316 to 5000 µg/plate with and without metabolic activation. Cytotoxicity (scarce background lawn and reduction of the number of revertants) was noted at concentrations ≥ 3160 µg/plate in both experiments. Hence, 3160 µg/plate were chosen as top concentration for the main study in the plate incorporation test and in the preincubation test, respectively.

ADDITIONAL INFORMATION ON CYTOTOXICITY: in the plate incorporation test and in the preincubation test, cytotoxicity (scarce background lawn and reduction of the number of revertants) was noted at the top concentration of 3160 µg/plate in all test strains both in the presence and absence of metabolic activation.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1. Test results of experiment I

Bacterial Reverse Mutation Assay, mean revertant colonies/plate (n=3 ± SD)
EXPERIMENT I (plate incorporation)
S9-Mix	Without
Concentration (per plate)	TA 98	TA 100	TA 102	TA 1535	TA 1537
SC (DMSO)	30 ± 6	142 ± 10	278 ± 12	15 ± 5	6 ± 3
Test item
10 µg	34 ± 4	119 ± 20	260 ± 10	14 ± 4	6 ± 3
31.6 µg	31 ± 3	123 ± 14	274 ± 12	16 ± 2	7 ± 2
100 µg	29 ± 2	115 ± 16	265 ± 3	16 ± 6	5 ± 4
316 µg	34 ± 6	122 ± 7	259 ± 7	12 ± 2	4 ± 2
1000 µg	30 ± 5	110 ± 6	262 ± 13	12 ± 3	6 ± 1
3160 µg	13 ± 2#	67 ± 10#	230 ± 3#	7 ± 1#	1 ± 1#
PC
SA (10 µg)	-	1045 ± 45	-	145 ± 27	-
2-NF (10 µg)	204 ± 15	-	-	-	-
9-AA (100 µg)	-	-	-	-	151 ± 12
MMS (1300 µg)	-	-	1019 ± 14	-	-
S9-Mix	With
Concentration (per plate)	TA 98	TA 100	TA 102	TA 1535	TA 1537
SC (DMSO)	26 ± 6	125 ± 11	262 ± 2	13 ± 2	7 ± 3
Test item
10 µg	36 ± 2	126 ± 6	268 ± 11	13 ± 2	7 ± 1
31.6 µg	28 ± 8	134 ± 31	272 ± 2	13 ± 2	8 ± 2
100 µg	26 ± 4	134 ± 5	280 ± 11	20 ± 6	9 ± 1
316 µg	23 ± 2	123 ± 4	274 ± 11	16 ± 8	5 ± 2
1000 µg	24 ± 3	102 ± 2	265 ± 9	21 ± 8	5 ± 1
3160 µg	17 ± 3#	87 ± 10#	221 ± 3#	5 ± 2#	1 ± 0
PC
2-AA (2 µg)	248 ± 13	-	1023 ± 34	-	152 ± 4
CA (1500 µg)	-	1000 ± 21	-	161 ± 7	-
SC = solvent control; PC = positive control substances; SD = standard deviation; SA: sodium azide; 2-NF: 2-nitro-fluorene; 9-AA: 9-amino-acridine; MMS: methylmethanesulfonate; 2-AA: 2-amino-anthracene; CA: cyclophosphamide # scarce background lawn

Table 2. Test results of experiment II

Bacterial Reverse Mutation Assay, mean revertant colonies/plate (n=3 ± SD)
EXPERIMENT I (preincubation)
S9-Mix	Without
Concentration (per plate)	TA 98	TA 100	TA 102	TA 1535	TA 1537
SC (DMSO)	30 ± 7	138 ± 19	292 ± 1	19 ± 4	5 ± 1
Test item
10 µg	22 ± 2	190 ± 3	275 ± 8	22 ± 2	6 ± 2
31.6 µg	29 ± 7	174 ± 4	280 ± 12	13 ± 4	5 ± 3
100 µg	39 ± 5	178 ± 19	276 ± 6	14 ± 7	4 ± 1
316 µg	42 ± 3	168 ± 7	280 ± 9	14 ± 4	5 ± 1
1000 µg	25 ± 2	165 ± 11	280 ± 6	19 ± 5	6 ± 1
3160 µg	16 ± 1#	81 ± 7#	144 ± 3#	6 ± 2#	1 ± 1#
PC
SA (10 µg)	-	1082 ± 28	-	216 ± 6	-
2-NF (10 µg)	120 ± 5	-	-	-	-
9-AA (100 µg)	-	-	-	-	268 ± 5
MMS (1300 µg)	-	-	1055 ± 48	-	-
S9-Mix	With
Concentration (per plate)	TA 98	TA 100	TA 102	TA 1535	TA 1537
SC (DMSO)	30 ± 8	132 ± 18	285 ± 4	18 ± 10	4 ± 3
Test item
10 µg	27 ± 2	113 ± 11	288 ± 9	15 ± 6	4 ± 2
31.6 µg	30 ± 9	128 ± 17	269 ± 25	12 ± 2	6 ± 3
100 µg	31 ± 10	127 ± 24	253 ± 6	13 ± 3	4 ± 2
316 µg	25 ± 5	123 ± 11	257 ± 11	15 ± 8	3 ± 2
1000 µg	25 ± 3	114 ± 5	259 ± 2	13 ± 2	4 ± 4
3160 µg	14 ± 2#	84 ± 10#	155 ± 12#	4 ± 1#	2 ± 1#
PC
2-AA (2 µg)	126 ± 3	-	1030 ± 7	-	222 ± 4
CA (1500 µg)	-	1065 ± 17	-	172 ± 9	-
SC = solvent control; PC = positive control substances; SD = standard deviation; SA: sodium azide; 2-NF: 2-nitro-fluorene; 9-AA: 9-amino-acridine; MMS: methylmethanesulfonate; 2-AA: 2-amino-anthracene; CA: cyclophosphamide # scarce background lawn

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative