Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Oral: LD50 > 300 - < 2000 mg/kg bw, female rat, OECD TG 420, 2016

Inhalation: LC50 (female): 2.72 (C.I. 1.88 – 3.55) mg/L, OECD TG 403, 2016

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, 2016

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:
2016
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 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
inspected: June 2015; signature: September 2015
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Recognised supplier
- Age at study initiation: 8 - 12 weeks.
- Weight at study initiation: 171-179 g (300 mg/kg including sighting test; sentinel); 179 g (2000 mg/kg sighting test; sentinel); The weight variation did not exceed ±20% of the mean weight in the definitive test (300 mg/kg bw).
- Fasting period before study: Overnight before dosing and three to four hours after dosing.
- Housing: Group housed in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes.
- 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 - 25
- Humidity (%): 30 - 70%
- Air changes (per hr): > 15 air changes per hour
- Photoperiod: 12 h light / 12 h dark

IN-LIFE DATES: From: To: 2015-12-16 to 2016-01-06
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
The specific gravity was determined and used to calculate the appropriate dose volume for the required dose level.
For the purpose of the 300 mg/kg dose level the test item was freshly prepared, as required, as a solution in arachis oil BP. Arachis oil BP was used because the test item did not dissolve/suspend in distilled water. For the purpose of the 2000 mg/kg dose level, the test item was used as supplied. The 300 mg/kg bw and 2000 mg/kg bw dose levels were treated stepwise. Singuarly and in the absence of mortality or evident toxicity a further group of 4 was tested in the appropriate dose level.
Doses:
300 mg/kg bw (initial sighting test and main study)
2000 mg/kg bw (initial sighting test)
No. of animals per sex per dose:
1 (sighting study) and 4 (main study) as applicable; total 5 per dose (in definitive test).
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 0 (the day of dosing) and on Days 7 and 14.
- Necropsy of survivors performed: yes
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
300 mg/kg bw (sentinel and definitive test): No mortality
2000 mg/kg bw (sentinel): Mortality.
Clinical signs:
300 mg/kg bw (sentinel and definitive test): No signs of systemic toxicity were noted.
2000 mg/kg bw (sentinel): Hunched posture on day of dosing.
Body weight:
300 mg/kg bw (sentinel and definitive test): All animals showed expected bodyweight gains over the study period.
2000 mg/kg bw (sentinel): Not applicable.
Gross pathology:
300 mg/kg bw (sentinel and definitive test): Abnormalities noted at necropsy in 1/5: Epithelial sloughing of the gastric mucosa and non glandular epithelium of the stomach. No abnormalities were noted at necropsy of 4/5.
2000 mg/kg bw (sentinel): Abnormalities noted at necropsy were dark liver, dark kidneys, gaseous stomach and epithelial sloughing of the gastric mucosa and non glandular epithelium of the stomach.
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 estimated to be in the range of 300 - 2000 mg/kg bw in female Wistar rats.
Executive summary:

The study was performed according to OECD TG 420 and EU Method B.1 bis Acute Toxicity (Oral) and in accordance with GLP to assess the acute oral toxicity of the test material following a single oral administration in the female Wistar strain rat by the fixed dose method. The test substance was administered by oral gavage in an initial sighting study at 300 mg/kg bw in a solution in arachis oil BP and following an absence of toxicity then at 2000 mg/kg bw. Subsequently, at a further group of four fasted females was given a single oral dose of test item, at a dose level of 300 mg/kg body weight. The animal treated at a dose level of 2000 mg/kg was found dead 1 day after dosing. There were no mortality at a dose level of 300 mg/kg. Hunched posture was noted during the day of dosing in the animal treated at a dose level of 2000 mg/kg. There were no signs of systemic toxicity and all animals showed expected gains in bodyweight over the study period at a dose level of 300 mg/kg. Abnormalities noted at necropsy of the animal treated at a dose level of 2000 mg/kg, that was found dead, were dark liver, dark kidneys, gaseous stomach and epithelial sloughing of the gastric mucosa and non-glandular epithelium of the stomach. Abnormalities noted at necropsy of one animal, treated at a dose level of 300 mg/kg, were epithelial sloughing of the gastric mucosa and non glandular epithelium of the stomach. No abnormalities were noted at necropsy of four animals treated at a dose level of 300 mg/kg. Under the conditions of this study, the oral LD50 was estimated to be in the range of 300 - 2000 mg/kg bw in the female Wistar rat. The test substance was classified as Acute Oral Toxicity: Category 4 according to Regulation (EC) 1272/2008.

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:
2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Guideline study performed under GLP. All relevant validity criteria were met. Minor deviation: Three males Group 3 were slightly outside ±20% (lower than) of the mean body weights of the Group 2 males. This was 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 males Group 3 were slightly outside ±20% (lower than) of the mean body weights of the Group 2 males. Not considered to impact the validity of the study.
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:
standard acute 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
- 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
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.
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: The test item was aerosolized using a metal 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: metal concentric jet nebulizer; the concentration within the exposure chamber was controlled by adjusting the rate of the infusion pump and air flow settings into the chamber. The chamber flow rate was maintained between 30 to 60 L/min providing 60 to 120 air changes per hour. 30 L/min in group 2 and 3 and 60 L/min in group 1.
- 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 20 °C and 30 to 70% humidity.

TEST ATMOSPHERE
- Brief description of analytical method used: The test atmosphere was sampled nine times during the exposure period. The sampling procedure involved 2 to 10 litres (dependent on target concentration) of test atmosphere being drawn through a glass fibre filter. Each filter was then submitted for chemical analysis by gas chromatography (GC). The test filter samples received were extracted with methanol to achieve the working concentration. Blank filters were accurately fortified with known amounts of test item and either analysed without further procedure or purged with nitrogen prior to analysis. A range of standard solutions were prepared in methanol from a stock solution of 1.023 mg/mL by serial dilution covering the concentration range 0.0512 to 0.1535 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 0.9996. The fortified samples of filters were found to have a recovery value of ± 10% of the fortification. The results indicate the accurate use of the test item and filters 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 5.0 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, body weight, 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.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
3.18 mg/L air
Based on:
test mat.
95% CL:
> 2.4 - < 3.96
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:
3.67 mg/L air
Based on:
test mat.
95% CL:
> 2.35 - < 4.99
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:
2.72 mg/L air
Based on:
test mat.
95% CL:
> 1.88 - < 3.55
Exp. duration:
4 h
Remarks on result:
other: Effect levels based on analytically confirmed: mean achieved concentrations for each exposure/sex groups
Mortality:
Mortalities are reported in table 3.
Clinical signs:
other: Common abnormalities noted during the study included increased respiratory rate, hunched posture, pilo-erection and wet fur. During exposure all organisms exhibited decreased respiratory rate. On removal from the chamber there were frequent instances of
Body weight:
In group 2, 0.95 mg/L: All males and one female exhibited body weight losses or showed no body weight gain on Day 1 post-exposure. Three female exhibited slight body weight losses from Days 3 to 7 post-exposure and one male exhibited a slight body weight loss during the final week of the recovery period. All other males and females gained weight during the remainder of the recovery period.
In group 3, 2.60 mg/L: All surviving males and one out of three surviving females exhibited body weight losses on Day 1 post-exposure. Body weight gains were noted for all surviving organisms during the remainder of the recovery period. These in general, were in the ranges expected and considered normal for this type of study.
In group 1: 5.17 mg/L: The surviving male showed body weight gain after initial day-1 exposure and subsequently exhibited body weight gains during the remainder of the recovery period. These in general, were in the ranges expected and considered normal for this type of study
Gross pathology:
In group 2, 0.95 mg/L: No macroscopic abnormalities in three males and two females; dark patches in lungs was seen in two males and three females.
In group 3, 2.60 mg/L: No macroscopic abnormalities in in four males and three female survivors. In the mortalities, gaseous distension of the small and/or large intestine and/or dark patches in lung was observed.
In group 1: 5.17 mg/L: Dark patches in lungs was observed in the single surviving male. In the mortalities, abnormally dark lungs and dark liver was observed along with instances of dark patches in the lungs.
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

5.00

2.25

76.7

2.21

2

1.04

2.19

77.8

2.21

3

3.05

2.17

78.2

2.21

 

Table 2. Mean achieved atmosphere concentrations:

Group Number

Atmosphere Concentration

Mean Achieved (mg/L)

Standard Deviation

Nominal (mg/L)

1

5.00

0.14

13.1

2

1.04

0.07

2.80

3

3.05

0.09

8.72

 

Table 3. Mortality data summary:

Group Number

Mean Achieved Atmosphere Concentration

(mg/L)

Deaths

Male

Female

Total

1

5.00

5/5

5/5

10/10

2

1.04

0/5

0/5

0/10

3

3.05

2/5

3/5

5/10

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 inhalation LC50 (male/female) was: 3.18 (C.I. 2.40 – 3.96) mg/L; LC50 (male): 3.67 (C.I. 2.35 – 4.99) mg/L and LC50 (female): 2.72 (C.I. 1.88 – 3.55) 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. Three 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: 5.17 mg/L, Group 2: 0.95 mg/L and Group 3: 2.60 mg/L. The characteristics of the achieved atmosphere where Mean Mass Median Diameter (particle size) and Inhalable Fraction <4 μm: Group 1: 3.48 μm and 56.9%; Group 2: 2.59 μm and 75.1% and Group 3: 2.46 μm and 74.5%. The Geometric Standard Deviation was Group 1: 2.25, Group 2: 1.91 and Group 3: 2.10, respectively. There was 4 male and 5 female mortalities in Group 1, no mortalities in Group 2 and 1 male and 2 female mortalities in Group 3. In Group 1, the surviving male indicated body weight gain during the recovery period. Within Group 2, all males and one female exhibited body weight losses or showed no body weight gain on Day 1 post-exposure. Three females exhibited slight body weight losses from Days 3 to 7 post-exposure and one male exhibited a slight body weight loss during the final week of the recovery period. However, in Group 2 all males and females gained weight during the recovery period. Within Group 3, All surviving males and one out of three surviving females exhibited body weight losses on Day 1 post-exposure. Body weight gains were noted for all survivors during the remainder of the recovery period. These in general, where in the ranges expected and considered normal for this type of study. In Group 2 clinical observations for all males and females appeared normal at day 2 post exposure. All survivors in Group 3 appeared normal at day 4 and 5 post exposure. During necropsy in surviving Group 1 male and two males and three females of Group 2: 0.95 mg/L there was evidence of dark patches in the lungs. No macroscopic abnormalities was observed in three males and two females in Group 2. No macroscopic abnormalities were observed in four males and three female survivors in Group 3: 2.60 mg/L. Under the conditions of this study, the inhalation LC50 (male/female) was: 3.18 (C.I. 2.40 – 3.96) mg/L; LC50 (male): 3.67 (C.I. 2.35 – 4.99) mg/L and LC50 (female): 2.72 (C.I. 1.88 – 3.55) mg/L within the RCCHan WIST rat. The test item was classified as Acute Inhalation Toxicity: Category 4 according to Regulation (EC) 1272/2008.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LC50
Value:
2 720 mg/m³
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:
2016
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
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Recognised supplier
- Age at study initiation: 8 - 12 weeks.
- Weight at study initiation: 211 - 283 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: From: To: 2016-05-11 to 2016-06-01
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: Not applicable.
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:
- Clinical observations: No signs of systemic toxicity were noted during the observation period.
- Dermal reactions: No signs of dermal irritation were noted.
Body weight:
Animals showed expected gains in body weight, except for three females which showed body weight loss (1/3) or no gain in body weight (2/3) during the first week but expected gain in body weight during the second week.
Gross pathology:
No abnormalities were noted at necropsy.

Applicant assessment indicates: as concluded within the study, since static or minor declines body weight were transient and/or not observed with significant clinical signs or gross pathology findings; that there was no evidence of 'significant toxicity' at the dose level employed during the test.

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 Acute Toxicity (Dermal) and 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 or abnormalities on necropsy. There was no mortality during the study. No signs of dermal irritation were noted. Animals showed expected gains in body weight, except for three females which showed body weight loss or no gain in body weight during the first week but expected gain in body weight during the second week. 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, 2016 - The study was performed according to OECD TG 420 and EU Method B.1 bis Acute Toxicity (Oral) and in accordance with GLP to assess the acute oral toxicity of the test material following a single oral administration in the female Wistar strain rat by the fixed dose method. The test substance was administered by oral gavage in an initial sighting study at 300 mg/kg bw in a solution in arachis oil BP and following an absence of toxicity then at 2000 mg/kg bw. Subsequently, at a further group of four fasted females was given a single oral dose of test item, at a dose level of 300 mg/kg body weight. The animal treated at a dose level of 2000 mg/kg was found dead 1 day after dosing. There were no mortality at a dose level of 300 mg/kg. Hunched posture was noted during the day of dosing in the animal treated at a dose level of 2000 mg/kg. There were no signs of systemic toxicity and all animals showed expected gains in bodyweight over the study period at a dose level of 300 mg/kg. Abnormalities noted at necropsy of the animal treated at a dose level of 2000 mg/kg, that was found dead, were dark liver, dark kidneys, gaseous stomach and epithelial sloughing of the gastric mucosa and non-glandular epithelium of the stomach. Abnormalities noted at necropsy of one animal, treated at a dose level of 300 mg/kg, were epithelial sloughing of the gastric mucosa and non glandular epithelium of the stomach. No abnormalities were noted at necropsy of four animals treated at a dose level of 300 mg/kg. Under the conditions of this study, the oral LD50 was estimated to be in the range of 300 - 2000 mg/kg bw in the female Wistar rat. The test substance was classified as Acute Oral Toxicity: Category 4 according to Regulation (EC) 1272/2008.

 

INHALATION:

Key study: OECD TG 403, 2016 - 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. Three 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: 5.17 mg/L, Group 2: 0.95 mg/L and Group 3: 2.60 mg/L. The characteristics of the achieved atmosphere where Mean Mass Median Diameter (particle size) and Inhalable Fraction <4 μm: Group 1: 3.48 μm and 56.9%; Group 2: 2.59 μm and 75.1% and Group 3: 2.46 μm and 74.5%. The Geometric Standard Deviation was Group 1: 2.25, Group 2: 1.91 and Group 3: 2.10, respectively. There was 4 male and 5 female mortalities in Group 1, no mortalities in Group 2 and 1 male and 2 female mortalities in Group 3. In Group 1, the surviving male indicated body weight gain during the recovery period. Within Group 2, all males and one female exhibited body weight losses or showed no body weight gain on Day 1 post-exposure. Three females exhibited slight body weight losses from Days 3 to 7 post-exposure and one male exhibited a slight body weight loss during the final week of the recovery period. However, in Group 2 all males and females gained weight during the recovery period. Within Group 3, All surviving males and one out of three surviving females exhibited body weight losses on Day 1 post-exposure. Body weight gains were noted for all survivors during the remainder of the recovery period. These in general, where in the ranges expected and considered normal for this type of study. In Group 2 clinical observations for all males and females appeared normal at day 2 post exposure. All survivors in Group 3 appeared normal at day 4 and 5 post exposure. During necropsy in surviving Group 1 male and two males and three females of Group 2: 0.95 mg/L there was evidence of dark patches in the lungs. No macroscopic abnormalities was observed in three males and two females in Group 2. No macroscopic abnormalities were observed in four males and three female survivors in Group 3: 2.60 mg/L. Under the conditions of this study, the inhalation LC50 (male/female) was: 3.18 (C.I. 2.40 – 3.96) mg/L; LC50 (male): 3.67 (C.I. 2.35 – 4.99) mg/L and LC50 (female): 2.72 (C.I. 1.88 – 3.55) mg/L within the RCCHan WIST rat. The test item was classified as Acute Inhalation Toxicity: Category 4 according to Regulation (EC) 1272/2008.

 

DERMAL:

Key study: OECD TG 402, 2016 - The study was performed according to OECD TG 402 and EU Method B.3 Acute Toxicity (Dermal) and in accordance with GLP to assess the acute dermal toxicity of the test substance in the Wistar strain rat. A group of five males and five females were 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 or abnormalities on necropsy. There was no mortality during the study. No signs of dermal irritation were noted. Animals showed expected gains in body weight, except for three females which showed body weight loss or no gain in body weight during the first week but expected gain in body weight during the second week. 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 meets classification criteria under Regulation (EC) No 1272/2008 for acute toxicity: inhalation category 4: H332

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

Categories Display