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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
multi-generation reproductive toxicity
Remarks:
based on test type
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Older published study, conducted prior to development of guidelines and GLP.
Justification for type of information:
Please see Analogue Approach

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
1951

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
Four successive generations of rats were exposed to the test substance in the diet from weaning, throughout breeding and lactation. The study was conducted prior to development of the guidelines.
GLP compliance:
no
Remarks:
conducted prior to GLP.
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
The test compound was stearyl citrate, a mixture of mono-, di-, and tri-alkyl esters of stearic acid with citric acid (12.5%, 75% and 12.5%, respectively). The test compound was a commercial product supplied by Best Foods, Inc. Identity was confirmed by analysis at the test facility.

Test animals

Species:
rat
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
The animals were weaned male and female rats, obtained from the stock colony at the University of South California School of Medicine.

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
The diet was made up of purified foodstuffs (casein, sucrose, margarine fat, fat soluble vitamin mixture, yeast, salt mixture and liver powder), with stearyl citrate added in appropriate quantities.
Details on mating procedure:
The rats were mated following 10 weeks exposure to the test diets (rats were 13 weeks old at the start of breeding).
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
From weaning, continuing during breeding and lactation.
Frequency of treatment:
Continuous dietary exposure.
Details on study schedule:
The parental generation were fed the test diets from weaning (3 weeks of age), continuing during breeding and lactation. When the offspring were weaned they were fed the same test diet as their parents, and the parental generation were discarded. The schedule was continued for four successive generations.
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 1.9 and 9.5% stearyl citrate
Basis:
nominal in diet
No. of animals per sex per dose:
Parental generation: 4-5 males and 10 females per group
Control animals:
yes, plain diet
Details on study design:
No further information available.
Positive control:
Not required.

Examinations

Parental animals: Observations and examinations:
Bodyweights at the start of treatment, after 5 weeks and after 10 weeks in parental and offspring generations. The percentage of successful pregnancies was determined (number of litters cast/number of rats bred), average time taken between cohabitation and birth.
Oestrous cyclicity (parental animals):
Not determined.
Sperm parameters (parental animals):
Not determined.
Litter observations:
Litter bodyweights were determined 3 days after birth and 21 days after birth, total rats weaned, average number of pups per litter and the number of pups that died between day 3 and day 21.
Postmortem examinations (parental animals):
Not determined.
Postmortem examinations (offspring):
Not determined.
Statistics:
No information available.
Reproductive indices:
Pregnancy rate.
Offspring viability indices:
Not determined.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

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
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined

Reproductive function / performance (P0)

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

Details on results (P0)

No toxicity was observed over the 4 generations of rats fed diets containing stearyl citrate. Fertility remained high in all groups, and was unaffected by treatment. The percentage of successful pregnancies was 93%, 98%, and 97% in the control, 1.9% and 9.5% groups respectively. A 100% fertility rate was obtained in the fourth generation in all three groups.

Effect levels (P0)

Key result
Dose descriptor:
NOAEL
Effect level:
95 000 ppm (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effect observed

Results: P1 (second parental generation)

General toxicity (P1)

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
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not specified

Reproductive function / performance (P1)

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

Effect levels (P1)

Key result
Dose descriptor:
NOAEL
Effect level:
95 000 ppm (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
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
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not examined
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings:
not examined
Other effects:
not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not specified

Details on results (F1)

Offspring from each successive generation went on to successfuly mate and produce offspring of their own. No signs of toxicity were reported, and the proportion of pups that died between day 3 and 21 was low (Table 1). The bodyweights of the offspring were slightly higher in the treated groups compared to the control group. The average weights of the 3 day old pups (of all 4 generations combined) were 6.9 g, 7.0 g and 7.6 g for the control, 1.9% and 2.5% groups respectively.

Effect levels (F1)

Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
95 000 ppm (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed

Results: F2 generation

General toxicity (F2)

Clinical signs:
no effects observed
Mortality / viability:
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
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not examined
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings:
not examined
Other effects:
not examined

Developmental neurotoxicity (F2)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F2)

Developmental immunotoxicity:
not specified

Effect levels (F2)

Key result
Dose descriptor:
NOAEL
Generation:
F2
Effect level:
95 000 ppm (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed

Overall reproductive toxicity

Key result
Reproductive effects observed:
not specified

Any other information on results incl. tables

Treatment Group – Stearyl Citrate Content

Parents

Litter data

Genera-tion

No. rats bred

No. Litters

Days from cohabitation to birth

Genera-tion

Average No./litter

Bodyweight (g)

No. pups weaned

No. pups died day 3-21

3 days

21 daysa

0%

1

10

9

28.0

2

9.4

6.9

34.0 (35)

35

0

2

14

13

27.3

3

8.3

6.6

34.3 (35)

60

3

3

19

17

27.7

4

7.7

7.2

33.6 (35)

65

5

4

16

16

26.6

5

7.1

6.9

34.5 (63)

72

7

1.9%

1

10

10

26.8

2

6.3

6.7

34.6 (21)

41

0

2

15

15

26.2

3

6.8

7.4

33.5 (42)

58

0

3

15

14

24.4

4

8.1

7.0

36.9 (56)

83

3

4

14

14

31.0

5

8.4

6.8

36.7 (56)

71

2

9.5%

1

11

11

30.2

2

7.6

7.0

34.5 (43)

61

2

2

23

23

26.2

3

8.5

7.2

35.5 (105)

117

0

3

16

14

26.5

4

8.5

8.5

41.0 (56)

67

0

4

14

14

27.4

5

9.4

7.6

37.1(84)

94

0

aaverage weights of 7-litter rats only, figures in parentheses give average number of rats

Table 2. Bodyweight data

Treatment Group – Stearyl Citrate Content

Generation

Males

Females

No.

Bodyweights (g)

No.

Bodyweights (g)

Start

After 5 weeks

After 10 weeks

Start

After 5 weeks

After 10 weeks

0%

1

5

37.8

164.0

265.0

10

37.3

135.7

182.5

2

4

34.0

156.6

243.6

21

35.7

125.2

166.3

3

16

37.4

168.2

253.5

19

35.8

122.7

163.1

4

18

36.9

161.7

289.3

22

36.5

126.1

179.0

1.9%

1

5

38.2

167.4

273.0

10

37.3

132.8

177.5

2

19

34.9

150.1

245.3

16

33.9

122.0

171.1

3

14

37.1

175.2

257.8

20

35.5

131.7

172.9

4

10

40.3

172.9

268.4

16

38.6

129.1

177.8

9.5%

1

4

36.5

160.5

267.3

10

37.6

131.5

177.4

2

19

36.6

153.7

246.0

26

32.5

125.3

165.4

3

10

33.2

162.2

237.1

19

32.7

124.7

166.2

4

13

43.8

174.9

273.3

19

39.5

132.0

178.8

Applicant's summary and conclusion

Conclusions:
There were no signs of toxicity and no effects on fertility were observed in four successive generations of rats fed diets containing 1.9% or 9.5% of the analogue substance stearyl citrate. The NOAEL can be considered to be 9.5% stearyl citrate.
Executive summary:

Groups of male and female rats were administered 0%, 1.9% or 9.5% of the structural related substance stearyl citrate in the diet continuously from weaning, during mating and lactation. The offspring were maintained on the same diet as their parents from weaning, during breeding and lactation. The schedule was continued for 4 successive generations.

No toxicity was observed over the 4 generations of rats. Fertility remained high in all groups, and was unaffected by treatment. The percentage of successful pregnancies was 93%, 98%, and 97% in the control, 1.9% stearyl citrate and 9.5% stearyl citrate groups respectively. A 100% fertility rate was obtained in the fourth generation in all three groups. Offspring from each successive generation went on to successfuly mate and produce offspring of their own. It is concluded that the structural related substance stearyl citrate is not toxic to reproduction, and the NOAEL can be considered to be 9.5% stearyl citrate (11875 ppm tristearyl citrate, equivalent to approximately 594 mg/kg bw/d). These conclusions are based upon fertility over 4 generations, lactation perfomance as judged by the 3 -day and 21 -day weights of offspring, as well as by their survival for the nursing period, and by the rate of growth of rats of the several generations over a 10 -week period follwing weaning.