3-(4,4-dimethylcyclohex-1-en-1-yl)propanal

Endpoint summary

Description of key information

Oral: LD50 > 300 - < 2000 mg/kg bw, female rat, OECD TG 420, 2009

Inhalation: LC50 >1 - ≤ 5 mg/L, female rat, OECD TG 436, 2012

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, 2010

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:

22-07-2009 to 23-12-2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Guideline study performed under GLP. All relevant validity criteria were met.

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: August 2008 ; signature: March 2009

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

Strain: HsdRccHan WIST

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Rationale for alternative/additional species to rat (if applicable): Not applicable.
- Source: Recognised supplier (documented in the full study report)
- Females (if applicable) nulliparous and non-pregnant: yes
- Rationale for use of males (if applicable): Not applicable.
- Age at study initiation: 8 - 12 weeks.
- Weight at study initiation: 166 - 180 g (300 mg/kg including sighting test; sentinel); 190 g (2000 mg/kg sighting test; sentinel); The weight variation did not exceed ±20% of the mean weight in the definitive test.
- 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.
- Historical data: The test laboratory possesses historical control data.
- 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.
- Microbiological status when known: No issues reported.
- Method of randomisation in assigning animals to test and control groups : Random allocation.

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: 22-07-2009 to 19-08-2009

Route of administration:

oral: gavage

Vehicle:

arachis oil

Details on oral exposure:

- Concentration in vehicle: 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. Singularly and in the absence of mortality or evident toxicity a further group of 4 was tested in the appropriate dose level.
The test item was formulated at concentrations of 30 mg/mL or dosed as supplied according to specific gravity of the test item. Formulations were prepared on the day of dosing.
- Amount of vehicle (if gavage): Test Item dose volume was 10 mL/kg (of bodyweight) in Arachis Oil BP (at 300 mg/kg bw) or dosed as supplied at 2.23 mL/kg (at 2000 mg/kg bw)
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed. Arachis oil BP was used because the test material did not dissolve/suspend in distilled water.
- Lot/batch no. (if required): See full study report.
- Purity: BP grade.

MAXIMUM DOSE VOLUME APPLIED: 300 mg/kg bw dose level: 10 mL/kg of test item in vehicle (Arachis Oil BP) ; 2000 mg/kg bw dose level: 2.23 mL/kg of test item as supplied.

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 in Arachis Oil BP (300 mg/kg bw) or 2.23 mL/kg as supplied (2000 mg/kg bw).

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Using the available information regarding the toxicity of the test item, 2000 mg/kg was chosen as the starting dose.

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 (5/5)
2000 mg/kg bw (sentinel): (1/1) humanely terminated day 1

Clinical signs:

other: 300 mg/kg bw (sentinel and definitive test): Hunched posture (4/5), noisy respiration (1/5 ; at 0.5 hours only). All animals appeared normal one to two days after dosing. No signs of toxicity (in 1/5) were noted. 2000 mg/kg bw (sentinel): hunched posture,

Gross pathology:

300 mg/kg bw (sentinel and definitive test): No abnormalities were noted at necropsy.
2000 mg/kg bw (sentinel): No abnormalities were noted at necropsy.

Other findings:

- Organ weights: Not reported.
- Histopathology: Not reported.
- Potential target organs: Not reported.
- Other observations: Not reported.

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 HsdRccHan WIST strain 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 item following a single oral administration in the female Wistar HsdRccHan WIST strain rats by the fixed dose method. The test item was administered by oral gavage in a solution in arachis oil BP in an initial sighting study at 2000 mg/kg bw and following a presence of toxicity then at 300 mg/kg bw. Subsequently, 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 humanely terminated on 1 day after dosing. Clinical signs were noted during the day of dosing in the animal treated at a dose level of 2000 mg/kg. There was no mortality at a dose level of 300 mg/kg. Hunched posture (4/5), noisy respiration (1/5 ; at 0.5 hours only) was observed. All animals appeared normal one to two days after dosing. All animals showed expected gains in bodyweight over the study period at a dose level of 300 mg/kg. No abnormalities were noted at necropsy of the animal treated at a dose level of 2000 mg/kg. No abnormalities were noted 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.

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:

27-09-2011 to 02-02-2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Guideline study performed under GLP. All relevant validity criteria were met.

Qualifier:

according to guideline

Guideline:

OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.52 (Acute Inhalation Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

inspected: July 2011; signature: August 2011

Test type:

acute toxic class 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
- Rationale for alternative/additional species to rat (if applicable): Not applicable.
- Source: Recognised supplier (documented in the full study report)
- Females (if applicable) nulliparous and non-pregnant: yes
- Rationale for use of males (if applicable): Used in accordance with the relevant guidelines, where applicable.
- Age at study initiation: 8 - 12 weeks.
- Weight at study initiation: 200 – 350 g (0.98 mg/L mean achieved atmosphere ; female: 217 – 225 g and male: 268 – 271 g
- Fasting period before study: None.
- Housing: Housed in groups of up to three by sex in solid-floor polypropylene cages with stainless steel lids, furnished with softwood flakes provided with environmental enrichment items.
- Historical data: The test laboratory possesses historical control data.
- 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.
- Microbiological status when known: No issues reported.
- Method of randomisation in assigning animals to test and control groups : Random allocation.

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: 27-09-2011 to 30-11-2011

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):

1.64 µm

Geometric standard deviation (GSD):

2.49

Remark on MMAD/GSD:

Group 1: 1.0 mg/L (Target aerosol concentration) or 0.98 mg/L Mean Achieved Concentration. The characteristics of the achieved atmosphere where Mean Mass Median Diameter (particle size) and fraction < 4 μm: Group 1: 1.64 μm and 83.8%. The Geometric Standard Deviation was 2.49

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: 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 and air flow settings into the chamber. The chamber flow rate was maintained 60 L/min providing 120 air changes per hour.
- 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. (Chamber temperature actual was: 19 °C ; Chamber relative humidity actual was: 11 – 13% during exposure).

TEST ATMOSPHERE
- Brief description of analytical method used: The test atmosphere was sampled five times during the exposure period. The sampling procedure involved pumping five litres of the chamber atmosphere through a glass impinger containing 40mL of methanol. After sampling the Dreschel head was flushed through with a further 40mL of methanol to remove any deposits. This gave an 80 mL sample to be submitted for chemical analysis. Preliminary investigations showed that when test atmosphere containing test item was passed through a series of three impingers containing methanol, no significant levels of test item were collected in the second or third impingers. The concentration of test item in the impingers was determined by GC using an external standard technique. During the testing 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 0.05 mg/mL by serial dilution covering the concentration range 0.00262 to 0.1049 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 > 0.99. 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 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 (as applicable) in table 1.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Remarks on duration:

In accordance with the guidelines.

Concentrations:

Following an appropriate equilibration period, a single group of six rats (three males and three females), was subjected to a single exposure to the test item for a period of four hours. Based on the results of the first test target concentration 1.0 mg/L (no mortality and/or observations at mean achieved atmosphere 0.98 mg/L), it was considered that no further testing was required in definitive test exposure. Full details are provided in table 2.

No. of animals per sex per dose:

3 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. The respiratory
tract was subjected to a detailed macroscopic examination for signs of irritancy or local toxicity.

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 1 - <= 5 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.

Mortality:

No mortalities during the study.

Clinical signs:

other: See "Other findings" for further information.

Body weight:

In group 1: 1.0 mg/L target concentration (0.98 mg/L mean achieved concentration): All males/females exhibited slight bodyweight losses on the first day post-exposure. Reasonable bodyweight development was noted in all animals during the remainder of the recovery period with the exception of one female animal which exhibited a slight bodyweight loss (-2 g) from Days 3 to 7 post-exposure, but which was gaining bodyweight at from Days 7 to 14 (+3 g).

Gross pathology:

In group 1: 1.0 mg/L target concentration (0.98 mg/L mean achieved concentration: Two males (#1 and #3) and one female (#2) showed no abnormalities. Dark and/or pale patches on the lungs were noted amongst one male (#2) and two of three females (#1 and #3).

Other findings:

- Other observations: The respiratory tract was subjected to macroscopic examination for signs of irritancy or local toxicity during necropsy.
- Clinical signs: In group 1.0 mg/L target concentration (0.98 mg/L mean achieved concentration): Signs of hunched posture and pilo-erection are commonly seen in males/females 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. These observations are considered to be associated with the restraint procedure and, in isolation, are not indicative of toxicity. No significant observations were apparent in any males/females during exposure. Immediately after the exposure, males/females showed decreased respiratory rate and ataxia. One hour after removal, little or no change in the condition of the animals was observed. One day after exposure, all males/females exhibited hunched posture and pilo-erection. Frequent occurrences of increased respiratory rate and isolated instances of decreased respiratory rate and laboured respiration (#3 female, day 1 only) were also apparent. All males/females appeared normal from Day 4 post-exposure. There was evidence of recovery. There were no clinical signs at the end of the observation period.

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	0.98	1.64	83.8	2.49

 

Table 2. Mean achieved atmosphere concentrations:

Group Number	Atmosphere Concentration
	Mean Achieved (mg/L)	Standard Deviation	Nominal (mg/L)
1	0.98	0.04	3.65

 

Table 3. Mortality data summary:

Group Number	Mean Achieved Atmosphere Concentration (mg/L)	Deaths
		Male	Female	Total
1	0.98	0/3	0/3	0/6

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 considered to be > 1.0 and ≤ 5.0 mg/L within the RCCHan WIST rat.

Executive summary:

The study was performed according to OECD TG 436 and EU Method B.52 in accordance with GLP to assess the acute inhalation toxicity of the test item. Following a sighting test at 2.22 mg/L mean achieved concentration, one group of six RccHanTM : WIST strain rats (three males and three 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 was Group 1: 0.98 mg/L. The characteristics of the achieved atmosphere where Mean Mass Median Diameter (particle size) and Inhalable Fraction < 4 μm: Group 1: 1.64 μm and 83.8%. The Geometric Standard Deviation was 2.49. There was no mortalities in Group 1. Common abnormalities noted during the study included decreased respiratory rate, ataxia, hunched posture, pilo-erection and wet fur. Frequent instances of increased respiratory rate and an isolated occurrence of laboured respiration were also noted. Animals recovered to appear normal on Day 4 post-exposure. All males/females exhibited slight bodyweight losses on the first day postexposure. Reasonable bodyweight development was noted in all animals during the remainder of the recovery period with the exception of one female animal which exhibited a slight bodyweight loss from Days 3 to 7 post-exposure, but which was gaining bodyweight at from Days 7 to 14 (+3 g). Two males (#1 and #3) and one female (#2) showed no abnormalities. Dark and/or pale patches on the lungs were noted amongst one male (#2) and two of three females (#1 and #3). As the mean achieved concentration was 98% of target and no deaths occurred, no further levels were required as it was considered that at the next dose level of 5 mg/L none of the males/females were considered to survive. This is further confirmed by the observations and the death noted at a mean achieved atmosphere concentration of 2.22 mg/L during the sighting study. Under the conditions of this study, the 4-hour inhalation LC50 (male/female) was considered to be > 1.0 and ≤ 5.0 mg/L within the RCCHan WIST rat.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

1 000 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:

09-05-2001 to 23-05-2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Guideline study performed under GLP. All relevant validity criteria were met.

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: September 2009 ; signature: November 2009

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

HsdRccHan : WIST

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Rationale for alternative/additional species to rat (if applicable): Not applicable.
- Source: Recognised supplier (documented in the full study report)
- Females (if applicable) nulliparous and non-pregnant: yes
- Rationale for use of males (if applicable): Conducted according to guideline requirements of the time.
- Age at study initiation: ca. 8 to 12 weeks age (nulliparous and non-pregnant)
- Weight at study initiation: > 200 g (actual: 234 – 251 g males and 207 – 225 g females
- Fasting period before study: Not applicable
- Housing: during acclimation and observation period: group housed by sex; during exposure: individually housed in polypropylene cages furnished with softwood bedding.
- Historical data: The test laboratory possesses historical control data.
- 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.
- Microbiological status when known: No issues reported.
- Method of randomisation in assigning animals to test and control groups : Random allocation.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25 °C (or 22 ± 3 °C)
- Humidity (%): 30 - 70%
- Air changes (per hr): > 15 air changes per hour
- Photoperiod: 12 h light / 12 h dark

IN-LIFE DATES: 16-12-2009 to 30-12-2009

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 by a semi-occlusive dressing and wrapped with a piece of elastic self-adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): After the 24-hour contact period the bandage was carefully removed, and the treated skin and surrounding hair wiped with cotton wool moistened with distilled water 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 (or dose volume 2.23 mL/kg)
- Concentration (if solution): See below.
- Constant volume or concentration used: 2.23 mL volume ; at a dose level of 2000 mg/kg bw test item.
- For solids, paste formed: Not applicable

VEHICLE
- Amount(s) applied (volume or weight with unit): Not applicable
- Concentration (if solution): Not applicable
- Lot/batch no. (if required): Not applicable
- Purity: Not applicable

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 per sex per dose (5 male/5 female)

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 days 2 to 15. Local effects were examined once daily days 2 to 15 after the completion of the 24-hour exposure period. Full details on the scoring and criteria (appears consistent with Draize for Erythema) 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: Very slight erythema (score = 1) was noted at the treatment site of two males and two females three days after dosing, two males and one f

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. Applicant assessment indicates 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 on the basis of absence of significant clinical toxicological effects and/or bodyweight increases in all males/females.

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 item in the Wistar HsdRccHan : WIST 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 was no mortality during the study. There were no signs of system toxicity or abnormalities on necropsy. It was considered there was no toxicologically significant effects on bodyweight. All males/females gained bodyweight during the study. Very slight erythema (score = 1) was noted at the treatment site of two males and two females three days after dosing, two males and one female four days after dosing and two males five and six days after dosing. Other skin reactions noted at the treatment site of seven animals (two males and five females) two to six days after dosing were loss of skin elasticity, brown discolouration of the epidermis, small superficial scattered scabs and slight desquamation. Treatment sites appeared normal three, four or seven days after dosing. No signs of dermal irritation were noted at the treatment site of three males. The dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar HsdRccHan : WIST rat. Applicant assessment indicates 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 on the basis of absence of significant clinical toxicological effects and/or bodyweight increases in all males/females.

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, 2009 : 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 item following a single oral administration in the female Wistar HsdRccHan WIST strain rats by the fixed dose method. The test item was administered by oral gavage in a solution in arachis oil BP in an initial sighting study at 2000 mg/kg bw and following a presence of toxicity then at 300 mg/kg bw. Subsequently, 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 humanely terminated on 1 day after dosing. Clinical signs were noted during the day of dosing in the animal treated at a dose level of 2000 mg/kg. There was no mortality at a dose level of 300 mg/kg. Hunched posture (4/5), noisy respiration (1/5 ; at 0.5 hours only) was observed. All animals appeared normal one to two days after dosing. All animals showed expected gains in bodyweight over the study period at a dose level of 300 mg/kg. No abnormalities were noted at necropsy of the animal treated at a dose level of 2000 mg/kg. No abnormalities were noted 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.

INHALATION:

Key study: OECD TG 403, 2012 : The study was performed according to OECD TG 436 and EU Method B.52 in accordance with GLP to assess the acute inhalation toxicity of the test item. Following a sighting test at 2.22 mg/L mean achieved concentration, one group of six RccHanTM : WIST strain rats (three males and three 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 was Group 1: 0.98 mg/L. The characteristics of the achieved atmosphere where Mean Mass Median Diameter (particle size) and Inhalable Fraction < 4 μm: Group 1: 1.64 μm and 83.8%. The Geometric Standard Deviation was 2.49. There was no mortalities in Group 1. Common abnormalities noted during the study included decreased respiratory rate, ataxia, hunched posture, pilo-erection and wet fur. Frequent instances of increased respiratory rate and an isolated occurrence of laboured respiration were also noted. Animals recovered to appear normal on Day 4 post-exposure. All males/females exhibited slight bodyweight losses on the first day postexposure. Reasonable bodyweight development was noted in all animals during the remainder of the recovery period with the exception of one female animal which exhibited a slight bodyweight loss from Days 3 to 7 post-exposure, but which was gaining bodyweight at from Days 7 to 14 (+3 g). Two males (#1 and #3) and one female (#2) showed no abnormalities. Dark and/or pale patches on the lungs were noted amongst one male (#2) and two of three females (#1 and #3). As the mean achieved concentration was 98% of target and no deaths occurred, no further levels were required as it was considered that at the next dose level of 5 mg/L none of the males/females were considered to survive. This is further confirmed by the observations and the death noted at a mean achieved atmosphere concentration of 2.22 mg/L during the sighting study. Under the conditions of this study, the 4-hour inhalation LC50 (male/female) was considered to be > 1.0 and ≤ 5.0 mg/L within the RCCHan WIST rat.

DERMAL:

Key study : OECD TG 402, 2010 : 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 item in the Wistar HsdRccHan : WIST 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 was no mortality during the study. There were no signs of system toxicity or abnormalities on necropsy. It was considered there was no toxicologically significant effects on bodyweight. All males/females gained bodyweight during the study. Very slight erythema (score = 1) was noted at the treatment site of two males and two females three days after dosing, two males and one female four days after dosing and two males five and six days after dosing. Other skin reactions noted at the treatment site of seven animals (two males and five females) two to six days after dosing were loss of skin elasticity, brown discolouration of the epidermis, small superficial scattered scabs and slight desquamation. Treatment sites appeared normal three, four or seven days after dosing. No signs of dermal irritation were noted at the treatment site of three males. The dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar HsdRccHan : WIST rat. Applicant assessment indicates 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 on the basis of absence of significant clinical toxicological effects and/or bodyweight increases in all males/females.

Justification for classification or non-classification

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

The substance meets classification criteria under Regulation (EC) No 1272/2008 for acute toxicity: inhalation Category 4.

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