Fatty acids, tall-oil, esters with ethylene glycol

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 Jul - 28 Jul 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Version / remarks:

adopted in 1981

Deviations:

no

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Version / remarks:

adopted in 2001

Deviations:

yes

Remarks:

only organogenesis covered

Test guideline 3

Qualifier:

according to guideline

Guideline:

EU Method B.31 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

only organogenesis covered

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C16-18, esters with ethylene glycol

EC Number:

292-932-1

EC Name:

Fatty acids, C16-18, esters with ethylene glycol

Cas Number:

91031-31-1

Molecular formula:

C18H36O3, C20H40O3, C34H66O2, C36H70O4, C38H74O2

Test animals

Species:

rat

Strain:

other: Sprague-Dawley, CD

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles Rover, Sulzfeld, Germany
- Age at study initiation: 8 weeks
- Weight at study initiation: 198 g
- Housing: animals were housed individually in Makrolon M3 cages with standard softwood bedding.
- Diet: pelleted Altromin Maintenance Diet 1324, Lot No. 170994/1340 (Altromin GmbH, Lage, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-24
- Humidity (%): 47-82
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.5% sodium carboxymethylcellulose and 0.25% Cremophor in aqua dest.

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: The test article was prepared daily before administration.

Analytical verification of doses or concentrations:

yes

Details on mating procedure:

- Impregnation procedure: purchased timed pregnant
- They were received at the testing facility on Day 0 of gestation.

Duration of treatment / exposure:

Day 6-15 of gestation

Frequency of treatment:

daily, 7 days/week

Duration of test:

Day 20 of gestation

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

300 mg/kg bw/day (actual dose received)

Doses / concentrations 3

Dose / conc.:

900 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

24 females

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: dose levels were based on the results of toxicological examinations (no further information, Henkel Report TBD 710070).

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least twice daily (working days) for mortality

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least twice daily (working days) for signs of reaction to treatment and/or symptoms of illness

BODY WEIGHT: Yes
- Time schedule for examinations: on Day 0, 6, 16 and 20 post coitum

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation Day 20
- Organs examined: Gross macroscopic examination of all maternal organs with emphasis on the uterus, uterine contents, and position of foetuses in the uterus.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

- External examinations: Yes: all per litter
- Visceral examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter

Statistics:

If the variables could be assumed to follow a normal distribution, the Dunnett-test, based on a pooled variance, was applied for the comparison between the treated groups and the control group. The Steel-test was applied when the data could not be assumed to follow a normal distribution. Fisher’s Exact test for 2x2 tables was applied if the variables could be dichotomized without loss of information (Bonferroni-Holm-corrected).

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Gross pathological findings:

no effects observed

Maternal developmental toxicity

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

no effects observed

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

Description (incidence and severity):

Only one dead fetus was observed in the middle- and one dead fetus in the high-dose group from 345 and 306 live foetuses, respectively.

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
No mortalities occurred during the study period. No compound-related symptoms were observed in the treatment groups in comparison to the control group. Body weight, body weight gains and corrected body weight were within expected ranges. No compound related differences were noted between the mean reproduction data of the test groups in comparison to the control group. At scheduled necropsy no macroscopic changes were noted in the dams of the treatment groups.

Effect levels (maternal animals)

Key result

Dose descriptor:

NOAEL

Effect level:

>= 900 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Maternal abnormalities

Key result

Abnormalities:

no effects observed

Results (fetuses)

Fetal body weight changes:

no effects observed

Description (incidence and severity):

no effects observed

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

External malformations:

no effects observed

Skeletal malformations:

no effects observed

Visceral malformations:

no effects observed

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No substance-related symptoms were observed in the treatment groups. Pre-implantation loss, post-implantation loss, mean number of resorptions, embryonic deaths and total foetuses were not affected by treatment. Only one dead foetus was observed in the middle- and one dead fetus in the high-dose group from 345 and 306 live foetuses, respectively. Mean foetal placental and uterus weights were not affected by treatment. Foetal sex ratio was comparable in all groups. No treatment-related foetal abnormalities were found at necropsy. The examined foetuses showed no treatment-related malformations. The figures of visceral variations in the test groups were considered to be similar to the control group.
The mean weight of live male and female foetuses in the mid-dose group was significantly increased (see Table 1 under "Any other information on results incl. tables"). The weights of live foetuses of the other treatment groups exhibited no significant differences on a litter and individual basis e.g. mean weight in comparison to the control group.
The figures of skeletal ossifications and variations showed no treatment-related deviations, significant evidences between treated groups and the control group are listed in Table 2 under "Any other information on results incl. tables". The statistically significant differences concerning increased or decreased figures of various findings were considered to be incidental. Furthermore, no dose-relationship in any finding was observed.

Effect levels (fetuses)

Key result

Dose descriptor:

NOAEL

Effect level:

>= 900 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: teratogenicity

Fetal abnormalities

Key result

Abnormalities:

no effects observed

Overall developmental toxicity

Key result

Developmental effects observed:

no

Any other information on results incl. tables

Table 1. Results of study.

Parameter	Group 1 0 mg/kg bw	Group 2 100 mg/kg bw	Group 3 300 mg/kg bw	Group 4 900 mg/kg bw
Number of corpora lutea [total/number of dams ± SD]	16.7 ± 1.9	17.1 ± 2.0	16.7 ± 1.9	16.5 ± 1.4
Implantations [total/number of dams ± SD]	15.3 ± 2.2	15.4 ± 2.7	14.9 ± 2.3	14.8 ± 1.5
Pre-implantation loss	34	39	44	38
Post-implantation loss	15	18	13	20
Embryonic death [total]	15	18	12	19
Embryonic resorptions [total/number of dams ± SD]	0.6 ± 1.1	0.7 ± 1.1	0.5 ± 0.7	0.7 ± 0.9
Foetal resorptions [total/number of dams ± SD]	0.0 ± 0.2	0.0 ± 0.2	0.0 ± 0.0	0.1 ± 0.4
Live foetuses	336	337	345	306
Dead foetuses	0	0	1	1
Malformed foetuses [total/number of dams ± SD]	0.0 ± 0.2	0.0 ± 0.0	0.0 ± 0.0	0.0 ± 0.2
Sex of foetuses (%males : % females)	47.3 : 52.7	51.6 : 48.4	50.3 : 49.4	52.1 : 47.6
Weights of live foetuses (mean ± SD) Males Females	4.3 ± 0.9 4.0 ± 0.8	4.2 ± 0.8 4.0 ± 0.7	5.0 ± 0.9* 4.6 ± 0.8*	4.5 ± 0.7 4.3 ± 0.7
Weights of placenta (mean ± SD)	0.6 ± 0.1	0.6 ± 0.1	0.6 ± 0.1	0.6 ± 0.1
Weights of uteri (mean ± SD)	88.7 ± 14.9	89.3 ± 19.2	97.9 ± 18.9	90.9 ± 11.7

*: Dunnett-test based on pooled variance, significant at level 5%

Table 2. Skeletal examination of foetuses (stage of development).

Parameter (%)	Group 1 0 mg/kg bw	Group 2 100 mg/kg bw	Group 3 300 mg/kg bw	Group 4 900 mg/kg bw
Foetal skeleton No abnormal findings	4.0	9.8	16.1##	10.0
Skull bones, single incompletely ossified	4.5	11.6#	10.0	5.0
Sternebrae incompletely ossified abnormally ossified	66.5 24.4	84.4## 17.9	51.1## 9.4##	66.3 17.5
Coccygeal vertebrae, 4 and more ossified	32.4	22.5	56.1##	47.5#
Pelvis, pubis: incompletely ossified	6.2	0.6#	0.6##	1.3

#/##: Fishers exact test (two-sided) significant at level 5% (#) or 1% (##), Bonferroni-Holm-corrected

Applicant's summary and conclusion

Conclusions:

The test substance had no effect on intrauterine development.