Sodium 2-(1-carboxylatoethoxy)-1-methyl-2-oxoethyl isooctadecanoate

Toxicity to reproduction

Administrative data

Endpoint:

three-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1970-01-07 to 1971-09-30

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1971

Report date:

1971

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

- Principle of test: Determine the effects of sodium stearoyl lactylates upon reproduction by a thee generation study in rats
- Short description of test conditions: Original parent rats (F0) were bred twice; the F1A pups were sacrificed at birth and part of each litter was examined either for skeletal abnormalities or for visceral changes. F1B pups were reared to weaning and pups from each litter were taken to constitute the next group of breeders. F1B rats were bred twice, and both F2A and F2B litters were reared to weaning. F2B pups were then distributed into new groups to breed the F3 generations. F3a pups were sacrificed at weaning and necropsied. F3b pups were treated similarly and some organs were weighed from two per sex per litter at necropsy.
The three-generation reproduction study was performed in 1971; no test guideline for such a test was available at that time.

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Reference substance 001

Cas Number:

25383-99-7

Specific details on test material used for the study:

- Appearance: off-white powder

SOURCE OF TEST MATERIAL
- Source and lot/batch no.of test material: 23 E 299

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

albino

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc.
- Age at study initiation: 28 days
- Fasting period prior to study: No
- Housing: Animals were individually housed in temperature-controlled quarters.
- Diet (e.g. ad libitum): Ad libitum, Purina Laboratory Chow-Meal
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: one week

Administration / exposure

Route of administration:

oral: feed

Vehicle:

other: USP cod liver oil (1%)

Details on exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): not specified
- Mixing appropriate amounts with (type of food): The experimental diets were prepared by grinding appropriate amounts of the test material with a small quantity of the basal ration (Purina Laboratory Chow-Meal) in a mortar and pestle and mixing the resultant blend in a Hobart-Dayton mixer with enough basal ration to make a 6000 g batch of the desired concentration.
- Storage temperature of food: not specified

VEHICLE
- Justification for use and choice of vehicle (if other than water): not specified
- Concentration in vehicle: All diets were fortified with USP cod liver oil at a concentration of 1%
- Amount of vehicle (if gavage): not applicable
- Lot/batch no. (if required): not specified
- Purity: not specified

Details on mating procedure:

- M/F ratio per cage: 1 M/1 F
- Length of cohabitation: 10 days
- Proof of pregnancy: Not specified

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Day 28 of F0 generation - end of study (weaning of F3 pups)

Frequency of treatment:

daily; ad libitum

Details on study schedule:

- F0 mated at 100 days old and 72 days into the study to create F1A (first mating) and F1B (second mating, 12 days after weaning). F0 sacrificed after weaning of F1B (21 days). FA1 sacrificed and 1/2 examined for visceral changes (preserved in Bouin's fixative), 1/2 for skeletal changes (cleared in KOH and stained with Alizarin Red S).
- F1B animals weighed, observed, weaned. Representative male and female selected (20 males, 20 females per dose group) and the remainder sacrificed. F1B began receiving test compounds, as appropriate, in the diet at full concentration at weaning. After 77 days, F1B rats mated.
- F1B animals mated to create F2A (First mating) and F2B (second mating 12 days after weaning F2A). F2A animals weighed, observed, weaned, sacrificed and gross necropsy performed.
- F2B animals weighed, observed, weaned, and selected at random to constitute groups for breeding. The remainder were sacrificed.
- F2B mated to create F3A (first mating) and F3B (second mating). F3A weighed, observed, weaned, sacrificed and performed gross necropsy. F2B parents sacrificed after weaning F3B pups. F3B weighed, observed, weaned, sacrificed, autopsy and histopathology performed.

Doses / concentrations

Dose / conc.:

20 000 ppm

Remarks:

Diet containing 2 % of sodium stearoyl lactylate.
However, during the first six weeks of the study, rats were fed 60% of these concentrations due to proportionately higher food consumption during that period.

No. of animals per sex per dose:

20

Control animals:

yes

Details on study design:

not specified

Positive control:

None

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Animals were checked for mortality and general physical condition

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly, time of sacrifice


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Mean weekly food intake measured

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

Oestrous cyclicity (parental animals):

Not specified

Sperm parameters (parental animals):

Not specified

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: not specified

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring: number and sex of pups, stillbirths, live births, presence of gross anomalies, weight gain

GROSS EXAMINATION OF DEAD PUPS: Yes, for skeletal or visceral abnormalities

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: not specified

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY: not specified

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals after offspring weaned.
- Maternal animals: All surviving animals after offspring weaned.

GROSS NECROPSY
- Gross necropsy consisted of gonad weights and litter data.

HISTOPATHOLOGY / ORGAN WEIGHTS
Gonad weights

Postmortem examinations (offspring):

GROSS NECROPSY
- Gross necropsy consisted of gonad weights and litter data.

HISTOPATHOLOGY / ORGAN WEIGHTS
- Gonad weights (i.e. testes in males and ovary in females) in all parent and foetal generations
- Heart, liver and kidney weights of F3 generations (occurred two days after weaning of F3 pups)
- Histopathology of F3 generations

Statistics:

Not specified

Reproductive indices:

not specified

Offspring viability indices:

Number of live/dead pups per litter

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Description (incidence and severity):

All P0 rats were in good condition throughout.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Description (incidence):

All P0 rats survived their portion of the study.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

At 19-20 weeks of study the treated P0 rats of either sex weighed 94-99 % as much as the controls.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

The mean food consumption paralleled body weights.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

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:

no effects observed

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

Description (incidence and severity):

The uterine implantation sites between treated and control female rats were comparable to one another. There was no significant differences observed in the number of gestation days between control (range of 21-22 days) and treated (range of 21 to 24 days) dams.

Details on results (P0)

There were no differences among control and test groups that could be ascribed to the feeding of sodium stearoyl lactylate.

Effect levels (P0)

Key result

Dose descriptor:

NOEL

Effect level:

20 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed

Target system / organ toxicity (P0)

Critical effects observed:

no

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

One control female died during the study on day 22 of gestation, having begun labor in very poor condition with wheezing and low body temperature.

An additional control was sacrificed in poor condition during week 23. This animal had born an F2a litter but did not conceive the second time.

Otherwise, the F1b rats were in a good condition.

Dermal irritation (if dermal study):

not examined

Mortality:

mortality observed, non-treatment-related

Description (incidence):

One control female died during the study on day 22 of gestation.

An additional control was sacrificed in poor condition during week 23. This animal had born an F2a litter but did not conceive the second time.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

At week 22 the males and females receiving the material in the diet weighed 92-101 % compared to the controls.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Food consumptions figures for the two groups were closely similar throughout the experiment.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

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:

no effects observed

Description (incidence and severity):

Mean testis and ovary weights of the treatment group were comparable to the control group.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

At necropsy ten male and female P1 animals contained kidney, liver and lung anomalies but these were neither consistent, frequent, or difference enough for control animals to show test material relationship.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Details on results:

Visceral findings showed nothing that could be related to dietary level of sodium stearoyl lactylate. Calculations that give the percentages of F1a pups which had zero, one, two or three or more anomalies per pup indicate no meaningful differences between the groups.

Reproductive function / performance (P1)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Description (incidence and severity):

Implantation sites of the treatment group were comparable to the control group. No meaningful discrepancies between total numbers of implantation sites and total numbers of pups per dam were found.

Effect levels (P1)

Key result

Dose descriptor:

NOEL

Effect level:

20 000 ppm

Sex:

male/female

Basis for effect level:

other: no adverse effects observed

Target system / organ toxicity (P1)

Critical effects observed:

no

Results: F1 generation

General toxicity (F1)

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

In one animal of the control group respiratory disease was observed. This animal died between week 16 and 18. Otherwise the F2b rats were in a good condition.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

One male from the control group died during the study. The death was attributed to respiratory disease.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Mean body weights of F2a and F2b pups at day 5 and at weaning were on par with control weights. Pups of the treatment group weighed 79% of the control weight.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Food consumptions figures for the two groups were closely similar throughout the experiment.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

The testis and ovary weights between treated and control groups were comparable.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Gross autopsy findings in F2b females were confined chiefly to lung abscesses in one female. Except for the testis weights, gross observations on the males were not made.

Histopathological findings:

not examined

Other effects:

not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not examined

Effect levels (F1)

Key result

Dose descriptor:

NOEL

Effect level:

20 000 ppm

Sex:

male/female

Basis for effect level:

other: no adverse effects observed

Target system / organ toxicity (F1)

Critical effects observed:

no

Results: F2 generation

General toxicity (F2)

Clinical signs:

no effects observed

Description (incidence and severity):

The F2 generation rats were in good condition. Slightly lower survival at five and twenty-one days which is believed to be because of an intercurrent infection of undetermined nature.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

Slightly lower survival at five and twenty-one days which is believed to be because of an intercurrent infection of undetermined nature.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

At week 20, the males and females receiving the material in the diet weighed 93 to 106 % of the controls.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No significant differences were noted.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

The gonadal weights between treated and control groups were comparable to one another.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No significant differences were shown between control and test rats. Five females showed lung and liver abnormalities and males were not observed. These are not considered different from abnormalities in controls.

Histopathological findings:

not examined

Other effects:

not examined

Developmental neurotoxicity (F2)

Behaviour (functional findings):

not examined

Developmental immunotoxicity (F2)

Developmental immunotoxicity:

not examined

Effect levels (F2)

Key result

Dose descriptor:

NOAEL

Effect level:

20 000 ppm

Sex:

male/female

Basis for effect level:

other: no adverse effects observed

Target system / organ toxicity (F2)

Critical effects observed:

no

Overall reproductive toxicity

Key result

Reproductive effects observed:

no

Any other information on results incl. tables

There was slightly lower survival at five and twenty-one days in all litters which was believed to be happenstance.

There were no other significant differences or effects observed for mortality, body weights, food intake, gross necropsy (gonad weights) in either of the P2 or F3 generations.

None of the histopathological observations made in F3 generation were believed to be related to the administration of the test substance sodium stearoyl lactylate under the conditions of this study.

The full tables of litter data for each generation are below.

F1A Litter Information

Observations	Control	Sodium Stearoyl Lactylate (2%)
Litters per group	18/20	16/20
Total live pups	207	178
Total Stillborn	1	0
Live pups per litter	11.5	11.1
Mean body weights (g) of live pups	5.53	5.94
Number of male pups	109	84
Number of female pups	98	94

 

F1B Litter Information

Observations	Control	Sodium Stearoyl Lactylate (2%)
Litters per group	12/20	18/20
Total live pups
Birth	151	221
Day 5	96	128
Weaning	79	106
Total Stillborn	0	0
Live pups per litter	12.6	12.3
Per cent survival at day 5	63.6	58.0
100x weaning survival/ 5 day survival	86.8	87.6
Mean body weights (g) of live pups at
Birth	5.90	6.14
Day 5	9.65	9.08
Weaning	41.5	37.7

 

F2A Litter Information

Observations	Control	Sodium Stearoyl Lactylate (2%)
Litters per group	19/19	17/20
Total live pups
Birth	221	196
Day 5	127	116
Weaning	81	58
Total Stillborn	1	2
Live pups per litter	11.6	11.5
Per cent survival at day 5	57.5	59.2
100x weaning survival/ 5 day survival	68.6	55.2
Mean body weights (g) of live pups at
Birth	5.96	6.18
Day 5	7.43	7.50
Weaning	33.9	33.8

 

F2B Litter Information

Observations	Control	Sodium Stearoyl Lactylate (2%)
Litters per group	17/19	19/20
Total live pups
Birth	210	219
Day 5	97	67
Weaning	63	55
Total Stillborn	0	1
Live pups per litter	12.4	11.5
Per cent survival at day 5	46.2	30.6
100x weaning survival/ 5 day survival	72.4	82.2
Mean body weights (g) of live pups at
Birth	5.91	6.08
Day 5	8.34	7.63
Weaning	39.5	31.1

 

F3A Litter Information

Observations	Control	Sodium Stearoyl Lactylate (2%)
Litters per group	14/20	18/20
Total live pups
Birth	143	193
Day 5	82	123
Weaning	68	113
Total Stillborn	0	0
Live pups per litter	10.2	10.2
Per cent survival at day 5	57.6	63.7
100x weaning survival/ 5 day survival	82.9	91.9
Mean body weights (g) of live pups at
Birth	5.73	5.87
Day 5	8.89	9.46
Weaning	37.5	33.5

 

F3B Litter Information

Observations	Control	Sodium Stearoyl Lactylate (2%)
Litters per group	16/20	17/20
Total live pups
Birth	167	194
Day 5	115	138
Weaning	101	127
Total Stillborn	1	0
Live pups per litter	11.1	11.4
Per cent survival at day 5	68.9	71.1
100x weaning survival/ 5 day survival	87.8	92.0
Mean body weights (g) of live pups at
Birth	6.03	6.20
Day 5	9.76	10.1
Weaning	36.7	36.4

 

Applicant's summary and conclusion

Conclusions:

There were no differences among control group and those fed sodium stearoyl lactylate that could be ascribed to treatment. Sodium stearoyl lactylate does not adversely effect reproduction in albino rats through three generations.

Executive summary:

In a three-generation reproduction study, sodium stearoyl lactylate was administered to 20 male and female Sprague-Dawley rats at dose levels of 0 and 20000 ppm in diet. The animals were placed on compound no later than one week after weaning. Original parent rats (F0 generation) were bred twice; the F1a pups were sacrificed at birth and part of each litter was examined either for skeletal abnormalities (cleared, stained specimens) or for visceral changes (Bouin’s fixative specimens). F1b pups were reared to weaning and pups from each litter were taken to constitute the next group of breeders. F1b rats were bred twice, and both F2a and F2b litters were reared to weaning. F2b pups were then distributed into new groups to breed the F3 generations (F3a and F3b). Mortality, body weights, food intake, gross necropsy (gonad weights) and litter data were determined. There were no differences among the control group and those fed sodium stearoyl lactylate that could be ascribed to treatment. Sodium stearoyl lactylate does not adversely affect reproduction in albino rats through three generations.