Reaction mass of Amines, hydrogenated tallow alkyl and azelaic acid and lithium hydroxide

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 April to 11 June 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-compliant, guideline study, available as an unpublished report.

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'.

Cross-reference

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

16 April to 11 June 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-compliant, guideline study, available as an unpublished report.

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

- Source: Charles River Deutschland, Sulzfeld, Germany. Animals were 8 to 10 weeks old and nulliparous and non-pregnant.
- Acclimatisation: The acclimatization period was at least 5 days before the start of treatment under laboratory conditions and animals were individually housed for the pilot study and group housed for the main study.
- Housing: Individual housing of animals in the pilot study and group housing of four animals per cage in the main study in labeled Makrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material and paper as cage-enrichment. The rats had free access to pelleted rodent diet and tap water.
- Environmental conditions: Temperature and relative humidity were set to achieve limits of 18 to 24 deg C, and 40 to 70% humidity. Rate of air exchange was at least 10 changes per hour and there was a 12 hour light/12 hour dark cycle.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

- Preparation: The preparations (w/w) were kept at room temperature protected from light and were dosed within 4 hours after adding the vehicle to the test substance. Homogeneity was obtained to visually acceptable levels.
- Dosing: Oral gavage, using plastic feeding tubes. The test item preparations were stirred on a magnetic stirrer during dosing. The concentration of the test substance in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.
- Fasting: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available.

Doses:

In the absence of any other toxicity information a "Pilot study" was conducted in which a single female rat was orally dosed with the test substance at 2000 mg/kg body weight. Based on the Pilot results, a fixed dose level of 2000 mg/kg body weight was then selected for the main study which was conducted in a stepwise approach on three animals. One animal at a time was treated at 2000 mg/kg body weight. When two animals were found dead, five animals were treated at a lower dose level of 300 mg/kg body weight.

No. of animals per sex per dose:

For the main study four female rats were treated at 2000 mg/kg body weight, and five female rats were subsequently treated at 300 mg/kg body weight.

Control animals:

no

Details on study design:

- Observations: Clinical observations were made on the day of dosing and then once daily until Day 15. Morbidity and viability checks were made twice daily and individual body weights were recorded on days 0, 8 and 15. At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

Statistics:

The Fixed Dose Procedure does not require any statistical analyses of the data.

Preliminary study:

In the preliminary study in one animal at 2000 mg/kg bw no mortality occurred. Hunched posture, piloerection and ptosis were noted between Days 1 and 12. The animal showed body weight loss between Days 1 and 8 and showed body weight gain again between Days 8 and 15. No findings were noted during macroscopic post mortem examination.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 mg/kg bw

Based on:

test mat.

Mortality:

At 2000 mg/kg, two animals were found dead, on Day 4 and 5 of the study.
At 300 mg/kg, no mortality occurred.

Clinical signs:

At 2000 mg/kg, hunched posture, piloerection and/or chromodacryorrhoea (snout) were noted for the animals between Days 1 and 4.
At 300 mg/kg, no clinical signs were noted for any of the animals.

Body weight:

At 2000 mg/kg, one of the surviving animals (pilot animal) showed bodyweight loss between Days 1 and 8 and showed body weight gain again between Days 8 and 15. The other surviving animal showed a normal body weight gain during the study period.
At 300 mg/kg, the mean body weight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

Gross pathology:

At 2000 mg/kg, abnormalities of the stomach (glandular mucosa: focus/foci, several, black-brown) were found in one of the animals found dead. Macroscopic post mortem examination of the other animal that died during the study and of the surviving animals at termination did not reveal any abnormalities. Autolysis was noted for both animals found dead. This was considered not toxicologically relevant.
At 300 mg/kg, no abnormalities were found at macroscopic post mortem examination of the animals.

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The highest dose level of Dilithium azelate that did not produce mortality in Wistar rats was established as 300 mg/kg body weight and the estimated minimum lethal dose level was 2000 mg/kg body weight. Based on the mortality rate at 2000 mg/kg body weight, it was concluded that the oral LD50 value of Dilithium azelate was within the range of 300-2000 mg/kg body weight. Based on these results Dilithium azelate should be classified as Category 4.

Executive summary:

The acute oral toxicity of Dilithium azelate to female Wistar rats was determined in a GLP-compliant, fixed-dose method study following OECD guideline 420 (Latour 2015). A preliminary study indicated no mortality of a single rat at 2000 mg/kg bw Dilithium azelate, therefore 2000 mg/kg bw was selected for the main study which was conducted in a stepwise approach on three further animals. Dilithium azelate was dosed by oral gavage and the rats were observed for mortality, systemic toxicity and bodyweight gain.When two animals were found dead at 2000 mg/kg bw, five animals were treated at a lower dose level of 300 mg/kg bw for 15 days. No deaths or significant signs of toxicity were observed at 300 mg/kg bw Dilithium azelate. The oral LD50 value is within the range 300 mg/kg bw - 2000 mg/kg bw Dilthium azelate. Based on these results Dilithium azelate should be classified as Category 4. The study is considered to be relevant and reliable for use for this endpoint.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Dilithium azelate

EC Number:

254-184-4

EC Name:

Dilithium azelate

Cas Number:

38900-29-7

Molecular formula:

C9H14O4.2Li

IUPAC Name:

dilithium azelate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Physical state: White powder
- Purity: 98.33%
- Substance identity: FLTL G14057
- Batch number: A058/99
- Expiration date: 2018-03-06
- Storage of test material: Room temperature in the dark

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

- Source: Charles River Deutschland, Sulzfeld, Germany. Animals were 8 to 10 weeks old and nulliparous and non-pregnant.
- Acclimatisation: The acclimatization period was at least 5 days before the start of treatment under laboratory conditions and animals were individually housed for the pilot study and group housed for the main study.
- Housing: Individual housing of animals in the pilot study and group housing of four animals per cage in the main study in labeled Makrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material and paper as cage-enrichment. The rats had free access to pelleted rodent diet and tap water.
- Environmental conditions: Temperature and relative humidity were set to achieve limits of 18 to 24 deg C, and 40 to 70% humidity. Rate of air exchange was at least 10 changes per hour and there was a 12 hour light/12 hour dark cycle.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

- Preparation: The preparations (w/w) were kept at room temperature protected from light and were dosed within 4 hours after adding the vehicle to the test substance. Homogeneity was obtained to visually acceptable levels.
- Dosing: Oral gavage, using plastic feeding tubes. The test item preparations were stirred on a magnetic stirrer during dosing. The concentration of the test substance in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.
- Fasting: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available.

Doses:

In the absence of any other toxicity information a "Pilot study" was conducted in which a single female rat was orally dosed with the test substance at 2000 mg/kg body weight. Based on the Pilot results, a fixed dose level of 2000 mg/kg body weight was then selected for the main study which was conducted in a stepwise approach on three animals. One animal at a time was treated at 2000 mg/kg body weight. When two animals were found dead, five animals were treated at a lower dose level of 300 mg/kg body weight.

No. of animals per sex per dose:

For the main study four female rats were treated at 2000 mg/kg body weight, and five female rats were subsequently treated at 300 mg/kg body weight.

Control animals:

no

Details on study design:

- Observations: Clinical observations were made on the day of dosing and then once daily until Day 15. Morbidity and viability checks were made twice daily and individual body weights were recorded on days 0, 8 and 15. At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

Statistics:

The Fixed Dose Procedure does not require any statistical analyses of the data.

Results and discussion

Preliminary study:

In the preliminary study in one animal at 2000 mg/kg bw no mortality occurred. Hunched posture, piloerection and ptosis were noted between Days 1 and 12. The animal showed body weight loss between Days 1 and 8 and showed body weight gain again between Days 8 and 15. No findings were noted during macroscopic post mortem examination.

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 mg/kg bw

Based on:

test mat.

Mortality:

At 2000 mg/kg, two animals were found dead, on Day 4 and 5 of the study.
At 300 mg/kg, no mortality occurred.

Clinical signs:

At 2000 mg/kg, hunched posture, piloerection and/or chromodacryorrhoea (snout) were noted for the animals between Days 1 and 4.
At 300 mg/kg, no clinical signs were noted for any of the animals.

Body weight:

At 2000 mg/kg, one of the surviving animals (pilot animal) showed bodyweight loss between Days 1 and 8 and showed body weight gain again between Days 8 and 15. The other surviving animal showed a normal body weight gain during the study period.
At 300 mg/kg, the mean body weight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

Gross pathology:

At 2000 mg/kg, abnormalities of the stomach (glandular mucosa: focus/foci, several, black-brown) were found in one of the animals found dead. Macroscopic post mortem examination of the other animal that died during the study and of the surviving animals at termination did not reveal any abnormalities. Autolysis was noted for both animals found dead. This was considered not toxicologically relevant.
At 300 mg/kg, no abnormalities were found at macroscopic post mortem examination of the animals.

Applicant's summary and conclusion

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The highest dose level of Dilithium azelate that did not produce mortality in Wistar rats was established as 300 mg/kg body weight and the estimated minimum lethal dose level was 2000 mg/kg body weight. Based on the mortality rate at 2000 mg/kg body weight, it was concluded that the oral LD50 value of Dilithium azelate was within the range of 300-2000 mg/kg body weight. Based on these results Dilithium azelate should be classified as Category 4.

Executive summary:

The acute oral toxicity of Dilithium azelate to female Wistar rats was determined in a GLP-compliant, fixed-dose method study following OECD guideline 420 (Latour 2015). A preliminary study indicated no mortality of a single rat at 2000 mg/kg bw Dilithium azelate, therefore 2000 mg/kg bw was selected for the main study which was conducted in a stepwise approach on three further animals. Dilithium azelate was dosed by oral gavage and the rats were observed for mortality, systemic toxicity and bodyweight gain.When two animals were found dead at 2000 mg/kg bw, five animals were treated at a lower dose level of 300 mg/kg bw for 15 days. No deaths or significant signs of toxicity were observed at 300 mg/kg bw Dilithium azelate. The oral LD50 value is within the range 300 mg/kg bw - 2000 mg/kg bw Dilthium azelate. Based on these results Dilithium azelate should be classified as Category 4. The study is considered to be relevant and reliable for use for this endpoint.