Ethyl 4-methyl-2-oxocyclohexanecarboxylate

Endpoint summary

Description of key information

Acute oral toxicity: OECD TG 423: LD50 > 2000 mg/kg bw
Acute dermal toxicity: OECD TG 402: LD50 > 2000 mg/kg bw

Acute inhalation toxicity: OECD 403: LC50 > 5 g/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:

06 April 2017 - 28 April 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

December 2001

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

December 2002

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

May 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Appendix to Director General Notification, No. 12-Nousan-8147. Agricultural Production Bureau, Ministry of Agriculture, Forestry and Fisheries of Japan (JMAFF)

Version / remarks:

November 2000, including the most recent revisions.

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

pH (1% in water, indicative range): 5.11 – 5.25
Specific gravity / density: 1.04 g/mL

Species:

rat

Strain:

other: Crl: WI(Han)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: approx. 8 weeks old
- Weight at study initiation: 145 - 156 g
- Fasting period before study: overnight (maximum of 20 hours)
- Housing: group housing up to 5 animals in polycarbonate cages containing sterilized sawdust as bedding material.
- Diet: pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany)
- Water: municipal tap-water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS (set to maintain)
- Temperature (°C): 21
- Humidity (%): 44-49
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 11 April 2017 - 28 April 2017

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg bw (since no vehicle was used, the dose volume was based on bodyweight measurement prior to dosing.

DOSAGE PREPARATION: the test Item was stored in the refrigerator and was allowed to warm to room temperature for at least 30 minutes before dosing. Adjustment was made for specific gravity of the test item. No correction was made for the purity/composition of the test item.

CLASS METHOD
- Rationale for the selection of the starting dose: The dose levels were based on the OECD test guidelines and were selected from the series 5 (lowest dose level), 50, 300 and 2000 (highest dose level) mg/kg body weight. The starting dose level should be the one that is likely to produce mortality in at least some of the animals and was selected based on available toxicity data of the test item.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Test groups consisted of 3 female animals. A first group was treated at a dose level of 2000 mg/kg. Based on the results, one additional group was dosed at 2000 mg/kg.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
* mortality: twice daily
* clinical observations: at least three times on the day of dosing (periodic intervals) and once daily thereafter.
* body weights (individually): on day 1, day 8 and day 15.
- Necropsy of survivors performed: yes
- Other observations: descriptions of all internal macroscopic abnormalities were recorded.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: Hunched posture, uncoordinated movements and piloerection were noted for all animals on Day 1.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Interpretation of results:

GHS criteria not met

Remarks:

Not classified according to Regulation (EC) No. 1272/2008

Conclusions:

Based on the results of an acute oral toxicity study, performed according to OECD guideline 423, the LD50 of FRET 13-0545 for female Wistar rats exceeded 2000 mg/kg bw. The test item is therefore not classified according to GHS and Regulation (EC) No. 1272/2008.

Executive summary:

In the acute oral toxicity study in rats, performed according to OECD TG 423, the oral LD50 value of FRET 13-0545 in Wistar rats was established to exceed 2000 mg/kg body weight.

According to the OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.

Based on these results, FRET 13-0545 does not have to be classified and has no obligatory labelling requirement for acute oral toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The acute oral toxicity result is of sufficient quality and adequate for this dossier.

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:

05 May 2017 - 14 September 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

September 2009

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Version / remarks:

August 1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

Version / remarks:

May 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Appendix to Director General Notification, No. 12-Nousan-8147. Agricultural Production Bureau, Ministry of Agriculture, Forestry and Fisheries of Japan (JMAFF)

Version / remarks:

November 2000, including the most recent revisions

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

Stability at higher temperatures: Yes, maximum temperature: 50°C, maximum duration: 1440 minutes
Vapour pressure: 1.18 Pa
pH (1% in water, indicative range): 5.11 – 5.25
Specific gravity / density: 1.04 g/mL

Species:

rat

Strain:

other: Crl: WI(Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9-11 weeks old
- Weight at study initiation: males: 251-300 g; females: 175-221 g
- Fasting period before study: not specified
- Housing: group housed (max 5 animals of the same sex and same exposure group together) in polycarbonate cages containing sterilized sawdust as bedding material
- Diet: pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany), ad libitum
- Water: municipal tap-water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS (set to maintain)
- Temperature (°C): 18-24 (actual: 20-21)
- Humidity (%): 40-70 (actual: 50-71)
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 8 May 2017 To: 14 September 2017

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Mass median aerodynamic diameter (MMAD):

> 1 - < 4 µm

Geometric standard deviation (GSD):

> 1.5 - < 3

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: exposure chamber based on the directed flow nose only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983)
- Exposure chamber volume: not indicated
- Method of holding animals in test chamber: polycarbonate restraining tubes
- Source and rate of air: at least 1 L/min (theoretical air flow in each animal port); mean total air flow: 22 L/min (first test group) and 28 L/min (second test group)
- System of generating aerosols: nebulization of the test item with pressurized air: the test item was transferred to a Collison nebulizer by means of a rotating pump (type VL500 digit, VERDER Lab Tec GmbH & Co. KG, Haan, Germany) in order to maintain a constant level of freshly available test item. The primary aerosol was subsequently diluted with pressurized air before it entered the exposure chamber
- Method of particle size determination: samples were drawn with a flow of 2 L/min. from the test atmosphere through a tube mounted in one of the free animal ports of the exposure chamber. The samples were collected with an 8 stage Marple personal cascade impactor containing fiber glass filters (TE-290-GF. Tisch Environmental, Cleves, Ohio, USA) and a fiber glass back-up filter (SEC- 290-F1, Westech, Upper Stondon, Bedfordshire, England). Amounts of test item collected were measured gravimetrically.
- Treatment of exhaust air: filtered and released to the exhaust of the fume hood
- Temperature and humidity in air chamber: 20.8-21.6 °C and 5-10%

TEST ATMOSPHEREn (see table 1 and 2 for MMAD and GSD)
- Brief description of analytical method used: Samples were drawn from the test atmosphere through a tube mounted in one of the free animal ports of the exposure chamber. Samples were drawn through a glass fiber filter. Sample volumes were measured by means of a dry gas meter. The collected amount of test item in the air sample was measured gravimetrically.
- Samples taken from breathing zone: yes, a total of 15 and 18 representative samples were taken for determination of the actual concentration during the first and second treatment group, respectively.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: trial generation results showed that the test item was not volatile and the test atmosphere consisted mainly of aerosol with a negligibly small (if any) vapor part. Trial generation results showed that it was not possible to generate a suitable test atmosphere sufficiently stable at the highest target concentration. Instead, the generation was performed at the technically maximum attainable concentration.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

5 mg/L for both test groups

No. of animals per sex per dose:

3 animals were initially exposed, 2 animals were additionally exposed

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
During exposure: mortality, behavioral signs of distress and effects on respiration at least three times a day
After exposure: mortality: twice daily; clinical signs: at least two times on the day of exposure and once daily thereafter
Body weights (individually): on day 1, 2, 4, 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: descriptions of all internal macroscopic abnormalities were recorded.

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

No mortality occurred.

Clinical signs:

other: During exposure, slow breathing was seen. After exposure, lethargy, hunched posture, labored respiration, chromodacryorrhea (nose) and ptosis were seen for the animals on days 1 and/or 2.

Body weight:

Overall body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study, except for one female which showed body weight loss up to Day 8. This female regained weight during the second week.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Other findings:

- Actual exposure concentrations: for the initially exposed group, the time weighted mean actual concentration was 5.0 ± 0.1 mg/L. The nominal concentration was 7.2 mg/L and the generation efficiency was 70%. For the additionally exposed group, the time weighted mean actual concentration was 5.1 ± 0.1 mg/L. The nominal concentration was 6.8 mg/L and the generation efficiency was 76%.
- Particle size: the Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice during each exposure period (see table 1 and 2).

Table 1 Aerodynamic particle size distribution in the test atmosphere (first test group)

Measurement 1:

Stage	Cut point (mm)		Mass sampled (mg)	Relative mass (%)	Cumulative mass (% of total sampled)
1	21.0		0.10	1.27	98.73
2	15.0		0.00	0.00	98.73
3	10.0		0.56	7.13	91.59
4	6.0		1.37	17.45	74.14
5	3.5		2.56	32.61	41.53
6	2.0		2.41	30.70	10.83
7	0.9		0.61	7.77	3.06
8	0.5		0.22	2.80	0.25
Back up	0.25		0.02	0.25	0.00
MMAD^1(mm):		3.7
gsd^2:		1.9

Measurement 2:

Stage	Cut point (mm)		Mass sampled (mg)	Relative mass (%)	Cumulative mass (% of total sampled)
1	21.0		0.03	0.37	99.63
2	15.0		0.08	0.99	98.65
3	10.0		0.57	7.02	91.63
4	6.0		1.25	15.39	76.23
5	3.5		2.76	33.99	42.24
6	2.0		2.56	31.53	10.71
7	0.9		0.71	8.74	1.97
8	0.5		0.12	1.48	0.49
Back up	0.25		0.04	0.49	0.00
MMAD^1(mm):		3.7
gsd^2:		1.9

¹ Mass Median Aerodynamic Diameter; ² Geometric standard deviation

 

Table 2 Aerodynamic particle size distribution in the test atmosphere (second test group)

Measurement 1:

Stage	Cut point (mm)		Mass sampled (mg)	Relative mass (%)	Cumulative mass (% of total sampled)
1	21.0		0.08	0.63	99.37
2	15.0		0.04	0.31	99.06
3	10.0		0.31	2.44	96.62
4	6.0		1.63	12.81	83.81
5	3.5		4.93	38.76	45.05
6	2.0		4.26	33.49	11.56
7	0.9		1.31	10.30	1.26
8	0.5		0.16	1.26	0.00
Back up	0.25		0.00	0.00	0.00
MMAD^1(mm):		3.7
gsd^2:		1.8

Measurement 2:

Stage	Cut point (mm)		Mass sampled (mg)	Relative mass (%)	Cumulative mass (% of total sampled)
1	21.0		0.03	0.22	99.78
2	15.0		0.00	0.00	99.78
3	10.0		0.35	2.58	97.20
4	6.0		1.78	13.11	84.09
5	3.5		5.30	39.03	45.07
6	2.0		4.45	32.77	12.30
7	0.9		1.49	10.97	1.33
8	0.5		0.18	1.33	0.00
Back up	0.25		0.00	0.00	0.00
MMAD^1(mm):		3.5
gsd^2:		1.7

¹ Mass Median Aerodynamic Diameter; ² Geometric standard deviation

Interpretation of results:

GHS criteria not met

Remarks:

Not classified according to Regulation (EC) No. 1272/2008

Conclusions:

In an acute inhalation study, performed according to OECD 403 and GLP, the LC50 of FRET 13-0545 in rats was established to exceed 5 mg/L. Based on these results, the test item is not classified according to GHS and Regulation (EC) No. 1272/2008.

Executive summary:

In an acute inhalation study, performed according to OECD 403 and GLP, FRET 13-0545 was administered as an aerosol by nose only inhalation for 4 hours to one group of three male and three female Wistar rats. This group was expanded with another two animals of each sex on a later time span to meet the guideline requirements.

For the initially exposed animals, the time-weighted mean actual concentration was 5.0 ± 0.1mg/L, and the MMAD was 3.7 μm (gsd 1.9)(twice). For the additionally exposed animals, the time-weighted mean actual concentration was 5.1 ± 0.1mg/L, the MMAD was 3.7 μm (gsd 1.8) and 3.5 μm (gsd 1.7) (measured twice).

The inhalation LC50, 4h value of FRET 13-0545 in Wistar rats was established to exceed 5 mg/L.

Based on these results FRET 13-0545 does not have to be classified and has no obligatory labelling requirement for acute inhalation toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015)

(including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

5 000 mg/m³ air

Quality of whole database:

The acute oral toxicity result is of sufficient quality and adequate for this dossier.

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:

05 July 2017 - 20 July 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

1987

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Version / remarks:

August 1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

May 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Appendix to Director General Notification, No. 12-Nousan-8147. Agricultural Production Bureau, Ministry of Agriculture, Forestry and Fisheries of Japan (JMAFF)

Version / remarks:

November 2000, including the most recent revisions.

Deviations:

no

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

pH (1% in water, indicative range): 5.11 – 5.25
Specific gravity / density: 1.04 g/mL (in this study, adjustment was made for the density of the test item)

Species:

rat

Strain:

other: Crl: WI(Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: approx. 10 weeks old
- Weight at study initiation: males: 273-299 g; females: 195-202 g.
- Fasting period before study: no
- Housing: Group housing (up to 5 animals of the same sex together) on arrival and individual housing during the study. Polycarbonate cages (Makrolon MIV type; height 18 cm.) containing sterilized sawdust were used.
- Diet: Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany), ad libitum throughout the study, except during designated procedures.
- Water: Municipal tap-water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-22
- Humidity (%): 45-72 (Temporary deviations from the maximum level of target humidity occurred. This study plan deviation did not affect the integrity of the study because laboratory historical data do not indicate an effect of these deviations.)
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 6 July 2017 To: 20 July 2017

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: 25 cm^2 for males, 18 cm^2 for females
- % coverage: 10% of the total body surface
- Type of wrap if used: The test item was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): using water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount applied: The dose volume for each animal was based on the body weight measurement prior to dosing. Dose volume (mL/kg body weight) was calculated as follows: Dose level (g/kg) / spec.gravity or density (g/mL).
- Constant volume or concentration used: no

Duration of exposure:

24 hours

Doses:

2000 mg/kg bodyweight

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: Mortality: twice daily; Clinical observations: at periodic intervals on the day of dosing (at least three times) and once daily thereafter; Body weights: on day 1 (pre-administration), day 8 and day 15.
- Necropsy of survivors performed: yes

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: Chromodacryorrhoea (snout) was noted for one male between Days 2 and 4 and three females on Day 1. No further clinical signs were noted.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Interpretation of results:

GHS criteria not met

Remarks:

Not classified according to Regulation (EC) No. 1272/2008

Conclusions:

Based on the results of an acute dermal toxicity study in rats, performed according to OECD guideline 402 and GLP principles, FRET 13-0545 is not classified according to GHS and Regulation (EC) No. 1272/2008.

Executive summary:

In an acute dermal toxicity study in rats, performed according to OECD guideline 402 and GLP principles, FRET 13-0545 was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours.

The dermal LD50 value of FRET 13-0545 in Wistar rats was established to exceed 2000 mg/kg body weight.

Based on these results, FRET 13-0545 does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The acute oral toxicity result is of sufficient quality and adequate for this dossier.

Additional information

Acute oral toxicity

In the acute oral toxicity study in rats, performed according to OECD TG 423, FRET 13-0545 was administered by oral gavage to two consecutive groups of three female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred. Hunched posture, uncoordinated movements and piloerection were noted for all animals on Day 1. The body weight gain shown by the animals over the study period was considered to be normal. No abnormalities were found at macroscopic post mortem examination of the animals.

The oral LD50value of FRET 13-0545 in Wistar rats was established to exceed 2000 mg/kg body weight.

Acute inhalation toxicity

In an acute inhalation study, performed according to OECD 403 and GLP, FRET 13-0545 was administered as an aerosol by nose only inhalation for 4 hours to one group of three male and three female Wistar rats. This group was expanded with another two animals of each sex on a later time span to meet the guideline requirements. Mortality and clinical signs were observed daily during the observation period and body weights were determined on Days 1, 2, 4, 8 and 15. Macroscopic examination was performed after terminal sacrifice (Day 15).

For the initially exposed animals, the time-weighted mean actual concentration was 5.0 ± 0.1 mg/L, the MMAD was 3.7 µm (gsd 1.9). For the additionally exposed animals, the time-weighted mean actual concentration was 5.1 ± 0.1 mg/L. the MMAD was 3.7 µm (gsd 1.8) and 3.5 µm (gsd 1.7).

No mortality occurred. During exposure, slow breathing was seen. After exposure, lethargy, hunched posture, labored respiration, chromodacryorrhea (nose) and ptosis were seen for the animals on Days 1 and/or 2. Overall body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study, except for one female which showed body weight loss up to Day 8. This female regained weight during the second week. No abnormalities were found at macroscopic post mortem examination of the animals.

The inhalation LC_{50, 4h}value of FRET 13-0545 in Wistar rats was established to exceed 5 mg/L.

Acute dermal toxicity

In an acute dermal toxicity study in rats, performed according to OECD guideline 402 and GLP principles, FRET 13-0545 was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred. Chromodacryorrhoea (snout) was noted for one male between Days 2 and 4 and three females on Day 1. No further clinical signs were noted. The body weight gain during the observation period was within the range expected for rats used in this type of study. No abnormalities were found at macroscopic post mortem examination of the animals.

The dermal LD50 value of FRET 13-0545 in Wistar rats was established to exceed 2000 mg/kg body weight.

Justification for classification or non-classification

According to the criteria outlined in Annex I of Regulation (EC) No 1272/2008/EC (CLP), the substance does not have to be classified for acute toxicity by the oral, dermal and inhalation route.