Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Oral Route

Key Study: Allen (1993)

The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2 000 mg/kg bodyweight.

Supporting Study: Walker (1992)

The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 5 000 mg/kg bodyweight.

Inhalation Route

Key Study: Griffiths (2007)

The acute inhalation median lethal concentration (4 hr LC50) of the test material, in the Sprague-Dawley Crl:CD (SD) IGS BR strain rat, was greater than 5.00 mg/L.

Dermal Route

Key Study: Walter (1992)

The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2 000 mg/kg bodyweight.

Supporting Study: Walker (1992)

The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2 000 mg/kg bodyweight.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
17 February 1992 to 10 March 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.
Qualifier:
according to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd., Manston, Kent, UK
- Age at study initiation: approximately five to eight weeks old
- Weight at study initiation: males 159 - 183g; females 140 - 156g
- Fasting period before study: overnight fast immediately before dosing
- Housing: the animals were housed in groups of up to five by sex in solid-floor polypropylene cages with sawdust bedding
- Diet and water: with the exception of an overnight fast immediately before dosing and for approximately two hours after dosing, free access to mains drinking water and food was allowed throughout the study
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 21°C
- Humidity (%):37 - 52%
- Air changes (per hr): approximately 15 changes per hour
- Photoperiod (hrs dark / hrs light): give 12 hours continuous light and 12 hours darkness
Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 5000 mg/ml
- Amount of vehicle (if gavage): 10 ml/kg

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg

DOSAGE PREPARATION: For the purpose of the study the test material was freshly prepared, as required, as a suspension at the appropriate concentration in arachis oil BP. The concentration, homogeneity and stability of the test material preparation were not determined.
Doses:
A preliminary range finding study was conducted on 1 male and 1 female rat using a dose level of 5000 mg/kg of test material delivered by gavage (500 mg/ml concentration) with a dose volume of 10 ml/kg.
This dose of 5000 mg/kg was then selected for the main study.
No. of animals per sex per dose:
1 male, 1 female: Range finding study
5 male, 5 female: Main finding study
Control animals:
no
Details on study design:
- Duration of observation period following administration: 0.5, 1, 2, and 4 hours then subsequently daily for 14 days
- Frequency of observations and weighing: weighed on days 0, 7 and 14
- Necropsy of survivors performed: rangefinding study, no; main study, yes
- Other examinations performed: clinical signs, body weight

All animals were dosed once only by gavage using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to its fasted bodyweight at the time of dosing.
Statistics:
Data evaluations included the relationship, if any, between the animals' exposure to the test material and the incidence and severity of all abnormalities including behavioural and clinical observations, gross lesions, bodyweight changes, mortality and any other toxicological effects.
Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test material was made.
Preliminary study:
Based upon a single dose of 5000 mg/kg by gavage in two rats (1 male, 1 female), there were no deaths or clinical signs of toxicity.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Mortality:
No deaths occurred in the range finding or main study.
Clinical signs:
No signs of systemic toxicity were noted during the study. Black-coloured staining of the fur was noted in 3 female rats during the day of dosing.
Body weight:
No toxicologically significant changes in body weight were noted during the study.
Gross pathology:
No abnormalities.
Interpretation of results:
other: Not classified according to EU criteria.
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 5000 mg/kg bodyweight.
Executive summary:

The test material was evaluated for acute oral toxicity in Sprague Dawley strain rats in accordance with OECD Test Method 401. A preliminary range finding study was conducted on two rats (1 male, 1 female) in a single oral dose of test material as a suspension in arachis oil at a dose level of 5000 mg/kg. No deaths or signs of systemic toxicity were noted. In the main study, five rats per sex received a single oral dose of test material as a suspension in arachis oil at a dose level of 5000 mg/kg.

Parameters examined during the two-week observation period included body weights and dermal reactions and systemic toxicity. All animals were examined for gross pathologic changes. All animals survived the 5000 mg/kg limit test and therefore no other dose level was tested. All animals were in apparent good health throughout the study and gained weight by study termination. It was therefore concluded that under the conditions of this study, the acute oral LD50 of the test material was greater than 5000 mg/kg.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
7 December 1992 and 31 December 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.
Qualifier:
according to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd., Manston, Kent, U.K.
- Age at study initiation: 5 - 8 weeks
- Weight at study initiation: males 142 - 161g, females 135 - 146g
- Fasting period before study: overnight fast immediately before dosing
- Housing: groups of five by sex in solid-floor polypropylene cages with sawdust bedding
- Diet and water: overnight fast before dosing and approximately two hours after dosing. Remainder of study ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 22°C
- Humidity (%): 50 - 73%. (On one occasion the humidity and temperature were outside the limits specified in the protocol (19°C and 70% respectively). This was considered not to have affected the purpose or integrity of the study.
- Air changes (per hr): 15/hour
- Photoperiod (hrs dark / hrs light): 12 h dark/12h light


Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
For the purpose of the study the test material was freshly prepared, as required, as a suspension at the appropriate concentration in arachis oil B.P. Homogeneity was assured by the use of a Silverson Homogeniser.
The concentration, homogeneity and stability of the test material preparations were not determined by analysis.
A preliminary range finding study was conducted using a single dose of 2000 mg/kg bodyweight (dose concentration: 200 mg/ml, dose volume: 10 ml/kg).
In the main study, all rats were given a single oral dose of test material, as a suspension in arachis oil B.P. at a dose level of 2000 mg/kg bodyweight (dose concentration: 200 mg/ml, dose volume: 10 ml/kg).
Doses:
Single oral dose by gavage of 2000 mg/kg suspension in arachis oil (range finding and main study). All animals were dosed once only by gavage using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to its fasted bodyweight at the
time of dosing.
No. of animals per sex per dose:
Range finding study: one male, one female
Main study: five male, five female
Control animals:
no
Details on study design:
- Duration of observation period following administration: 0.5, 1, 2 and 4 h after dosing and subsequently once daily for 14 days
- Frequency of observations and weighing: Individual bodyweights were recorded prior to dosing on Day 0 and on Days 7 and 14
- Necropsy of survivors performed: range finding study, no; main study, yes
- Other examinations performed: clinical signs, body weight
Statistics:
Data evaluations included the relationship, if any, between the animals' exposure to the test material and the incidence and severity of all abnormalities including behavioural and clinical observations, gross lesions, bodyweight changes, mortality and any other toxicological effects.
Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test material was made.
Preliminary study:
In the rangefinding study, no deaths or signs of toxicity were noted. A main study was therefore conducted on five male and five female rats using the same dosage.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortalities.
Clinical signs:
No signs of systemic toxicity.
Black-coloured staining of the fur was noted in all animals during the study.
Body weight:
All animals showed expected gain in bodyweight during the study.
Gross pathology:
No abnormalities were noted at necropsy.
Interpretation of results:
other: Not classified according to EU criteria.
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg
bodyweight.
Executive summary:

The test material was assessed in an acute oral toxicity study conducted according to OECD Test Method 401. Following a range-finding study, a group of ten fasted animals (five males and five females) was given a single oral dose of test material, as a suspension in arachis oil B.P. at a dose level of 2000 mg/kg bodyweight. The animals were observed for fourteen days after the day of dosing and then subjected to gross pathological examination.

There were no deaths. No signs of systemic toxicity were noted during the study. All animals showed expected gain in bodyweight during the study. No abnormalities were noted at necropsy.

The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was therefore found to be greater than 2000 mg/kg bodyweight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

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:
1 October 2006 to 25 October 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate, Kent
- Age at study initiation: approximately eight to twelve weeks old
- Weight at study initiation: within the weight range of 200g to 350g
- Housing: housed in groups of five by sex in solid-floor polypropylene cages with stainless steel lids, furnished with softwood flakes (Datesand Ltd., Cheshire, UK) and provided with environmental enrichment items: wooden chew blocks (B & K Universal Ltd, Hull, UK) and cardboard "fun tunnels" (Datesand Ltd., Cheshire, UK).
- Diet (e.g. ad libitum): With the exception of the exposure period, free access to food (EU Rodent Diet 5LF2, BCM IPS Limited, London, UK) was allowed throughout the study.
- Water (e.g. ad libitum): free access to mains drinking water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 °C
- Humidity (%):30 - 70%
- Air changes (per hr): at least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): give twelve hours continuous light and twelve hours darkness
Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
Atmosphere Generation
A dust atmosphere was produced from the test material using a SAG 410 Solid Aerosol Generator (TOPAS GmbH, Dresden, Germany) located adjacent to the exposure chamber. The SAG 410 was connected to a metered compressed air supply.
Compressed air was supplied by means of an oil free compressor and passed through a water trap and respiratory quality filters before it was introduced to the SAG 410.
The cylindrical exposure chamber had a volume of approximately 30 litres (dimensions: 28 cm diameter x 50 cm high). The concentration within the chamber was controlled by adjusting the test material feed rate from the SAG 410. The extract from the exposure chamber passed through a 'scrubber' trap and was connected with a high efficiency filter to a metered exhaust system. The chamber was maintained under negative pressure.
Homogeneity of the test atmosphere within the chamber was not specifically determined during this study. Chambers of the same design (ADG Developments Ltd, Hitchin, Herts, UK) have been fully validated and shown to produce evenly distributed atmospheres in the animals' breathing zone with a wide variety of test materials (Green J D et al, 1984).
Prior to the start of the study, test material atmospheres were generated within the exposure chamber. During this characterisation period air flow settings and test material input rates were varied to achieve the required atmospheric concentrations.

Exposure Procedure
Each rat was individually held in a tapered, polycarbonate restraining tube fitted onto a single tier of the exposure chamber and sealed by means of a rubber '0' ring. Only the nose of each animal was exposed to the test atmosphere.
Following an appropriate equilibration period a single group of ten rats (five males and five females) was exposed to an atmosphere of the test material for a period of four hours. A target concentration of 5.0 mg/L was used for the exposure. As the mean achieved concentration was 100% of target and no deaths occurred, no further levels were required.

Exposure Chamber Temperature and Relative Humidity
The temperature and relative humidity inside the exposure chamber were measured by an electronic thermometer/humidity meter (Hanna Instruments Ltd, Beds., UK) located in a vacant port in the animals' breathing zone of the chamber and recorded every thirty minutes throughout the four-hour exposure period.

Exposure Chamber Oxygen Concentration
Oxygen levels within the exposure chamber were measured by an electronic oxygen analyser (Servomex (UK) Ltd, Crowborough, East Sussex) located in a sampling port in the animals breathing zone during each exposure period. The test atmosphere was generated to contain at least 19% oxygen.

Exposure Chamber Atmosphere Concentration
The actual chamber concentration was measured at regular intervals during the exposure period. The gravimetric method used glass fibre filters placed in a filter holder. The holder was temporarily sealed in a vacant port in the exposure chamber in the animals' breathing zone and a suitable, known volume of exposure chamber air was drawn through the filter using a vacuum pump.
Each filter was weighed before and after sampling in order to calculate the weight of collected test material. The difference in the two weights, divided by the volume of atmosphere sampled, gave the actual chamber concentration.
The nominal chamber concentration was calculated by dividing the mass of test material used by the total volume of air passed through the chamber.

Particle Size Distribution
The particle size of the generated atmosphere inside the exposure chamber was determined three times during the exposure period using a Marple Personal Cascade Impactor (Westech IS Ltd, Beds., UK). This device consisted of six impactor stages (9.6, 6.6, 3.5, 1.8, 0.87 and 0.33 µm cut points) with stainless steel collection substrates and a back up glass fibre filter, housed in an aluminium sampler. The sampler was temporarily sealed in a sampling port in the animals' breathing zone and a suitable, known volume of exposure chamber air was drawn through it using a vacuum pump.
The collection substrates and backup filter were weighed before and after sampling and the weight of test material, collected at each stage, calculated by difference.
The mean amount for each stage was used to determine the cumulative amount below each cut-off point size. In this way, the proportion (%) of aerosol less than 9.6, 6.6, 3.5, 1.8, 0.87 and 0.33 µm was calculated.
The resulting values were converted to probits and plotted against Log10 cut-point size. From this plot, the Mass Median Aerodynamic Diameter (MMAD) was determined (as the 50% point) and the geometric standard deviation was calculated. In addition the proportion (%) of aerosol less than 4 µm (considered to be the inhalable fraction) was determined.
Analytical verification of test atmosphere concentrations:
yes
Remarks:
gravimetric method
Duration of exposure:
4 h
Concentrations:
The mean achieved atmosphere concentration was 5.00 ± 0.20 mg/L. The mean mass median aerodynamic diameter was 3.07 µm, and the inhalable fraction (% < 4 µm) was 60.3 while the geometric standard deviation was 2.78.
No. of animals per sex per dose:
Five males and five females
Control animals:
no
Details on study design:
Observations

Clinical Signs
All animals were observed for clinical signs at hourly intervals during exposure, immediately on removal from the restraining tubes at the end of exposure, one hour after termination of exposure and subsequently once daily for fourteen days. Any evidence of overt toxicity was recorded at each observation.

Bodyweight
Individual bodyweights were recorded prior to treatment on the day of exposure and on Days 7 and 14.

Necropsy
At the end of the fourteen day observation period the animals were killed by intravenous overdose of sodium pentobarbitone. All animals were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded. The respiratory tract was subjected to a detailed macroscopic examination for signs of irritancy or local toxicity.
Statistics:
Data evaluations included the relationship, if any, between the animals' exposure to the test material and the incidence and severity of all abnormalities including behavioural and clinical observations, necropsy findings, bodyweight changes, mortality and any other toxicological effects.
Using the mortality data obtained, an estimate of the acute inhalation median lethal concentration (LC50) of the test material was made.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
other: No deaths occurred
Mortality:
No deaths occurred
Clinical signs:
other: Signs of hunched posture and pilo-erection are commonly seen in animals for short periods on removal from the chamber following 4-hour inhalation studies. Wet fur is commonly recorded both during and for a short period after exposure. Fur staining by the
Body weight:
Variations in bodyweight gain are frequently seen for female animals of this strain and age during this type of study and, in isolation, are considered not to be significant.
Apart from one female animal that showed slight bodyweight loss during Week 2, normal bodyweight development was noted during the study.
Gross pathology:
The following macroscopic abnormalities were detected amongst animals at necropsy:
Lungs — fluid filled, abnormally dark, dark patches.

Exposure Chamber Concentration

The test atmosphere was sampled at approximately fifteen minute intervals during the exposure period and the actual concentration of the test material calculated. The mean values obtained were:

 

Atmosphere Concentration

Mean Achieved (mg/L)

Standard Deviation

Nominal (mg/L)

 

5.00

0.20

8.24

 

The chamber flow rate was maintained at 55 L/min providing 110 air changes per hour.

The theoretical chamber equilibration time (T99) was 3* minutes (Silver, 1946).

* = Atmospheres of the test material were generated prior to animal insertion, therefore, the equilibration period was carried out for a total of 16 minutes.

 

Particle Size Distribution

The particle size analysis of the atmosphere drawn from the animals' breathing zone was as follows:

Mean Achieved

Atmosphere Concentration (mg/L)

Mean Mass Median

Aerodynamic Diameter

(µm)

Inhalable Fraction

(% <4 µm)

Geometric Standard Deviation

 

5.00

3.07

60.3

2.78

 

Interpretation of results:
other: Not classified according to EU criteria.
Conclusions:
No deaths occurred in a group of ten rats exposed to a mean achieved atmosphere concentration of 5.00 mg/L for four hours. It was therefore considered that the acute inhalation median lethal concentration (4 hr LC50) of the test material, in the Sprague-Dawley Crl:CD (SD) IGS BR strain rat, was greater than 5.00 mg/L.
Executive summary:

A study was performed to assess the acute inhalation toxicity of the test material. The method used followed that described in the OECD Guidelines for Testing of Chemicals No. 403 "Acute Inhalation Toxicity". A group of ten Sprague-Dawley strain rats (five male and five female) were exposed to a dust atmosphere for four hours using a nose only exposure system, followed by a fourteen day observation period. Common abnormalities noted during the study included increased respiratory rate, hunched posture, pilo-erection, fur staining by the test material and wet fur. There were frequent instances of laboured respiration. Animals recovered to appear normal from Days 6 to 8 post-exposure. One female rat showed slight bodyweight loss during Week 2 while normal bodyweight development was noted in the remaining 9 rats. Macroscopic abnormalities noting during the study were fluid filled lungs, as well as abnormally dark and dark patched lungs.

No deaths occurred, therefore the acute inhalation median lethal concentration (4h LC50) of the test material in the Sprague-Dawley strain rat was greater than 5.00 mg/L.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
Value:
5 000 mg/m³

Acute toxicity: via dermal route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 September 1992 to 30 September 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd., Manston , Kent, UK
- Age at study initiation: 10 - 14 weeks
- Weight at study initiation: males: 208 - 231g; females: 211 - 235 g
- Fasting period before study: None
- Housing:suspended polypropylene cages furnished with softwood sawdust, individually for 24 h exposure, followed by groups of 5 by sex for remainder of study.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 22 °C
- Humidity (%): 60 - 75%. On two occasions the humidity was above the limit specified in the protocol (70%). This was considered not to have affected the purpose or integrity of the study.
- Air changes (per hr): ca. 15/hr
- Photoperiod (hrs dark / hrs light): 12 h continuous light, 12 h continuous dark
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: 5 cm x 4 cm. The appropriate amount of the test material was applied uniformly to the shorn skin which had previously been moistened with arachis oil B.P.
- % coverage: ca. 10% of total body surface
- Type of wrap if used: surgical gauze measuring 7 cm x 4 cm was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage (HYPERTIE). The bandage was further secured with a piece of BLENDERM wrapped around each end.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): cotton wool moistened with acetone
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg


Duration of exposure:
24 h
Doses:
2000 mg/kg bodyweight
No. of animals per sex per dose:
5 male, 5 female
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observed 0.5, 1, 2, and 4 hours after dosing and subsequently once daily for 14 days for deaths, overt signs of toxicity and any adverse dermal reactions. Individual bodyweights were recorded prior to application of the test material on Day 0 and on Days 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, histopathology
At the end of the study the animals were killed by cervical dislocation and subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.
Statistics:
Data evaluations include the relationship, if any, between the animals' exposure to the test material and the incidence and severity of all abnormalities (behavioural, clinical observations, gross lesions, bodyweight changes, mortality and other toxicological effects). Using the mortality data obtained, an estimate of the acute dermal median lethal dose (LD50) of the test material was made.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No deaths.
Clinical signs:
No signs of systemic toxicity were noted. Black coloured staining of fur was commonly noted.
Body weight:
All animals showed expected gain in bodyweight during the study.
Gross pathology:
No abnormalities were noted at necropsy.
Interpretation of results:
other: Not classified according to EU criteria.
Conclusions:
The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg bodyweight.
Executive summary:

A study was performed to assess the acute dermal toxicity of the test material in the Sprague-Dawley strain rat. The method used followed that described in the OECD Guidelines for Testing of Chemicals (1981) No. 402 "Acute Dermal Toxicity". A group of ten animals (five males and five females) was given a single 24-hour, semi-occluded dermal application to intact skin at a dose level of 2000 mg/kg bodyweight. The animals were observed for fourteen days after the day of treatment and then subjected to gross pathological examination. There were no deaths. No signs of systemic toxicity or skin irritation were noted during the study. All animals showed expected gain in bodyweight during the study. No abnormalities were noted at necropsy.

The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg bodyweight.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
3 March 1992 to 17 March 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River UK Ltd., Manston Kent, UK
- Age at study initiation: 10-14 weeks
- Weight at study initiation: male: 212-236 g; female: 202-218 g
- Fasting period before study: none
- Housing: individually in solid floor polypropylene cages containing with softwood sawdust bedding during 24 h exposure, then in groups of 5 by sex in cages with polypropolyene grid floors suspended over trays lined with absorbent paper.
- Diet: ad libitum throughout study
- Water: ad libitum throughout study
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22 °C
- Humidity (%): 52-53 %
- Air changes (per hr): 15/h
- Photoperiod (hrs dark / hrs light): 12h dark/12h light

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: 5 cm x 4 cm. The appropriate amount of the test material was applied uniformly to the shorn skin which had previously been moistened with arachis oil B.P.
- % coverage: ca. 10% of total body surface
- Type of wrap if used: surgical gauze measuring 7 cm x 4 cm was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage (HYPERTIE). The bandage was further secured with a piece of BLENDERM wrapped around each end.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): cotton wool moistened with arachis oil
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg

Duration of exposure:
24h
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 male and 5 female
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observed after 0.5, 1, 2, and 4 h and subsequently once per day for 14 d for deaths, overt signs of toxicity and any adverse dermal reactions. Individual bodyweights were recorded prior to application of the test material on Day 0 and on Days 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, organ weights, histopathology
At the end of the study the animals were killed by cervical dislocation and subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.
Statistics:
Data evaluations included the relationship, if any, between the animals' exposure to the test material and the incidence and severity of all abnormalities including behavioural and clinical observations, gross lesions, bodyweight changes, mortality and any other toxicological effects.
Using the mortality data obtained, an estimate of the acute dermal median lethal dose (LD50) of the test material was made.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No deaths.
Clinical signs:
No signs of systemic toxicity or skin irritation were noted during the study. Black-coloured staining of the treatment area and fur was noted in all animals up to three days after dosing.
Body weight:
All animals showed expected bodyweight gain during the study.
Gross pathology:
No abnormalities were noted at necropsy.
Interpretation of results:
other: Not classified according to EU criteria.
Conclusions:
The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg bodyweight.
Executive summary:

A study was performed to assess the acute dermal toxicity of the test material in the Sprague-Dawley strain rat. The method used followed that described in the OECD Guidelines for Testing of Chemicals (1981) No. 402 "Acute Dermal Toxicity". A group of ten animals (five males and five females) was given a single 24-hour, semi-occluded dermal application to intact skin at a dose level of 2000 mg/kg bodyweight. The animals were observed for fourteen days after the day of treatment and were then subjected to gross pathological examination. There were no deaths. No signs of systemic toxicity or skin irritation were noted during the study. All animals showed expected gain in bodyweight during the study. No abnormalities were noted at necropsy.

The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg bodyweight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Additional information

The substance is not acutely harmful or toxic by either oral or dermal exposure routes. These are considered the most relevant routes of exposure for the substance.

In the absence of any other factors key studies have been selected based on aniline content of the batches tested. Batches with known aniline content are judged to be key and in cases where two batches of known aniline content have been tested the one with the higher content is considered key.

Key Study Oral Route: Allen (1993)

The test material was assessed in an acute oral toxicity study conducted according to OECD Test Method 401. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

Following a range-finding study, a group of ten fasted animals (five males and five females) was given a single oral dose of test material, as a suspension in arachis oil B.P. at a dose level of 2 000 mg/kg bodyweight. The animals were observed for fourteen days after the day of dosing and then subjected to gross pathological examination.

There were no deaths. No signs of systemic toxicity were noted during the study. All animals showed expected gain in bodyweight during the study. No abnormalities were noted at necropsy.

The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was therefore found to be greater than 2 000 mg/kg bodyweight.

Supporting Study Oral Route: Walker (1992)

The test material was evaluated for acute oral toxicity in Sprague Dawley strain rats in accordance with OECD Test Method 401. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

A preliminary range finding study was conducted on two rats (1 male, 1 female) in a single oral dose of test material as a suspension in arachis oil at a dose level of 5 000 mg/kg. No deaths or signs of systemic toxicity were noted. In the main study, five rats per sex received a single oral dose of test material as a suspension in arachis oil at a dose level of 5 000 mg/kg.

Parameters examined during the two-week observation period included body weights and dermal reactions and systemic toxicity. All animals were examined for gross pathologic changes. All animals survived the 5 000 mg/kg limit test and therefore no other dose level was tested. All animals were in apparent good health throughout the study and gained weight by study termination. It was therefore concluded that under the conditions of this study, the acute oral LD50 of the test material was greater than 5 000 mg/kg.

 

Key Study Inhalation Route: Griffiths (2007)

A study was performed to assess the acute inhalation toxicity of the test material. The method used followed that described in the OECD Guidelines for Testing of Chemicals No. 403 "Acute Inhalation Toxicity". The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

A group of ten Sprague-Dawley strain rats (five male and five female) were exposed to a dust atmosphere for four hours using a nose only exposure system, followed by a fourteen day observation period. Common abnormalities noted during the study included increased respiratory rate, hunched posture, pilo-erection, fur staining by the test material and wet fur. There were frequent instances of laboured respiration. Animals recovered to appear normal from Days 6 to 8 post-exposure. One female rat showed slight bodyweight loss during Week 2 while normal bodyweight development was noted in the remaining 9 rats. Macroscopic abnormalities noting during the study were fluid filled lungs, as well as abnormally dark and dark patched lungs.

No deaths occurred therefore the acute inhalation median lethal concentration (4h LC50) of the test material in the Sprague-Dawley strain rat was greater than 5.00 mg/L.

 

Key Study Dermal Route: Walter (1992)

A study was performed to assess the acute dermal toxicity of the test material in the Sprague-Dawley strain rat. The method used followed that described in the OECD Guidelines for Testing of Chemicals (1981) No. 402 "Acute Dermal Toxicity". The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

A group of ten animals (five males and five females) was given a single 24-hour, semi-occluded dermal application to intact skin at a dose level of 2 000 mg/kg bodyweight. The animals were observed for fourteen days after the day of treatment and then subjected to gross pathological examination. There were no deaths. No signs of systemic toxicity or skin irritation were noted during the study. All animals showed expected gain in bodyweight during the study. No abnormalities were noted at necropsy.

The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2 000 mg/kg bodyweight.

Supporting Study Dermal Route: Walker (1992)

A study was performed to assess the acute dermal toxicity of the test material in the Sprague-Dawley strain rat. The method used followed that described in the OECD Guidelines for Testing of Chemicals (1981) No. 402 "Acute Dermal Toxicity". The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

A group of ten animals (five males and five females) was given a single 24-hour, semi-occluded dermal application to intact skin at a dose level of 2 000 mg/kg bodyweight. The animals were observed for fourteen days after the day of treatment and were then subjected to gross pathological examination. There were no deaths. No signs of systemic toxicity or skin irritation were noted during the study. All animals showed expected gain in bodyweight during the study. No abnormalities were noted at necropsy.

The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2 000 mg/kg bodyweight.

Justification for classification or non-classification

Based upon the high LD50 values for oral, inhalation and dermal exposure, and the absence of other major significant effects, the registered substance does not need to be classified for acute toxicity according to Regulation (EC) No 1272/2008.