Reaction products of 12-hydroxyoctadecanoic acid with ethane-1,2-diamine and hexane-1,6-diamine and 1,3-phenylenedimethanamine

Endpoint summary

Description of key information

The acute oral, inhalation and dermal toxicity studies indicate that the substance is of low  toxicity if swallowed (rat LD0 > or = 2000 mg/kg bw), applied onto the skin (rat LD0 > or = 2000 mg/kg bw) or inhaled (rat LC0> or = 5140 mg/m3)

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:

From 2012-07-19 to 2012-11-12

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

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

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: breeder: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: approximately 8 weeks old on the day of treatment
- Mean body weight at study initiation: 217 g (range: 202 g to 227 g)
- Fasting period before study: yes, during the night before treatment
- Housing: the animals were housed by three from the same group in polycarbonate cages with stainless steel lids
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: at least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h.

IN-LIFE DATES: 04 September 2012 to 04 October 2012.

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 and 200 mg/ml
- Justification for choice of vehicle: The vehicle used in this study was selected from the results of solubility assays. The solubility assay first started at the concentration of 200 mg/mL, and the first choice vehicle was drinking water treated by reverse osmosis. As unsatisfactory solubility of the test item was obtained in this vehicle (i.e. heterogeneous preparation at the concentration of 200 mg/mL was obtained), another vehicle was chosen from the following organic solvents (in order of preference): 0.5% methylcellulose aqueous solution and corn oil. A homogenous suspension was obtained at the concentration of 200 mg/mL in corn oil.

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg bw

CLASS METHOD (if applicable):
- Rationale for the selection of the starting dose:
Since no relevant toxicity data were available for the estimation of a lethal dose-level and any existing data were taken into account, the starting dose level was 300 mg/kg for ethical reasons.

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

3 per treatment step.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Clinical observations: frequently during the hours following treatment; then, at least once a day.
- Body weight: just before treatment, then on day of treatment (day 1) and on days 8 and 15.
- Necropsy of survivors performed: yes

Statistics:

none

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No unscheduled deaths occurred during the study.

Clinical signs:

Piloerection was observed in 3 out of 6 females given 2000 mg/kg bw 4 hours after treatment on day 1 and on day 2.
No clinical signs were observed in other animals.

Body weight:

A slightly lower body weight gain was noted in one or two females per group (30 g to 37 g vs. 44 g ± 4.2 g in control data base) between day 1 and day 8. A lower body weight gain was also noted in one or two females per group (3 g to 10 g vs. 20 g ± 5.7 g in control data base) between day 8 and day 15.
When considering the day 1 to day 15 period, the body weight change was lower in two females from group 1 treated at the dose-level of 300 mg/kg (40 g and 45 g) and in 4/6 females treated at the dose-level of 2000 mg/kg (between 49 g and 57 g).

Gross pathology:

No abnormalities were noted.

Other findings:

None

Table 7.2.1/1: Mean body weight changes (g) in treated animals during the observation period compared to laboratory historical control data

Sex	Female
Group	Laboratory Historical control data	1	2	3
Dose-level (mg/kg bw)	0	300	2000	2000
Body weight
Day 1	219	225	218	210
Day 8	264	264	254	253
Day 15	284	273	270	269
Body weight change
Day 1	+44	+39	+36	+43
Day 8	+20	+10	+16	+16
Day 15	+64	+48	+52	+60

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The oral LD50 of the test substance was higher than 2000 mg/kg in rats.
Therefore, the substance is not classified according to Regulation (EC) No. 1272/2008 and its subsequent amendments on classification, labeling and packaging (CLP) of substances and mixtures.

Executive summary:

The substance was tested for acute oral toxicity according to OECD 423 guideline and in compliance with Good Laboratory Practices.

 The test item was administered once by gavage to 3 groups of 3 fasted female rats under a dosage-volume of 10 mL/kg bw. The test item was prepared in corn oil. Since no relevant toxicity data were available for the estimation of a lethal dose-level and any existing data have been taken into account, the starting dose-level was 300 mg/kg bw for ethical reasons. After the first assay, the next higher dose-level of 2000 mg/kg bw was tested. Then, as no toxicity was observed at this higher dose-level, the results were confirmed in other females.

Each animal was observed at least once a day for mortality and clinical signs for 15 days. Body weight was recorded before treatment then on days 1, 8 and 15. All surviving animals were necropsied at the end of the observation period.

No unscheduled deaths occurred during the study.

Piloerection was observed in 3 out 6 females given 2000 mg/kg bw 4 hours after treatment on day 1 and on day 2. No clinical signs were observed in other animals.

When compared to laboratory historical control data, a slightly lower body weight gain was noted in one or two females per group between day 1 and day 8 and between day 8 and day 15. When considering the day 1 to day 15 period, the body weight change was lower in two females treated at the dose-level of 300 mg/kg bw and in 4 out of 6 females treated at the dose-level of 2000 mg/kg bw compared to laboratory historical control data.

At necropsy, there were no macroscopic findings related to the test item administration.

The acute oral LD50 was found greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Study complete and sufficient to fulfill the endpoint requirements.

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:

From 2013-09-24 to 2013-12-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan (UK) Ltd.
- Age at study initiation: 10 weeks
- Weight at study initiation: 247 to 277 g (males) and 176 to 200 g (females)
- Fasting period before study:no
- Housing: The animals were housed three of one sex per cage
- Diet: Ad libitum, standard rodent diet (Rat and Mouse n°1 Maintenance Diet). This diet contained no added antibiotic or other chemotherapeutic or prophylactic agent.
- Water: Ad libitum, potable water taken from the public supply.
- Acclimation period: 10 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19 to 23°C
- Humidity: 40 to 70%
- Air changes (per hr):Each animal room was supplied with filtered fresh air, which was passed to atmosphere and not re-circulated.
- Photoperiod (hrs dark / hrs light): 12 h continuous light and 12 h continuous dark/24 h.

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: flow through nose only chamber. This system was an aluminium alloy construction comprising a base unit, a single animal exposure section with 20 ports and a top section incorporating a central aerosol inlet.
- Method of holding animals in test chamber: Rats were held in polycarbonate tubes with their snouts protruding from the end of the tubes into the exposure chamber.
- Source and rate of air: From in-house compressed air system(breathing quality), Generator flow: 19 L/minute
- System of generating particulates/aerosols:Rotative Brush Generator mechanism designed to produce and maintain test atmospheres
containing dust by suspending material brushed from the surface of a lightly compressed powder in a stream of dry air. The concentration of dust in the air was altered by selecting different size pistons and by changing the delivery rate. A regulated flow of compressed air conducted the aerosol to the inhalation chamber.
- Temperature, humidity, pressure in air chamber:
Chamber air temperature was measured using an electronic thermometer probe placed in the breathing zone of the animals via an unused exposure port. The mean chamber temperature value was 23.4°C.
- Humidity: Chamber relative humidity was measured using an electronic hygrometer probe inserted into the breathing zone of the animals via an unused exposure port. The mean chamber relative humidity was 38.8%
Pressure in air chamber was not reported.
- Air flow rate: Airflow were 19 L/minute.
- Method of particle size determination: Particle size analysis of generated atmospheres was performed using a 8-stage cascade impactor (Marple 298). A measured volume of air was drawn at a rate of 2 litres/minute from an unused exposure port on the exposure chamber through the cascade impactor and measured using a wet-type gas meter in line with a pump. Three samples were collected during the exposure, however the first
sample was not calculated as the total capture was below the pre-determined minimum capture due to generation problems around the time of the sample. The collection substrates and the backup filter were weighed before and after sampling and the weight of test item, collected at each stage, was calculated by this difference. The total amount collected for each stage was used to determine the cumulative amount below each cut-off point size. In this way, the proportion (%) of aerosol less than < 0.52, 0.93, 1.55, 3.5, 6.0 , 9.8, 14.8 and 21.3 µm (aerodynamic diameter) was calculated. From this data, the Mass Median Aerodynamic Diameter (MMAD), and Geometric Standard Deviation (GSD) were calculated. The mean MMAD was 2.5 µm and the geometric standard deviation (GSD) was 2.37.
- Treatment of exhaust air: Extract airflow was drawn by in-house vacuum system at a flow rate of 20 L/minute. the airflow was filtered locally.

TEST ATMOSPHERE
- Brief description of analytical method used: A measured volume of air was drawn at a rate of 2 litres/minute from an unused exposure
port on the exposure chamber through a glass microfibre filter, mounted in an open face filter holder and measured using a wet-type gas meter in line with a pump. Thirteen samples were collected during the exposure. The filters were weighed before and after sampling.. The difference in the pre- and post-sampling weights, divided by the volume of atmosphere sampled, was equal to the actual achieved test atmosphere concentration. The mean achieved test atmosphere concentration was 5.14 +/- 1.66 mg/L

- Samples taken from breathing zone: yes, samples were collected from a vacant animal exposure port.

VEHICLE (if applicable)
- none

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

Target concentration: 5.0 mg/L
Mean achieved atmosphere concentration: 5.14 mg/L ; Standard deviation: 1.66

No. of animals per sex per dose:

3/sex/dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 d

OBSERVATIONS:
- Morbidity/Mortality: Animals were checked hourly during exposure, immediately following exposure and then at 1 h and 2 hrs post-exposure. During the 14-day observation period, the animals were observed twice daily (early and late in the working day) for morbidity and/or mortality.

- Clinical Signs: All animals were observed for clinical signs at hourly intervals during exposure, as soon as practically possible following removal from restraint at the end of exposure, 1 h and 2-hrs after exposure and subsequently twice daily for 14 d.

- Bodyweight: Individual bodyweights were recorded prior to treatment, on the day of exposure (Day 1) prior to dosing and on Days 2, 4, 8 and 15.

- Necropsy: At the end of the 14-d observation period, the animals were euthanised by exsanguination under anaesthesia (injection of pentobarbital solution) and gross macroscopic examination was performed. All animals were subject to a gross necropsy which consisted of opening the cranial, thoracic and abdominal cavities.any macroscopic abnormalities in appareance of the organs were recorded.

Statistics:

None

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.14 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

There were no unscheduled deaths.

Clinical signs:

other: Immediately after the exposure 2/3 females were noted to have a partially closed right eye, this remained evident in 1 female at 1 hour after the exposure. Furthermore, at 1 hour after exposure 1 female was noted to be underactive with irregular breathing

Body weight:

Bodyweight losses were evident in all males and females on the day following the exposure. This loss of bodyweight is not unusual following exposures of this duration as food and water were unavailable during the exposure period, and is therefore considered not to be test substance related.
Recovery from the bodyweight loss was observed at the next weighing occasion, Day 4, after which body weight gains continued for the remainder of the study.

Gross pathology:

The macroscopic examination performed after a single administration followed by a 14 day observation period revealed enlargement of the tracheobronchial lymph nodes in all animals.

Table 7.2.2/1: Body weights - individual and group mean values (g)

Group	Male					Female
Animal number	A1	A2	A3	Mean	SD	A4	A5	A6	Mean	SD
. Day -7	246	227	241	238	9.8	166	176	184	175	9.0
. Day 1	277	247	269	264	15.5	176	190	200	189	12.1
. Day 2	269	237	248	251	16.3	172	182	189	181	8.5
. Day 4	275	249	262	262	13.0	177	188	201	189	12.0
. Day 8	290	260	280	277	15.3	183	198	212	198	14.5
. Day 15	304	278	297	293	13.5	192	210	218	207	13.3

SD : Standard Deviation

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The LC50 of the test substance was in excess of 5.14 mg/L for male and female rats.
Therefore, the substance is not classified according to Regulation (EC) No. 1272/2008 and its subsequent amendments on classification, labeling and packaging (CLP) of substances and mixtures.

Executive summary:

The substance was tested for acute inhalation toxicity according to the OECD 436 guideline and in compliance with Good Laboratory Practice.

A group of three male and three female Wistar rats was exposed, nose-only, to an atmosphere of the test item using a flow-through exposure system. The animals were exposed for four hours to a target concentration of 5 mg/L followed by a fourteen day observation period. Each animal was observed for mortality and clinical signs at hourly intervals during exposure, then one hour and two hours after exposure and at least twice a day during the 14 -day observation period. Bodyweights were recorded before treatment then on the day of exposure (day 1) and on days 2, 4,8 and 15. All surviving animals were necropsied at the end of the observation period.

The mean achieved atmosphere concentration was 5.14 mg/L and the mean mass median aerodynamic diameter was 2.5 µm. No deaths occurred during the study. Immediately after the exposure two out of three females were noted to have a partially closed right eye, this was still present in one female at one hour after the exposure. Furthermore, at one hour after exposure one female was noted to be underactive with irregular breathing, the irregular breathing persisted and remained evident at two hours after the exposure. From two hours after exposure and for the remainder of the observation period all males were considered to be clinically normal; females were considered normal from Day 2.

Bodyweight losses were evident in all males and females on the day following the exposure. Recovery from the bodyweight loss was observed at the next weighing occasion, Day 4, after which body weight gains continued for the remainder of the study.

Enlargement of the tracheobronchial lymph nodes was noted in all animals.

The 4 -hour inhalation LC50 was found to be greater than 5.14 mg/L (ie 5140 mg/m3).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

5 140 mg/m³

Quality of whole database:

Study complete and sufficient to fulfill the endpoint requirements.

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:

From 2013-05-22 to 2013-07-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: breeder: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: approximately 8 weeks old on the day of treatment
- Mean body weight at study initiation: the males had a mean body weight of 401 g (range: 391 g to 409 g) and the females had a mean body weight of 192 g (range: 182 g to 201 g)
- Fasting period before study: no
- Housing: the animals were housed by five from the same sex and group in polycarbonate cages with stainless steel lids
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: at least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h.

IN-LIFE DATES: 11 June 2013 to 28 June 2013.

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: 10% of body surface, dorsal site
- Type of wrap if used: hydrophilic gauze pad + adhesive hypoallergenic aerated semi-occlusive dressing

REMOVAL OF TEST SUBSTANCE
- Removal of dressing: 24h post-exposure
- Washing: at 24h post-exposure, with a moistened cotton pad

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): The quantity of test item applied to each animal was adjusted according to the body weight recorded on the day of dose application.
- Constant volume: no

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw.

No. of animals per sex per dose:

Ten Sprague-Dawley rats (five males and five nulliparous and non pregnant females).

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Clinical observations: frequently during the hours following treatment; then, at least once a day.
- Body weight: just before treatment on day 1; then on days 8 and 15.
- Necropsy of survivors performed: yes

Statistics:

None

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No unscheduled deaths occurred during the study.

Clinical signs:

No clinical signs were observed in any animals.

Body weight:

The mean body weight gain was unaffected by the test item treatment.

Gross pathology:

There were no macroscopic findings at necropsy.

Other findings:

On the application site, 4/5 females showed a very slight to well defined erythema on day 3 or from day 3 up to day 4 or 6, associated with dryness of the skin from day 3 or 5 up to day 6 or 7. In one of these four females, scabs were also observed from day 3 to day 5. Moreover, 3/5 males presented a very slight erythema on the application site on day 3 or days 3 to 4. These local reactions were considered to be related to the test substance.
No cutaneous reactions were noted in other animals.

Table 7.2.3/1: Mean body weight changes (g) in treated animals during the observation period compared to laboratory historical control data

 

Sex	Female		Male
Group	Laboratory Historical control data	1	Laboratory Historical control data	2
Dose-level (mg/kg bw)	0	2000	0	2000
Body weight (mean (± SD))
. Day 1	236 (± 8.9)	192 (± 7.2)	362 (± 12.0)	401 (± 6.8)
. Day 8	253 (± 12.0)	223 (± 9.9)	394 (± 15.3)	435 (± 11.7)
. Day 15	273 (± 16.3)	247 (± 9.5)	441 (± 21.5)	474 (± 16.7)
Body weight change (mean (± SD))
. Days 1-8	+17 (± 11.0)	+31 (± 7.9)	+32 (± 9.1)	+34 (± 7.9)
. Days 8-15	+20 (± 7.1)	+24 (± 6.5)	+47 (± 7.5)	+39 (± 5.1)
. Days 1-15	+37 (± 16.3)	+55 (± 6.7)	+79 (± 15.6)	+73 (± 12.0)

SD: standard deviations.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The dermal LD50 of the test substance was higher than 2000 mg/kg bw in rats.
Therefore, the substance is not classified according to Regulation (EC) No. 1272/2008 and its subsequent amendments on classification, labeling and packaging (CLP) of substances and mixtures.

Executive summary:

The substance was tested for acute dermal toxicity according to OECD 402 guideline and in compliance with Good Laboratory Practices.

The test item was applied in its original form to the skin of five female then five male Sprague‑Dawley rats at the dose-level of 2000 mg/kg bw. The application site was covered by a semi‑occlusive dressing for 24 hours.

Each animal was observed at least once a day for mortality and clinical signs for 15 days. From day 2, any local reactions at the treatment site were also noted. Body weight was recorded the day before treatment, then on days 1, 8 and 15.

On completion of the observation period, the animals were sacrificed and then submitted to a macroscopic post-mortem examination.

No unscheduled deaths occurred during the study and no clinical signs were observed in any animals.

Local reactions such as erythema, dryness of the skin and scabs were transiently noted in 7 out 10 animals given 2000 mg/kg bw. These were considered to be treatment- related.

When compared to historical control data, the mean body weight gain was considered to be unaffected by the test item treatment in males and females. At necropsy, there were no macroscopic findings related to the test item administration.

The acute dermal LD50 was found greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Study complete and sufficient to fulfill the endpoint requirements.

Additional information

Oral route:

In an acute oral toxicity study performed according to OECD 423 guideline and in compliance with Good Laboratory practices, groups of 3 females rats received by gavage a single dose of 2000 or 300 mg/kg bw of the test substance.

The starting dose-level was 300 mg/kg bw for ethical reasons. After the first assay, the next higher dose-level of 2000 mg/kg bw was tested. Then, as no toxicity was observed at this higher dose-level, the results were confirmed in other females. Animals were then observed for 14 days following exposure.

No mortalities were observed throughout the study. Piloerection was observed in 3 out 6 females given 2000 mg/kg bw during the day of dosing and one day after. No clinical signs were observed in other animals. Compared to historical control data, the body weight gain was lower in 2 females given 300 mg/kg bw and in 4 out 6 females given 2000 mg/kg bw. At necropsy, there were no macroscopic findings related to the test item administration.

The acute oral LD0 was found equal or greater than 2000 mg/kg bw (Gerbeix, 2013a).

 

Dermal route:

 

In an acute dermal toxicity study performed according to OECD 402 guideline and in compliance with Good Laboratory Practices,the test substance was applied in its original form to the skin of five female then five male rats at the dose-level of 2000 mg/kg bw. The application site was covered by a semi‑occlusive dressing for 24 hours. Each animal was observed at least once a day for mortality and clinical signs for 15 days. From day 2, any local reactions at the treatment site were also noted. Body weight was recorded the day before treatment, then on days 1, 8 and 15. On completion of the observation period, the animals were sacrificed and then submitted to a macroscopic post-mortem examination.

No unscheduled deaths occurred during the study and no clinical signs were observed in any animals.

Local reactions such as erythema, dryness of the skin and scabs were transiently noted in 7 out 10 animals given 2000 mg/kg bw. These were considered to be treatment- related. When compared to historical control data, the mean body weight gain was considered to be unaffected by the test item treatment in males and females.At necropsy, there were no macroscopic findings related to the test item administration.

The acute dermal LD50 was found greater than 2000 mg/kg bw (Leclère, 2013a).

 

Inhalation:

 

In an acute inhalation toxicity study performed according to OECD 436 guideline and in compliance with Good Laboratory Practices,a group of three male and three female Wistar rats were exposed, nose-only, to an atmosphere of the test item using a flow-through exposure system. The animals was exposed for four hours to a target concentration of 5 mg/L followed by a fourteen day observation period.

Each animal was observed for mortality and clinical signs at hourly intervals during exposure, then one hour and two hours after exposure and at least twice a day during the 14 -day observation period. Bodyweights were recorded before treatment then on the day of exposure (day 1) and on days 2, 4, 8 and 15. All surviving animals were necropsied at the end of the observation period.

The mean achieved atmosphere concentration was 5.14 mg/L and the mean mass median aerodynamic diameter was 2.5 µm. No deaths occurred during the study. Immediately after the exposure two out of three females were noted to have a partially closed right eye, this was still present in one female at one hour after the exposure. Furthermore, at one hour after exposure one female was noted to be underactive with irregular breathing, the irregular breathing persisted and remained evident at two hours after the exposure. From two hours after exposure and for the remainder of the observation period all males were considered to be clinically normal; females were considered normal from Day 2.

Bodyweight losses were evident in all males and females on the day following the exposure. Recovery from the bodyweight loss was observed at the next weighing occasion, Day 4, after which body weight gains continued for the remainder of the study.

Enlargement of the tracheobronchial lymph nodes was noted in all animals.

The 4 -hour inhalation LC50 was found to be greater than 5.14 mg/L (ie 5140 mg/m3) (Beebe, 2013)

Justification for selection of acute toxicity – oral endpoint
Study performed according to the OECD guideline 423 and GLP compliant.

Justification for selection of acute toxicity – inhalation endpoint
Study performed according to the OECD guideline 436 and GLP compliant.

Justification for selection of acute toxicity – dermal endpoint
Study performed according to the OECD guideline 402 and GLP compliant.

Justification for classification or non-classification

No mortalities were noted in rats either after treatment by oral or dermal routes with a single dose of 2000 mg/kg body weight or exposure by inhalation for 4 hours to 5.14 mg/L.

On the basis of these results and according to regulation (EC) No. 1272/2008 and its subsequent amendments on classification, labeling and packaging (CLP) of substances and mixtures, no classification is warranted with respect to acute oral, dermal and inhalation toxicity.