Linseed oil, ester with pentaerythritol

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

no

GLP compliance:

yes

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Glycerides, C16-18 and C18-unsatd.

EC Number:

266-948-4

EC Name:

Glycerides, C16-18 and C18-unsatd.

Cas Number:

67701-30-8

IUPAC Name:

Glycerides, C16-18 and C18-unsatd.

Details on test material:

- Name of test material (as cited in study report): Pine nut oil. Under the SDA nomenclature, the name of this substance is 'Glycerides, C16-18 and C18 unsatd.'
- Substance type: Vegetable oil
- Analytical purity: yes, analyses for contamination and microbiological analyses were preformed
- Composition of test material, percentage of components: Pinolenic acid 15.3%, mono unsaturated fatty acid 26.5%, poly unsaturated fatty acids 66%, oleic acid 25.1%, linoleic acid 45.5%, saturated fatty acids 7.1%
- Lot/batch No.: 7284/20071217-076
- Storage condition of test material: 119 days at 15 deg. C or 4 days at room temperature

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Breeding centre Charles River Deutchland, Sulzfeld, Germany)
- Age at study initiation: 6 weeks
- Housing: in group of five in Macrolon cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.9-22.0 deg. C
- Humidity (%): 31-84%
- Air changes (per hr): 15times/h
- Photoperiod (hrs dark / hrs light): 12 h dark / 12 h light

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: accuracy of the diet preparations ranged from 87% to 95% of nominal value, which was considered as an acceptable level of accuracy

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Chemical analyses revealed that experimental diets were homogenously prepared; accuracy of the diet ranged from 87%-95% of nominal value

Duration of treatment / exposure:

98 and 100 days for female and male rats, respectively

Frequency of treatment:

Daily

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0%
Basis:
nominal in diet

Doses / concentrations 2

Remarks:

Doses / Concentrations:
1%
Basis:
nominal in diet

Doses / concentrations 3

Remarks:

Doses / Concentrations:
5%
Basis:
nominal in diet

Doses / concentrations 4

Remarks:

Doses / Concentrations:
15%
Basis:
nominal in diet

No. of animals per sex per dose:

10

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale: No higher dose level than 15% was selected as nonspecific effects of high fat intake in rats can be expected to appear at dose levels above 15%

Positive control:

None

Examinations

Observations and examinations performed and frequency:

All animals were observed twice daily for mortality/viability and at least once daily for clinical observations and signs for toxicity, feed intake and body weight were evaluated once a week

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Other examinations:

None

Statistics:

Where the variables were assessed to follow a normal distribution, a Dunnett-test based on a pooled variance estimate, was applied to compare the treated groups and the control group of each gender. The Steel-test was applied where the data could not be assumed to follow a normal distribution. The exact Fishertest was applied to frequency data. All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance. Group means were calculated for continuous data.

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:

no effects observed

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

not specified

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

HAEMATOLOGY: Statistically significant changes in haematology parameters were considered to be of no
toxicological relevance as they occurred in the absence of a dose related response and/or were of a very slight size. These changes included lower white blood cell counts (WBC) and protrombin times in males fed 15% KPNO, higher relative neutrophil and lower relative lymphocyte count in females fed 1% and 5% KPNO but not in females of the 15% KPNO dose group, and lower haemoglobin and haematocrit levels in females fed 1% KPNO but not in females fed 5% or 10% KPNO in the diet.

CLINICAL CHEMISTRY: Higher alkaline phosphatase activity levels (ALP) in females fed 15% KPNO, lower total bilirubin levels in males fed 5% and 15% KPNO, lower total protein levels in females fed 1%, 5% and 15% KPNO, lower cholesterol levels in males fed 15%, and in females fed 1%, and 15% KPNO, lower calcium levels in males and females fed 1%, 5% and 15% KPNO, lower inorganic phosphate levels in males fed 15% and increased creatinine and sodium levels in the females of 5% KPNO group. Although some statistical significant changes in biochemical parameters were observed it is important to note that compared to rats on a normal fat diet, most changes fall within the historical control values.


URINALYSIS


NEUROBEHAVIOUR


ORGAN WEIGHTS

Effect levels

1

Key result

Dose descriptor:

NOAEL

Effect level:

8 866 mg/kg bw/day (nominal)

Sex:

male

Basis for effect level:

other: see 'Remark'

2

Key result

Dose descriptor:

NOAEL

Effect level:

10 242 mg/kg bw/day (actual dose received)

Sex:

female

Basis for effect level:

other: see 'Remark'

Target system / organ toxicity

Key result

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Under the study conditions, the NOAEL for subchronic oral toxicity was determined to be 8866 and 10242 mg/kg bw/day for male and female rats, respectively.

Executive summary:

A study was conducted to determine the subchronic oral toxicity of glycerides, C16-18 and C18-unsatd. (in the form of pine nut oil) according to OECD Guideline 408, in compliance with GLP. The test substance was administered through the diet to three groups, each of ten male and ten female Wistar Crl:Wi(Han) strain rats, for up to 98 and 100 consecutive days, at dose levels of 0, 1, 5 and 15% per day. In Week 12, a functional observation test was carried out. At Week 13, ophthalmoscopic examinations were conducted. Clinical signs, bodyweight development, food intake and mortality/viability were monitored during the study. Haematology and blood chemistry were evaluated for all animals at the end of the treatment period. All animals were subjected to gross necropsy and macroscopic examination and a complete histopathological examination was performed. No toxicological significant changes were observed. Under the study conditions, the NOAEL for subchronic oral toxicity was determined to be 8866 and 10242 mg/kg bw/day for male and female rats, respectively (Gerrit, 2009).