1,2,3-Propanetricarboxylic acid, 2-hydroxy-, tri-C12-13-branched alkyl esters

Toxicity to reproduction

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

Test material information

Constituent 1

Reference substance name:

Stearyl citrate

IUPAC Name:

Stearyl citrate

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 or test system 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 days^a
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.