Rape oil, reaction products with diethanolamine

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

migrated 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:

other: see 'Remark'

Remarks:

Read across study conducted with the structural analogue amides, C12-18(even-numbered) and C18(unsatd.), N,N-bis(hydroxyethyl) hence maximum reliability rating of 2 assigned according to ECHA guidance, although study was conducted according to the OECD Guideline 414 (Teratogenicity) and the EU Directives 87/302/EEC and 91/325/EEC. Compliance to GLP has been claimed through the GLP regulations according to the Chemical Act (FRG), March 14, 1990.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Official Journal of European Community L 133, May 30, 1988; 87/302/EEC

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Official Journal of European Community L 180, March 01, 1991; 91/325/EEC

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Amides, C12-18 and C18-unsatd., N,N-bis(hydroxyethyl)

EC Number:

292-477-9

EC Name:

Amides, C12-18 and C18-unsatd., N,N-bis(hydroxyethyl)

Cas Number:

90622-74-5

IUPAC Name:

90622-74-5

Details on test material:

- Name of test material (as cited in study report): (Amides, coco, N,N-bis (hydroxyethyl))
- Physical state: Yellow liquid
- Analytical purity: 90-95 %
- Lot/batch No.: 3111196
- Expiration date of the lot/batch: July, 1993
- Stability under test conditions: Stable in arachidis oil, DAB 9
- Storage condition of test material: Room temperature

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Wiga D-Sulzfeld
- Age at study initiation: 8-10 wk
- Weight at study initiation: 209 g (mean)
- Housing: Single animal in Makrolon Type M3 cage (Ebeco) with standard softwood bedding
- Diet: Pelleted Altromin Maintenance Diet 1324, ad libitum
- Water: Tap water, ad libitum
- Acclimation period: 5 d


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 43-66
- Air changes (per h): 10-15
- Photoperiod (h dark/h light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: Arachidis oil, DAB 9

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Test material was suspended in Arachidis oil, DAB 9 such that the required dose per kg body weight was contained in 5 mL.

VEHICLE
- Concentration in vehicle: 0, 20, 60 and 200 mg/mL

Analytical verification of doses or concentrations:

no

Details on mating procedure:

- Impregnation procedure: Purchased timed pregnant

Duration of treatment / exposure:

From Day 6 up to Day 15 post coitum

Frequency of treatment:

Once daily

Duration of test:

20 d

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 100, 300 and 1,000 mg/kg/d
Basis:
actual ingested
(Dosing on the basis of body weight of Day 6 post coitum)

No. of animals per sex per dose:

30

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on the results of toxicological examinations done before (Report No. 486 = TBD 830034, June 27, 1983) (details not reported)

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily

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

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation Day 20
- Organs examined: All maternal organs, with emphasis on the uterus and uterine contents

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
- Other: Position of fetus in the uterus

Fetal examinations:

External examinations: Yes: [all per litter]
- Soft tissue examinations: Yes: [approximately half per litter]
- Skeletal examinations: Yes: [approximately half per litter]
- Head examinations: Yes: [approximately half per litter]

(See Table 1 for exact number of fetuses examined)

Statistics:

The following statistical methods were used:
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).

Indices:

- Pre-implantation loss (%) = [(Number of corpora lutea - number of implantations)/number of corpora lutea] X 100
- Post-implantation loss (%) = [(Number of implantations - number of live fetuses)/number of implantations] X 100
- Sex ratio (%) = [(number of males/females)/number of fetuses] X 100

Historical control data:

None

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
Mortality: No mortality at any dose level.

Clinical symptoms: Salivation and propulsion of the head in all dose groups. Additionally, the highest dose group showed a severe salivation. These symptoms were noted variable in the individual groups during the application period.

Body weight: No treatment-related effects on body weight gain were observed in the dams.

Necropsy: No macroscopic changes were observed in the survived dams except for one dam at 100 mg/kg/d, which showed greenish-brownish fluid in the uterine horn.

Placenta and uterus weight: No significant differences between the control and the treatment groups.

Reproduction data: Pre-implantation loss was not affected by the treatment. The post-implantation loss and total embryonic deaths were significantly increased in all treatment groups. However, these findings were considered to be incidental because the values in the 100 mg/kg/d group were significantly greater than other two higher dose groups and in each group there was one single female with a high incidence of embryonic death.

Effect levels (maternal animals)

1

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day

Basis for effect level:

other: maternal toxicity

2

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day

Basis for effect level:

other: developmental toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Body weight: The weights of live fetuses exhibited no significant differences on a litter and individual basis.

Sex ratios: The sex ratio of the fetuses was not affected by the treatment.

External examinations: No macroscopic findings were observed at external examination of fetuses which were considered to be an effect of the treatment. 1 dead fetus in control and 7 dead fetuses (4 out of 7 partly mummified) in 100 mg/kg/d group were recorded. One fetus showed a stump tail at 300 mg/kg/d and paleness was observed in one fetus at 1,000 mg/kg/d. These singular findings are normal observations in the animal strain used.

Visceral examination: No treatment-related abnormalities.

Skeletal examination:
(i) Retardations: No significant finding at 100 mg/kg/d. Two sternebrae were non-ossified in 19 and 29 fetuses (statistically significant) at 300 and 1,000 mg/kg/d, respectively. Statistically significant increase in the number of fetuses with incomplete ossification of skull bones (17 fetuses) and decrease in the number of fetuses with incomplete ossification of 13th rib (0 fetus) was observed at 1,000 mg/kg/d. The increased "incomplete ossified skull bones" was essentially due to only 2 dams. The other statistically significant differences were considered to be incidental because these retardation effects were not accompanied by weight retardation and were within the normal range of variation for this strain.
(ii) Variations: No variations in any group.
(iii) Malformations: One fetus with stump tail and missing vertebrae coccigycae at 300 mg/kg/d (not considered to be treatment-related).

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Any other information on results incl. tables

Table 2. Summary of performance of mated females

Treatment dose (mg/kg/d)	0	100	300	1,000
No. of mated females	30	30	30	30
No. of pregnant females	30	29	28	29
No. of females with premature litter	1	0	2	3
No. of mortalities	0	0	0	0
No. of females with live fetuses at termination	29	29*	26	26

* One dam out of these was not included because the weights of fetuses were not determined

Applicant's summary and conclusion

Conclusions:

Based on the results, the structurally similar amides, C12-18(even-numbered) and C18(unsatd.), N,N-bis(hydroxyethyl) was not found to be cumulative toxic to pregnant rats and did not reveal any embryotoxic or teratogenic potential at dose levels up to 1,000 mg/kg/d.

Executive summary:

A study was carried out to assess the effects of structurally similar amides, C12-18(even-numbered) and C18(unsatd.), N,N-bis(hydroxyethyl) on

embryonic and foetal development in pregnant Sprague-Dawley CD rats following the procedures indicated by the OECD guideline 414 (Teratogenicity) and the EU Directives 87/302/EEC and 91/325/EEC.

The test material was administered to groups of 30 female rats orally by gavage at dose levels of 0, 100, 300 and 1,000 mg/kg/d, once daily from Day 6 to day 15 of gestation inclusive. Control animals were dosed with the vehicle alone (arachidis oil, DAB 9). Clinical condition and reaction to treatment were recorded at least once daily. Body weights were reported on Day 0, 6, 16 and 20 of gestation. All surviving females were sacrificed on Day 20 of gestation and the foetuses were removed by caesarean section. At necropsy the females were examined macroscopically. Live foetuses were weighed, sexed and were examined for visceral and skeletal abnormalities.

No deaths or treatment-related changes in body weight gain and necropsy findings were observed in dams at any dose level. Treatment-related symptoms observed in all treatment groups were salivation and propulsion of the head. Additionally, the highest dose group showed severe salivation. Apart from the control (1 dead foetus) and the 100 mg/kg/day group (7 dead foetuses), all females had viable foetuses. Pre-implantation loss and mean numbers of resorptions were not affected by treatment. The figures of post-implantation loss, embryonic deaths and total foetuses showed some deviations which were considered to be non-treatment-related. Mean foetal placental and uterus weights were not affected by the 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 visceral and skeletal abnormalities/variations. One foetus in 300 mg/kg/d group showed a stump tail and missing vertebrae coccigycae. Further, the figures of skeletal ossifications showed some deviations in the two highest dose groups. However, all these effects were assessed to be non-treatment-related.

Based on the results the test substance was found not to be cumulative toxic to pregnant rats and did not reveal any embryotoxic or teratogenic potential to Sprague-Dawley CD rats at dose levels up to 1,000 mg/kg bw/day.