Reaction products resulting from the esterification of C18 unsaturated fatty acid with glycerol and glycerol oligomers

Repeated dose toxicity: oral

Administrative data

Endpoint:

chronic toxicity: oral

Type of information:

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

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Justification of read-across: Both chemicals are of comparable structures and can be characterized as esters of polyglycerol and fatty acids and undgo the same metabolism (Refer to Read Across Justification presented in Section 13). Justification of reliability of 2: scientifically well-performed study

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: CARCINOGENIC RISKS IN FOOD ADDITIVES AND PESTICIDES (1960), Ministry of Health, UK

Deviations:

not specified

GLP compliance:

no

Remarks:

Studies were performed prior to implementation of GLP

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Polyglycerol polyricinoleate

IUPAC Name:

Polyglycerol polyricinoleate

Constituent 2

Reference substance name:

PGPR

IUPAC Name:

PGPR

Constituent 3

Reference substance name:

29894-35-7

EC Number:

608-428-5

Cas Number:

29894-35-7

IUPAC Name:

29894-35-7

Test material form:

liquid: viscous

Details on test material:

PGPR is prepared by the esterification of condensed castor oil fatty acids (primarily ricinoleic acid)

Test animals

Species:

other: rat and mouse

Strain:

other: rat: Wistar; mouse: C57BL

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Unilever Colworth Laboratory, Sharnbrook, Bedford, UK
- Age at study initiation: 32-46 days (rat); 6-8 weeks (mouse)
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

rat study: 2-year
mouse study: 80-week

Frequency of treatment:

feed; continuously

Doses / concentrations

Remarks:

Doses / Concentrations:
0 and 5% (w/w, in diet)
Basis:
nominal in diet

No. of animals per sex per dose:

rat study: 30/sex/group
mouse study: 25/sex/group

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale:
Rat study: The single dose level used for this study was chosen by comparing the likely human intake and the no-effect levels found in earlier chronic studies. Dosage levels for carcinogenicity studies should include a dose that is as high as can be administered without reducing the lifespan of the test animals. Previous studies showed no toxicity in rats fed 9% PGPR in diet for 45 wk. When testing food additives it is recognized that addition to the diet of levels above 5% can disturb the nutritional balance of the diet.
Mouse study: A single dose level was chosen for this study by comparing the likely human intake and the no-effect levels found in earlier subchronic and chronic rat feeding studies. PGPR was added to a semipurified diet at a level of 5% (by weight) by substituting part of the groundnut oil fed to control animals.

- Rationale for animal assignment (if not random): random

Positive control:

no

Examinations

Observations and examinations performed and frequency:

Rat Study:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- three times weekly and evaluated as a weekly amount.

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 data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood and how many animals: after 80 wk from four rats of each sex fed PGPR or the control diet, and on all surviving animals at study termination
- Anaesthetic used for blood collection: Yes
- Animals fasted: No data
- Parameters: erythrocyte and leucocyte counts, haemoglobin concentrations and value, red cell fragility and prothrombin time.

CLINICAL CHEMISTRY: No data


URINALYSIS: yes

NEUROBEHAVIOURAL EXAMINATION: No

Mouse Study:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- twice weekly and evaluated as a weekly amount.

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 data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood and how many animals: all surviving animals at study termination
- Anaesthetic used for blood collection: Yes
- Animals fasted: No data
- Parameters: erythrocyte and leucocyte counts and haemoglobin concentrations

CLINICAL CHEMISTRY: No data


URINALYSIS: yes

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

Rat Study:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Each animal was subjected to gross examination at autopsy. The following organs were weighed and the organ/body weight ratios determined: adrenals, heart, kidney, spleen, liver, testes, thyroid and pituitary. These organs, together with the lung, ovary, uterus, thymus, stomach, intestine, caecum, bladder, lymph nodes, skin, mammary gland, tongue and any macroscopic abnormality were removed, fixed and processed for histological examination.

Mouse Study:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Each animal was subjected to gross examination at autopsy (at test termination and for those animals dying during the test). The following organs were weighed: heart, kidney, liver and testes. These organs, together with lung, spleen, adrenals, skin, stomach, intestine, thyroid, thymus, mammary gland and lymph nodes, together with any macroscopic abnormality were removed, fixed and processed for histological examination.

Other examinations:

In rat study, Liver function was examined after 84 and 103 week using the bromosulfothalein excretion test. Kidneyfunction was assessed at the same times by measuring urine concentration.

Statistics:

All records were examined separately for each sex by analysis of variance to assess the significance of any inter-group differences. Organ weightswere also analysed if justified by an analysis of covariance on final body weight.
In the analysis of variance, the F-ratio test was used to test whether the treatments differed. Student's t-tests were then used to compare every treatment mean individually against the control.

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:

no effects observed

Clinical biochemistry findings:

not specified

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

ORGAN WEIGHTS
Rat study:
Organ weight measurements showed that kidneys from male and female rats and livers from female (but not male) rats fed PGPR were heavier than those fed the control diet.

Mouse study:
Organ weight measurements revealed that livers and kidneys from female mice fed PGPR were heavier than those from mice fed the control purified diet.

Effect levels

Key result

Dose descriptor:

NOAEL

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

no NOAEL identified

Target system / organ toxicity

Key result

Critical effects observed:

not specified

Any other information on results incl. tables

Rats in both treatment and control groups occasionally showed clinical signs of respiratory disease, which in some cases was accompanied by loss of body weight and reduced food consumption. The incidence of respiratory disease was most evident from wk 50 onwards. Bronchiectasis and emphysema were the main cause of death among animals in both test and control groups. The incidence of respiratory disease was comparable between treatment and control groups.

Mean weights of liver and kidney of rats and mice.

Species	Organ	Mean organ weights (g)
		Males		Females
		PGPR diet	Purified diet (control)	PGPR diet	Purified diet (control)
Rats	Liver	12.18	11.45	9.15 *	8.67
	Kidney	3.59 *	3.19	2.52 *	2.37
Mice	Liver	2.81	2.54	3.55 *	2.26
	Kidney	0.70	0.68	0.66 *	0.53

 * significantly different from control (P=0.05)

Applicant's summary and conclusion

Conclusions:

The treatment of 5% target chemical in the diet (approx. 2500 mg/kg bw/day) is not considered likely to induce adverse effects in rats and mice.

Executive summary:

The repeated dose toxicity of the target substance was assessed based on the analogue approach using PGRP as a read-across supporting substance.

The long-term toxicity potential of PGPR was evaluated in rats and mice. Groups of 60 male and 60 female rats were given purified diets containing 5% of either PGPR or groundnut oil for 2 years. Groups of 25 male and 25 female mice were given purified diets containing 5% of either PGPR or groundnut oil for 80 weeks. No carcinogenic effect of PGPR was observed. In addition, dietary PGPR had no adverse effect on growth, food consumption, longevity and haematology. Organ weight analysis revealed an increase in liver and kidney weight in both male and female rats and female mice. Histological analysis of tissues revealed no treatment related adverse effects. Based on the results, it was concluded that treatment of 5% test item in the diet (approx. 2500 mg/kg bw/day) did not induce adverse effects in rats and mice. Likewise, treatment with the target chemical of the same level is not considered likely to induce adverse effects.