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Toxicological information

Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
January 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study (OECD 423)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
yes
Remarks:
The temperature and relative humidity recorded in the animal room were sometimes outside of the target ranges specified in the study plan
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
yes
Remarks:
(see above)
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): AZDN
- Physical state: solid (white powder)
- Analytical purity: 99.2%
- Lot/batch No.: 6089
- Expiration date of the lot/batch: March 2010
- Storage conditions of test material: below 15°C and protected from light

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Breeder: Janvier, Le Genest-Saint-Isle, France.
- Age at study initiation: approximately 9 weeks old for the first assay and 8 weeks old for the next assays
- Weight at study initiation: 215 +/- 23 g
- Fasting period before study: approximately 18 hours
- Housing: polycarbonate cages with stainless steel lid (48 cm x 27 cm x 20 cm)
- Diet (e.g. ad libitum): conventional laboratory diet (SSNIFF R/M-H pelleted maintenance diet)
- Water (e.g. ad libitum): drinking water filtered by a FG Millipore membrane (0.22 micron), provided ad libitum.
- Acclimation period: at least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2°C
- Humidity (%): 30 to 70%
- Air changes (per hr): 12 cycles/hour
- Photoperiod (hrs dark / hrs light): 12h/12h (7:00 - 19:00)

IN-LIFE DATES: from 21 January 2010 to 17 February 2010

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: sesame oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 or 200 mg/kg
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: standard vehicle used for specific routes of administration
- Lot/batch no. (if required): 067K0069

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: according to information available on the test item, for animal welfare reasons, the starting dose-level of 300 mg/kg was chosen.
Doses:
300 mg/kg and 2000 mg/kg
No. of animals per sex per dose:
3 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: the animals were observed frequently during the hours following administration of the test item, for detection of possible treatment-related clinical signs. Thereafter, observation of the animals was made at least once a day. The animals were weighed individually just before administration of the test item on day 1 and then on days 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight.
Statistics:
no

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Mortality:
Dose-level of 300 mg/kg: none.
Dose-level of 2000 mg/kg: Due to signs of poor clinical conditions observed (hypoactivity, sedation, piloerection, tremors, dyspnea, hypersensitivity to touch and tonic clonic convulsions), the animals were sacrificed on day 1 for ethical reasons.
Clinical signs:
Dose-level of 300 mg/kg (three females then confirmation on three other females) :
Hypoactivity, piloerection and rhinorrhea were observed in all the animals on days 1 and/or 2.
Rhinorrhea persisted in 2/6 animals on day 3. Tremors and hypersensitivity to noises and being touched were also noted in 3/6 animals on day 2.

Dose-level of 2000 mg/kg (three females)
Hypoactivity then sedation, piloerection, tremors, dyspnea, hypersensitivity to touch and tonic clonic convulsions were observed in all the animals within 4h30 of treatment. In view of these signs of poor clinical conditions, all three animals were sacrificed on day 1 for ethical reasons.
Body weight:
When compared to CIT historical control data, a lower body weight gain (30 g vs. 41 g ± 9 g in control data base) was noted between day 1 and day 8 in 1/6 females given 300 mg/kg, returning to normal thereafter.
Gross pathology:
Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.
Other findings:
None.

Applicant's summary and conclusion

Interpretation of results:
Toxicity Category IV
Remarks:
Migrated information Criteria used for interpretation of results: other: Directive 67/548/EEC and EU CLP No 2008/1272.
Conclusions:
Under the experimental conditions of this study, the oral LD50 of the test item AZDN (batch No. 6089, purity: 99.2%) was comprised between 300 and 2000 mg/kg in rats.

According to the classification criteria laid down in Council Directive 67/548/EEC (and subsequent adaptations), the test item AZDN should be assigned the symbol Xn, the indication of danger "Harmful" and the risk phrase R 22: "Harmful if swallowed".
Executive summary:
The acute oral toxicity of the test item, AZDN, was evaluated in rats according to OECD (No. 423, 17th December 2001) and Commission Regulation (EC) (No. 440/2008, B1tris, 30 May 2008) guidelines. The test item was prepared in sesame oil and was administered by oral route (gavage), under a volume of 10 mL/kg, to groups of three fasted female Sprague-Dawley rats. Rats were exposed to 300 or 2000 mg/kg bw. Clinical signs, mortality and body weitht gain were checked for a period of up to 14 days following the single administration of the test item. All animals were subjected to necropsy.

Dose-level of 300 mg/kg (three females then confirmation on three other females): No mortality was recorded. Hypoactivity, piloerection and rhinorrhea were observed in all the animals on days 1 and/or 2. Rhinorrhea persisted in 2/6 animals on day 3. Tremors and hypersensitivity to noises and being touched were also noted in 3/6 animals on day 2.   When compared to CIT historical control data, a lower body weight gain (vs.±in control data base) was noted in 1/6 females between day 1 and day 8, returning to normal thereafter.  

Dose-level of 2000 mg/kg (three females): Hypoactivity then sedation, piloerection, tremors, dyspnea, hypersensitivity to touch and tonic clonic convulsions were observed in all the animals within 4h30 of treatment. Lateral recumbency was also noted in 1/3 animals 3 hours after treatment. In view of these signs of poor clinical conditions, all three animals were sacrificed on day 1 for ethical reasons. At necropsy, no apparent abnormalities were observed in any animal.

Under the experimental conditions of this study, the oral LD50of the test item AZDN (batch No. 6089, purity: 99.2%) was comprised between 300 and 2000 mg/kg in rats. According to the classification criteria laid down in Council Directive 67/548/EEC (and subsequent adaptations),the test item AZDN should be assigned the symbol Xn, the indication of danger "Harmful" and the risk phrase R 22: "Harmful if swallowed", and AZDN in classified in category 4 according to Regulation EC no.1272/2008 (CLP).