Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Oral: measured LD50 > 300 and < 2000 mg/kg bw and the estimated LD50 cut-off was considered to be 1000 mg/kg bw, female rat, OECD TG 423, 2015

Inhalation: measured LC50 > 6.43 mg/L (mean achieved concentration), male/female rat, OECD TG 403, 2015

Dermal: measured LD50 > 2000 mg/kg bw and the estimated LD50 cut-off value was considered to be > 5000 mg/kg bw, male/female rat, OECD TG 402, 2015

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:
18-11-2014 to 25-02-2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met.
Justification for type of information:
Information as to the availability of the in vivo study is provided in 'attached justification'.
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries, Test Data for Registration of Agricultural Chemicals, Acute oral toxicity (2-1-1), 12 Nousan No 8147, Agricultural Production Bureau, November 24, 2000
GLP compliance:
yes (incl. QA statement)
Remarks:
inspected: July 2014; signature: September 2014
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Remarks:
RccHan: WIST
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Recognised supplier
- Females (if applicable) nulliparous and non-pregnant: Yes.
- Age at study initiation: 8 - 9 weeks.
- Weight at study initiation: 161 - 182 g
- Fasting period before study: Overnight before dosing and approximately four hours after dosing.
- Housing: Group housed in groups of up to three same sex in suspended solid-floor polycarbonate cages furnished with softwood flakes and aspen chew block as cage enrichment..
- Diet (e.g. ad libitum): Certified diet from recognised supplier, provided ad libitum (except for fasting period).
- Water (e.g. ad libitum): ad libitum (except for fasting period)
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 23
- Humidity (%): 40 - 70%
- Air changes (per hr): Not reported. Positive pressure, filtered air environment.
- Photoperiod: 12 h light / 12 h dark

IN-LIFE DATES: From: To: 2014-11-20 to 2014-12-16
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: The test item was formulated at concentrations of 30 or 200 mg/mL in the vehicle and administered at a volume of 10 mL/kg body weight. Formulations were prepared on the day of dosing.
- Amount of vehicle (if gavage): The concentration of the test item in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed.
- Lot/batch no. (if required): See full study report.
- Purity: See full study report.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw.

DOSAGE PREPARATION (if unusual): Not applicable. The test item was prepared in the vehicle. It was administered to the animals under a volume of 10 mL/kg. The volume administer was adjusted according to bodyweight on day of treatment.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: 300 mg/kg bw based on guideline recommendations. In the absence of data regarding the toxicity of the test item, 300 mg/kg was chosen as the starting dose.
Doses:
300 mg/kg bw in corn oil vehicle (starting dose); 2000 mg/kg bw
No. of animals per sex per dose:
3 initial females (sighting study) and further 3 females (main study); total of up to 6 per dose according to sequential guideline testing strategy.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were made 0.5, 1, 2, and 4 hours after dosing and then daily for fourteen days. Morbidity and mortality checks were made twice daily. Individual body weights were recorded on Day 1 (the day of dosing) and on Days 8 and 15.
- Necropsy of survivors performed: yes
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 mg/kg bw
Based on:
test mat.
Mortality:
300 mg/kg bw: No mortality
2000 mg/kg bw: Two females mortalites (humane termination).
Clinical signs:
other: 300 mg/kg bw: No signs of systemic toxicity were noted. 2000 mg/kg bw: Surviving female signs comprised of piloerection and hunched posture on day 3. Recovery complete by day 11. In the non-survivors, clinical signs prior to humane termination comprised o
Gross pathology:
300 mg/kg bw: No abnormalities were noted at necropsy.
2000 mg/kg bw: No abnormalities were noted at necropsy amongst survivors. In the non-survivors, post humane termination: congestion (characterised by darkened tissues) of the lungs, pallor of the kidneys liver and spleen was observed.
Other findings:
- Organ weights: Not reported.
- Histopathology: Not reported. No macropathological abnormalities amongst survivors.
- Potential target organs: Not applicable.
- Other observations: Not applicable.
Interpretation of results:
Category 4 based on GHS criteria
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study the oral LD50 was established to be > 300 and < 2000 mg/kg bw in female Wistar rats.
Executive summary:

The study was performed according to OECD TG 423, EU Method B.1 tris, US EPA OPPTS 870.1100 and Japan Acute Oral Toxicity (2-1-1) (2000) guidelines in accordance with GLP to assess the acute oral toxicity of the test item by the acute toxic class method following a single oral administration in the female Wistar RccHan: WIST strain rat. The test item was administered by oral gavage in corn oil vehicle at a dose volume of approximately 10 mL/kg in an initial step at 300 mg/kg in three females. In the absence of significant toxicity, at this dose level the test item was then administered again in a further three females. In the absence of toxicity the test item was finally administered at 2000 mg/kg bw in corn oil by oral gavage to a group of three females. At 300 mg/kg bw there was no mortality, no significant clinical signs and bodyweight gains were as expected. There was no abnormalities at necropsy. At 2000 mg/kg bw, two of three females were humanely terminated at day 2 due to poor clinical condition. Clinical signs prior to death comprised of unsteady gait, decreased activity, unresponsive behaviour, cold to touch, piloerection, hunched posture and reduced body tone, flattened posture, unresponsive behaviour, flattened posture, lachrymation (both eyes), splayed hindlimbs, shallow breathing and partially closed eye lids (both eyes). Signs were seen from Day 2 for both females. A loss in body weight was noted for both of the decedents. Macroscopic examination of the animals revealed congestion (characterised by darkened tissues) of the lungs, pallor of the kidneys liver and spleen. The survivor at 2000 mg/kg bw comprised clinical signs of piloerection and hunched posture. These signs were first noted on Day 3. Recovery of the surviving animal, as judged by external appearance and behaviour, was complete by Day 11. Survivors had expected bodyweight gain and there were no abnormalities at necropsy. Under the conditions of this study the oral LD50 was established to be > 300 and < 2000 mg/kg bw in the female Wistar rat.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
discriminating dose
Value:
300 mg/kg bw
Quality of whole database:
The available information as a whole meets the tonnage driven information requirements of REACH.

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:
25-03-2015 to 16-07-2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met with acceptable minor deviations not considered to impact the validity of the study.
Justification for type of information:
Information as to the availability of the in vivo study is provided in 'attached justification'.
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
yes
Remarks:
three samples (total n=9) within 4 hour period were outside ±20% (two lower and one higher) the mean achieved concentration; not considered to impact the study; see "overall remarks, attachments" for further information
Qualifier:
according to guideline
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Deviations:
yes
Remarks:
See above
GLP compliance:
yes (incl. QA statement)
Remarks:
inspected: June 2015; signature: September 2015
Test type:
traditional method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Recognised supplier
- Females (if applicable) nulliparous and non-pregnant: Yes.
- Age at study initiation: 8 - 12 weeks; females were nulliparous and non pregnant.
- Weight at study initiation: 200 - 350 g
- Fasting period before study: None.
- Housing: Housed in groups of up to five by sex in solid-floor polypropylene cages with stainless steel lids, furnished with softwood flakes provided with environmental enrichment items.
- Diet (e.g. ad libitum): Certified diet from recognised supplier, provided ad libitum (except for exposure period period)
- Water (e.g. ad libitum): ad libitum (except for exposure period period)
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25
- Humidity (%): 30 - 70%
- Air changes (per hr): > 15 air changes per hour
- Photoperiod: 12 h light / 12 h dark

IN-LIFE DATES: 25-03-2015 to 18-06-2015
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
air
Remarks:
Oxygen concentration (%) was monitored during the study by an electronic oxygen analyzer. The test atmospheres were generated to contain at least 19% oxygen.
Mass median aerodynamic diameter (MMAD):
2.86 µm
Geometric standard deviation (GSD):
2.23
Remark on MMAD/GSD:
MMAD and GSD associated with the highest concentration level: 6.43 mg/L
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: The test item was aerosolized using a glass concentric jet nebulizer located at the top of the exposure chamber. The nebulizer was connected to a glass syringe attached to an infusion pump, which provided a continuous supply of test item under pressure, and to a metered compressed air supply.
- Exposure chamber volume: approximately 30 litres (dimensions: 28 cm diameter x 50 cm high)
- Method of holding animals in test chamber: Prior to the day of exposure each rat was acclimatized (for approximately 2 hours) to a tapered polycarbonate restraining tube. During the exposure period, 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 ‘O’ ring. Only the nose of each animal was exposed to the test atmosphere.
- Source and rate of air: filtered air; chamber flow rate was maintained at 60 L/min providing 120 air changes per hour.
- Method of conditioning air: 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 nebulizer.
- System of generating particulates/aerosols: glass concentric jet nebulizer; the concentration within the exposure chamber was controlled by adjusting the rate of the infusion pump.
- Method of particle size determination: Particle size was determined using a cascade impactor. The device consisted of six impactors stages. The mean amount for each stage was used to determine the cumulative amount below each cut-off point size to calculate the proportional (%) aerosol of defined size ranges. 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.
- Treatment of exhaust air: 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. A schematic of the dynamic (continuous flow) system is presented in the full study report.
- Temperature, humidity, in air chamber: The temperature and relative humidity inside the exposure chamber were measured by an electronic thermometer/humidity meter: 19 to 21 °C and 30 to 70% humidity. The relative humidity within the exposure chamber during the exposure was found to be lower (28 – 29 %) during the second exposure period than the range specified in the inhalation test guidelines (30 – 70 %). The decreased humidity may well have been due to the nature of the test item, the decreased humidity was therefore considered to be an accurate representation within the exposure chamber and is considered not to have affected the purpose or validity of the study.

TEST ATMOSPHERE
- Brief description of analytical method used: The test atmosphere was sampled nine times during each exposure period. The sampling procedure involved two liters or five liters of test atmosphere being drawn through a series of two glass impingers (dependent on concentration) containing Methanol (each made up to 80mL). The samples were then submitted for chemical analysis by gas chromatography (GC). The test impinger samples received were diluted with methanol to achieve the working concentration. The solvent (methanol) and a blank impinger (control) were analysed. Neither the solvent nor the blank impinger produced a signal that interfered with the signal due to the test item. A range of standard solutions were prepared in methanol from a stock solution of 1.030 mg/mL by serial dilution covering the concentration range 0 to 0.1545 mg/mL. Full details of the analytical method are provided in the full study report. The linearity of the analytical system used for sample analyses was demonstrated with a good relationship between peak areas measured and working standard concentrations. The regression coefficients calculated were found to be 0.9999. The fortified samples of impingers were found to have a recovery value of ± 10% of the fortification. the chromatographic run. In conclusion, the results indicate the accurate use of the test item and impingers during this study. Working solutions of the test item were shown to be sufficiently stable to complete the analysis without deterioration of the analyte. Full details of the analytical method are provided in the full study report.
- Samples taken from breathing zone: yes

TEST ATMOSPHERE
- Particle size distribution: The particle size of the generated atmosphere inside the exposure chamber was determined three times during each exposure period using a cascade impactor. The particle size distribution for each group is reported in table 1.
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): The Mass Median Aerodynamic Diameter (MMAD) was determined and is reported for each group in table 1.
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
Following an appropriate equilibration period three groups were subjected to a single exposure to the test item for a period of four hours. Based on the expected toxicity of the test item, a target concentration of 2.0 mg/L was used for the first exposure. Further concentrations were selected after consideration of the results of the previous exposure. Full details are provided in table 2.
No. of animals per sex per dose:
5 per sex per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 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 up to fourteen days. Any evidence of mortality or overt toxicity was recorded at each observation. Individual body weights were recorded on arrival, prior to treatment on the day of exposure and on Days 1, 3, 7 and 14 or after mortality.
- Necropsy of survivors performed: yes (and in the event of any mortalities)
- Other examinations performed: clinical signs, bodyweight, organ weights, and any other relevant toxicological effects were reported.
Statistics:
Using the mortality data obtained, an estimate of the acute inhalation median lethal concentration (LC50) was calculated using validated data analysis software which utilized Log-Normal (Probit) regression models in order to calculate the LC50 values. LC50 values and 95% confidence limits were calculated for males and females separately. Where appropriate.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 6.43 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
other: Effect levels based on analytically confirmed: mean achieved concentrations for each exposure/sex groups
Sex:
male
Dose descriptor:
LC50
Effect level:
> 6.43 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
other: Effect levels based on analytically confirmed: mean achieved concentrations for each exposure/sex groups
Sex:
female
Dose descriptor:
LC50
Effect level:
> 6.43 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
other: Effect levels based on analytically confirmed: mean achieved concentrations for each exposure/sex groups
Mortality:
There were no mortalities during the study as reported in table 3.
Clinical signs:
other: Common abnormalities noted during the study included increased respiratory rate, or decreased respiratory rate, noisy respiration, hunched posture, pilo-erection and wet fur. One female also demonstrated sneezing. All organisms in group 2 (6.43 mg/L mean
Body weight:
In group 2, 6.43 mg/L: All males and females exhibited body weight losses on Day 1 post-exposure. Body weight gains were noted in all males/females during the remainder of the recovery period.
In group 1, 2.14 mg/L: All males and four female animals exhibited body weight losses on Day 1 post-exposure. All male animals exhibited body weight gains during the remainder of the recovery period. One female animal showed no body weight gain from Days 1 to 3 post-exposure, however, body weight gains were noted in all females during the remainder of the recovery period.

All males/females were gaining bodyweight at the end of the recovery period.
Gross pathology:
In group 2, 6.43 mg/L: Abnormally red lungs or dark patches on the lungs were noted in two males (out of five) and two females (out of five) at necropsy.
In group 1, 2.14 mg/L: Dark patches on the lungs were noted in two males (out of five) and one female (out of five) at necropsy.
Other findings:
- Other observations: The respiratory tract was subjected to a detailed macroscopic examination for signs of irritancy or local toxicity during necropsy.

Table 1. Characteristics of the achieved atmosphere:

Group Number

Mean Achieved Atmosphere Concentration (mg/L)

Mean Mass Median Aerodynamic Diameter (µm)

Inhalable Fraction

(% <4 µm)

Geometric Standard Deviation

1

2.14

2.41

77.7

1.96

2

6.43

2.86

66.3

2.23

 

Table 2. Mean achieved atmosphere concentrations:

Group Number

Atmosphere Concentration

Mean Achieved (mg/L)

Standard Deviation

Nominal (mg/L)

1

2.14

0.04

6.87

2

6.43

1.92

21.7

 

Table 3. Mortality data summary:

Group Number

Mean Achieved Atmosphere Concentration

(mg/L)

Deaths

Male

Female

Total

1

2.14

0/5

0/5

0/10

2

6.43

0/5

0/5

0/10

The target concentrations were group 1: 2.00 mg/L and group 2: 5.00 mg/L, respectively

Interpretation of results:
GHS criteria not met
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study, the inhalation LC50 (male/female) was: > 6.43 mg/L within the RCCHan WIST rat.
Executive summary:

The study was performed according to OECD TG 403 and EU Method B.2 in accordance with GLP to assess the acute inhalation toxicity of the test item. Two groups of ten RccHanTM : WIST strain rats (five males and five females) were exposed to an aerosol atmosphere of the test item. The groups were exposed for four hours using a nose only exposure system, followed by a fourteen day observation period. The mean achieved atmosphere concentrations were as follows: Group 1: 2.14 mg/L and Group 2: 6.43 mg/L. The characteristics of the achieved atmosphere where Mean Mass Median Diameter (particle size) and Inhalable Fraction <4 μm: Group 1: 2.41 μm and 77.7% and Group 2: 2.86 μm and 66.3%. The Geometric Standard Deviation was Group 1: 1.96 and Group 2: 2.23, respectively. There were no male or female mortalities in Group 1 or Group 2. In Group 1, all males and four female animals exhibited body weight losses on Day 1 post-exposure. All male animals exhibited body weight gains during the remainder of the recovery period. One female animal showed no body weight gain from Days 1 to 3 post-exposure, however, body weight gains were noted in all females during the remainder of the recovery period.. Within Group 2, all males and females exhibited body weight losses on Day 1 post-exposure. Body weight gains were noted in all males/females during the remainder of the recovery period. In Group 1, One day after exposure, all animals exhibited increased respiratory rate, hunched posture and pilo-erection and one female animal also exhibited sneezing, which persisted for three days. All animals subsequently appeared normal on Day 5 post-exposure. In Group 2, on removal from the chamber all animals exhibited noisy respiration, there were frequent instances of increased respiratory rate and isolated occurrences of decreased respiratory rate. One hour post-exposure, all animals exhibited noisy respiration, all male animals exhibited increased respiratory rate and all females exhibited decreased respiratory rate. One day after exposure, all animals exhibited hunched posture and pilo-erection and there were frequent instances of increased respiratory rate. One male animal also exhibited decreased respiratory rate and noisy respiration. All males and females appeared normal from days 5 to 6 post-exposure. During necropsy in Group 1, dark patches on the lungs were noted in two males (out of five) and one female (out of five). In Group 2, abnormally red lungs or dark patches on the lungs were noted in two males (out of five) and two females (out of five) at necropsy. Under the conditions of this study, the inhalation LC50 (male/female) was > 6.43 mg/L within the RCCHan WIST rat.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating conc.
Value:
6 430 mg/m³ air
Quality of whole database:
The available information as a whole meets the tonnage driven information requirements of REACH.

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:
08-07-2015 to 12-11-2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met.
Justification for type of information:
Information as to the availability of the in vivo study is provided in 'attached justification'.
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)
Remarks:
inspected: June 2015; signature: September 2015
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Remarks:
RccHan : WIST
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Recognised supplier
- Females (if applicable) nulliparous and non-pregnant: Yes.
- Age at study initiation: 8 - 12 weeks.
- Weight at study initiation: Males: 234 - 260 g ; Females: 201 - 212 g; the weight variation did not exceed ±20% of the mean weight for each sex.
- Fasting period before study: Not applicable
- Housing: suspended solid floor polypropylene cages furnished with woodflakes; the initial two animals were housed individually throughout the study. The further group of eight animals (four male and four female) were housed individually during the 24 Hour exposure period and in groups of four, by sex, for the remainder of the study.
- Diet (e.g. ad libitum): Certified diet from recognised supplier, provided ad libitum.
- Water (e.g. ad libitum): ad libitum.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25
- Humidity (%): 30 - 70%
- Air changes (per hr): > 15 air changes per hour
- Photoperiod: 12 h light / 12 h dark

IN-LIFE DATES:08-07-2015 to 18-08-2015
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: the day before treatment the back and flanks were clipped free of hair. Dorsal area application.
- % coverage: Approximately 10% of total body surface
- Type of wrap if used: The area of application was covered piece of surgical gauze was placed over the treatment area and semi occluded with a piece of self-adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated skin and surrounding hair wiped with cotton wool moistened with arachis oil BP to remove any residual test item
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg
- Concentration (if solution): Not applicable.
- Constant volume or concentration used: Dose volume was ca. 2.16 mL/kg. Dose level was adjusted by bodyweight.
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 per sex per dose (5 male/5 female); treated sequentially following application to 1 sentinel per sex per dose .
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations and mortality checks were conducted at approximately 0.5, 1, 2, and 4 hours and subsequently once daily for 14 days. Local effects were examined once daily for 14 days after the completion of the 24-hour exposure period. Full details on the scoring and criteria (consistent with Draize) are given in the full study report. Individual bodyweights were recorded prior to application of the test item on Day 0 and on Days 7 and 14.
- Necropsy of survivors performed: yes
Statistics:
No statistical analyses were performed.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality was observed.
Clinical signs:
other: - Clinical observations: No signs of systemic toxicity were noted during the observation period. - Dermal reactions: Crust formation was noted at the test site of one male 2 to 5 days after dosing. No signs of dermal irritation were noted at the test site
Gross pathology:
No abnormalities were noted at necropsy.
Other findings:
- Organ weights: Not reported.
- Histopathology: Not reported. No macropathological abnormalities.
- Potential target organs: Not applicable.
- Other observations: Not applicable.
Interpretation of results:
GHS criteria not met
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study the dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar rat. Under the conditions of this study and under the Globally Harmonized Classification System of Classification and Labelling of Chemicals (GHS), the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.
Executive summary:

The study was performed according to OECD TG 402 and EU Method B.3 in accordance with GLP to assess the acute dermal toxicity of the test substance in the Wistar strain rat. A group of ten animals (five males and five females) was given a single, 24 hour, semi-occluded dermal application of the undiluted test item to intact skin at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy. There were no signs of system toxicity. There was no mortality during the study. No signs of dermal irritation were noted. Crust formation was noted at the test site of one male 2 to 5 days after dosing. Animals showed expected gains in body weight, except for two females which showed no gain in body weight during the first week but expected gain in body weight during the second week. All animals gained weight during the study. There was no abnormalities on necropsy. Under the conditions of this study, the dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar rat. Under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
The available information as a whole meets the tonnage driven information requirements of REACH.

Additional information

ORAL: Key Study: OECD TG 420, 2015 - The study was performed according to OECD TG 423, EU Method B.1 tris, US EPA OPPTS 870.1100 and Japan Acute Oral Toxicity (2-1-1) (2000) guidelines in accordance with GLP to assess the acute oral toxicity of the test item by the acute toxic class method following a single oral administration in the female Wistar RccHan: WIST strain rat. The test item was administered by oral gavage in corn oil vehicle at a dose volume of approximately 10 mL/kg in an initial step at 300 mg/kg in three females. In the absence of significant toxicity, at this dose level the test item was then administered again in a further three females. In the absence of toxicity the test item was finally administered at 2000 mg/kg bw in corn oil by oral gavage to a group of three females. At 300 mg/kg bw there was no mortality, no significant clinical signs and bodyweight gains were as expected. There was no abnormalities at necropsy. At 2000 mg/kg bw, two of three females were humanely terminated at day 2 due to poor clinical condition. Clinical signs prior to death comprised of unsteady gait, decreased activity, unresponsive behaviour, cold to touch, piloerection, hunched posture and reduced body tone, flattened posture, unresponsive behaviour, flattened posture, lachrymation (both eyes), splayed hindlimbs, shallow breathing and partially closed eye lids (both eyes). Signs were seen from Day 2 for both females. A loss in body weight was noted for both of the decedents. Macroscopic examination of the animals revealed congestion (characterised by darkened tissues) of the lungs, pallor of the kidneys liver and spleen. The survivor at 2000 mg/kg bw comprised clinical signs of piloerection and hunched posture. These signs were first noted on Day 3. Recovery of the surviving animal, as judged by external appearance and behaviour, was complete by Day 11. Survivors had expected bodyweight gain and there were no abnormalities at necropsy. Under the conditions of this study the oral LD50 was established to be > 300 and < 2000 mg/kg bw in the female Wistar rat. The test item was classified as Acute Oral Toxicity: Category 4 according to Regulation (EC) 1272/2008.

INHALATION: Key Study: OECD TG 403, 2015 - The study was performed according to OECD TG 403 and EU Method B.2 in accordance with GLP to assess the acute inhalation toxicity of the test item. Two groups of ten RccHanTM : WIST strain rats (five males and five females) were exposed to an aerosol atmosphere of the test item. The groups were exposed for four hours using a nose only exposure system, followed by a fourteen day observation period. The mean achieved atmosphere concentrations were as follows: Group 1: 2.14 mg/L and Group 2: 6.43 mg/L. The characteristics of the achieved atmosphere where Mean Mass Median Diameter (particle size) and Inhalable Fraction <4 μm: Group 1: 2.41 μm and 77.7% and Group 2: 2.86 μm and 66.3%. The Geometric Standard Deviation was Group 1: 1.96 and Group 2: 2.23, respectively. There were no male or female mortalities in Group 1 or Group 2. In Group 1, all males and four female animals exhibited body weight losses on Day 1 post-exposure. All male animals exhibited body weight gains during the remainder of the recovery period. One female animal showed no body weight gain from Days 1 to 3 post-exposure, however, body weight gains were noted in all females during the remainder of the recovery period.. Within Group 2, all males and females exhibited body weight losses on Day 1 post-exposure. Body weight gains were noted in all males/females during the remainder of the recovery period. In Group 1, One day after exposure, all animals exhibited increased respiratory rate, hunched posture and pilo-erection and one female animal also exhibited sneezing, which persisted for three days. All animals subsequently appeared normal on Day 5 post-exposure. In Group 2, on removal from the chamber all animals exhibited noisy respiration, there were frequent instances of increased respiratory rate and isolated occurrences of decreased respiratory rate. One hour post-exposure, all animals exhibited noisy respiration, all male animals exhibited increased respiratory rate and all females exhibited decreased respiratory rate. One day after exposure, all animals exhibited hunched posture and pilo-erection and there were frequent instances of increased respiratory rate. One male animal also exhibited decreased respiratory rate and noisy respiration. All males and females appeared normal from days 5 to 6 post-exposure. During necropsy in Group 1, dark patches on the lungs were noted in two males (out of five) and one female (out of five). In Group 2, abnormally red lungs or dark patches on the lungs were noted in two males (out of five) and two females (out of five) at necropsy. Under the conditions of this study, the inhalation LC50 (male/female) was > 6.43 mg/L within the RCCHan WIST rat.

DERMAL: Key Study: OECD TG 402, 2015 - The study was performed according to OECD TG 402 and EU Method B.3 in accordance with GLP to assess the acute dermal toxicity of the test substance in the Wistar strain rat. A group of ten animals (five males and five females) was given a single, 24 hour, semi-occluded dermal application of the undiluted test item to intact skin at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy. There were no signs of system toxicity. There was no mortality during the study. No signs of dermal irritation were noted. Crust formation was noted at the test site of one male 2 to 5 days after dosing. Animals showed expected gains in body weight, except for two females which showed no gain in body weight during the first week but expected gain in body weight during the second week. All animals gained weight during the study. There was no abnormalities on necropsy. Under the conditions of this study, the dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar rat. Under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.

Justification for classification or non-classification

The substance meets classification criteria under Regulation (EC) No 1272/2008 for acute toxicity: oral: category 4: H302

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for acute toxicity: inhalation

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for acute toxicity: dermal