Reaction mass of 2-hydroxyethyl oleate, oleic acid, and 2-hydroxyethyl palmitate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

The effects of the substance were investigated after oral administration, via gavage, in female rats during pregnancy and on embryo-foetal development. No animals died during the study. No clinical signs were noted during the study and no signs of reaction to treatment were observed during the dosing period. Animals killed at termination did not show any relevant macroscopic changes. No abnormalities were detected at the external examination of foetuses. No abnormalities were detected at the visceral examination of foetuses. On the basis of the results obtained in this study, the NOAEL for maternal toxicity and for developmental toxicity is considered to be >900 mg/kg bw/day (the highest dose employed).

Effect on fertility: via oral route

Endpoint conclusion:

no study available

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Effects on developmental toxicity

Description of key information

In a reliable study in which a structurally similar substance was investigated after oral administration, via gavage, in female rats during pregnancy and on embryo-foetal development. Three groups, each of 24 mated female Sprague Dawley SD rats, received the substance via oral gavage at the fixed inclusion levels of 100, 300 and 900 mg/kg bw/day during the gestation period, starting from Day 6 through Day of gestation. A fourth similarly constituted group received the vehicle only and acted as a control. Body weight and daily clinical signs were recorded during the in vivo phase. All females were caesarean-sectioned on Day 20 post coitum and subjected to post mortem examination. The number of corpora lutea, implantations, early and late intrauterine deaths, live and dead foetuses, uterus weight, foetal weight and sex were recorded. All foetuses were examined for external abnormalities. The foetuses in each litter was examined for fixed-visceral and skeletal abnormalities. No animals died during the study. No clinical signs were noted during the study and no signs of reaction to treatment were observed during the dosing period. Animals killed at termination did not show any relevant macroscopic changes.No abnormalities were detected at the external examination of foetuses. No abnormalities were detected at the visceral examination of foetuses. On the basis of the results obtained in this study, the NOAEL for maternal toxicity and for developmental toxicity is considered to be >900 mg/kg bw/day.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Read-across to K2 study therefore K2 is the maximum Klimisch value.

Justification for type of information:

Read-across approach - see read-across justification in section 13.

Reason / purpose for cross-reference:

read-across source

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Remarks:

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

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

Dose / conc.:

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

Dose / conc.:

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

Dose / conc.:

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

No. of animals per sex per dose:

24 P 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).

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).

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Other effects:

no effects observed

Description (incidence and severity):

At scheduled necropsy no macroscopic changes were noted in the dams of the treatment groups.

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

no effects observed

Dead fetuses:

no effects observed

Description (incidence and severity):

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.

Other effects:

no effects observed

Description (incidence and severity):

Mean foetal placental and uterus weights were not affected by treatment.

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.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 900 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

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.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 900 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: teratogenicity

Key result

Abnormalities:

no effects observed

Key result

Developmental effects observed:

no

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

Conclusions:

The maternal and developmental NOAEL is considered to be >900 mg/kg bw/day based on the absence of an adverse effect on intrauterine development.

Executive summary:

The effects of the substance were investigated after oral administration, via gavage, in female rats during pregnancy and on embryo-foetal development. Three groups, each of 24 mated female Sprague Dawley SD rats, received the substance via oral gavage at the fixed inclusion levels of 100, 300 and 900 mg/kg bw/day during the gestation period, starting from Day 6 through Day of gestation. A fourth similarly constituted group received the vehicle only and acted as a control. Body weight and daily clinical signs were recorded during the in vivo phase. All females were caesarean-sectioned on Day 20 post coitum and subjected to post mortem examination. The number of corpora lutea, implantations, early and late intrauterine deaths, live and dead foetuses, uterus weight, foetal weight and sex were recorded. All foetuses were examined for external abnormalities. The foetuses in each litter was examined for fixed-visceral and skeletal abnormalities. No animals died during the study. No clinical signs were noted during the study and no signs of reaction to treatment were observed during the dosing period. Animals killed at termination did not show any relevant macroscopic changes. No abnormalities were detected at the external examination of foetuses. No abnormalities were detected at the visceral examination of foetuses. On the basis of the results obtained in this study, the NOAEL for maternal toxicity and for developmental toxicity is considered to be >900 mg/kg bw/day.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

900 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Sufficient to address requirements.

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the findings of a reliable sub-acute developmental toxicity study conducted on a structurally similar substance, classification of the substance is not justified.