Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 June 2015 to 14 July 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to current OECD guideline and in compliance with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
GLP compliance:
yes (incl. certificate)
Remarks:
Two certificates, signed 2013-08-30 and 2015-06-16
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Velaz Prague, Czech Republic
- Age at study initiation: At least 8-12 weeks; female animals were non-pregnant and nulliparous
- Weight at study initiation: not reported
- Fasting period before study: not reported
- Housing: The animals were housed in plastic cages suspended on stainless steel racks, up to 1-2 animals per cage
- Diet: A laboratory food Altromin (Altromin Spezialfutter GmbH, Germany) was offered in recommended amounts each day approximately at the
same time after dosing.
- Water: The animals received tap water for human consumption. Supply of drinking was unlimited.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2° C
- Humidity (%): 55 ± 10 %
- Air changes (per hr): room equipped with central air conditioning (no further information available)
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg bw
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
1 in sighting study, 4 in main study (females)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed individually immediately after the administration of the test item and then
0.5, 1, 2, and 4 hours later.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: Observations included changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems, and somatomotor activity and behaviour pattern. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Results and discussion

Preliminary study:
The purpose of the sighting study was to allow selection of the appropriate starting dose for the main test. The starting dose for the sighting study could be selected from the fixed dose levels of 5, 50, 300, or 2000 mg/kg. Available information indicated test item is likely to be non-toxic. One female rat was dosed. There was no test item related mortality, but signs of toxicity were observed during 24 hours, therefore the animal was observed for another 24 hour period. After this time, all observed signs were no longer recorded. The Sighting Study was completed and the Main Test started with the dose of 2000 mg/kg.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
One female died 24 hours after dosing.
Clinical signs:
Half an hour after dosing in 4/4 animals reduced activity were noted. The animals were lethargic during the next 4 hours. Lacrimation was observed in 2/4 animals. One animal died after 24 hours after dosing. In the remaining animals, all observed signs were no longer noted. Neither visible change of health nor negative reactions were recorded during the 14-day observation period.
Body weight:
The body weight gain of animals was variable, from weight increase through no gain to a slight decrease after week 1. No body weight losses compared to week 1 were observed 2 weeks after administration.
Gross pathology:
No macroscopic findings were reported at necropsy.
Other findings:
None reported.

Any other information on results incl. tables

The test item α-methylenebenzyl acetate was administered to 5 females at the limit dose of 2000 mg/kg. There was no test item related mortality, but signs of toxicity such as lacrimation, reduced activity, lethargy and mild spasm in hind legs were observed in the Sighting Study during the first 24 hours. Therefore, the animal was observed for a further 24 hours. After this time, all observed signs were no longer recorded. The Main Test started with the dose of 2000 mg/kg. Four additional animals were dosed in the Main Study with the dose of 2000 mg/kg. One female died 24 hours after dosing. In other animals, the same clinical signs were observed. Within 48 hours, all signs were no longer observed. No body weight losses were observed between one and two week after administration of the test item. Neither visible change of health nor negative reactions were registered during the 14- day observation period. No macroscopic changes were noticed.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The key study for acute oral toxicity was conducted according to OECD TG 420 and in compliance with GLP, and reports an LD50 value of >2000 mg/kg bw (Hameln, 2015).