Fatty acids, C16-18 (even numbered), iron(III) salts

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 October 2018 to 08 March 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C16-18 (even numbered), iron(III) salts

EC Number:

947-665-1

Molecular formula:

C54H105FeO6, C48H93FeO6 and C42H81FeO6.

IUPAC Name:

Fatty acids, C16-18 (even numbered), iron(III) salts

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Nitika Pharmaceutical Specialities Pvt Ltd. / Bx IRST7H780L
- Expiration date of the lot/batch: August 2023
- Purity test date: September 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Inoriginal container in dry cool place.
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/vehicle:
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: none
- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST (if different from that of starting material)

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable)

OTHER SPECIFICS:
- measurement of pH, osmolality, and precipitate in the culture medium to which the test chemical is added:
- other information:

Test animals

Species:

rat

Strain:

other: RCCHan:WIST

Details on species / strain selection:

The preferred species is the rat. It has been historically shown to be a suitable model for repeated dose toxicity studies and is recommended by the regulatory authorities. The results may be of value in predicting the toxicity of the test item to humans.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Commercial laboratory animal supplier.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 to 9 weeks
- Weight at study initiation: 278.62 to 284.42 g
- Fasting period before study: no data
- Housing: Rats were housed in groups of 2 or 3 rats/cage/sex during the study period in sterile solid floor polypropylene rat cages (size: 41 × 28.2 × 18 cm). Each cage was fitted with a stainless steel (grille) top having provision for keeping rodent pellet feed and a polypropylene water bottle with a stainless steel drinking nozzle. Cages were placed in multi-tier racks (5 x 5) to ensure similar environmental conditions for all groups.
- Diet: Teklad Certified Global 16% protein rodent maintenance diet, Envigo USA, ad libitum
- Water: Reverse Osmosis (RO) water ad libitum (RO water filtration system) in polypropylene bottles.
- Acclimation period: 7 days

DETAILS OF FOOD AND WATER QUALITY: Every feed consignment received is accompanied by a certificate of analysis of nutrient content from the feed supplier. The quality of feed and water is monitored in compliance with JRF/TOX/SOP-2077.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24°C
- Humidity (%): 64 to 66%
- Air changes (per hr): 21
- Photoperiod (hrs dark / hrs light): The photoperiod was 12 hours light and 12 hours darkness, fluorescence light hours being 06.00 - 18.00 hours.

IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.5% carboxy-methyl-cellulose (CMC) with 1% tween 80

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): at least twice a week.
- Mixing appropriate amounts with (Type of food): Teklad Certified Global 16% protein rodent maintenance diet
- Storage temperature of food: no data

VEHICLE
- Justification for use and choice of vehicle (if other than water): after consultation with the Sponsor,
- Concentration in vehicle: The test item was mixed with 1% tween 80 (i.e., 1% of total volume to be prepared) followed by mixing with 0.5% CMC. The final volume was made up with 0.5% CMC.
- Amount of vehicle (if gavage):
- Lot/batch no. (if required):
- Purity:

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Element : Iron (Fe)
Wavelength : 238.204 nm
RF power : 1300 Watts
Pump (Flow rate): 1.5 mL/minute

Gas Flow Rate
Plasma : 15.0 L/minute
Auxillary : 0.2 L/minute
Nebulizer : 0.8 L/minute

Duration of treatment / exposure:

14 days

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (actual dose received)

Remarks:

G1 - control (vehicle)

Doses / concentrations 2

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Remarks:

G2 - Low dose

Doses / concentrations 3

Dose / conc.:

300 mg/kg bw/day (actual dose received)

Remarks:

G3 - mid dose

Doses / concentrations 4

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

Remarks:

G4 - High dose

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on the results of the present study, the dose levels were chosen for the definitive reproduction/developmental toxicity screening study:
- Rationale for animal assignment (if not random):
- Fasting period before blood sampling for clinical biochemistry: no blood sample taken.
- Rationale for selecting satellite groups:
- Post-exposure recovery period in satellite groups:
- Section schedule rationale (if not random):
- Other:

Positive control:

No

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
- Cage side observations included; mortality, morbidity and clinical

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: The body weight was recorded for individual rats on days 1 (before commencement of treatment), and on days 4, 8, 11 and 14. Rats were also weighed on the day of necropsy (fasted body weight).

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
A weighed amount of feed was offered to rats in each cage on days 1, 4, 8 and 11 and the leftover feed was measured on days 4, 8, 11 and 14.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: No
- Time schedule for examinations:
- Dose groups that were examined:

HAEMATOLOGY: No
- Time schedule for collection of blood:
- Anaesthetic used for blood collection: Yes (identity) / No / Not specified
- Animals fasted: Yes / No / Not specified
- How many animals:
- Parameters checked in table [No.?] were examined.

CLINICAL CHEMISTRY: No
- Time schedule for collection of blood:
- Animals fasted: Yes / No / Not specified
- How many animals:
- Parameters checked in table [No.?] were examined.

URINALYSIS: No
- Time schedule for collection of urine:
- Metabolism cages used for collection of urine: Yes / No / Not specified
- Animals fasted: Yes / No / Not specified
- Parameters checked in table [No.?] were examined.

NEUROBEHAVIOURAL EXAMINATION: No
- Time schedule for examinations:
- Dose groups that were examined:
- Battery of functions tested: sensory activity / grip strength / motor activity / other:

IMMUNOLOGY: No
- Time schedule for examinations:
- How many animals:
- Dose groups that were examined:
- Parameters checked in table [No.?] were examined.

OTHER:

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
The thoracic and abdominal cavities were opened and a thorough examination of the organs was carried out to detect any abnormalities.
Organs were weighed from all rats. Adherent adipose tissue from organs was trimmed off and the wet weight of organs recorded for the liver, kidneys, adrenals, testes, epididymis, prostate, cowper’s gland, levator ani plus bulbocavernosus muscles (LABC), seminal vesicles with coagulating glands, thyroid with parathyroid, thymus, heart, brain, spleen, uterus with cervix, and ovaries. The thyroid with parathyroid were weighed after overnight fixation. Paired organs were weighed together and a combined weight presented. The organ weight ratio as percentage of body weight was calculated. The tissues (or any organs showing gross lesions, none present in this study) were weighed and fixed in 10% neutral buffered formalin.

HISTOPATHOLOGY: No
Histopathology was not performed, hence, preserved organs/tissues will be discarded during report finalisation.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Neuropathological findings:

no effects observed

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical signs

food consumption and compound intake

gross pathology

mortality

organ weights and organ / body weight ratios

Target system / organ toxicity

Key result

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

Fatty acids, C16-18 (even numbered), iron(III) salts at 100 mg/kg b. wt./day
- No mortality or clinical signs were observed.
- No treatment related effects on body weight, body weight gain, food consumption and organ weights (absolute and relative) were observed.
- External and internal examination of rats of either sex did not reveal any abnormality.

Fatty acids, C16-18 (even numbered), iron(III) salts at 300 mg/kg b. wt./day
- No mortality or clinical signs were observed.
- No treatment related effects on body weight, body weight gain, food consumption and organ weights (absolute and relative) were observed.
- External and internal examination of rats of either sex did not reveal any abnormality.

Fatty acids, C16-18 (even numbered), iron(III) salts at 1000 mg/kg b. wt./day
- No mortality or clinical signs were observed.
- No treatment related effects on body weight, body weight gain, food consumption and organ weights (absolute and relative) were observed.
- External and internal examination of rats of either sex did not reveal any abnormality.