4,8-Cyclododecadien-1-one

Endpoint summary

Description of key information

Oral: measured LD50 > 2000 mg/kg bw and the estimated LD50 cut-off was considered to be > 2000 and < 5000 mg/kg bw, female rat, OECD TG 423, 2014

Inhalation: LC50 (male/female): > 4.53 mg/L mean achieved atmosphere concentration, OECD TG 403, 2014

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, 2014

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:

05-12-2013 to 27-12-2013

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 (2000)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

inspected: November 2012; signature: February 2013

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Crl:CD (SD)

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: 214 g to 262; The weight variation did not exceed ±20% of the mean weight at the start of treatment.
- Fasting period before study: Overnight before dosing and four hours after dosing.
- Housing: Group housed in groups of up to three in polycarbonate cages with a stainless steel mesh lid with a quantity of autoclaved softwood bark-free fibre bedding.
- Diet (e.g. ad libitum): Certified diet from recognised supplier, provided ad libitum (except for overnight fasting period and four hours post doing).
- 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): The room was kept at positive pressure with respect to the outside by its own supply of filtered fresh air, which was passed to atmosphere and not re-circulated. The air changes was monitored periodically.
- Photoperiod: 12 h light / 12 h dark

IN-LIFE DATES: From: To: 2013-12-10 to 2013-27-12

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: Formulated at concentration of 200 mg/mL in the vehicle and administered at a volume of 10 mL/kg body weight
- Amount of vehicle (if gavage): Administered at a volume of 10 mL/kg body weight
- Justification for choice of vehicle: Not applicable.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bodyweight

DOSAGE PREPARATION (if unusual): Not applicable.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Assessment of available infornation in accordance with the study guideline.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6 females per dose (tested sequentially in accordance with the guideline).

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Cages were checked at least twice daily for any mortalities. Observations were made after dosing and at frequent intervals for the remainder of Day 1. On subsequent days, observations were made once in the morning and again at the end of the experimental day (with the exception of Day 15 - morning only). The nature and severity, where appropriate, of the clinical signs and the time were recorded at each observation. Individual body weights were recorded on Day 1 (prior to dosing) and on Days 8 and 15 or at mortality. Individual weekly body weight changes and group mean body weights were calculated.
- Necropsy of survivors performed: yes

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

One female (J4) dosed at 2000 mg/kg was terminated due to poor clinical condition on Day 2.

Clinical signs:

other: 2000 mg/kg bw: Clinical signs prior to humane sacrifice (female J4 on Day 2 ) comprised of unsteady gait, tremors, uncoordinated gait, piloerection, hunched posture, flat posture, shallow breathing and reduced body temperature. These signs were seen from

Gross pathology:

Macroscopic examination of the humanely sacrificed female due to poor condition on day 2 revealed pallor of the lungs, liver and kidneys and yellow fluid content in the small and large intestines. Macroscopic examination at study termination on Day 15 revealed pallor of the kidneys in one female (J1). No abnormalities were revealed in any other surviving females.

Interpretation of results:

not classified

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study the oral LD50 was established to exceed 2000 mg/kg bw in female Sprague-Dawley Crl:CD (SD) rats. Applicant assessment indicates, under the conditions of this study and according to the OECD TG 423 criteria, the LD50 cut-off value was considered to be greater than 2000 and less than 5000 mg/kg body weight.

Executive summary:

The study was performed according to OECD TG 423 and EU Method B.1 tris, US EPA OPPT 870.1100 and Japan Guidelines for Acute Toxicity Oral Class Method and in accordance with GLP. The objective of the study was to assess the acute oral toxicity of the test material following a single oral administration in the female Sprague-Dawley Crl:CD (SD) strain rat by the acute toxic class method. The test item was formulated in corn oil vehicle at 200 mg/ml and then administered by single oral gavage in an initial group of three females at 2000 mg/kg. As the results of this group indicated the acute lethal oral dose of the test substance to be greater than 2000 mg/kg body weight a further group of three females was dosed at 2000 mg/kg bw in accordance with the guidelines. There was one female humane sacrifice in the second group on day 2. Clinical signs prior to humane sacrifice mortality comprised unsteady gait, tremors, uncoordinated gait, piloerection, hunched posture, flat posture, shallow breathing and reduced body temperature. These signs were seen from approximately 30 minutes after dosing. Macroscopic examination of the female revealed pallor of the lungs, liver and kidneys and yellow fluid content in the small and large intestines. Clinical signs of reaction to treatment for the remaining females receiving 2000 mg/kg comprised of unsteady gait, tremors, uncoordinated gait and piloerection, loose faeces, hunched posture, reduced activity, shallow breathing, elevated gait and urine staining. These signs were first noted approximately 30 minutes after dosing. Recovery of surviving females, as judged by external appearance and behaviour, was complete by Day 4 or 6. Slightly low body weight gain was recorded during the second week of the observation period for the two surviving rats of the second cohort. All other females were considered to have achieved satisfactory body weight gains throughout the study. Macroscopic examination at study termination on Day 15 revealed pallor of the kidneys in one female. No abnormalities were revealed in any other surviving females. Under the conditions of this study the oral LD50 was established to exceed 2000 mg/kg bw in female Sprague-Dawley Crl:CD (SD) rat. Applicant assessment indicates, under the conditions of this study and according to the OECD TG 423 criteria, the LD50 cut-off value was considered to be greater than 2000 and less than 5000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

2 000 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:

23-07-2014 to 09-10-2014

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 (bodyweights of females were lower than 20% of the mean group body weights) but not considered to affect the purpose or 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:

Due to availability, the following females were outside the weight range specified in the study plan (200-350g): 7f: 193g, 8f: 190g, 9f: 193g. This deviation is considered not to affect the purpose or validity of the study. Since were at the lower range.

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: March 2014 ; signature: May 2014

Test type:

traditional method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

RccHanTM : WIST

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: 190-350 g; due to availability, the following females were outside the weight range specified in the study plan (200-350 g): 7f: 193g, 8f: 190g and 9f: 193g. With the exception of these females all body weights were within 20% of the mean weight for each sex, the deviation in relation to body weight for these three females was considered to not impact on the integrity of study. The applicant indicates: this is since the females were at the lower ranges of bodyweight.
- Fasting period before study: None.
- Housing: groups of up to 5 by sex in solid-floor polypropylene cages with stainless steel lids, furnished with softwood flakes and cage enrichment (wooden chew blocks and cardboard fun tunnels).
- Diet (e.g. ad libitum): Certified diet from recognised supplier, provided ad libitum (except for exposure period period)
- Water (e.g. ad libitum): mains drinking water, ad libitum (except for exposure period period)
- Acclimation period: at least 5 days

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

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

clean 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):

2.68 µm

Geometric standard deviation (GSD):

2.1

Remark on MMAD/GSD:

MMAD/GSD relates to: 35.1 mg/L (nominal), 4.53 mg/L (mean achieved concentration) dose

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. The chamber flow rate was maintained at 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: 20-22°C, 28-29% 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 drawn through a glass impinger containing Methanol (made up to 90mL). The sample 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 1.041 mg/mL by serial dilution covering the concentration range 0.0 to 0.1562 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 0.999. 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 (if not tabulated)
- 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:

35.1 mg/L (nominal), 4.53 mg/L (mean achieved concentration)

No. of animals per sex per dose:

5 per sex per dose. Full details are provided in table 2.

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 (where possible). Alternatively, an estimate of the LC50 of the test item was made based on the available information.

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 4.53 other: mg/l

Based on:

test mat.

Mortality:

1 female mortality occurred (humane sacrifice day 2) ; mortalities are reported in table 3.

Clinical signs:

other: Common abnormalities noted during the study included: During exposure one female (10f) animal exhibited decreased respiratory rate. On removal from the chamber all females exhibited body tremors and increased activity. All males/females also demonstrated

Body weight:

All animals exhibited body weight losses on the first day post-exposure. Body weight gains were noted in all surviving animals during the remainder of the recovery period.

Gross pathology:

Abnormally red lungs with dark patches were seen in 1 male (1m). 1 female exhibited pale liver and kidneys (10f). The female (8f) that was humanely sacrificed during the course of the study had dark liver and kidneys. No abnormalities were noted in any other males or females.

Other findings:

- Other observations: The respiratory tract was subjected to a detailed macroscopic examination for signs of irritancy or local toxicity during necropsy.

Additional comments regarding test atmosphere generation:

It is noted within the study that one sample (taken at 60 minutes of exposure) was lower than 20% of the mean achieved atmosphere concentration. As all other samples taken during the exposure were within the accepted 20 % of the mean achieved atmosphere concentration and only one animal mortality occurred this slight deviation to the test guideline was considered not to have affected the purpose or validity of the study. Rerunning the group was not considered appropriate within the study under the guideline. The results obtained (in terms of observations) would be considered to be similar on another group exposed at the same generation rates. It was considered that the test atmosphere consistency would not be able to be improved and the this was further confirmed by the large nominal concentration when compared to the mean achieved concentration.

 

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	4.53	2.68	70.7	2.10

 

Table 2. Mean achieved atmosphere concentrations:

Group Number	Mean Achieved Atmosphere Concentration (mg/L)	Standard Deviation	Nominal (mg/L)
1	4.53	0.48	35.1

 

Table 3. Mortality data summary:

Group Number	Mean Achieved Atmosphere Concentration (mg/L)	Mortalities
		Female	Male	Total
1	4.53	1/5	0/5	1/10

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study, the inhalation 4h-LC50 (male/female) was considered to be > 4.53 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. A single group 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: 4.53 mg/L based on a nominal concentration of 35.1 mg/L. The characteristics of the achieved atmosphere where Mean Mass Median Diameter (particle size) and Inhalable Fraction <4 μm were: 2.68 μm and 70.7% with geometric Standard Deviation 2.10. There was no male and one female mortalities in the 4.54 mg/L mean achieved atmosphere concentration. The single female mortality was a humane sacrifice. Observation changes noted during the study in the surviving animals included decreased respiratory rate, increased respiratory rate, ataxia, hunched posture, pilo-erection and wet fur. Occasional instances of body tremors, occasional body tremors and increased activity. Surviving animals recovered to appear normal from Days 7 to 8 post-exposure. Observations noted in the female that was humanely sacrificed during the study included decreased respiratory rate, ataxia, body tremors, occasional body tremors, increased activity, dehydration, lethargy, ptosis, splayed gait, hunched posture, pilo-erection and wet fur. All males and females exhibited body weight losses on the first day post-exposure. Body weight gains were noted in all survivors during the remainder of the recovery period. During necropsy abnormally red lungs with dark patches were seen in one male out of nine survivors. A single female survivor showed pale liver and kidney. No other macroscopic abnormalities were observed. In the single female mortality dark liver and dark kidney was seen at necropsy. A single mortality occurred in the group of five females and five males at the 4.53 mg/L mean achieved atmosphere concentration. Under the conditions of this study, the inhalation 4h-LC50 (male/female) was considered to be > 4.53 mg/L within the RCCHan WIST rat.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

4 530 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:

29-05-2015 to 19-06-2014

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: March 2014 ; signature: May 2014

Test type:

fixed dose procedure

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: Female: 208 - 230 g; Male: 228 – 270 g; The weight variation did not exceed ±20% of the mean weight per sex at the start of treatment.
- 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

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): dose volume: 2.06 ml/kg
- Concentration (if solution): 2000 mg/kg
- Constant volume or concentration used: 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); 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: Very slight erythema was noted at the test site of four females (maximum score = 1; in 4 females at day 1 only). No other signs of dermal

Gross pathology:

No abnormalities were noted at necropsy

Interpretation of results:

not classified

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.

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 was no mortality during the study. There were no signs of system toxicity or abnormalities on necropsy. All males and females showed expected gains in bodyweight over the study period, except for one female which showed no gain in body weight during the first week with expected gain in body weight during the second week. However, this was still considered to be within the historical range for this strain. Very slight erythema (score = 1) was noted at the test site of four females on day 1 only. No other signs of dermal irritation were noted. 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.

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: OECD TG 423, 2014 - The study was performed according to OECD TG 423 and EU Method B.1 tris, US EPA OPPT 870.1100 and Japan Guidelines for Acute Toxicity Oral Class Method and in accordance with GLP. The objective of the study was to assess the acute oral toxicity of the test material following a single oral administration in the female Sprague-Dawley Crl:CD (SD) strain rat by the acute toxic class method. The test item was formulated in corn oil vehicle at 200 mg/ml and then administered by single oral gavage in an initial group of three females at 2000 mg/kg. As the results of this group indicated the acute lethal oral dose of the test substance to be greater than 2000 mg/kg body weight a further group of three females was dosed at 2000 mg/kg bw in accordance with the guidelines. There was one female humane sacrifice in the second group on day 2. Clinical signs prior to humane sacrifice mortality comprised unsteady gait, tremors, uncoordinated gait, piloerection, hunched posture, flat posture, shallow breathing and reduced body temperature. These signs were seen from approximately 30 minutes after dosing. Macroscopic examination of the female revealed pallor of the lungs, liver and kidneys and yellow fluid content in the small and large intestines. Clinical signs of reaction to treatment for the remaining females receiving 2000 mg/kg comprised of unsteady gait, tremors, uncoordinated gait and piloerection, loose faeces, hunched posture, reduced activity, shallow breathing, elevated gait and urine staining. These signs were first noted approximately 30 minutes after dosing. Recovery of surviving females, as judged by external appearance and behaviour, was complete by Day 4 or 6. Slightly low body weight gain was recorded during the second week of the observation period for the two surviving rats of the second cohort. All other females were considered to have achieved satisfactory body weight gains throughout the study. Macroscopic examination at study termination on Day 15 revealed pallor of the kidneys in one female. No abnormalities were revealed in any other surviving females. Under the conditions of this study the oral LD50 was established to exceed 2000 mg/kg bw in female Sprague-Dawley Crl:CD (SD) rat. Applicant assessment indicates, under the conditions of this study and according to the OECD TG 423 criteria, the LD50 cut-off value was considered to be greater than 2000 and less than 5000 mg/kg body weight.

INHALATION: OECD TG 403, 2014 - 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. A single group 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: 4.53 mg/L based on a nominal concentration of 35.1 mg/L. The characteristics of the achieved atmosphere where Mean Mass Median Diameter (particle size) and Inhalable Fraction <4 μm were: 2.68 μm and 70.7% with geometric Standard Deviation 2.10. There was no male and one female mortalities in the 4.54 mg/L mean achieved atmosphere concentration. The single female mortality was a humane sacrifice. Observation changes noted during the study in the surviving animals included decreased respiratory rate, increased respiratory rate, ataxia, hunched posture, pilo-erection and wet fur. Occasional instances of body tremors, occasional body tremors and increased activity. Surviving animals recovered to appear normal from Days 7 to 8 post-exposure. Observations noted in the female that was humanely sacrificed during the study included decreased respiratory rate, ataxia, body tremors, occasional body tremors, increased activity, dehydration, lethargy, ptosis, splayed gait, hunched posture, pilo-erection and wet fur. All males and females exhibited body weight losses on the first day post-exposure. Body weight gains were noted in all survivors during the remainder of the recovery period. During necropsy abnormally red lungs with dark patches were seen in one male out of nine survivors. A single female survivor showed pale liver and kidney. No other macroscopic abnormalities were observed. In the single female mortality dark liver and dark kidney was seen at necropsy. A single mortality occurred in the group of five females and five males at the 4.53 mg/L mean achieved atmosphere concentration. This was the maximum achievable concentration within the study using a nominal concentration of 35.1 mg/L. Under the conditions of this study, the inhalation 4h-LC50 (male/female) was considered to be > 4.53 mg/L within the RCCHan WIST rat.

DERMAL: OECD TG 402, 2014 - 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 was no mortality during the study. There were no signs of system toxicity or abnormalities on necropsy. All males and females showed expected gains in bodyweight over the study period, except for one female which showed no gain in body weight during the first week with expected gain in body weight during the second week. However, this was still considered to be within the historical range for this strain. Very slight erythema (score = 1) was noted at the test site of four females on day 1 only. No other signs of dermal irritation were noted. 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.

Justification for classification or non-classification

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

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

The substance meets classification criteria under Regulation (EC) No 1272/2008 for Specific Target Organ Toxicity: Single Exposure – Category 3 (narcotic effects): H336

Ataxia, tremors, uncoordinated gait and lethargy was observed in studies at Acute Oral and Acute Inhalation: GHS Category 4 dose levels. All effects were transient in nature.