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Toxicological information

Genetic toxicity: in vitro

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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline-study OECD 471. Not in full compliance with GLP but according top good scientific practice and laboratory involved was subject to regular GLP inspections at the time of performance. Study according to relevant guideline.

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1990
Report date:
1990
Reference Type:
review article or handbook
Title:
Unnamed
Year:
1994
Reference Type:
other: SCF (scientific Committee on Food)
Title:
Unnamed
Year:
1999
Reference Type:
publication
Title:
Unnamed
Year:
1993

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
The bacterial reverse mutation test is able to identify substances that cause point mutations, by substitution, addition or deletion of one or a few
DNA base-pairs. Mutagenic substances can induce reversion in histidine-(for Salmonella typhimurium) or tryptophan-(for Escherichia coli)
deficient strains which are then able to grow and form colonies in a histidine- or tryptophan limited medium, while non-reverted strains cannot.
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
N-(hydroxymethyl)methacrylamide
EC Number:
213-086-1
EC Name:
N-(hydroxymethyl)methacrylamide
Cas Number:
923-02-4
Molecular formula:
C5H9NO2
IUPAC Name:
N-(hydroxymethyl)-2-methylprop-2-enamide
Details on test material:
- Name of test material (as cited in study report): N-Methylolmethacrylamide 15 % aqueous solution
- Substance type: organic
- Physical state at room temperature: liquid
- Storage condition of test material: 4 to 6 °C

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
S9-Mix fraction of Aroclor 1254-induced rat livers
Test concentrations with justification for top dose:
0, 100, 500, 2500, 5000 and 7500 µg/plate (all 4 strains), and at 2500, 5000, 7500 and 10.000 µg/plate (TA 100 only)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: distilled water
- Justification for choice of solvent/vehicle: The solvent was chosen because of its relative nontoxicity for the bacteria.
Controlsopen allclose all
Negative solvent / vehicle controls:
yes
Remarks:
dissolved in DMSO
Positive controls:
yes
Remarks:
for TA 1537
Positive control substance:
other: 9-aminoacridine chloride monohydrate, without S9-mix
Remarks:
9-aminoacridine chloride monohydrate (purity: no data, supplier: no data) dissolved in DMSO; concentration: 100 µg/plate
Negative solvent / vehicle controls:
yes
Remarks:
dissolved in DMSO
Positive controls:
yes
Remarks:
TA 98
Positive control substance:
other: 4-nitro-o-phenylendiamine, without metabolic activation
Remarks:
4-nitro-o-phenylenediamine (purity: no data, supplier: no data) dissolved in DMSO; concentration: 10 µg/plate
Negative solvent / vehicle controls:
yes
Remarks:
dissolved in DMSO
Positive controls:
yes
Remarks:
for TA 1535, TA 100
Positive control substance:
other: N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), without metabolic activation
Remarks:
N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) (purity: no data, supplier: no data ) dissolved in DMSO; concentration: 5 µg/plate
Negative solvent / vehicle controls:
yes
Remarks:
dissolved in DMSO
Positive controls:
yes
Remarks:
for TA 1535, TA 1537, TA 98, TA 100
Positive control substance:
other: 2-aminoanthracene, with metabolic activation
Remarks:
2-aminoanthracene (purity: no data, supplier: no data ) dissolved in DMSO; concentration: 10 µg/plate
Untreated negative controls:
yes
Remarks:
solvent control, sterility control
Remarks:
+/- S9 mix; determination of spontaneous mutation rate
Details on test system and experimental conditions:
NUMBER OF REPLICATIONS: 3 test plates per dose or control standard plate test and preincubation test with and without S9-mix
Evaluation criteria:
A test article is considered positive if either a dose related and reproducible increase in the number of revertants or a significant and reproducible
increase for at least one test concentration is induced.
A test article producing neither a dose related and reproducible increase in the number of revertants nor a significant and reproducible positive
response at any one of the test points is considered non-mutagenic in this system.

A significant response is described as follows:
A test article is considered mutagenic if the number of reversions is at least twice as high (control) in one of the tester strains. It exists a dose-response relationship and the results are reproducible.

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
other: weakly positive when reevaluated by SCF negative
Additional information on results:
A weakly bacteriotoxic effect was observed only in the preincubation assay at doses greater 5000 µg/plate without S-9 mix using TA 1537 and TA 98.
Remarks on result:
other: other: Salmonella typhimurium TA 1535, TA 100, TA 1537, TA 98
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Results:
In the standard plate test, no induction of point mutations by 

N-Methylolmethacrylamide could be detected in any of the 4 stains, with or 

without S9-mix, although none were tested up to the cytotoxic dose range. In the 

preincubation test. There was no evidence of mutagenic activity in stains 

TA98, TA1535 and TA1537, with and without S9-mix. A week bacteriotoxic effect 

was seen only in strains TA98 and TA1537 at >= 5000 µg/plate without S9-mix. 

Strain TA 100 showed weak mutagenic activity in the preincubation test at 

5000 - 10000 µg/plate in the presence of S9-mix. At 5000 µg/plate the number of 

his+ revertants increased by a factor of 1.4 - 1.6, and at 10000 µg/plate by a factor of 1.9. In the absence of S9-mix, in one of the two 

experiments with TA100, there was a slight but not dose-dependent increase (by a 

factor of 1.4 -  1.7) in the number of his+ revertants at 2500 - 10000 µg/plate. 

The inverstigators concluded that 15 % aqueous N-Methylolmethacrylamide is 

weakly mutagenic in the Ames test but a reevaluation of the SCF (Scientific Committee on Food) classified the substance under the above mentioned conditions as negative in the Ames test.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

The inverstigators concluded that 15 % aqueous N-Methylolmethacrylamide is weakly mutagenic in the Ames test in one of the four strains (TA 100) tested. A reevaluation of the SCF (Scientific Committee on Food) classified the substance under the above mentioned conditions as negative in the Ames test according to OECD guideline 471.
Executive summary:

In a reverse gene mutation assay in bacteria (Ames test), strains TA1535, TA1537, TA98, TA100 of Salmonella typhimurium were exposed to N-Methylolmethacrylamide 15 % aqueous solution at concentrations of up to 7500 µg/plate in the presence and absence of mammalian metabolic activation S9 -mix all tester stains and up to 10000 µg/plate tester stain TA 100

 

In the standard plate test, no induction of point mutations by N-Methylolmethacrylamide could be detected in any of the 4 stains, 

with or without S9-mix, although none were tested up to the cytotoxic dose range. In the preincubation test. There was no evidence of mutagenic activity in stains TA98, TA1535 and TA1537, with and without S9-mix. A week bacteriotoxic 

effect was seen only in strains TA98 and TA1537 at >= 5000 µg/plate without S9-mix. Strain TA 100 showed weak mutagenic 

activity in the preincubation test at 5000 - 10000 µg/plate in the presence of S9-mix. At 5000 µg/plate the number of  his+ revertants increased by a factor of 1.4 - 1.6, and at 10000 µg/plate by a factor of 1.9. In the absence of S9-mix, in one of the two experiments with TA100, there was a slight but not 

dose-dependent increase (by a factor of 1.4 -  1.7) in the number of his+ revertants at 2500 - 10000 µg/plate.

Appropriate reference mutagens were used as positive controls.

The positive controls induced the appropriate responses in the corresponding strains.

This study is classified as acceptable. This study satisfies the requirement for tests for in vitro mutagenicity (bacterial reverse gene mutation) data.

Conclusion:

Under the conditions of this study according to OECD 471, the test substance, N-Methylolmethacrylamide 15 % aqueous solution showed a weak mutagenic activity in only one on the tester strains TA100 in the bacterial reverse mutation test with Salmonella typhimurium. A reevaluation of the SCF (Scientific Committee on Food) classified the substance under the above mentioned conditions as negative in the Ames test according to OECD guideline 471.

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