Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23 October to 20 November 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Proprietary study conducted according to relevant guidelines and GLP, with no deviations.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
The test material, batch no. PK280-100, described as a very viscous amber liquid with an expiry date of 03 October 2014, was stored at room temperature. The active component content was 100%.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
The test animals were 8-10 week old healthy female Wistar Crl: WI(Han) (full barrier, SPF) rats. They were nulliparous and non-pregnant. Body weights on the day of administration ranged from 160 - 179 g. The rats were group housed in IVC cages (type III H) on Altromin saw fibre bedding, in an air-conditioned room maintained at a temperature of 22±3°C and relative humidity of 55±10%. There was a 12 hour light/dark cycle, and 10 air changes per hour. The rats were allowed free access to food (Altromin 1324 maintenance diet for rats and mice) and tap water, sulphur acidified to a pH value of approximately 2.8. Prior to test substance administration, food was withheld from the animals for 16 to 19 hours (access to water was allowed). Food was returned to the animals approximately 4 hours post-dosing. The rats were acclimatised to the laboratory conditions for at least five days.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
sterile water
Details on oral exposure:
Aqua ad injectionem (sterile water) was used as the vehicle. Homogeneity of the test material in the vehicle was maintained by vortexing the test solution for 30 minutes prior to each administration. The dosing volume was 10 mL/kg bw.
Doses:
Step 1 and 2: 2000 mg/kg bw
Step 3 and 4: 300 mg/kg bw
No. of animals per sex per dose:
3 female rats per Step
Control animals:
no
Details on study design:
The acute toxic class method was followed. The starting dose was 2000 mg/kg bw. No test material related mortality was recorded for any animal of Step 1, therefore a second step was performed at a dose of 2000 mg/kg bw. Mortality was noted at Step 2, therefore a third step was performed at 300 mg/kg bw. Based on the results obtained at Step 3, a fourth step was performed at 300 mg/kg bw. Surviving animals were observed for 14 days post-dosing. Clinical examinations were made several times on the day of dosing, at least once during the first 30 minutes and with special attention during the first 4 hours post-dose. Animals were observed for clinical signs once daily thereafter, until the end of the observation period. Body weights were recorded on Day 1 (prior to administration), and on Days 8 and 15 of the observation period. At the end of the 14 day observation period, survivors were sacrificed and subject to gross necropsy.
Statistics:
Not required.

Results and discussion

Preliminary study:
Not applicable.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
500 mg/kg bw
Based on:
test mat.
Remarks on result:
other: cut-off value
Mortality:
No mortalities were observed in the Step 1 rats dosed with 2000 mg/kg bw. All three rats in Step 2 (2000 mg/kg bw) were found dead between approximately 1 and 4 hours post-dosing. No mortalities were observed in the Step 3 and 4 rats (300 mg/kg bw).
Clinical signs:
Clinical signs observed in Step 1 rats within 4 hours of dosing included slightly to moderately reduced spontaneous activity, recumbency, bradykinesia, slight piloerection, hunched posture and half-eyelid closure. No signs of toxicity were noted from the day after dosing through to the end of the observation period.
Clinical signs observed in Step 2 rats within 4 hours of dosing (prior to death) included slightly to moderately reduced spontaneous activity, recumbency, bradykinesia, slight to moderate piloerection, kyphosis, catalepsis and half-eyelid closure.
Clinical signs observed in Step 3 rats within 4 hours of dosing included slightly reduced spontaneous activity, bradykinesia, slight piloerection, kyphosis and half-eyelid closure. No signs of toxicity were noted from the day after dosing through to the end of the observation period.
Clinical signs observed in Step 4 rats within 4 hours of dosing included moving the bedding, slight wasp waist, slight piloerection, kyphosis, moderately reduced spontaneous activity and half-eyelid closure. Slightly reduced spontaneous activity was observed between 4 hours post dosing and day 2. No signs of toxicity were observed from day 3 until the end of the observation period.
Body weight:
Bodyweight loss was noted in one Step 3 female (300 mg/kg bw) during week 2 of the observation period, all other surviving rats gained weight during the observation period.
Gross pathology:
Two rats in Step 2 (2000 mg/kg bw) were found to have stomachs distended with gas at necropsy. No other findings were noted.
Other findings:
No other findings reported.

Any other information on results incl. tables

No further information.

Applicant's summary and conclusion

Interpretation of results:
Toxicity Category IV
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on the results of the study, the LD50 cut-off value is considered to be 500 mg/kg bw.
Executive summary:

The acute oral toxicity of AAI_C18_PEHA_ACETATES was evaluated in female Wistar Crl: WI(Han) rats, according to the Acute Toxic Class method (OECD 423). The test material was dissolved in sterile water and administered to two groups of 3 females each at a dose of 2000 mg/kg bw, by gavage. A further two groups of 3 females each were administered a dose of 300 mg/kg bw by gavage. Animals were observed regularly within the first 4 hours after dosing for signs of toxicity and mortality. Daily observations were made for 14 days thereafter, and gross necropsy was performed on decedents or on survivors at the end of the observation period. Body weights were recorded on Day 1 (prior to administration), and on Days 8 and 15 of the observation period. There were no deaths in Step 1 animals administered 2000 mg/kg bw, however all 3 rats administered 2000 mg/kg bw in Step 2 died spontaneously on the day of dosing. No deaths were observed in the Step 3 and 4 animals administered 300 mg/kg bw. Relevant clinical findings in animals administered 2000 mg/kg bw included reduced spontaneous activity, piloerection, half eyelid-closure, bradykinesia, recumbency, catalepsis and kyphosis. All symptoms recovered by Day 2 for surviving animals. Relevant clinical findings in animals administered 300 mg/kg bw included reduced spontaneous activity, piloerection, half eyelid-closure, bradykinesia, moving the bedding, wasp waist and hunched posture. All symptoms recovered by Day 3. There were no effects on weight gain during the observation period, except for one animal which showed weight loss during the second week. No treatment-related macroscopic findings were observed.

Based on the results of the study, the LD50 cut-off value is considered to be 500 mg/kg bw.