Acetic acid, 2,2-difluoro-2-[[2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy]-, ammonium salt (1:1)

Endpoint summary

Description of key information

Two acute toxicity studies conducted in rats in compliance with GLP and according to testing standards are available on cC6O4 ammonium salt with the following results:

- an oral toxicity study according to OECD guideline 423 and EU method B.1 : 300 < rat LD50 < 2000 mg/kg

- a dermal toxicity study according to OECD guideline 402 and EU method B.3 : rat LD50 > 2000 mg/kg.

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:

2009-03-09 to 2009-12-14

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)

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

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Italy S.r.l., 33049 San Pietro al Natisone (UD), Italy
- Age at study initiation: 6 to 7 weeks old
- Weight at study initiation: 150 to 174 grams
- Fasting period before study: Overnight prior to dosing (Day –1)
- Housing: Polycarbonate cages measuring 59x38.5x20 cm, with stainless steel mesh lid and floor
- Diet (e.g. ad libitum): 4 RF 18 ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 2°C
- Humidity (%): 55% ± 15%
- Air changes (per hr): Approximately 15 to 25 air changes per hour
- Photoperiod (hrs dark / hrs light): Artificial (fluorescent tubes), daily light/dark cycle of 12/12 hours

IN-LIFE DATES: From: 2009-03-11 To: 2009-04-03

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

distilled

Details on oral exposure:

VEHICLE
- Concentration in vehicle: (160mg(dry salt)/ml) and 24 mg(dry salt)/ml
- Amount of vehicle (if gavage): Dose volume of 12.5 ml/kg of body weight for each animal.
- Justification for choice of vehicle: no justification given
- Lot/batch no. (if required): n.a.
- Purity: distilled water

MAXIMUM DOSE VOLUME APPLIED: Dose volume of 12.5 ml/kg of body weight for each animal.

DOSAGE PREPARATION (if unusual): n.a.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: 2000 mg(dry salt)/kg body weight, as limit dose for acute oral toxicity category IV (GHS/CLP)

Doses:

300 and 2000 mg (dry salt)/kg body weight

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Allocation (Day-1), Days 1, 2, 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

no statistics applied

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Mortality occurred in the 3 female animals dosed at 2000 mg(dry salt)/kg body weight (step 1); animals were found dead approximately 3 hours after dosing. No mortality was observed in the female animals dosed at 300 mg (dry salt)/kg body weight.

Clinical signs:

other: 2000 mg (dry salt)/kg body weight: Lethargy, difficult breathing, piloerection, tremors and salivation were observed prior to death with a various incidence. At 300 mg (dry salt)/kg body weight clinical signs were limited to piloerection observed at 2 and

Gross pathology:

At necropsy examination, performed on the early decedent animals dosed at 2000 mg(dry salt)/kg (step 1), abnormal colour of the liver (dark), abnormal contents (brown gelatinous) or areas (dark) in the stomach and red staining on the muzzle were recorded in all animals. In addition, abnormal colour (red) of the thymus was recorded in two animals.
No abnormalities were observed at necropsy examination performed at the end of the observation period on the animals dosed at 300 mg(dry salt)/kg body weight (steps 2 and 3).

Other findings:

none reported

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

cC6O4 ammonium salt has a toxic effect on the rat following oral administration of a single dose at a level of 2000 mg(dry salt)/kg.
The mortality pattern of the test item, cC6O4 ammonium salt, demonstrates the LD50 to be greater than 300 mg(dry salt)/kg but less than 2000 mg(dry salt)/kg body weight.
These results meet the criteria for European CLP Regulation classification as : Acute oral toxicity - Category 4.

Executive summary:

The acute toxicity of cC6O4 ammonium salt was investigated after a single oral administration (12.5 ml/kg in water) to female Sprague Dawley rats followed by a 14-day observation period. The animals were sacrificed at the end of the observation period and subjected to necropsy examination.

A first sub-group of 3 female animals was initially dosed at 2000 mg(dry salt)/kg body weight (step 1).

Mortality occurred in all animals between 2 and 4 hours after dosing. Lethargy, difficult breathing, piloerection, tremors and salivation were observed prior to death with various incidence.

At necropsy examination, performed on the early decedent animals, dark colour of the liver, brown gelatinous contents or dark areas in the stomach and red staining on the muzzle were recorded in all animals. In addition, red colour of the thymus was recorded in two animals.

A second and third subgroup, each composed of 3 females, were then dosed at 300 mg(dry salt)/kg body weight (steps 2 and 3). No mortality was recorded in any animal. Clinical signs were limited to piloerection observed on the day of dosing.

A very slight body weight loss was detected in two animals on Day 15, without any toxicological significance. Body weight changes were within the expected range for this strain and age of animals.

No abnormalities were observed at necropsy examination performed at the end of the observation period in these animals.

These results indicate that the test item, cC6O4 ammonium salt, has a toxic effect on the rat following oral administration of a single dose at a level of 2000 mg(dry salt)/kg. The mortality pattern demonstrates the LD50 to be greater than 300 mg(dry salt)/kg but less than 2000 mg(dry salt)/kg body weight. Based on criteria in the European Directives concerning the classification, packaging and labelling of dangerous substances (Council Regulation (EC) No. 1272/2008 and subsequent revisions), the classification is the following:

Classification : Acute oral toxicity - Category 4

Signal word : Warning

Hazard statement (Oral) : H302: Harmful if swallowed

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

> 300 - < 2 000 mg/kg bw

Quality of whole database:

Reliable guideline study

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

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:

2009-03-09 to 2009-12-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes

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: Harlan Italy S.r.l., 33049 San Pietro al Natisone (UD), Italy
- Age at study initiation: 6 to 8 weeks old
- Weight at study initiation: 183.7 to 194.1 grams
- Fasting period before study: Overnight prior to dosing (Day –1)
- Housing: Polycarbonate cages measuring 59x38.5x20 cm, with stainless steel mesh lid and floor
- Diet (e.g. ad libitum): 4 RF 18 ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 2°C
- Humidity (%): 55% ± 15%
- Air changes (per hr): Approximately 15 to 25 air changes per hour
- Photoperiod (hrs dark / hrs light): Artificial (fluorescent tubes), daily light/dark cycle of 12/12 hours

IN-LIFE DATES: From: 2009-02-26 To: 2009-03-26

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: 2.5x2.5 cm
- % coverage: 10%
- Type of wrap if used: A strip of synthetic film was placed over the treated site and the whole assembly held in place by encircling the trunk of the animal with a length of elastic adhesive bandage, this forming a semi-occlusive barrier.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treatment area was cleaned by gentle swabbing of the skin with cotton wool soaked with lukewarm water.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 367.4 mg - 388.2 mg of dry salt
- Concentration (if solution): n.a.
- Constant volume or concentration used: yes
- For solids, paste formed: A volume of 0.5 ml or 1.0 ml of sterile water was added and mixed to a paste at the dosing procedure

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.5 ml or 1.0 ml of sterile water
- Concentration (if solution): n.a.
- Lot/batch no. (if required): n.a.
- Purity: distilled water

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Allocation (Day-1), Days 1, 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

no statistics applied

Preliminary study:

not applicable

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

none observed

Clinical signs:

other: No systemic toxic effect were observed in male or female animals during the observation period. External examination of the treated site showed a slight oedema on Day 2 in all animals. Recovery from this sign occurred in all animals by Day 3. Desquamation

Gross pathology:

No abnormalities were found in any animals at necropsy performed at termination of the study.

Other findings:

none reported

Interpretation of results:

GHS criteria not met

Conclusions:

These results indicate that the test item, cC6O4 ammonium salt, has no toxic effect on the rat following dermal exposure over a 24 hour period at a level of 2000 mg/kg. The lack of mortality demonstrates the LD50 to be greater than 2000 mg/kg, thus there results no classification according to the European CLP regulation.

Executive summary:

The acute toxicity of cC6O4 ammonium salt was investigated following administration of a single dermal dose to the rat. No mortality occurred following dosing and no significant clinical signs were observed. These results indicate that the test item, cC6O4 ammonium salt, has no toxic effect on the rat following dermal exposure over a 24 hour period at a level of 2000 mg/kg. The lack of mortality demonstrates the LD50 to be greater than 2000 mg/kg. European Directives concerning the classification, packaging and labelling of dangerous substances and preparations (Regulation (EC) no. 1907/2006, 67/548/EEC and subsequent revisions) would indicate the following:

Classification : Not required

Signal word : None indicated

Hazard statement : None indicated

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

> 2 000 mg/kg bw

Quality of whole database:

Reliable guideline study

Additional information

An oral toxicity study according to OECD guideline 423 and EU method B.1 and a dermal toxicity study according to OECD guideline 402 and EU method B.3 were performed to test the acute toxicity of cC6O4 ammonium salt.

The studies were performed under GLP conditions.

In the oral toxicity study, the acute toxicity of cC6O4 ammonium salt was investigated after a single oral administration (12.5 ml/kg in water) to female Sprague Dawley rats followed by a 14-day observation period. (Doses: 300mg/kg b.w.(dry salt) , 2000 mg/kg b.w.(dry salt)).

The animals were sacrificed at the end of the observation period and subjected to necropsy examination. The results of the test indicate that the test item, cC6O4 ammonium salt, caused death on all the treated rats following oral administration of a single dose at a level of 2000 mg/kg b.w. (dry salt).

No mortality was recorded in any animal dosed at 300 mg/kg b.w. (dry salt).

These results are consistent with the toxicity observed in the dose-range finding study preliminary to the rat in vivo micronucleus study where the dose of 2000 mg/kg cause death of one out of 2 dosed Wistar rats, within a 48-hr observation period. No death was observed at up to the dose of 1750 m/kg bw after a 48-hr observation period.

In the dermal toxicity study, the acute toxicity of cC6O4 ammonium salt was investigated following administration of a single dermal dose to the rat. No mortality occurred following dosing and no significant clinical signs were observed. These results indicate that the test item, cC6O4 ammonium salt, has no toxic effect on the rat following dermal exposure over a 24 hour period at a level of 2000 mg/kg b.w. (dry salt). The lack of mortality demonstrates the LD50 to be greater than 2000 mg/kg b.w. (dry salt).

Justification for classification or non-classification

Basing on the results above reported, REGULATION (EC) No 1272/2008 (EU Regulation on Classification, Labelling and Packaging of substances and mixtures) would indicate the following:

 

Oral Toxicity

Classification : Acute oral toxicity - Category 4

Signal word : Warning

Hazard statement (Oral) : H302: Harmful if swallowed

 

Dermal toxicity

Classification : Not required

Signal word : None indicated

Hazard statement : None indicated