Registration Dossier

Diss Factsheets

Administrative data

Description of key information

In an acute oral toxicity study in female rats, conducted in accordance with OECD 423 (2001), EU Method B.1 tris (2008), EPA OPPTS 870.1100 (2002) and according to GLP principles, a LD50 oral of >2000 mg/kg was determined. In an acute inhalation study in 5 male and 5 female rats, conducted in accordance with OECD 403 (2009) and according to GLP principles, a 4h-LC50 of >5.09 mg/L was determined. In an acute dermal toxicity study in 5 male and 5 female rats, performed in accordance with OECD 402 (1987), EU Method B.3 ( 2008), EPA OPPTS 870.1200 (1998) and according to GLP principles, a LD50 of >2000 mg/kg bw was determined. No mortalities occurred in these studies.

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:
December 7, 2010 - February 21, 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study has been performed in accordance with OECD 423 (2001), EU Method B.1 tris (2008), EPA OPPTS 870.1100 (2002) and according to GLP principles.
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
(2001)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
(2008)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
(2002)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Age at study initiation: 8 - 10 weeks
- Weight at study initiation:
Step 1/animals no. 1-3: 170-180 g
Step 2/animals no. 4-6: 153-159 g
- Fasting period before study: 16 to 19 hours
- Housing: the animals were kept in groups in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 040810)
- Diet: free access to Altromin 1324 maintenance diet for rats and mice (lot no. 1013). Free access again from 4 hrs post dosing.
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 55 ± 10%
- Air changes (per hr): 10 per hr
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light

Date of Dose Administration:
Step 1, animals no. 1, 2 and 3: December 22, 2010
Step 2, animals no. 4, 5 and 6: December 29, 2010
Route of administration:
oral: gavage
Vehicle:
cotton seed oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 0.4 g/mL
- Amount of vehicle: 5 mL
- Justification for choice of vehicle: due to its non-toxic characteristics
- Lot/batch no.: Sigma, lot no. MKBB7604, expiry date 30/01/2011
- Purity: not applicable

DOSE VOLUME APPLIED: 5 mL/kg

DOSAGE PREPARATION: the test item was weighed out into a tared plastic vial on a precision balance. Homogeneity was maintained by vortexing. The dosages were made shortly before administration.

CLASS METHOD
- Rationale for the selection of the starting dose: no details provided
Doses:
2000 mg/kg
No. of animals per sex per dose:
3 females twice (6 females in total)
Control animals:
other: not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of weighing: the animals were weighed on day 1 (prior to the administration), on day 8 (1 week thereafter) and on day 15 (2 weeks thereafter)
- Frequency of observations: a careful clinical examination was made several times on the day of dosing (at least once during the 1st 30 minutes and with special attention given during the 1st 4 hours post-dose). Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. According to the OECD Guideline.
- Necropsy of survivors performed: yes
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortalities occurred
Mortality:
- No mortalities occurred
Clinical signs:
other: - At 1 hour after administration, slight piloerection was observed in animal #2 and #3 - In animal #1, prone position and slight piloerection was observed at 3 and 4 hours after administration, until day 2 - At 3 and 4 hours after administration, prone po
Gross pathology:
- No abnormalities were observed
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an acute oral toxicity study in female rats, conducted in accordance with OECD 423 (2001), EU Method B.1 tris (2008), EPA OPPTS 870.1100 (2002) and according to GLP principles, a LD50 oral of >2000 mg/kg was determined.
Executive summary:

The acute oral toxicity in female rats has been studied in accordance with OECD 423 (2001), EU Method B.1 tris (2008), EPA OPPTS 870.1100 (2002) and according to GLP principles. The substance was dosed in cotton seed oil at 2 g/kg bw in 6 female rats, in 2 steps. At 1, 3 and 4 hours after administration, slight reduced spontaneous activity, prone position and slight to moderate piloerection was observed in all animals of the 1st group, until day 2. No mortalities occurred. Necropsy did not show any abnormality. The acute oral toxicity (LD50) was determined to be >2000 mg/kg. Based on this result, the substance does not need to be classified for acute toxicity by the oral route.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The study has Klimisch score 1

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
November 2, 2010 - July 28, 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study has been performed in accordance with OECD 403 (2009) and according to GLP principles.
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
(2009)
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: SAGE LABS, Boyertown, PA, USA (received on January 18, 2011)
- Age at study initiation: 9 to 10 weeks
- Weight at study initiation: 272-293 g (males) and 188-210 g (females)
- Fasting period before study: no data
- Housing: singly housed in suspended stainless steel caging with mesh floors
- Diet: Purina Rodent Chow #5012
- Water: tap water (ad libitum, except during exposure)
- Acclimation period: 15 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23 °C and 20-21 °C (during exposure)
- Humidity (%): 27-53% and 28-34% (during exposure)
- Air changes (per hr): 11 per hr
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light
Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: a nose-only inhalation chamber (ADG Developments Ltd.)
- Method of holding animals in test chamber: individually in polycarbonate holding tubes
- Source and rate of air: approximately 28.4 liters per minute of filtered air was supplied by an air compressor (Airgas) to the dust generator
- Method of conditioning air: an additional 3.3 liters per minute of compressed mixing air, supplied using air from a compressed air tank (Airgas), was introduced into the chamber, creating a vortex at the chamber inlet
- System of generating particulates/aerosols: the unground substance was aerosolized using a modified Wright Dust Generator driven by a variable speed motor (Dayton)
- Method of particle size determination: an eight-stage ACFM Westech Ambient Particle Sizing Sampler was used to assess the particle size distribution of the test atmosphere. Samples were withdrawn from the breathingzone of the animals at 2 intervals.
- Treatment of exhaust air: not applicable
- Temperature, humidity, pressure in air chamber: the exposure tube temperature and relative humidity ranges during exposure were 20 degrees Celsius and 33-36%, respectively

TEST ATMOSPHERE
- Samples taken from breathing zone: yes
- MMAD (Mass median aerodynamic diameter): 3.92 µm

CLASS METHOD
- Rationale for the selection of the starting concentration: not reported
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
5.09 mg/L (time weighted average)
No. of animals per sex per dose:
5
Control animals:
other: not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Body weights: prior to exposure and on days 1, 3, 7 and 14
Clinical signs: upon removal from the exposure tube and at least once daily thereafter
- Necropsy of survivors performed: yes
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5.09 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
other: Time weighted average chamber concentration; no mortalities occurred and no clinical signs were observed during the study
Mortality:
- There were no mortalities during the study
Clinical signs:
other: - There were no signs of treatment observed during the study
Body weight:
- No abnormalities were observed
Gross pathology:
- No abnormalities were observed
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an acute inhalation study in rats, conducted in accordance with OECD 403 (2009) and according to GLP principles, rats (5 males and 5 females) were exposed by the nose only. The MMAD of the substance was determined to be 3.92 µm. No mortalities occurred during the study. There were also no signs of treatment observed during the study. Necropsy did not show any abnormality. Based on the results of this study, the 4h-LC50 was determined to be >5.09 mg/L and
the substance does not need to be classified for acute toxicity by the inhalation route.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
Value:
5 090 mg/m³ air
Quality of whole database:
The study has Klimisch score 1

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:
December 19, 2010 - January 3, 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study has been performed in accordance with OECD 402 (1987), EU Method B.3 ( 2008), EPA OPPTS 870.1200 (1998) and according to GLP principles.
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
(1987)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
(2008)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
(1998)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: Wistar strain, Crl:WI (Han)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Age at study initiation:
Males: 8-9 weeks
Females: 14 weeks
- Weight on the day of administration:
Males: 273-287 g
Females: 217-230 g
- Housing: Individually in IVC cages, type III H, polysulphone
- Diet: ad libitum (Altromin 1324)
- Water: ad libitum (tap water, sulphur acidified to a pH value of approx. 2.8)
- Acclimation period: 10 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
occlusive
Vehicle:
water
Remarks:
(aqua ad injectionem (B. Braun Melsungen, lot no. 0195A191, expiry date: 04/2013))
Details on dermal exposure:
TEST SITE
- Approximately 24 hours before the test, the fur was removed from the dorsal area of the trunk by using an electric clipper.
- Area of exposure: approx. 10% of the total body surface
- Dressing: gauze and non-irritating tape, which was fixed with an additional dressing

REMOVAL OF TEST SUBSTANCE
- Washing: yes, using water
- Time after start of exposure: 24 hours

VEHICLE AND WATER USED FOR REMOVAL OF TEST SUBSTANCE
Aqua ad injectionem (B. Braun Melsungen, lot no. 0195A191, expiry date: 04/2013)
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of weighing: The animals were weighed on day 1 (prior to the application) and on days 8 and 15
- Frequency of observations: A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). Thereafter, the animals were observed for clinical signs once daily until the end of the observation period.
- Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Necropsy of survivors performed: yes
- Other: Signs of erythema and oedema were assessed using the scoring system laid down in OECD 404 (2002)
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortalities and no clinical signs of toxicity occurred
Mortality:
No mortalities occurred
Clinical signs:
other: No clinical signs of toxicity occurred
Gross pathology:
Macroscopic examination of the animals did not reveal any abnormalities.
Other findings:
- Eschar was observed in 1 male, on day 9 only
- Eschar was observed in 4 females, in the period day 5-9
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an acute dermal toxicity study with rats, performed in accordance with OECD 402 (1987), EU Method B.3 ( 2008), EPA OPPTS 870.1200 (1998) and according to GLP principles, a LD50 of >2000 mg/kg bw was determined.
Executive summary:

The acute dermal toxicity of the substance was determined in the rat, in accordance with OECD 402 (1987), EU Method B.3 ( 2008), EPA OPPTS 870.1200 (1998) and according to GLP principles. The substance was administered to five Wistar rats of each sex by a single dermal application of 2000 mg/kg bw for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice. No mortality and no clinical signs of toxicity occurred. Body weight gain was observed in all animals. Eschar was observed in 1 male, on day 9 only and in 4 females, in the period day 5-9. Macroscopic examination of the animals did not reveal any abnormalities. The dermal LD50 value of the substance in Wistar rats was established to be >2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The study has Klimisch score 1

Additional information

Acute toxicity: oral

The acute oral toxicity in female rats has been studied in accordance with OECD 423 (2001), EU Method B.1 tris (2008), EPA OPPTS 870.1100 (2002) and according to GLP principles. The substance was dosed in cotton seed oil at 2 g/kg bw in 6 female rats, in 2 steps. At 1, 3 and 4 hours after administration, slight reduced spontaneous activity, prone position and slight to moderate piloerection was observed in all animals of the 1st group, until day 2. No mortalities occurred. Macroscopic examination of the animals did not reveal any abnormalities. The acute oral toxicity (LD50) was determined to be >2000 mg/kg bw.

 

Acute toxicity: inhalation

In an acute inhalation study in rats, conducted in accordance with OECD 403 (2009) and according to GLP principles, rats (5 males and 5 females) were exposed by the nose only. The MMAD of the substance was determined to be 3.92 µm. No mortalities occurred during the study. There were also no signs of treatment observed during the study. Macroscopic examination of the animals did not reveal any abnormalities. Based on the results of this study, the 4h-LC50 was determined to be >5.09 mg/L.

 

Acute toxicity: dermal

The acute dermal toxicity of the substance was determined in the rat, in accordance with OECD 402 (1987), EU Method B.3 ( 2008), EPA OPPTS 870.1200 (1998) and according to GLP principles. The substance was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg bw for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice. No mortality and no clinical signs of toxicity occurred. Body weight gain was observed in all animals. Eschar was observed in 1 male, on day 9 only and in 4 females, in the period day 5-9. Macroscopic examination of the animals did not reveal any abnormalities. The dermal LD50 value of the substance in Wistar rats was established to be >2000 mg/kg bw.

Justification for selection of acute toxicity – oral endpoint
Only one study available

Justification for selection of acute toxicity – inhalation endpoint
Only one study available

Justification for selection of acute toxicity – dermal endpoint
Only one study available

Justification for classification or non-classification

The substance does not need to be classified for acute toxicity in accordance with the CLP Regulation as the acute oral and dermal toxicity is >2000 mg/kg bw and as the acute inhalation toxicity is >5.09 mg/L.