Ammonium undecafluorohexanoate

Endpoint summary

Description of key information

Oral (OECD 420), rat: LD50 > 300 mg/kg bw < 2000 mg/kg bw

Dermal (OECD 402), rat: LD50 > 2000 mg/kg bw

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03 - 24 Apr 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Version / remarks:

adopted in 2001

Deviations:

no

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

no

Specific details on test material used for the study:

- storage: at room temperature in a shaded and air-tight container in a desiccator

Species:

rat

Strain:

other: Crl:CD(SD)

Remarks:

SPF

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hino Breeding Centers (Charles River Laboratories), Hino, Japan
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 to 9 weeks
- Weight at study initiation: 189.1 g (first sighting study), 193.2 g (second sighting study) and 196.3 – 200.0 g (main study)
- Fasting period before study: animals were fasted overnight prior to administration (17 – 19 h)
- Housing: Group housing of 4 females per cage in stainless steel cages (260 x 380 x 180 mm) with mesh-floor
- Diet: MF pelleted diet (Oriental Yeast), ad libitum
- Water: chlorinated tap water (chloric level maintained from 3 to 5 ppm by addition of sodium hypochloride (Purelox) to tap water), ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 25
- Humidity (%): 40 - 70
- Air changes (per hr): 10 -15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 27 Mar 2018 To: 24 Apr 2018

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

Administration /exposure:
VEHICLE
- Concentration in vehicle: 20% (w/v)
- Amount of vehicle: 10 mL/kg bw
- Lot/batch No.: PC171219

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION:
Dosing formulation was prepared freshly on each administration day. The test substance was weighed and mixed with purified water to be dissolved. The solution was filled up to the prescribed volume with purified water to prepare the dosing formulation. The dosing formulations were administered within one hour after the preparation.

Doses:

300 mg/kg bw
(300 mg/kg bw for 1st sighting, 2000 mg/kg bw for 2nd sighting)

No. of animals per sex per dose:

300 mg/kg bw: 5 (incl. sighting study)
2000 mg/kg bw: 4

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were observed for mortality and clinical signs continuously for 10 min after the administration, 30 min and 3 h after the administration and once daily during Days 1 to 14 after the administration. Individual body weights were recorded before dose administration and 1, 7 and 14 days post-dose.
- Necropsy of survivors performed: yes

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

300 mg/kg bw: 0/4 (main study), 0/1 (1st sighting study)
2000 mg/kg bw: 1/1 (2nd sighting study) within 24 h after administration

Clinical signs:

300 mg/kg bw: No clinical signs of toxicity were observed up to the end of the 14 day observation period
2000 mg/kg bw: decreased spontaneous locomotion, decreased respiratory rate, incomplete eyelid opening and moist hair (abdomen) in 1/1 animals 3 h after administration. Lacrimation was observed 5 h after administration.

Body weight:

One animal dosed at 300 mg/kg bw showed a low body weight gain of 4.5 g one day after administration. The effect was not considered toxicologically significant, because no abnormalities were observed in the general clinical observation or necropsy.

Gross pathology:

300 mg/kg bw: Necropsy and histopathological examination revealed no substance-related findings.
2000 mg/kg bw: edematous change of limiting ridge of the forestomach was observed

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

CLP: Acute Oral (4), H302

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

10 Feb - 2 Mar 2004

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:

adopted in 2001

Deviations:

yes

Remarks:

Due to a calculation error animals were inadvertently dosed at 3000 mg/kg bw instead of 300 mg/kg bw in first step.

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted in 1996

Deviations:

yes

Remarks:

Due to a calculation error animals were inadvertently dosed at 3000 mg/kg bw instead of 300 mg/kg bw in first step.

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

adapted in 2002

Deviations:

not specified

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

Crl:(WI) BR (outbred, SPF-Quality)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: approx. 10 weeks
- Weight at study initiation: 206 - 232 g, body weight did not exceed ± 20% of the sex mean
- Fasting period before study: overnight (for a maximum of 20 h) prior to dosing until 3-4 h after administration of the test substance
- Housing: groups of 3 animals in Macrolon cages (Type IV; height 18 cm) on sawdust bedding
- Diet: standard pelleted laboratory animal diet (VRF1, Altromin, Lage, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15.4-21.7
- Humidity (%): 29-66
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 2.79 mL/kg bw

CLASS METHOD
- Rationale for the selection of the starting dose: The first set of animals was inadvertently dosed at 3000 mg/kg bw instead of 300 mg/kg bw (due to a calculation error).

Doses:

3000 mg/kg bw (step 1; inadvertently dosed by calculation error instead of 300 mg/kg bw)
2000 mg/kg bw (step 2)

No. of animals per sex per dose:

3 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed from Day 1 to 15 twice daily for mortality/viability and once daily and for clinical signs. Body weights were determined before administration (Day 1) and on Days 8 and 15.
- Necropsy of survivors performed: yes

Statistics:

No statistical analysis was performed.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 3 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: The LD50 was converted from 3000 mg/kg bw/day (as given in the study report) to 600 mg/kg bw/day, since the study was conducted with a 20% solution of the test substance.

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: according to OECD 423

Mortality:

No mortality occurred during the study period.

Clinical signs:

Hunched posture was noted in all animals on day 1 and/or 2.

Body weight:

The body weight gain shown by the animals over the study period was considered to be similar to that expected of untreated animals of the same age and strain.

Gross pathology:

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

Interpretation of results:

study cannot be used for classification

Conclusions:

In this acute oral toxicity study in rats no animal died up to the highest dose tested. Therefore, a LD50 value of > 3000 mg/kg bw was determined. Since the study was conducted with a 20% solution of the test substance, the LD50 was converted to 600 mg/kg bw.
This study cannot be used for classification purposes, as a dilution of the test substance was used. As the undiluted substance was shown to be corrosive to the skin, generating additional acute oral toxicity data is not required.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Quality of whole database:

The available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.5, of Regulation (EC) No.1907/2006.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 Mar - 17 Apr 2018

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:

adopted in 1987

Deviations:

no

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

- storage: at room temperature in a shaded and air-tight container in a desiccator

Species:

rat

Strain:

other: Crl:CD (SD)

Remarks:

SPF

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hino Breeding Centers (Charles River Laboratories), Hino, Japan
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 7 weeks (males) and 9 weeks (females)
- Weight at study initiation: 250.4 – 254.4 g (males) and 218.5 – 232.5 g (females)
- Housing: Individually in stainless steel cages (260 x 380 x 180 mm) with mesh-floor
- Diet: MF pelleted diet (Oriental Yeast), ad libitum
- Water: chlorinated tap water (chloric level maintained from 3 to 5 ppm by addition of sodium hypochloride (Purelox) to tap water), ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 25
- Humidity (%): 40 - 70
- Air changes (per hr): 10 -15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 27 Mar 2018 To: 17 Apr 2018

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: 5 x 5 cm clipped skin of the dorsal area
- Type of wrap if used: The test material was applied on a non-woven gauze which was covered and fixed by elastic adhesive bandage (SILKYTEX5, ALCARE).

REMOVAL OF TEST SUBSTANCE
- Washing: Residual test substance was removed using purified water and absorbent cotton.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount applied: 2000 mg/kg bw
- Concentration: 20% (w/v)
- Dose volume: 10 mL/kg bw

VEHICLE
- Amount applied: 10 mL/kg bw
- Lot/batch No.: PC171219

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were observed for clinical signs and mortality continuously for 10 min after the administration, 30 min and 3 h after the administration and once daily during Days 1 to 14 after the administration. Individual body weights were recorded prior to administration as well as 1, 7 and 14 days post-dose.
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occured during the study period.

Clinical signs:

2000 mg/kg bw: All animals showed slight decreased spontaneous locomotion after the application and 3 h after administration. All effects were reversible within Day 2 of the observation period.

Body weight:

Body weight gains were within the expected ranges in males and females of this species and strain during the study period.

Gross pathology:

2000 mg/kg bw: Pelvic dilation of the right kidney was observed in 1/5 males.

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008

Conclusions:

CLP: not classified

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

30 Mar - 13 Apr 2004

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:

adopted in 1987

Deviations:

yes

Remarks:

occlusive instead of semi-occlusive dressing

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted in 1992

Deviations:

yes

Remarks:

occlusive instead of semi-occlusive dressing

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Version / remarks:

adopted in June 1996

Deviations:

not specified

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

Crl:(WI) BR (outbred, SPF-Quality)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: approx. 11 weeks
- Weight at study initiation: 405-462 g (range males) and 254-298 g (range females)
- Housing: individually in Macrolon cages (Type III, height 15 cm) on saw dust bedding
- Diet: standard pelleted laboratory animal diet (VRF 1, Altromin, Lage, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.5-24.2
- Humidity (%): 29-67
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: 5x7 cm clipped skin on the dorsal area of the trunk
- % coverage: 10%
- Type of wrap if used: The test substance was held in contact with the skin with a dressing, consisting of a surgical gauze patch, successively covered with aluminium 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: The skin was cleaned of residual test substance using water.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount applied: 1.86 mL/kg bw
- Constant volume or concentration used: no

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed from Day 1 to Day 15 twice daily for mortality and once daily for clinical signs. Body weights were determined before administration (Day 1) and on Days 8 and 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.

Remarks on result:

other: The LD50 was converted from 2000 mg/kg bw/day (as given in the study report) to 400 mg/kg bw/day, since the study was conducted with a 20% solution of the test substance.

Mortality:

No mortality occurred during the study period.

Clinical signs:

Flat or hunched posture was noted 5/5 males and 4/5 females and chromodacryorrhea was noted in three males. The animals had recovered from the symptoms between Days 2 and 4.

Body weight:

The body weight gain in males and females were within the range expected for this species and strain.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.
Cysts in ovaries, found in one female, are commonly noted among rats of this age and strain and was therefore considered not toxicologically relevant.

Other findings:

Skin reactions:
Scales and/or scrabs were observed in the treated skin-area of 4/5 males and females, respectively during the observation period.

Interpretation of results:

study cannot be used for classification

Conclusions:

In this acute dermal toxicity study in rats no animal died up to the highest dose tested. Therefore, a LD50 value of > 2000 mg/kg bw was dertermined. Since the study was conducted with a 20% solution of the test substance, the LD50 was converted to 400 mg/kg bw.
This study cannot be used for classification purposes, as a dilution of the test substance was used. As the undiluted substance was shown to be corrosive to the skin, generating additional acute dermal toxicity data is not required.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.5, of Regulation (EC) No. 1907/2006.

Additional information

Acute oral toxicity:

Two experimental studies are available to evaluate the acute oral toxicity of ammonium undecafluorohexanoate.

The first study (Teunissen, 2004) the acute oral toxicity of a 20% dilution of the test substance was assessed according to OECD guideline 423, EU Method B.1 tris and in compliance with GLP (Teunissen, 2004). In a first step, a total dose of 3000 mg/kg bw of the test substance was administered to 3 female Wistar rats. Due to a calculation error the first set of animals was inadvertently dosed at 3000 mg/kg bw instead of 300 mg/kg bw. Animals were observed from Day 1 until Day 15 twice daily for mortality and once daily for clinical signs of toxicity. The individual body weights were recorded prior to dosing (Day 1) and on Day 7 and 15. Macroscopic examination was performed at terminal sacrifice. Since no mortality and no severe clinical signs occurred in first step, another 3 females were treated with 2000 mg/kg bw of the test substance in a second step. No mortality was observed at this dose level until the end of the study period. Hunched posture was noted in animals of both dose groups on Day 1 and/or 2.The body weight (gain) was not affected by the administration of the test substance. Based on the results of this study, the LD50 value for acute oral toxicity was determined to be > 3000 mg/kg bw in rats. Since the study was conducted with a 20% solution of the test substance, the LD50 value was converted to > 600 mg/kg bw.

This study cannot be used for classification purposes, as a dilution of the test substance was used.

A second study was performed using test material based on 100% mass formulation. Ammonium undecafluorohexanoate was tested for acute oral toxicity according to OECD guideline 420 (fixed dose method) and in compliance with GLP (Tsubokura, 2018a). The test substance was administered as a 20% (w/v) formulation dissolved in water by oral gavage to female Crl:CD (SD) rats.

In the first of 2 sighting studies, 1 animal was administered 300 mg/kg bw.  No mortality was observed and no clinical signs were reported. In the 2nd sighting study, 1 animal was administered 2000 mg/kg bw. This animal showed decreased spontaneous locomotion, decreased respiratory rate, incomplete eyelid opening and moist hair from three hours after administration. Five hours after administration lacrimation was observed. The animal was found dead within 24 hours after administration. Based on the result of the sighting studies, 300 mg/kg bw was selected as the dose for the main study. Four animals were administered 300 mg/kg bw and observed for a period of 14 days. No mortality occurred during the observation period. No clinical signs were reported and no toxicologically relevant effects on body weight were noted. Necropsy of animals dosed at 300 mg/kg bw showed no treatment-related findings, while the animal administered 2000 mg/kg had an edematous change of limiting ridge of the forestomach. The acute oral LD50 value in female rats was > 300 < 2000 mg/kg bw.

Conclusion acute oral toxicity

In conclusion, the results obtained in the first study with diluted test material supports the findings observed in the second study with undiluted test material. Based on all available data on acute oral toxicity the acute oral LD50 value of ammonium undecafluorohexanoate in female rats was > 300 < 2000 mg/kg bw. Ammonium undecafluorohexanoate is therefore classified as Acute Tox. 4.

Acute dermal toxicity:

There are two study reports available assessing the acute dermal toxicity potential of ammonium undecafluorohexanoate, both according to OECD guideline 402 (fixed dose method) and compliant with GLP (NOTOX B.V., 2004 and Tsubokura, 2018b).

In the first study (Teunissen, 2014) the acute dermal toxicity of a 20% dilution of the test substance was assessed in a limit test performed in 5 male and 5 female Wistar rats. The test substance was applied as a single dose of 2000 mg/kg bw to a clipped dorsal area of the trunk of the animals and held in contact with the skin with an occlusive dressing for 24 hours. Animals were observed from Day 1 until Day 15 twice daily for mortality and once daily for clinical signs of toxicity. The individual body weights were recorded prior to dosing (Day 1) and on Day 7 and 15. Macroscopic examination was performed at terminal sacrifice. None of the animals died and no clinical signs were observed. The body weight (gain) was not affected by the administration of the test substance. In one female animal cysts were noted in the ovaries but as no other parameters or animals were affected this is an incidental finding. No further pathological findings were observed at necropsy. Based on the results of this study, the LD50 value for acute dermal toxicity was determined to be > 2000 mg/kg bw in rats. Since the study was conducted with a 20% solution of the test substance, the LD50 value was converted to > 400 mg/kg bw.

This study cannot be used for classification purposes, as a dilution of the test substance was used.

A second study was performed using test material based on 100% mass formulation. Ammonium undecafluorohexanoate was applied as a 20% formulation dissolved in water to the clipped skin of the dorsal area of 5 male and 5 female Crl:CD (SD) rats. The skin was covered with a semi-occlusive wrap for a treatment period of 24 hours. After exposure ended, residual test substance was removed with water and absorbent cotton. No mortality was observed throughout the 14-day study period. Slightly decreased spontaneous locomotion in all animals was observed after administration and 3 h after administration. Body weight gains remained unaffected and were within the ranges expected for this species and strain. At terminal necropsy pelvic dilation of the right kidney was observed in a single male animal, which was considered to be a spontaneous lesion with no toxicological relevance. The acute dermal value LD50 in male and female rats was > 2000 mg/kg bw.

Conclusion acute dermal toxicity

In conclusion, based on the available data on acute dermal toxicity the acute dermal LD50 of ammonium undecafluorohexanoate in male and female rats was > 2000 mg/kg bw.

Justification for classification or non-classification

The available data on acute oral and dermal toxicity of the test substance meet the criteria for classification as Acute tox. oral 4 (H302) according to Regulation (EC) No. 1272/2008.