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Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
24-Feb-2021 to 19-Mar-2021
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2021
Report date:
2021

Materials and methods

Test guideline
Qualifier:
no guideline required
Principles of method if other than guideline:
Preliminary 14 day dose range finding study designed to allow selection of appropriate dose levels for a subsequent 90-day repeated dose toxicity study with the reproduction/developmental toxicity screening test by oral gavage
GLP compliance:
no
Remarks:
Preliminary dose range finding study
Limit test:
yes

Test material

Constituent 1
Reference substance name:
3-methyldecane; benzene; cyclooctane; nonane
EC Number:
941-364-9
Molecular formula:
Not applicable to UVCB substance
IUPAC Name:
3-methyldecane; benzene; cyclooctane; nonane
Test material form:
other: Low visocosity, liquid hydrocarbon
Details on test material:
Batch number: 204377961
Physical description: Colourless or pale yellow coloured liquid
Purity: 100% UVCB
Expiry date: 20 August 2022 if kept under storge conditions
Source and site of characterisation: Repsol Refinery, Carretera de la Calzada s/n, Apartado de Correos 12, 13500 Puertollano, Ciudad Real, Spain
Storage conditions: Store under controlled humidity conditions and temperature (18-25 °C) in a sealed container, protecting the product from sunlight (opaque container or in a cabinet)

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Crl: WI (Han)
Sex:
female
Details on test animals or test system and environmental conditions:
Test animals
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at initiation of dosing: 6-7 weeks
- Weight at study initiation:126-142 g
- Housing:Animals were group housed (up to 3 animals of the same dosing group together) in polycarbonate cages (Macrolon, MIV type, height 18 cm)
- Diet:Ad libitum access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany)
- Water:ad libitum
- Acclimation period:9 days prior to the commencement of dosing

Environmental Conditions:
- Temperature: 19 to 21°C
- Humidity: 49 to 63%

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Remarks:
Specific gravity: 0.92
Details on oral exposure:
Preparation of dosing solutions:
Test item dosing formulations (w/w) will be homogenized to visually acceptable levels at appropriate concentrations to meet dose level requirements.
The dosing formulations will be prepared at least weekly, filled out in daily portions and stored in the refrigerator protected from light. Dosing formulations will be removed from the refrigerator at least 30 minutes before dosing and will be stirred at room temperature. The dosing formulations will be kept at room temperature until dosing. If practically possible, the dosing formulations will be continuously stirred until and during dosing.

Dosing Details:
Oral gavage using plastic catheter attached to plastic disposable syringe.
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
14 days
Frequency of treatment:
once daily
Doses / concentrationsopen allclose all
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
Group 1
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
Group 2
No. of animals per sex per dose:
3 females per dose
Control animals:
no
Details on study design:
The dose levels were selected based on the results of an acute oral toxicity study in rats (LD50 >2000 mg/kg; registration dossier, echa.europe.eu).

Examinations

Observations and examinations performed and frequency:
Mortality:
- Twice daily throughout the study

Cage Side Clinical Observation:
- At least 3 times daily up to the day prior to necropsy: 0-15 min, 45-75 min, 2.5-3.5 hr post-dose

Detailed Clinical Observation:
- Weekly and on the day of necropsy

Body Weight:
- Day 1 prior to dosing and Days 4, 8, 11 and 14 (body weight gain will be calculated against the body weight of Day 1 of treatment of each period).

Food Consumption:
- over Days 1-4, 4-8, 8-11, and 11-14
- Animals showing pain, distressed or discomdort which was considered not transient in nature or was likely to become more severe, were sacrificed for humane reasons based on OECD guideline document on humane endpoints.
Sacrifice and pathology:
Animals surviving until the scheduled euthanasia were deeply anaesthetised using isoflurane and subsequently exsanguinated

Animals were subjected to an external, thoracic and abdominal examination. Terminal bodyweight, liver and kidney weights were recorded. Organ weight relative to body weight were calculated against the terminal body weight for scheduled intervals. Macroscopic observations were recorded, but not retained.

Results and discussion

Results of examinations

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Group 1: slight salivation was observed in 3/3 females on Days 13 and 14
Group 2: slight salivation was observed in 3/3 females on 1 to 3 days of treatment
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
Endocrine findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Kidney and liver weights were slightly increased in Group 2 (1000 mg/kg bw)
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Details on results:
Mortality
Group 1: no mortality.
Group 2: no mortality.

Clinical observations
Group 1: slight salivation was observed in 3/3 females on Days 13 and 14.
Group 2: slight salivation was observed in 3/3 females on 1 to 3 days of treatment.

Body weight
Group 1: normal.
Group 2: normal.

Food consumption
Group 1: normal.
Group 2: normal.

Organ weights
Group 1: Kidney and liver weights were considered normal.
Group 2: Kidney and liver weights were slightly increased

Macroscopic examination
Group 1: no abnormalities noted.
Group 2: no abnormalities noted.

Effect levels

Dose descriptor:
NOAEL
Remarks on result:
not determinable

Applicant's summary and conclusion

Conclusions:
Based on the results of the Dose Range Finder, selected dose levels for the 90-Day Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test by Oral Gavage (Test Facility Study No. 20276349) are 100, 300 and 1000 mg/kg/day.
Executive summary:

The objective of this study was to determine the highest tolerated dose level of Petroleum gas oil fraction, co-processed with renewable hydrocarbons of plant and/or animal origin (EC 941-364-9) and to determine the peak effect of occurrence of clinical signs after dosing when given orally by gavage in non-pregnant rats. This study was performed as a Dose Range Finding study for the subsequent 90-Day Repeated Dose Toxicity study with the reproduction/developmental toxicity screening test by oral gavage.

Based on the results of the Dose Range Finder, selected dose levels for the 90-Day Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test by Oral Gavage are 100, 300 and 1000 mg/kg/day.