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Administrative data

Description of key information

Acute oral : the oral LD50 of AZDN was comprised between 300 and 2000 mg/kg in rats (GLP guideline study, OECD 423).
Acute inhalation : No reliable study (data waiving).
Acute dermal : the dermal LD50 of the test item AZDN was higher than 2000 mg/kg in rats (GLP guideline study, OECD 402).

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
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)
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
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

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
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.
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).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
discriminating dose
Value:
300 mg/kg bw

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
03 March 2010 to
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: compliant to GLP and testing guideline; adequate coherence between data, comments and conclusions
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France.
- Age at study initiation: 8 weeks old
- Weight at study initiation: 371 +/- 9 g for the males and 235 +/- 7 g for the females
- Fasting period before study: none
- Housing: polycarbonate cages with stainless steel lid (48 cm x 27 cm x 20 cm)
- Diet (e.g. ad libitum): SSNIFF R/M-H pelleted maintenance diet
- Water (e.g. ad libitum): drinking water filtered by a FG Millipore membrane (0.22 micron)
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 30 to 70%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h (7:00 - 19:00)

IN-LIFE DATES: From 03 March 2010 to 17 March 2010
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: 5 cm x 7 cm for the males and 5 cm x 6 cm for the females
- % coverage: 10% of the total body surface of the animals
- Type of wrap if used: gauze pad held in contact with the skin by means of an adhesive hypoallergenic aerated semi-occlusive dressing and a restraining bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): any residual test item was removed using a dry cotton pad
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg
- For solids, paste formed: yes (substance moistened with 2 mL of purified water)

Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 animals per sex per dose
Control animals:
other: historical control animals
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, and then at least once a day until day 15. 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
Statistics:
no
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Sex:
male
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
None.
Clinical signs:
None.
Body weight:
When compared to historical control data, a lower body weight gain was noted in all the males (16 to 35 g vs. 47 g ± 7 g in historical data base) and 1/5 females (7 g vs. 25 g ± 11 g in historical data base) between day 1 and day 8; returning to normal thereafter.
Gross pathology:
Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: other: Regulation EC 1272/2008 (CLP)
Conclusions:
Under the experimental conditions of this study, the dermal LD50 of the test item AZDN (batch No. 6089, purity: 99.2%) was higher than 2000 mg/kg in rats.
Executive summary:

The acute dermal toxicity of the test item, AZDN, was evaluated in rats according to OECD (No. 402, 24th February 1987) and Commission Regulation (EC) (No. 440/2008, Part B.3, 30 May 2008) guidelines.

The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.

Methods

The test item was applied to the skin of one group of ten Sprague-Dawley rats (five males and five females).

The application was performed with the test item in its original form at the dose-level of 2000 mg/kg.

The test site was then covered by a semi-occlusive dressing for 24 hours.

Clinical signs, mortality and body weight gain were checked for a period of 14 days following the single application of the test item.

All animals were subjected to necropsy.

 

Results

No mortality, no clinical signs and no cutaneous reactions were observed during the study.

When compared to historical control data, a lower body weight gain was noted in all the males (16 to 35 g vs.47 ± 7 g in historical data base) and 1/5 females (7 g vs. 25 g ± 11 g in historical data base) between day 1 and day 8; returning to normal thereafter.

No apparent abnormalities were observed at necropsy in any animal.

 

Conclusion

Under the experimental conditions of this study, the dermal LD50 of the test item AZDN (batch No. 6089, purity: 99.2%) was higher than 2000 mg/kg in rats.

According to the classification criteria laid down in Council Directive 67/548/EEC (and subsequent adaptations), concerning the potential toxicity by dermal route, the test item should not be classified.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw

Additional information

Acute oral (Pelcot, CIT, 2010) :

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.

At 300 mg/kg (three females then confirmation on three other females):No mortality was recorded but some clinical signs (hypoactivity, piloerection and rhinorrhea) in all the animals on days 1 and/or 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.  

At 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 LD50 of the test item AZDN (batch No. 6089, purity: 99.2%) was comprised between 300 and 2000 mg/kg in rats.

Acute dermal (Pelcot, CIT, 2010) :

The test item was applied to the skin of one group of ten Sprague-Dawley rats (five males and five females). The application was performed with the test item in its original form at the dose-level of 2000 mg/kg. The test site was then covered by a semi-occlusive dressing for 24 hours. Clinical signs, mortality and body weight gain were checked for a period of 14 days following the single application of the test item. All animals were subjected to necropsy.

No mortality, no clinical signs and no cutaneous reactions were observed during the study.

When compared to historical control data, a lower body weight gain was noted in all the males (16 to 35 gvs.47 ± 7 g in historical data base) and 1/5 females (7 g vs. 25 g ± 11 g in historical data base) between day 1 and day 8; returning to normal thereafter.

No apparent abnormalities were observed at necropsy in any animal.

Under the experimental conditions of this study, the dermal LD50 of the test item AZDN (batch No. 6089, purity: 99.2%) was higher than 2000 mg/kg in rats.

Justification for classification or non-classification

Acute oral :

According to the classification criteria laid down in Council Directive 67/548/EEC, AZDN should be assigned the symbol Xn, the indication of danger "Harmful" and the risk phrase R 22: "Harmful if swallowed".

According to Regulation EC no.1272/2008 (CLP), AZDN is classified in category 4 for acute oral toxicity.

Justification : 300< LD50 < 2000 mg/kg bw.

Acute inhalation : no valid data to justify the current DSD classification (19 ATP): Xn R20

Acute dermal :

According to the classification criteria laid down in Council Directive 67/548/EEC, AZDN is not classified.

According to Regulation EC no.1272/2008 (CLP), AZDN is not classified for acute dermal toxicity.

Justification : LD50>2000 mg/kg bw.