Mecrilate

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian germ cell study: gene mutation

Remarks:

Point mutations and small deletions

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study is in line with OECD TG 477 and met GLP requirements.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1984

Report date:

1984

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 477 (Genetic Toxicology: Sex-linked Recessive Lethal Test in Drosophila melanogaster)

Deviations:

yes

Remarks:

Age of the insects was not specified

GLP compliance:

yes (incl. QA statement)

Type of assay:

Drosophila SLRL assay

Test material

Test material information

Constituent 1

Reference substance name:

Mecrilate

EC Number:

205-275-2

EC Name:

Mecrilate

Cas Number:

137-05-3

Molecular formula:

C5H5NO2

IUPAC Name:

methyl 2-cyanoprop-2-enoate

Test material form:

other: liquid

Details on test material:

Two bottles of a clear, colorless liquid, designated Super Bonder 430, Metal Bonding-Instant Adhesive A5050 202 (94.2 % methyl cyanoacrylate monomer) were submitted by the sponsor.
Upon arrival in the laboratory, the test agent was assigned Laboratory Number 340, stored at room temperature and one bottle of the test material was shipped on August 16, 1983, to Dr. Stanley Zimmering, Brown University, Division of Biology and Hedicine, Providence, Rhode lsland, U.S.A.
Under the conditions of this assay the test material is assumed to be stable.

Test animals

Species:

Drosophila melanogaster

Strain:

other: not applicable

Sex:

male/female

Details on test animals or test system and environmental conditions:

Stocks of Canton-S (wild-type males) and Basc females are maintained in Dr. Stanley Zimmering's laboratory, Division of Biology and Medicine, Brown University, Providence, Rhode lsland, U.S.A.

Administration / exposure

Route of administration:

other: oral (feed) Series I / inhalation Series II-V

Vehicle:

Series I: Acetone (Fischer, Lot 793661, ACS),
Series II-V: unchanged (no vehicle)

Details on exposure:

see "Any other information on materials and methods"

Duration of treatment / exposure:

see "Any other information on materials and methods"

Frequency of treatment:

see "Any other information on materials and methods"

Post exposure period:

see "Any other information on materials and methods"

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0.03, 0.045, 0.06 mL
Basis:
nominal conc.
Series IV (inhalation)

Doses / concentrations 2

Remarks:

Doses / Concentrations:
0.03, 0.04 mL
Basis:
nominal conc.
Series V (inhalation)

No. of animals per sex per dose:

see "Any other information on materials and methods"

Control animals:

yes

Positive control(s):

Dimethylnitrosamine DMN (Eastman, Lot C/B, 99.9 %)

Examinations

Tissues and cell types examined:

not applicable

Details of tissue and slide preparation:

not applicable

Evaluation criteria:

For the sex-linked recessive lethal test in Drosophila melanogaster to be considered valid, the following criteria must be met:
A. Demonstration of toxicity of the chemical for the male Drosphila melanogaster, unless this is not possible due to a limited solubility of the test compound or the test compound at any concentration is not lethal.
B. The frequency of lethals in the solvent controls is within the normal range.
C. Confirmation of sensitivity and responsiveness of the tester system to detect mutagenic activity.
If the above criteria are met, a chemical will be considered as providing evidence of a mutagenic effect if an increment of 0.2 % above the lethal frequency in the solvent control is obtained.

Statistics:

- Kastenbaum, M.A. and K.O. Bowrnan (1966), the Minimum Significant Number if Successes in a Binomial Sample. Publication ORNL-30-09, Oak Ridge National Laboratory, Oak Ridge, Tennessee.
- Marolin, B. H., Collins, B.J. and Mason, J.M. (1983), Statistical Analysis and Sample-Size Determinations for Mutagenicity Experiments with Binomial Responses. Environmental Mutagenesis 5, 705-716.

Results and discussion

Test results

Key result

Sex:

male

Genotoxicity:

negative

Toxicity:

yes

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

RESULTS AND DISCUSSION

As shown in Table 1, the induced mortality rates at 0.03 and 0.045 mL of Super Bonder 430 were 3 % and 46 %, respectively (Series IV).

The percent mortality observed for Treatment Series V confirms the finding of the first assay which indicated that the test material was toxic at 0.045 mL test concentration.

As previously stated, the physical properties of the test agent precluded accurate volume determination. It would appear that this limitation contributed to the differences in exposure times required to achieve comparable percent mortalities. It was also noted that efficiency in preparation of the treatment vials was markedly increased by familiarity with the procedure, which may account for the relatively short time required to reach toxicity in Treatment Series V. The overall evaluation of the data was, therefore, based on the percent mortality rather than on the length of time required to achieve comparable toxic effects.

Treatment Series IV

Also presented in Table 1 is the number of lethals per total number of cultures for each brood. In Treatment Series IV a total of nine lethals was recovered; however, six lethals scored from one male in Brood 3 occured as a cluster of mutations. Mutations which occur in clusters greater than two are considered outside the limits of the Poisson distribution; accordingly, all F1 tests from this male were excluded from the calculations. Only the corrected values are shown in Table 1. No statistically significant differences in percent lethals were noted between the test groups and the negative air control group.

Treatment Series V

Data presented in Table 1 for Treatment Series V indicates that all observed lethal cultures occured as single mutations. These data further indicate that no increase in the occurrence of lethal cultures accompanied exposure of the flies to the test material.

Combined Results Treatment Series IV and V

Data from both experiments were analyzed statistically using the Kastenbaum-Bowman Test. Based on this analysis, the results indicate that neither Treatment Series was different from the control.

Positive control results, which are included in Table 1, confirm the metabolism of dimethylnitrosamine into a strong mutagen by Drosophila males.

Combined data from Treatment Series IV and V are presented in Table 2. While volumes for the highest test concentrations in Treatment Series IV and V were slightly different, results achieved with 0.045 and 0.04 mL of Super Bonder 430 were comparable; therefore, overall findings for these doses were combined. An analysis of these data confirms the results of the two independent studies and indicates that inhalation exposure of male Drosophila to two levels of Super Bonder 430 failed to provide evidence of a mutagenic response.

CONCLUSION

For the sex-linked recessive lethal test in Drosophila melanogaster to be considered valid, the following criteria must be met:

A. Demonstration of toxicity of the chemical for the male Drosphila melanogaster, unless this is not possible due to a limited solubility of the test compound or the test compound at any concentration is not lethal.

B. The frequency of lethals in the solvent controls is within the normal range.

C. Confirmation of sensitivity and responsiveness of the tester system to detect mutagenic activity.

If the above criteria are met, a chemical will be considered as providing evidence of a mutagenic effect if an increment of 0.2 % above the lethal frequency in the solvent control is obtained.

Criteria (A) and (B) were satisfied by the test conditions used with the test material. Criterion (C) was met as judged by results following treatment with the indirect acting mutagen, dimethylnitrosamine.

The overall results indicate that the test material failed to induce an increment of 0.2 % above the lethal frequency in the solvent control.

Under the conditions of these studies it is concluded that the Drosophila Recessive Lethal Assay of the experimental agent, Super Bonder 430, provides no evidence of a mutagenic effect in this test system.

TABLE 1

Treatment	Conc. [mL]	Exposure	No. P1 males mated	% mortality	Brood 1	Brood 2	Brood 3	Total	% Lethals
Series IV	0.03	2 h	52	3	1/1302	0/1333	1/1346	2/3981	0.05
	0.045	6 h	42	46	1/1335	1/1352	0/1367	2/4054	0.05
	air control	2.5 h	40	0	0/1250	4/1352	0/1454	4/4056	0.1
Series V	0.03	20 min	51	14	2/1344	1/1383	1/1353	4/4080	0.1
	0.04	30 min	30	47	0/809	0/1056	2/1440	2/3305	0.06
	air control	4 h	40	0	1/1250	3/1288	0/1268	4/3806	0.1
Pos. Control	10 mM DMN	1 d	50	0	132/974	NA	NA	NA	13.6
Solvent Control	5 % Sucrose	1 d	41	2	0/999	NA	NA	NA	0

TABLE 2 - combined results

Treatment	Conc. [mL]	Brood 1	Brood 2	Brood 3	Total	% Lethals
Series IV & V	0.03	3/2646	1/2716	2/2699	6/8061	0.07
	0.04 - 0.045	1/2144	1/2408	2/2807	4/7359	0.05
	air control	1/2500	7/2640	0/2722	8/7862	0.10

Applicant's summary and conclusion

Conclusions:

The overall results indicate that the test material failed to induce an increment of 0.2 % above the lethal frequency in the solvent control.
Under the conditions of these studies it is concluded that the Drosophila Recessive Lethal Assay of the experimental agent, Super Bonder 430, provides no evidence of a mutagenic effect in this test system.