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Toxicological information

Genetic toxicity: in vivo

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Administrative data

Endpoint:
in vivo mammalian germ cell study: cytogenicity / chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1972
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Study without detailed documentation; Oral gavage dose volume, purity not reported.

Data source

Reference
Reference Type:
publication
Title:
Detection of chemical mutagens by the dominant lethal assay in the mouse
Author:
Epstein S.S., Arnold E., Andrea J., Bass W., and Bishop Y
Year:
1972
Bibliographic source:
Toxicology and Applied Pharmacology 23, 288-325 (1972)

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 478 (Genetic Toxicology: Rodent Dominant Lethal Test)
Deviations:
yes
Remarks:
-Oral gavage dose volume, purity not reported.
GLP compliance:
no
Remarks:
Study pre-dates GLP.
Type of assay:
rodent dominant lethal assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
not specified
Details on test material:
- Name of test material (as cited in study report): Heliotropine

Test animals

Species:
mouse
Strain:
other: ICR/Ha Swiss
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Age at study initiation: males were 8-10 weeks old at the study initiation; females were 8-10 weeks old when mated
- Weight at study initiation: Not reported
- Assigned to test groups randomly: Not reported
- Fasting period before study: Not reported
- Housing: housed in suspended mesh caged in air-conditioned rooms with automated light-darkness cycles.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Not reported

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not reported
- Humidity (%): Not reported
- Air changes (per hr): Not reported
- Photoperiod (hrs dark / hrs light): Not reported

Administration / exposure

Route of administration:
other: Oral gavage (daily for 5 days) or intraperitoneal (single dose).
Vehicle:
- Vehicle(s)/solvent(s) used: The study stated that agents under test were freshly prepared in triacprylin or distilled water for single administration to male mice by intraperitoneal injection or by gavage; however, it is unclear which vehicle was used in this study.
Details on exposure:
The study stated that agents under test were freshly prepared in triacprylin or distilled water for single administration to male mice by intraperitoneal(ip) injection or by gavage (further details were not reported). No details were available regarding the day(s) of dosing relative to start of mating periods.
Duration of treatment / exposure:
IP: Single administration
Oral gavage: Once per day for 5 successive days
Frequency of treatment:
IP injection: Single administration
Oral gavage administration: Once per day for 5 successive days
Post exposure period:
Each treated male was mated with 3 females per week for 8 weeks. Females were sacrificed 13 days after the midweek of their caging and presumptive mating, without being checked for vaginal plugs.
Doses / concentrationsopen allclose all
Dose / conc.:
124 other: mg/kg
Remarks:
nominal conc.
IP injection for males only
Dose / conc.:
620 other: mg/kg
Remarks:
nominal conc.
IP injection for males only
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
oral gavage administration for males only
No. of animals per sex per dose:
124 mg/kg: 7 males per dose; 21 females per dose (each mated with 3 females per week for 8 weeks)
640, 1000 mg/kg: 9 males per dose; 27 females per dose (each mated with 3 females per week for 8 weeks)
Control group: 10 males; 30 females per dose (each mated with 3 females per week for 8 weeks)
Control animals:
yes, concurrent vehicle
Positive control(s):
A total of 174 chemicals including, but not limited to, pharmaceuticals, food additives, and pesticides in the dominant lethal assay. Some substances, including acetylaminofluorene and benzoyl peroxide, produced positive results (and therefore can be considered positive controls by default).

Examinations

Tissues and cell types examined:
At autopsy, each female was scored for pregnancy, and for numbers of total implants and early fetal deaths were analyzed. Corpora lutea counts were omitted.
Details of tissue and slide preparation:
Information not reported
Evaluation criteria:
See below for information on evaluation criteria.
Statistics:
Analaysis of variance performed for the complete experiment and its replicates.

Results and discussion

Test results
Key result
Sex:
male
Genotoxicity:
negative
Toxicity:
not examined
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
On the basis of these procedures, heliotropin was classified as a test agent meeting screening criteria but judged non-significant by analysis of variance. See Table 1.

Any other information on results incl. tables

Table 1. Agent producing early fetal deaths and/or pre-implantation losses beyond control limits but with differences not significant by analysis of variance

Agent

Route and frequency of administration

Total weeks of mating

Dose (mg/kg)

No. of males (No. of deaths)

Parameters selected in screen (mating week)

% pregnancy

Implants per pregnancy

Early deaths per pregnancy

% pregnant females with early deaths

Heliotropine*

IP

8

124

7

-

-

-

-

 

 

 

620

9

-

-

1.80 (2)

-

 

 

8

124

7

20 (8)

-

-

-

 

 

 

620

9

19 (8)

-

1.00 (1)

-

 

 Oral

8

1000

9

-

-

-

-

*Compound meeting only less stringent screening criteria.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
Heliotropine (piperonal) produced early fetal deaths beyond control limits, but judged non-significant by analysis of variance and is therefore considered to be negative for genotoxicity.
Executive summary:

Compared with control values, oral administration of heliotropin (piperonal) for 5 consecutive days had no effect on the incidence of pregnancies, early fetal deaths, or pre-implantation losses at any of the timepoints evaluated. Intraperitoneal administration of heliotropin (piperonal) (620 mg/kg body weight) resulted in a slight increase in early fetal deaths only during the first or second week after dosing; this effect was not statistically significant relative to control values and was therefore considered not adverse or reflective of potential genotoxicity. Based on these findings, the no-observed-adverse-effect level (NOAEL) for this study can be considered to be an oral dose level of 1,000 mg/kg body weight/day.