Naphthenic acids, reaction products with diethylenetriamine

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015-2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Naphthenic acids, reaction products with diethylenetriamine

EC Number:

268-610-1

EC Name:

Naphthenic acids, reaction products with diethylenetriamine

Cas Number:

68131-13-5

Molecular formula:

not applicable

IUPAC Name:

Naphthenic acids, reaction products with diethylenetriamine

Test animals

Species:

rat

Strain:

Sprague-Dawley

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Duration of treatment / exposure:

from gestation day 6 to gestation day 19 / per gavage

Frequency of treatment:

once per day

Duration of test:

until the cesarean section on gestation day 20

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

80 mg/kg bw/day

Remarks:

The treatment at 250 mg/kg/day was reduced to 80 mg/kg/day due to the apparent toxicity. The period of treatment at 250 mg/kg/day was at most four days at the beginning of the treatment.

Doses / concentrations 2

Dose / conc.:

40 mg/kg bw/day

Remarks:

The treatment at 120 mg/kg/day was reduced to 40 mg/kg/day due to the apparent toxicity. The period of treatment at 120 mg/kg/day was at most four days at the beginning of the treatment.

Doses / concentrations 3

Dose / conc.:

20 mg/kg bw/day

Remarks:

The treatment at 60 mg/kg/day was reduced to 20 mg/kg/day due to the apparent toxicity. The period of treatment at 60 mg/kg/day was at most four days at the beginning of the treatment.

No. of animals per sex per dose:

24

Control animals:

yes, concurrent vehicle

Details on study design:

The dose selection was based on the dose-range-finding study, in which the animals were treated at volume of 10 mL/kg. In the presented main study the animals were treated at volume of 4 mL/kg, so that the test-item concentration of the applied formulation was in creased. In the presented main study, the animals exhibited clinical signs indicative of higher toxicity when compared to the dose-range finding study. After max. four days of exposure, the dose levels were reduced. The increased toxicity observed in the presented main study is likely to be associated with the local effects of the test-material, which in turn is likely to be associated with increased test-item concentration of the applied formulation.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

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:

not examined

Gross pathological findings:

effects observed, treatment-related

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Maternal developmental toxicity

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

effects observed, non-treatment-related

Description (incidence and severity):

one animal out of 21 pregnant animals at mid dose

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

Changes in pregnancy duration:

no effects observed

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed

Changes in number of pregnant:

no effects observed

Effect levels (maternal animals)

Key result

Dose descriptor:

NOAEL

Effect level:

20 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

body weight and weight gain

food consumption and compound intake

Maternal abnormalities

Key result

Abnormalities:

no effects observed

Results (fetuses)

Fetal body weight changes:

effects observed, treatment-related

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined

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

Description (incidence and severity):

not relevant

External malformations:

no effects observed

Skeletal malformations:

no effects observed

Visceral malformations:

no effects observed

Effect levels (fetuses)

Key result

Dose descriptor:

NOAEL

Effect level:

80 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

fetal/pup body weight changes

Fetal abnormalities

Key result

Abnormalities:

no effects observed

Overall developmental toxicity

Key result

Developmental effects observed:

yes

Lowest effective dose / conc.:

80 mg/kg bw/day

Treatment related:

yes

Relation to maternal toxicity:

developmental effects as a secondary non-specific consequence of maternal toxicity effects

Any other information on results incl. tables

Summary of Clinical Signs, Physical examination and Mortality
The animals were treated at dose of 0, 60, 120 and 250 mg/kg bw from the gestation day 5. After four days of treatment, the dose levels were reduced to 0, 20, 40, and 80 due to the severe maternal toxicity.
	Group No.	G1	G2	G3	G4
Observations	Dose (mg/kg/day)	0	60/20	120/40	250/80
	Total No. of rats found sperm positive	24	24	24	24
Clinical signs and Physical examination
Slight/moderate/severe salivation		0	0	7	11
Reddish nasal discharge		0	0	2	7
Reddish eye discharge		0	0	0	5
Moderate gasping		0	0	0	1
Mortality		0	0	0	1

Summary of maternal group mean body weight (g)
The animals were treated at dose of 0, 60, 120 and 250 mg/kg bw from the gestation day 5. After four days of treatment, the dose levels were reduced to 0, 20, 40, and 80 due to the severe maternal toxicity.
Group No.	Dose (mg/kg/day)	No. of Dams	Group mean body weight (g) on day
				0	3	5	8	11	14	17	20
G1	0	21	Mean	225.49	238.50	243.67	253.85	269.83	285.16	317.43	353.16
			SD	14.85	15.10	14.59	14.94	15.62	17.02	18.76	23.12
G2	60/20	21	Mean	228.50	241.63	247.57	250.37	266.63	279.29	306.02	342.75
			SD	15.70	18.69	18.47	21.70	22.56	25.22	34.11	42.34
G3	120/40	21	Mean	225.64	238.91	244.96	244.03	249.79*	259.49*	282.75*	310.79*
			SD	17.34	17.91	19.09	24.23	27.24	33.90	45.87	54.99
G4	250/80	21	Mean	226.53	240.94	246.50	242.10	247.44*	256.26*	278.65*	307.31*
			SD	15.18	14.96	15.21	18.88	24.63	28.59	39.17	47.22

*: Significantly different from vehicle control group.

Summary of Food Intake (g/rat /day)
The animals were treated at dose of 0, 60, 120 and 250 mg/kg bw from the gestation day 5. After four days of treatment, the dose levels were reduced to 0, 20, 40, and 80 due to the severe maternal toxicity.
Period of treatment (days of gestation)	Group No.		G1	G2	G3	G4
	Dose (mg/kg/day)		0	60/20	120/40	250/80
	No. of Dams		21	21	21	21
Intermittent food intake
0-3		Mean	16.64	17.24	17.34	17.45
		SD	1.58	2.30	1.89	1.45
3-5		Mean	18.56	18.98	19.28	19.14
		SD	1.63	1.92	2.29	1.73
5-8		Mean	16.01	13.97	13.17*	11.38*
		SD	1.66	3.75	4.43	3.80
8-11		Mean	18.46	17.13	14.62*	12.66*
		SD	1.46	3.19	4.47	5.94
11-14		Mean	19.95	18.49	16.65*	14.81*
		SD	1.59	3.39	5.68	4.58
14-17		Mean	22.28	20.83	18.92	17.91*
		SD	2.58	5.11	6.64	6.19
17-20		Mean	21.64	21.89	18.16	18.06
		SD	2.35	4.66	6.06	5.89

*: Significantly different from vehicle control group.

 

Summary of Maternal Data
The animals were treated at dose of 0, 60, 120 and 250 mg/kg bw from the gestation day 5. After four days of treatment, the dose levels were reduced to 0, 20, 40, and 80 due to the severe maternal toxicity.
	Group No.	G1	G2	G3	G4
Parameters	Dose (mg/kg/day)	0	60/20	120/40	250/80
	No. of Dams	21	21	21	21
Gravid uterine weight (g)	Mean	79.12	66.92	64.69	70.21
	SD	19.23	18.14	25.18	20.02
Number of Corpora lutea	Mean	16.52	15.48	15.90	17.05
	SD	2.52	2.50	2.21	2.04
Number of Implantations	Mean	14.43	12.86	13.86	14.81
	SD	3.64	3.53	2.92	2.23
Early Resorptions	Mean	0.67	0.76	1.62	0.95
	SD	0.80	1.04	3.20	1.63
Late Resorptions	Mean	0.19	0.48	0.52	0.62
	SD	0.68	0.98	1.78	1.91
Pre-implantation Loss	Mean	2.10	2.62	2.05	2.24
	SD	1.61	2.16	1.75	1.61
Post-implantation Loss	Mean	0.86	1.24	2.14	1.57
	SD	1.28	1.34	3.58	2.29
Dams with any Resorption	Total	10	14	11	11
Dams with only implantation site	Total	0	0	1	0
(complete resorptions)

 

Summary of Litter Data
The animals were treated at dose of 0, 60, 120 and 250 mg/kg bw from the gestation day 5. After four days of treatment, the dose levels were reduced to 0, 20, 40, and 80 due to the severe maternal toxicity.
	Group No.	G1	G2	G3	G4
Parameters	Dose (mg/kg/day)	0	60/20	120/40	250/80
	No. of Dams	21	21	21	21
No. of litters		21	21	20	21
Total no. of fetuses		285	244	246	278
Mean litter size		13.6	11.6	12.1	13.2
Dead fetuses	Total	0	0	0	0
	%	0	0	0	0
Total live fetuses
a.  Number		285	244	246	278
b.  Weight (g)	Mean	3.83	3.72	3.38	3.13*
	SD	0.33	0.56	0.74	0.79
Live male fetuses
a.  Number		137	129	121	140
b.  Weight (g)	Mean	3.96	3.82	3.47	3.24*
	SD	0.33	0.59	0.78	0.79
Live female fetuses
a.  Number		148	115	125	138
b.  Weight (g)	Mean	3.71	3.62	3.30	3.02*
	SD	0.35	0.51	0.69	0.77
Sex Ratio - Male : Female		1:1.08	1:0.89	1:1.03	1:0.99

*: Significantly different from vehicle control group.

Applicant's summary and conclusion

Conclusions:

NA-DETA is not a developmental toxicant. No classification is warranted.

Executive summary:

The developmental toxicity of NA-DETA was investigated according to the OECD Guideline 414 in SD rats. The animals were treated daily from gestation day 5 to 19 and the cesarean section was performed on gestation day 20. The initial dose levels of 0, 60, 120 and 250 mg/kg/day induced clinical signs indicative of severe toxicity (including mortality) so that the dose levels were reduced to 0, 20, 40 and 80 mg/kg bw.

The NOAEL of maternal toxicity was 60/20 mg/kg/day due to the clinical signs, reduced body weight and reduced food intake in the mid and high dose groups. No effect on the maternal reproductive performance was found in all treated animals.

The NOAEL of developmental toxicity was 120/40 mg/kg/day due to the reduced mean fetal weight in high dose group. No effect on litter size was found in all treated groups.

The NOAEL of teratogenicity was 250/80 mg/kg/day as no effect was found upon skeletal and visceral observation.