Fatty acids, C16-18, reaction products with diethanolamine

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Version / remarks:

2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C16-18, reaction products with diethanolamine

EC Number:

293-014-3

EC Name:

Fatty acids, C16-18, reaction products with diethanolamine

Cas Number:

91032-08-5

Molecular formula:

C22H45NO3 to C52H101NO5

IUPAC Name:

Fatty acids, C16-18, reaction products with diethanolamine

Test material form:

solid

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Envigo RMS srl, San Pietro al Natisone (UD), Italy
- Age at arrival: 9 weeks (females), 11 weeks (males)
- Weight at arrival: 320-339 g (males), 190-214 g (females)
- Fasting period before study: no
- Housing: during pre-pairing phase and after mating: max. 5 of one sex to a cage, in polisulphone solid bottomed cages; during mating: 1 m + 1 f in clear polisulphone cages with a stainless steel mesh lid and floor
- Diet (e.g. ad libitum): laboratory rodent diet (4 RF 21, Mucedola S.r.l.) ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: approximately 3 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±2
- Humidity (%): 55±15
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.5% carboxymethyl cellulose

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The formulations were prepared daily.

VEHICLE
- Concentration in vehicle: 10, 30 and 100 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Formulations of the test item were prepared as suspensions in 0.5% CMC. Concentration and homogeneity of the low and high dose level were assessed by taking six analytical aliquots in different positions. For the intermediate levels, only concentration was assessed by taking two different analytical aliquots.
28 hour stability at room temperature and a 8 day stability at +4°C were verified in the range from 10 to 100 mg/mL.
The results of the analyses were within the acceptability limits (85-115% for concentration and CV < 10% for homogeneity).

Details on mating procedure:

- Impregnation procedure: cohoused
- M/F ratio per cage: 1:1
- Length of cohabitation: ???
- Further matings after two unsuccessful attempts: no
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

from day 6 through day 19 post coitum

Frequency of treatment:

daily

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.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

24

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: based on information from previous study (reproduction/developmental toxicity screening test)
- Rationale for animal assignment (if not random): computerised stratified randomisation to give approximately equal initial group mean body weights

Examinations

Maternal examinations:

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: days 0, 6, 9, 12, 15, and 20 post coitum

FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
6, 9, 12, 15 and 20 post coitum

WATER CONSUMPTION: No

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day #20
- Organs examined: abnormalities

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: gross evaluation of placentae

Fetal examinations:

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

Statistics:

For continuous variables the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data. Statistical analysis of non-continuous variables was carried out by means of the Kruskal-Wallis test and intergroup differences between the control and treated groups assessed by a non-parametric version of the Williams test.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

no effects observed

Description (incidence and severity):

No signs of toxicological significance were noted during the study and no signs of reactions to treatment were observed during the dosing period. The scab on the neck seen in one high dose female during the last day of study was considered of no relevance.

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No relevant differences were noted in body weight and body weight gain of females during the study, between control and treated groups. No importance was attributed to the slight statistically significant increase in body weight gain on gestation Day 9 in group 4 females.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No relevant changes were detected in food consumption between treated and control females

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):

No toxicological relevant changes were seen in uterus weight in treated groups compared to controls.

Gross pathological findings:

no effects observed

Description (incidence and severity):

No macroscopic changes were observed at post mortem examination in treated animals, when compared to the controls. The only macroscopic change, consisting of scab on dorsal neck, is suggested to be incidental.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

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

Changes in pregnancy duration:

not examined

Changes in number of pregnant:

no effects observed

Description (incidence and severity):

Four females in the low dose group and one each in the mid- and high dose groups were found not pregnant at necropsy. Since the mating occurred
before the start of treatment, the incidence of non pregnant females is unrelated to treatment.
One female in each control, low and mid-dose groups had unilateral implantation. The number of females with live foetuses on gestation Day 20 was 24 in the control group, 20 in the low dose and 23 in the mid- and high dose groups.

Effect levels (maternal animals)

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Basis for effect level:

other: no adverse effects observed up to the limit dose

Maternal abnormalities

Key result

Abnormalities:

no effects observed

Results (fetuses)

Fetal body weight changes:

no effects observed

Description (incidence and severity):

A total of 5 small foetuses (< 2.7 g) were detected; 1 out of 317 in the control, 2 out of 280 in the low dose, 1 out of 304 in the mid-dose group and 1 out of 322 in the high dose group.
Since the number of litters affected was similar in the control and treated groups, the finding was considered incidental. No other abnormalities were detected at the external examination of foetuses.

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

Changes in postnatal survival:

not examined

External malformations:

no effects observed

Skeletal malformations:

no effects observed

Description (incidence and severity):

The skeletal findings did not give rise to concern. Anomalies (AN) and variations (VA) were observed, but they were distributed in a similar way in dose groups and control. Therefore they were considered not treatment related. The only malformation (Forepaw Flexure) was limited to control (one fouetus) and low dose group (3 foetuses/2 litters) and, hence, not dose related. Due to the low incidence and the presence only in the low dose group, this finding was considered incidental.

Visceral malformations:

no effects observed

Description (incidence and severity):

The visceral examination does not give rise to concern. Anomalies (AN) and variations (VA) were observed, but their incidence was similar in the treated groups and the control, and in some cases even more pronounced in the control.
Extreme pelvic dilatation (malformation) was seen in three foetuses/three litters in controls and in one foetus each in the low and high dose group. Due to the lower incidence observed in treated groups respect to controls, this change was considered not related to treatment.
Ureters extremely enlarged and/or kinked were seen in treated and control groups, with similar or lower incidence respect to controls and therefore this finding was considered not treatment-related.
A pulmonary cyst was reported for one foetus in the mid-dose group. Due to the lowincidence and lack of dose-response relationship, this change was considered not related to treatment.

Effect levels (fetuses)

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed up to the limit dose

Fetal abnormalities

Abnormalities:

no effects observed

Overall developmental toxicity

Developmental effects observed:

no

Any other information on results incl. tables

FATE OF FEMALES

	0 mg/kg bw/d	100 mg/kg bw/d	300 mg/kg bw/d	1000 mg/kg bw/d
Initial group size	24	24	24	24
Not pregnant	0	4	1	1
Unilateral implantation	1	1	1	0
With live foetuses at gestation Day 20	24	20	23	23

 

BODY WEIGHT (g) OF FEMALES WITH LIVE FOETUSES ON GESTATION DAY 20 – GROUP MEAN DATA

Dose level [mg/kg bw/d]		days
		0	6	9	12	15	20
0	(n) Mean SD	24 234.86 8.55	24 264.87 8.56	24 274.20 10.33	24 288.52 9.93	24 306.45 12.43	24 379.71 21.44
10	(n) Mean SD	20 235.79 8.03	20 265.86 8.59	20 274.87 9.07	20 289.63 9.71	20 307.29 12.52	20 381.31 27.30
300	(n) Mean SD	23 235.98 11.70	23 266.94 12.76	23 277.94 12.98	23 291.47 13.02	23 310.26 15.29	23 383.07 23.35
1000	(n) Mean SD	23 236.99 10.17	23 263.97 8.70	23 276.26 9.53	23 290.95 10.77	23 311.93 10.59	23 389.81 16.25

 

BODY WEIGHT GAIN PER DAY (g) OF FEMALES WITH LIVE FOETUSES ON GESTATION DAY 20 – GROUP MEAN DATA (mean daily body weight gain over the previous period starting from gestation day 0)

Dose level [mg/kg bw/d]		days
		6	9	12	15	20
0	(n) Mean SD	24 5.001 0.771	24 3.112 1.349	24 4.772 1.203	24 5.977 2.077	24 14.652 2.805
10	(n) Mean SD	20 5.013 0.654	20 3.002 1.548	20 4.920 1.519	20 5.887 2.021	20 14.804 3.328
300	(n) Mean SD	23 5.159 1.125	23 3.668 1.036	23 4.508 1.350	23 6.266 1.505	23 14.561 2.275
1000	(n) Mean SD	23 4.497 1.237	23 4.098* 1.379	23 4.895 1.469	23 6.994 1.670	23 15.576 1.796

 

* = mean value of group is significantly different from control at p < 0.05

LITTER DATA AND SEX RATIOS - GROUP MEAN DATA

Dose level [mg/kg bw/d]		Corpora lutea	Implan-tations	Uterine deaths			Viable young			% males	Implantation loss [%]			Litter weight [g]	Mean foetal weight [g]
				Early	Late	Total	Total	M	F		Pre	Post	Total
0	Mean SD (n)	13.88 3.04 24	13.83 3.03 24	0.63 1.01 24	0.00 0.00 24	0.63 1.01 24	13.21 3.20 24	6.65 2.35 23	6.83 2.01 24	47.72 14.08 23	0.28 1.37 24	5.44 9.24 24	5.71 9.16 24	47.25 12.06 24	3.58 0.23 24
10	Mean SD (n)	14.40 3.70 20	14.20 3.99 20	0.20 0.70 20	0.00 0.00 20	0.20 0.70 20	14.0 4.14 20	6.63 2.27. 19	8.11 1.70 19	45.89 9.49 18	3.06 8.73 20	3.44 11.74 20	5.66 15.69 20	49.62 14.38 20	3.60 0.29 20
300	Mean SD (n)	13.61 2.50 23	13.57 2.52 23	0.35 0.57 23	0.00 0.00 23	0.35 0.57 23	13.22 2.33 23	6.65 2.69 23	6.57 2.45 23	50.07 16.57 23	0.33 1.61 23	2.33 3.82 23	2.64 4.54 23	47.26 8.19 23	3.59 0.21 23
1000	Mean SD (n)	14.74 2.03 23	14.61 1.90 23	0.57 0.79 23	0.04 0.21 23	0.61 0.84 23	14.00 2.07 23	7.04 2.02 23	6.96 2.23 23	50.35 14.50 23	0.78 2.06 23	4.23 6.01 23	4.96 6.35 23	52.02 7.03 23	3.73 0.25 23

 

EXTERNAL EXAMINATION OF FOETUSES – GROUP INCIDENCE

Dose level [mg/kg bw/d]	Organ	Cat	Observation	No. foetuses			No. litters
				Observed	Affected	%	Observed	Affected	%
0	Whole foetus		No abnormalities detected	317	316	99.68	24	-	-
	Whole foetus	AN	small	317	1	0.32	24	1	4.17
10	Whole foetus		No abnormalities detected	280	278	99.29	20	-	-
	Whole foetus	AN	small	280	2	0.71	20	2	10.00
300	Whole foetus		No abnormalities detected	304	303	99.67	23	-	-
	Whole foetus	AN	small	304	1	0.33	23	1	4.35
1000	Whole foetus		No abnormalities detected	322	321	99.69	23	-	-
	Whole foetus	AN	small	322	1	0.31	23	1	4.35

 

SKELETAL EXAMINATION OF FOETUSES – GROUP INCIDENCE

Dose level [mg/kg bw/d]	Organ	Cat	Observation	No. foetuses			No. litters
				Observed	Affected	%	Observed	Affected	%
0	Forepaw(s)	AN	Metacarpal(s) no ossification 4^th	164	62	37.80	24	17	70.83
	Forepaw(s)	MA	Flexure	164	1	0.61	24	1	4.17
	Hindpaw(s)	AN	Metatarsal(s) no ossification 4^th	164	1	0.61	24	1	4.17
	Hindpaw(s)	MA	Metatarsal(s) incomplete ossification 4th	164	4	2.44	24	4	16.67
	Lumbar vertebrae	AN	Centrum dumb-bell shaped	164	1	0.61	24	1	4.17
	Lumbar vertebrae	AN	Asymmetrical dumb-bell shaped	164	1	0.61	24	1	4.17
	Pelvic girdle	AN	Pubis incomplete ossification	164	4	2.44	24	3	12.50
	Pelvic girdle	AN	Ischium incomplete ossification	164	2	1.22	24	2	8.33
	Ribs	VA	rudimentary 14th	164	32	19.51	24	16	66.67
	Ribs	VA	short 14th	164	1	0.61	24	1	4.17
	Sacral vertebrae	AN	Arch(es) incomplete ossification	164	2	1.22	24	2	8.33
	Skull	AN	Temporal incomplete ossification	164	27	16.46	24	18	75.00
	Skull	VA	Interparietal incomplete ossification	164	1	0.61	24	1	4.17
	Skull	VA	Supraoccipital incomplete ossification	164	5	3.05	24	5	20.83
	Sternebrae	AN	Asymmetrical ossification	164	6	3.66	24	6	25.00
	Sternebrae	AN	Asymmetrical ossification 5th	164	8	4.88	24	5	20.83
	Sternebrae	AN	No ossification 6th	164	6	3.66	24	4	16.67
	Sternebrae	AN	Dumb-bell shaped 5th	164	1	0.61	24	1	4.17
	Sternebrae	VA	Incomplete ossification 5th	164	41	25.00	24	16	66.67
	Sternebrae	VA	Incomplete ossification 6th	164	56	34.15	24	13	54.17
	Sternebrae	VA	No ossification 5th	164	9	5.49	24	6	25.00
	Sternebrae	VA	Incomplete ossification	164	20	12.20	24	10	41.67
	Thoracic vertebrae	AN	Centrum no ossification	164	2	1.22	24	2	8.33
	Thoracic vertebrae	AN	Centrum bipartite	164	1	0.61	24	1	4.17
	Thoracic vertebrae	VA	Centrum incomplete ossification	164	20	12.20	24	13	54.17
	Thoracic vertebrae	VA	Centrum dumb-bell shaped	164	11	6.71	24	8	33.33
	Thoracic vertebrae	VA	Centrum asymmetrical dumb-bell shaped	164	4	2.44	24	4	16.67
100	Forepaw(s)	AN	Metacarpal(s) no ossification 4^th	145	53	36.55	20	13	65.00
	Forepaw(s)	MA	Flexure	145	3	2.07	20	2	10.00
	Lumbar vertebrae	AN	Centrum dumb-bell shaped	145	1	0.69	20	1	5.00
	Ribs	VA	14 ribs	145	1	0.69	20	1	5.00
	Ribs	VA	rudimentary 14th	145	21	14.48	20	14	70.00
	Ribs	VA	short 14th	145	1	0.69	20	1	5.00
	Skull	AN	Temporal incomplete ossification	145	25	17.24	20	15	75.00
	Skull	VA	Supraoccipital incomplete ossification	145	2	1.38	20	2	10.0
	Skull	VA	Interparietal incomplete ossification	145	1	0.69	20	1	5.00
	Sternebrae	AN	No ossification 6th	145	2	1.38	20	2	10.0
	Sternebrae	AN	Asymmetrical ossification	145	4	2.76	20	3	15.00
	Sternebrae	AN	Asymmetrical ossification 5th	145	5	3.45	20	4	20.00
	Sternebrae	VA	Incomplete ossification 5th	145	35	24.14	20	15	75.00
	Sternebrae	VA	No ossification 5th	145	4	2.76	20	4	20.00
	Sternebrae	VA	Incomplete ossification	145	13	8.97	20	11	55.00
	Sternebrae	VA	Incomplete ossification 6th	145	52	35.86	20	15	75.00
	Thoracic vertebrae	AN	Centrum bipartite	145	1	0.69	20	1	5.00
	Thoracic vertebrae	AN	Centrum no ossification	145	1	0.69	20	1	5.00
	Thoracic vertebrae	VA	Centrum asymmetrical dumb-bell shaped	145	4	2.76	20	4	20.00
	Thoracic vertebrae	VA	Centrum dumb-bell shaped	145	27	18.62	20	16	80.00
	Thoracic vertebrae	VA	Centrum incomplete ossification	145	12	8.28	20	7	35.00
300	Forepaw(s)	AN	Metacarpal(s) no ossification 4^th	150	44	29.33	23	15	65.22
	Lumbar vertebrae	VA	Centrum incomplete ossification	150	2	1.33	23	28.70
	Ribs	AN	Wavy	150	1	0.67	23	1	4.35
	Ribs	VA	rudimentary 14th	150	21	14.00	23	11	47.83
	Skull	AN	Temporal incomplete ossification	150	31	20.67	23	13	56.52
	Skull	VA	Supraoccipital incomplete ossification	150	1	0.67	23	1	4.35
	Skull	VA	Interparietal incomplete ossification	150	6	4.00	23	4	17.39
	Sternebrae	AN	Asymmetrical ossification 5th	150	12	8.00	23	8	34.78
	Sternebrae	AN	No ossification 6th	150	1	0.67	23	1	4.35
	Sternebrae	AN	Asymmetrical ossification	150	8	5.33	23	7	30.43
	Sternebrae	VA	Incomplete ossification	150	9	6.00	23	5	21.74
	Sternebrae	VA	No ossification 5th	150	3	2.00	23	3	13.04
	Sternebrae	VA	Incomplete ossification 6th	150	36	24.00	23	17	73.91
	Sternebrae	VA	Incomplete ossification 5th	150	25	16.67	23	13	56.52
	Thoracic vertebrae	AN	Centrum bipartite	150	1	0.67	23	1	4.35
	Thoracic vertebrae	VA	Centrum incomplete ossification	150	8	5.33	23	6	26.09
	Thoracic vertebrae	VA	Centrum asymmetrical dumb-bell shaped	150	1	0.67	23	1	4.35
	Thoracic vertebrae	VA	Centrum dumb-bell shaped	150	27	18.00	23	15	65.22
1000	Forepaw(s)	AN	Metacarpal(s) no ossification 4^th	164	52	31.71	23	15	65.22
	Hindpaw(s)	VA	Metatarsal(s) incomplete ossification 4th	164	5	3.05	23	3	13.04
	Pelvic girdle	AN	Pubis incomplete ossification	164	1	0.61	23	1	4.35
	Pelvic girdle	AN	Ischiumincomplete ossification	164	1	0.61	23	1	4.35
	Ribs	VA	Rudimentary 14th	164	20	12.20	23	11	47.83
	Ribs	VA	14 ribs	164	2	1.22	23	1	4.35
	Ribs	VA	Short 14^th	164	2	1.22	23	2	8.70
	Skull	AN	Temporal incomplete ossification	164	32	19.51	23	16	69.57
	Skull	VA	Parietalincomplete ossification	164	2	1.22	23	1	4.35
	Skull	VA	Supraoccipital incomplete ossification	164	4	2.44	23	3	13.04
	Skull	VA	Interparietal incomplete ossification	164	4	2.44	23	3	13.04
	Sternebrae	AN	Asymmetrical ossification	164	3	1.83	23	2	8.70
	Sternebrae	AN	Asymmetrical ossification 5th	164	8	4.88	23	6	26.09
	Sternebrae	AN	No ossification	164	1	0.61	23	1	4.35
	Sternebrae	VA	Incomplete ossification 5th	164	30	18.29	23	14	60.87
	Sternebrae	VA	Incomplete ossification	164	8	4.88	23	5	21.74
	Sternebrae	VA	Incomplete ossification 6th	164	33	20.12	23	13	56.52
	Sternebrae	VA	No ossification 5th	164	2	1.22	23	2	8.70
	Thoracic vertebrae	AN	Centrum no ossification	164	1	0.61	23	1	4.35
	Thoracic vertebrae	VA	Centrum dumb-bell shaped	164	14	8.54	23	11	47.83
	Thoracic vertebrae	VA	Centrum incomplete ossification	164	11	6.71	23	8	34.78
	Thoracic vertebrae	VA	Centrum asymmetrical dumb-bell shaped	164	6	3.66	23	6	26.09

 

VISCERAL EXAMINATION OF FOETUSES – GROUP INCIDENCE

Dose level [mg/kg bw/d]	Organ	Cat	Observation	No. foetuses			No. litters
				Observed	Affected	%	Observed	Affected	%
0	Abdomen	VA	Haemorrhagic	153	23	15.03	24	8	33.33
	Great vessels	AN	Innominate artery absent	153	1	0.65	24	1	4.17
	Great vessels	VA	Innominate artery short	153	7	4.58	24	5	20.83
	Heart	AN	Pericardial haemorrhage	153	23	15.03	24	11	45.83
	Heart	AN	Pericardial fluid	153	10	6.54	24	6	25.00
	Heart	AN	Atrium enlarged / slight	153	2	1.31	24	2	8.33
	Heart	VA	Ventricle enlarged / slight	153	8	5.23	24	6	25.00
	Kidneys	AN	Ectopic	153	16	10.46	24	10	41.67
	Kidneys	AN	Pelvic dilatation / moderate	153	5	3.27	24	5	20.83
	Kidneys	AN	Haemorrhagic	153	1	0.65	24	1	4.17
	Kidneys	MA	Pelvic dilatation /extreme	153	3	1.96	24	3	12.50
	Kidneys	VA	Pelvic dilatation / slight	153	6	3.92	24	5	20.83
	Liver	AN	Mottled	153	6	3.92	24	2	8.33
	Testis	AN	Displaced	153	5	3.27	24	4	16.67
	Thoracic cavity	AN	Haemorrhage	153	4	2.61	24	3	12.50
	Ureter	AN	Enlarged/ moderate	153	9	5.88	24	8	33.33
	Ureter	AN	Kinked/ moderate	153	1	0.65	24	1	4.17
	Ureter	MA	Kinked/extreme	153	2	1.31	24	2	8.33
	Ureter	MA	Enlarged/ extreme	153	4	2.61	24	4	16.67
	Ureter	VA	Enlarged/ slight	153	19	12.42	24	12	50.00
	Ureter	VA	Kinked/ slight	153	3	1.96	24	2	8.33
100	Abdomen	VA	Haemorrhagic	135	8	5.93	19	4	21.05
	Great vessels	VA	Innominate artery short	135	1	0.74	19	1	5.26
	Great vessels	VA	Innominate artery longer	135	1	0.74	19	1	5.26
	Heart	AN	Pericardial fluid	135	8	5.93	19	3	15.79
	Heart	AN	Pericardial haemorrhage	135	2	1.48	19	1	5.26
	Heart	VA	Ventricle enlarged / slight	135	3	2.22	19	3	15.79
	Kidneys	AN	Ectopic	135	8	5.93	19	7	36.84
	Kidneys	AN	Pelvic dilatation / moderate	135	1	0.74	19	1	5.26
	Kidneys	MA	Pelvic dilatation /extreme	135	1	0.74	19	1	5.26
	Kidneys	VA	Pelvic dilatation / slight	135	5	3.70	19	5	26.32
	Testis	AN	Displaced	135	7	5.19	19	6	31.58
	Thoracic cavity	AN	Haemorrhage	135	1	0.74	19	1	5.26
	Ureter	AN	Enlarged/ moderate	135	5	3.70	19	4	21.05
	Ureter	AN	Kinked/ moderate	135	1	0.74	19	1	5.26
	Ureter	MA	Enlarged/ extreme	135	2	1.48	19	2	10.53
	Ureter	VA	Enlarged/ slight	135	23	17.04	19	11	57.89
	Ureter	VA	Kinked/ slight	135	6	4.44	19	5	26.32
300	Abdomen	VA	Haemorrhagic	147	25	17.01	23	9	39.13
	Great vessels	VA	Innominate artery longer	147	1	0.68	23	1	4.35
	Heart	AN	Pericardial haemorrhage	147	16	10.88	23	7	30.43
	Heart	AN	Atrium enlarged / slight	147	1	0.68	23	1	4.35
	Heart	VA	Pericardial fluid	147	15	10.20	23	9	39.13
	Heart	VA	Ventricle enlarged / slight	147	7	4.76	23	7	30.43
	Kidneys	AN	Pelvic dilatation / moderate	147	4	2.72	23	4	17.39
	Kidneys	AN	Ectopic	147	13	8.84	23	9	39.13
	Kidneys	VA	Pelvic dilatation / slight	147	5	3.40	23	4	17.39
	Lungs	AN	Lobe abnormal shape	147	1	0.68	23	1	4.35
	Lungs	MA	Pulmonary cyst	147	1	0.68	23	1	4.35
	Testis	AN	Displaced	147	6	4.08	23	5	21.74
	Ureter	AN	Enlarged/ moderate	147	3	2.04	23	2	8.70
	Ureter	MA	Enlarged/ extreme	147	2	1.36	23	2	8.70
	Ureter	MA	Kinked / extreme	147	2	1.36	23	2	8.70
	Ureter	VA	Kinked/ slight	147	7	4.76	23	6	26.09
	Ureter	VA	Enlarged/ slight	147	16	10.88	23	9	39.13
1000	Abdomen	VA	Haemorrhagic	158	10	6.33	23	7	30.43
	Great vessels	VA	Innominate artery short	158	4	2.53	23	3	13.04
	Great vessels	VA	Innominate artery longer	158	1	0.63	23	1	4.35
	Heart	AN	Atrium enlarged / slight	158	1	0.63	23	1	4.35
	Heart	AN	Pericardial haemorrhage	158	22	13.92	23	11	47.83
	Heart	AN	Pericardial fluid	158	7	4.43	23	4	17.39
	Heart	VA	Ventricle enlarged / slight	158	3	1.90	23	2	8.70
	Kidneys	AN	Haemorrhagic	158	1	0.63	23	1	4.35
	Kidneys	AN	Ectopic	158	13	8.23	23	10	43.48
	Kidneys	AN	Pelvic dilatation / moderate	158	2	1.27	23	2	8.70
	Kidneys	MA	Pelvic dilatation / extreme	158	1	0.63	23	1	4.35
	Testis	AN	Displaced	158	2	1.27	23	1	4.35
	Ureter	AN	Kinked/ moderate	158	1	0.63	23	1	4.35
	Ureter	AN	Enlarged/ moderate	158	11	6.96	23	6	26.09
	Ureter	MA	Kinked / extreme	158	2	1.27	23	2	8.70
	Ureter	MA	Enlarged/ extreme	158	1	0.63	23	1	4.35
	Ureter	VA	Enlarged/ slight	158	13	8.23	23	9	39.13
	Ureter	VA	Kinked/ slight	158	7	4.43	23	4	17.39

AN = anomaly

VA = variant

MA = malformation

Applicant's summary and conclusion

Conclusions:

Neither clinical signs nor signs of reaction to treatment were noted in treated females. No significant differences were noted in body weight, food consumption, gravid uterus weight, litter data and macroscopic observation of treated females when compared to controls. No changes were noted at the skeletal and visceral examinations of the foetuses which were considered treatment-related.
On the basis of these results the dosage of 1000 mg/kg/day could be considered the NOAEL in this study.

Executive summary:

In a prenatal developmental toxicity study according to OECD Guideline 414 (2001) Fatty acids, C16-18, reaction products with diethanolamine (98.81% a.i.) was administered to 24 female Sprague-Dawley rats in 0.5% carboxymethyl cellulose by gavage at dose levels of 0, 100, 300, or 1000 mg/kg bw/day from days 6 through 19 of gestation.

Body weight, daily clinical signs and food consumption were recorded during thein vivophase. 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. Approximately one half of the foetuses in each litter was examined for fixed-visceral and skeletal abnormalities.

No animals died during the study. Four females in the low dose group and one each in the mid- and high dose groups were found not pregnant at necropsy. One female in control, low and mid-dose groups had unilateral implantation. The number of females with live fetuses on gestation Day 20 was 24 in the control group, 20 in the low dose and 23 in the mid- and high dose groups.

No signs of toxicological significance were noted during the study and no signs of reactions to treatment were observed during the dosing period.

No relevant differences were noted in body weight and body weight gain of females during the study, between control and treated groups.

No relevant changes were detected in food consumption between treated and control females.

No relevant changes were seen in terminal body weight, uterus weight and absolute weight gain, in treated animals compared to controls.

Litter data and sex ratios were not affected by treatment.

No macroscopic changes were observed at post mortem examination in treated animals, when compared to the controls.

A total of 5 small foetuses (<2.7 g) were detected: 1 in each control, mid- and high dose groups and 2 in the low dose group. No other abnormalities were detected at the external examination of foetuses. No changes were noted at the skeletal examination of the foetuses which were considered treatment-related. No changes were noted at the visceral examination of the foetuses which were considered treatment-related.

On the basis of these results the dosage of 1000 mg/kg/day could be considered the NOAEL in this study.