Registration Dossier

Administrative data

Endpoint:
three-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1950-MM-DD to 1960-MM-DD
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Sound scientific publication. This study was conducted during the late 1950s/early1960s and the methods represented the state of the art which existed during this period. They nevertheless allow an adequate assessment of the potential reproductive and chronic toxicity of PGPR.
Cross-reference
Reason / purpose for cross-reference:
reference to same 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: This study was conducted during the late 1950s/early1960s and the methods represented the state of the art which existed during this period. They nevertheless allow an adequate assessment of the potential reproductive and chronic toxicity of PGPR.
Principles of method if other than guideline:
A three-generation reproduction study was conducted on PGPR to investigate the potential adverse effects on reproduction. Breeding was conducted using a continuous breeding protocol in which pairs of animals were maintained until each female had produced five litters (or it became evident that breeding had ceased) and this was conducted over three generations. The main focus of the study design was to observe any effect on breeding. Parameters measured in each of the three generations included number of litters per dam, average litter size, average weaning weights of males and females, litters per group showing 100% survival and total survival (%) at day 21.
GLP compliance:
no
Remarks:
Studies were performed prior to implementation of GLP
Limit test:
yes

Test material

Constituent 1
Reference substance name:
Polyglycerol polyricinoleate
IUPAC Name:
Polyglycerol polyricinoleate
Constituent 2
Reference substance name:
29894-35-7
EC Number:
608-428-5
Cas Number:
29894-35-7
IUPAC Name:
29894-35-7
Constituent 3
Reference substance name:
PGPR
IUPAC Name:
PGPR
Test material form:
not specified
Details on test material:
ADMUL WOL, a brand of polyglycerol polyricinoleate (PGPR). PGPR is a very powerful emulsifier which was primarily developed for the production of water-in-oil emulsions for tin-greasing in the baking trade. However, its main use is in chocolate where in addition to its action as an emulsifier, it also has valuable properties as a viscosity modifier and thus improves the moulding properties of the molten chocolate.
Specific details on test material used for the study:
ADMUL WOL tradename/brand of polyglycerol polyricinoleate (PGPR)

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
Colworth Wistar rats (obtained from the breeding facility at the Unilever Colworth Laboratory, Sharnbrook, Bedford, UK) were used in these stu-
dies.
TEST ANIMALS
- Source: Colworth Wistar rats (obtained from the breeding facility at the Unilever Colworth Laboratory, Sharnbrook, Bedford, UK) were used in these studies.
- Housing: Each pair occupied a single cage. Breeding was conducted using a continuous breeding protocol in which pairs of animals were maintained until each female had produced five litters (or it became evident that breeding had ceased) and this was conducted over three generations.)
- Diet (e.g. ad libitum): yes
- Water (e.g. ad libitum): yes
- Acclimation period: n.a.

ENVIRONMENTAL CONDITIONS
no inforamtion

IN-LIFE DATES: during late 1950's/early 1960's

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): Fresh mixes were prepared each week, and PGPR was incorporated into the ground diet first by hand,
then well mixed in a mechanical mixer.
- Mixing appropriate amounts with (Type of food): commercial pelleted stock diet (Spital)
- Storage temperature of food: n.a.

VEHICLE
none
Details on mating procedure:
The first-generation parents were selected from five litters which were assigned randomly into two groups: a control (11 males and 17 females) and a treatment group fed 1.5% PGPR (six males and 13 females). The dietary level of 1.5% PGPR was chosen to provide an intake by the pregnant and lactating rat which was in excess of 150 times the intake of a person consuming 5 mg PGPR/kg body weight/day.

All rats were weaned at 23 days and mated at 121 days.
Breeding was continuous and the males were only separated from the females when it was apparent that the female was pregnant. Each pair occupied a single cage and they were maintained until the female had produced five litters or until such time as it became evident that breeding had ceased.
In all instances the first litters were discarded after weaning and second-generation breeders were randomly selected (two males and two females)
from each of the second and fourth litters. By selecting from two first-generation litters the number of animals was increased to 52 of each sex in
the control and 32 of each sex in the PGPR group. The third-generation breeders were selected in a similar manner, by which the control and the PGPR groups were increased to 92 and 44 rats of each sex, respectively. The large control groups were considered necessary to get an indication of
the variations in breeding results that occur within a normal breeding colony. The control rats were fed a pelleted stock diet and the PGPR group were given the same diet ground with 1.5% PGPR.
The main focus of the study design was to observe any effect on breeding. Parameters measured in each of the three generations included number of litters per dam, average litter size, average weaning weights of males and females, litters per group showing 100% survival and total survival
(%) at day 21.
Analytical verification of doses or concentrations:
not specified
Frequency of treatment:
daily
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
The PGPR group were given the same diet ground added with 1.5% PGPR.
Basis:
nominal in diet
The dietary level of 1.5% PGPR was chosen to provide an intake by the pregnant and lactating rat which was in excess of 150 times the intake of a person consuming 5 mg PGPR/kg body weight/day.
Remarks:
Doses / Concentrations:
The control rats were fed a pelleted stock diet
Basis:

No. of animals per sex per dose:
1st generation parents (P) were selected from five litters which were assigned randomly into two groups
Treatment group: 6 males and 13 females
Control group: 11 males and 17 females
In all instances the first litters were discarded after weaning and second-generation breeders were randomly selected (two males and two females) from each of the second and fourth litters. By selecting from two first-generation litters the number of animals was increased to 52 of each sex in the control and 32 of each sex in the PGPR group. The third-generation breeders were selected in a similar manner, by which the control and the PGPR groups were increased to 92 and 44 rats of each sex, respectively.
Control animals:
yes, plain diet

Examinations

Statistics:
A Student's t-test was conducted in which the two groups were compared

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Growth was monitored from weaning to mating during the first 4 months of each generation. Weight females recorded at weaning and mating. Males weight at weaning and day 65.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Growth was monitored from weaning to mating during the first 4 months of each generation. Weight females recorded at weaning and mating. Males weight at weaning and day 65.
Organ weight findings including organ / body weight ratios:
not specified
Histopathological findings: non-neoplastic:
not specified
Other effects:
not specified
Description (incidence and severity):
Test substance intake: During the three-generation study the breeding females consumed PGPR at levels of > 2 g/kg body weight (based on a food intake level of up to 40 g/day during lactation and the inclusion of PGPR in the diet of 1.5%)

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed

Effect levels (P0)

Key result
Dose descriptor:
NOAEL
Effect level:
> 2 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Remarks on result:
other: Generation: for all three generations (migrated information)

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Description (incidence and severity):
There were no deaths during the experimental period and no evidence of abnormal behaviour or functional disorder associated with the consumption of PGPR throughout the three generations of the study.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
The body weight data for test and control rats indicate that they both grew in a similar, normal pattern.
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified

Effect levels (F1)

Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
> 2 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
sexual maturation
clinical signs
mortality
body weight and weight gain
Remarks on result:
other: There was no effect of PGPR on the suckling pups receiving PGPR from their mothers' milk.

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
Rats fed 1.5% (w/w) PGPR showed no evidence of a cumulative effect on breeding performance over three generations. Growth was comparable to controls throughout the three generations and there were no deaths or clinical signs associated with the consumption of PGPR. The only significant change in breeding performance was a reduction in the percentage of animals weaned in the second generation, but as this occurred in the control group to a similar extent it was concluded that this was due to an unknown environmental factor and was not treatment related. A histological examination of selected tissues from those rats continued for 1 year failed to show any lesions which could be ascribed to the consumption of PGPR.
Executive summary:

A publication is available where a series of toxicology studies were conducted in the 1950s and 1960s to investigate the toxicity of ADMUL WOL, a brand of polyglycerol polyricinoleate (PGPR). Included as part of these investigations was a three-generation reproduction study in rats. The control rats received a commercial pelleted stock diet and the treated rats were given the same diet ground with 1.5% (w/w) PGPR. A continuous breeding protocol was adopted, in which the breeding pairs were maintained until the female had produced five litters or when it became evident that breeding had ceased. The main focus of the study design was to observe any effect on breeding. The parameters measured in each of the three generations included number of litters per dam, average litter size, average weaning weights of males and females, litters per group showing 100% survival and total survival (%) at day 21.

Growth was comparable to controls throughout the three generations and there were no deaths or clinical signs associated with the consumption of PGPR. In conclusion, rats fed 1.5 % (w/w) PGPR showed no evidence of a cummulative effect on breeding performance over three generations.

During the three-generation study the breeding females consumed PGPR at levels of greater than 2 g/kg body weight (based on a food intake level of up to 40 g/day during lactation and the inclusion of PGPR in the diet at the fixed level of 1.5%). At this level of PGPR consumption the breeding performance of the treated rats was similar to those rats fed the control diet. There was no effect of PGPR on the suckling pups receiving PGPR from their mother's milk.