Neodecanoic acid, iron salt

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-09-04 to 2017-11-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Version / remarks:

adopted 17 December 2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 17 December 2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Neodecanoic acid, iron salt

EC Number:

257-446-6

EC Name:

Neodecanoic acid, iron salt

Cas Number:

51818-55-4

Molecular formula:

C10H20O2.xFe

IUPAC Name:

λ²-iron(2+) bis(2-ethyl-2,5-dimethylhexanoate)

Test material form:

solid

Details on test material:

- CAS Number: 51818-55-4
- EC Number: 257-446-6
- Molecular formula: C30H57FeO6
- Molecular weight: 569.6 g/mol
- Purity: 100%
- Storage conditions: Refrigerated under Nitrogen (2-8 °C)

Specific details on test material used for the study:

- Storage: 2 to 8 °C, protected from light, under nitrogen
- Batch Number: Ln11013894
- Retest Date : 31 July 2019
- Purity: 100%
- Appearance:brown viscous liquid


TREATMENT OF TEST MATERIAL PRIOR TO TESTING
Dose levels were expressed gravimetrically and in terms of test article received (without regard to purity or active content). The test article was prepared in corn oil. The formulated concentrations were calculated from the selected dose level and the dose volume of 10 mL/kg. All formulations were used within two hours of preparation. Dose containers were repeatedly inverted prior to administration to ensure homogeneity.

Test animals

Species:

rat

Strain:

other: Female (nulliparous, non-pregnant) Crl:WI(Han) strain rats

Sex:

female

Details on test animals or test system and environmental conditions:

- Environmental Conditions, Diet, Water and Bedding
The animals were kept in the following conditions except for short periods of time where experimental procedures dictated otherwise.
- Housing
The animals were housed in groups of up to five in cages that conformed to the 'Code of Practice for the Housing and Care of Animals Bred, Supplied or Used for Scientific Purposes’ (Home Office, London, 2014).
Bedding was provided on a weekly basis to each cage by use of clean European soft wood bedding (Datesand Ltd., Manchester, UK). The bedding had been analysed for specific contaminants.
No contaminants were present in bedding at levels which might have interfered with achieving the objective of the study. Results are retained on file at Covance.
- Water
Mains water was provided ad libitum via water bottles. The water was periodically analysed for specific contaminants.
No contaminants were present in water at levels which might have interfered with achieving the objective of the study. Results are retained on file at Covance.
- Diet
Throughout the study the animals had access to 5LF2 EU Rodent Diet 14%, which was freely available to the animals at all times, except for a period of fasting from the evening of the day prior to dosing (Day-1) until approximately 3 hours after dosing. Each batch of diet had been analysed for specific constituents and contaminants by the manufacturer.
No contaminants were present in diet at levels which might have interfered with achieving the objective of the study. Results are retained on file at Covance.
- Environment
The animal rooms were designed to permit 15 to 20 air changes per hour. The target temperature and humidity ranges were 20 to 24 Deg C and 45 to 65% respectively. Daily recordings of maximum and minimum temperature and humidity were made.
Fluorescent lighting was controlled automatically to give a cycle of 12 hours light and 12 hours dark.
- Environmental Enrichment
In order to enrich both the environment and the welfare of the animals, they were provided with wooden Aspen chew blocks and rodent retreats.
Environmental enrichment materials were removed during the period of fasting.
- Animal Identification
A number written on the tail in indelible ink on the day before dosing individually identified the rats. A colour-coded card on each cage gave information including study number, group number, animal numbers and sex.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Doses:

In the preliminary study a female fasted rat was given the test article as a single dose on Day 1 by oral gavage at a dose level of 300 mg/kg. As no deaths were observed at this dose, a subsequent dose of 2000 mg/kg was administered to a single female fasted rat by oral gavage. The animal treated at 2000 mg/kg was humanely killed approximately 3 hours after dosing due the severity of the clinical signs. A further four female fasted rats were therefore given a single oral dose of test article at a dose level of 300 mg/kg body weight. No deaths were noted in animals treated at 300 mg/kg.

No. of animals per sex per dose:

4 females in the main study

Control animals:

no

Details on study design:

- Health Monitoring:
All animals were observed at the beginning and the end of the working day for signs of ill health or overt toxicity. Any animal which showed marked signs of ill health was isolated.
- Clinical Signs:
Treated rats were observed closely for clinical signs of reaction to treatment. Clinical signs were recorded prior to dose immediately post-dose, at approximately 15 and 30 minutes post-dose, hourly between 1 and 4 hours post-dose (inclusive), twice daily on Days 2, 3 and 4 and once daily from the fifth to last day of the observation period. Individual records of clinical signs and times of death were maintained for each treated rat.
Animals showing severe and enduring clinical signs and those considered moribund were killed by exanguination under deep isoflurane anaesthesia.
- Body Weights:
Rats were weighed on Day-1 (day before dosing) and on Days 1, 4, 8 and 15. Decedent carcass weights were also recorded prior to necropsy.
- Necropsy:
Rats surviving treatment were killed on Day 15. Each animal was given isoflurane anaesthesia. Once a suitable deep plane of anaesthesia had been established, the animal was exsanguinated by the severing of major blood vessels. After exsanguination a full macroscopic necropsy was performed and all lesions were recorded. The necropsy procedure included inspection of external surfaces and orifices, all viscera and tissue within the abdominal, thoracic and cranial cavities, free-hand sectioning of the liver and kidneys and examination of representative sections of mucosal surfaces of the stomach, small and large intestines.
Animals dying during the observation period were subject to a similar necropsy with the least possible delay. Carcasses were stored in a refrigerator until necropsy commenced.
- Preservation of Tissues for Histopathology:
Representative samples of macroscopic changes found at necropsy were retained in 10% neutral buffered. However, the Sponsor confirmed that microscopic examination was not required.

Results and discussion

Preliminary study:

In the preliminary study a female rat was given the test article as a single dose on Day 1 by oral gavage at a dose level of 300 mg/kg. As no deaths were observed at this dose, a subsequent dose of 2000 mg/kg was administered to a single female fasted rat by oral gavage.
The animal treated at 2000 mg/kg was humanely killed approximately 3 hours after dosing due the severity of the clinical signs. A further four female fasted rats were therefore given a single oral dose of test article at a dose level of 300 mg/kg body weight. No deaths were noted in animals treated at 300 mg/kg.

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No deaths were noted in animals treated at 300 mg/kg. No clinical signs were seen in animals treated at 300 mg/kg.

Mortality:

No deaths were noted in animals treated at 300 mg/kg. The animal treated at 2000 mg/kg was humanely killed approximately 3 hours after dosing due to the severity of the clinical signs.

Clinical signs:

other: No clinical signs were seen in animals treated at 300 mg/kg. Clinical signs noted in the animal treated at 2000 mg/kg were ataxia, decreased activity, laboured breathing, hypothermia, prone posture and loss of righting reflex. The clinical signs developed

Gross pathology:

The only abnormality noted at necropsy of the animal that was killed in extremis was dark contents in the stomach. nNo abnormalities were noted at necropsy of animals killed at the end of the study.

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The test article was classified as Category 4 in respect of its acute oral toxicity according to CLP Regulation 1272/2008.

Executive summary:

In an acute oral toxicity study conducted according to OECD guideline 420, groups of fasted female rats were given Neodecanoic acid, iron salt (100% purity) as a single oral dose in corn oil. In a preliminary study, a female rat was given the test article as a single dose at a dose level of 300 mg/kg bw. As no deaths were observed, a subsequent dose of 2000 mg/kg bw was administered to a further single female by oral gavage. The animal treated at 2000 mg/kg was humanely killed approximately 3 hours after dosing due to severity of the clinical signs. A further four female rats were therefore given a single oral dose of 300 mg/kg bw. No mortality occurred and no clinical signs were recorded. All surviving rats gained weight during the first and second weeks of the 14-day observation period. No abnormalities were noted at necropsy of animals killed at the end of the study. Based on the result, the LD50 is greater than 300, but lower than 2000 mg/kg bw. Thus, classification as Acute Tox 4, H302 is warranted in accordance with CLP Regulation 1272/2008.