Naphthenic acids, reaction products with diethylenetriamine

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

The commercial naphthenic acid was investigated for its repeated dose toxicity according to the OECD Guideline 422. The applied doses were 0, 100, 300 and 900 mg/kg bw.

All animals survived to scheduled termination except two females of 900 mg/kg bw group. The terminal body weight of males and females of 900 mg/kg bw were reduced. Reduced red blood cell count for males of 900 mg/kg bw and increased white blood cell count for females of 900 and 300 mg/kg bw were observed. Liver weight was increased for animals of 900 mg/kg bw and kidney weight was increased for males only in this group. The histopathological findings comprised hepatocellular hypertrophy, hyaline-droplet nephropathy, cortial lymphoid depletion in thymus, epithelial hypertrophy and cytoplasmic vacuolation of the thyroid gland and cytoplasmic vacuolation in the adrenal cortex. The NOAEL was 100 mg/kg bw for parental animals.

With respect to the reproductive/developmental toxicity investigation, reduced live pubs at birth and reduced survival rate from birth to PND 4 was found at doses of 300 and 900 mg/kg bw and reduced pub weights at dose of 900 mg/kg bw. The NOAEL for reproductive/developmental toxicity was 100 mg/kg bw. 

The introduced study is proposed to be used to derive the reproduction toxicity of NA-DETA by read-across.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

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

Adequacy of study:

key study

Study period:

2012

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Rationale for read-across: naphthenic acid is presumed to be the toxic metabolite of NA-DETA Rationale for assigning reliability of 2: read-across, scientifically well performed study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

corn oil

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Male dosing was for 28-29 days
Females were dosed for 39-53 days

Frequency of treatment:

daily

Remarks:

Doses / Concentrations:
0, 100, 300, 900 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

12

Control animals:

yes, concurrent vehicle

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

unspecific toxic signs in females of 900 mg/kg bw

Body weight and weight changes:

no effects observed

Description (incidence and severity):

no statistically significant effect

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

no statistically significant effect

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

alterations found in liver, kidney, thyroid, thymus and adrenal

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

reduced survival of pubs from birth to lactation day 4

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

reduced at 900 mg/kg bw

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

reduced for pubs of 900 mg/kg bw

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

100 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Number of pubs born; Percentage of pups surviving from birth to termination PND 4

Reproductive effects observed:

not specified

Parental toxicity data:

Table 1. Results of Assessments of hematology and clinical chemistry parameters which were statistically different from control values.^1
Parameter Measured	Corn Oil Control	100 mg/kg/day	300 mg/kg/day	900 mg/kg/day
Males, data taken at terminal sacrifice
Red Blood Cell Count (10^6/ul)^2	9.22 ± 0.54	9.28 ± 0.28	8.91 ± 0.34	8.78 ± 0.22*
Hemoglobin (g/dL)^2	15.7 ± 0.72	15.8 ± 0.48	15.2 ± 0.54	14.7 ± 0.44*
Hematocrit (%)^2	48.1 ± 2.4	48.6 ± 1.6	46.5 ± 1.5	45.0 ± 1.8**
Platelet (10^3/ul)^2	854 ± 151	885 ± 84	803 ± 144	976 ± 87**
Leukocytes, absolute (10^3/ul)^2	0.02 ± 0.02	0.03 ± 0.02	0.02 ± 0.02	0.04 ± 0.03*
RDW (%)^2	11.4 ± 0.4	11.5 ± 0.4	11.6 ± 0.4	12.5 ± 0.6**
HDW (g/dL)^2	2.58 ± 0.10	2.68 ± 0.12	2.76 ± 0.16*	2.77 ± 0.27*
Females, data taken at termination (lactation day 4)
White blood cell count^2	5.15 ± 1.30	6.89 ± 1.58	7.68 ± 2.24*	7.59 ± 1.85*
APTT (seconds)^2	16.8 ± 1.9	15.9 ± 2.3	15.8 ± 3.1	13.9 ± 1.4 *
Lymphocytes, absolute (10^3/ul),	3.32 ± 0.61	4.50 ± 1.42	5.11 ± 1.75*	4.96 ± 1.60*
Monocytes, absolute (10^3/ul)	0.11 ± 0.10	0.24 ± 0.21	0.21 ± 0.12	0.35 ± 0.23*

 

1. Parameters not affected by treatment included:

a. Males – white blood cell count, mean corpuscular volume (fL), mean corpuscular hemoglobin (pg), mean corpuscular hemoglobin content (g/dL), prothrombin time (sec), APTT (sec), reticulocytes (%), reticulocytes, absolute (10³/ul), MPV (fL), neutrophils (%), lymphocytes (%), monocytes (%), eosinophils (%), basophils (%), leucocytes(%), neutrophils, absolute (103/ul), lymphocytes, absolute (10³/ul), monocytes, absolute (10³/ul), eosinophils, absolute (10³/ul), basophils, absolute (10³/ul).

b. Females – red blood cell count (10⁶/ul), Hemoglobin content (g/dL), hematocrit (%), mean corpuscular volume (fL), mean corpuscular hemoglobin (pg), mean corpuscular hemoglobin content (g/dL), platelet count (10³/ul), prothrombin time (sec), reticulocytes (%), reticulocytes, absolute (103/ul), ), MPV (fL), neutrophils (%), lymphocytes (%), monocytes (%), eosinophils (%), basophils (%), leucocytes(%), neutrophils, absolute (10³/ul), eosinophils, absolute (10³/ul), basophils, absolute (10³/ul), Leukocytes absolute (10³/ul), RDW (%), HDW (g/dL)

 

2. Data given as mean + SD

* = p < 0.05, ** = p < 0.01

 

Table 2. Statistically significant changes in terminal body weights and organ weights.The data are given as mean + SD.
Parameter	Sham Control	Corn Oil Control	100 mg/kg/day	300 mg/kg/day	900 mg/kg/day
Males
Final Body Weight	467 ± 27	454 ± 45	448 ± 45	439 ± 34	412 ± 28
Liver	15.61 ± 1.43	13.46 ± 2.01	13.98 ± 2.04	15.69 ± 1.83*	19.94 ± 2.08**
Kidney	3.51 ± 0.25	3.21 ± 0.20*	3.38 ± 0.39	3.53 ± 0.33	3.77 ± 0.46**
Heart	1.46 ± 0.09	1.46 ± 0.21	1.41 ± 0.14	1.43 ± 0.13	1.32 ± 0.13
Thyroid/parathyroid	0.019 ± 0.002	0.019 ± 0.001	0.020 ± 0.002	0.020 ± 0.002	0.020 ± 0.002
Epididymis (LT)	0.57 ± 0.14	0.60 ± 0.05	0.60 ± 0.04	0.66 ± 0.05*	0.63 ± 0.06
Epididymis (RT)	0.62 ± 0.04	0.62 ± 0.06	0.61 ± 0.03	0.66 ± 0.04	0.65 ± 0.06
Females
Final body Weight	335 ± 25	313 ± 23	301 ± 30	294 ± 24	289 ± 24
Liver	13.6 ± 2.0	11.7 ± 1.5	12.1 ± 1.1	13.3 ± 1.5	17.9 ± 2.4**
Kidney	2.39 ± 0.17	2.07 ± 0.18*	2.11 ± 0.15	2.05 ± 0.25	2.17 ± 0.19
Heart	1.21 ± 0.23	1.10 ± 0.10	1.08 ± 0.10	1.07 ± 0.11	1.01 ± 0.13
Lungs	1.36 ± 0.13	1.40 ± 0.13	1.26 ± 0.12*	1.20 +± 0.12**	1.20 ± 0.07**
Uterus/Vagina	1.07 ± 0.19	1.00 ± 0.14	0.86 ± 0.08*	0.88 ± 0.11*	0.85 ± 0.12*

* = p < 0.05, ** = p < 0.01

 

Table 3. Summary of microscopic findings
Doses, mg/kg/day	Males				Females
	Corn Oil	100	300	900	Corn Oil	100	300	900
N	12	12	12	12	9	12	10	10
Kidney
Hyaline Droplets	0	3	10**	11**	0	0	0	0
Minimal	0	3	9**	9**	0	0	0	0
Mild	0	0	1	2
Nephropathy	0	0	2	9**	0	0	0	0
Minimal	0	0	2	5*	0	0	0	0
Mild	0	0	0	4
Liver
Hypertrophy, hepatocellular, centrilobular	0	0	0	8**	0	0	0	10**
Minimal	0	0	0	8**	0	0	0	10**
Vacuolation, hepatocellular	2	1	2	0	0	1	0	2
Minimal	1	1	2	0	0	1	0	2
Mild	1	0	0	0	0	0	0	0
Thymus
Depletion, lymphoid, cortex	0	0	0	0	0	1	0	5
Minimal	0	0	0	0	0	1	0	4
Mild	0	0	0	0	0	0	0	1
Thyroid
Hypertrophy, epithelial	0	6	9*	11**	0	3	4	8**
Minimal	0	6	9**	11**	0	3	4	6*
Mild	0	0	0	0	0	0	0	2
Vacuolation, cytoplasmic	0	6	9**	10**	0	3	4	8**
Minimal	0	6	9**	10**	0	3	4	8**
Adrenal Cortex
Vacuolation, cytoplasmic	0	2	3	2	0	0	0	2
Minimal	0	2	3	2	0	0	0	1
Mild	0	0	0	0	0	0	0	1
Heart
Cardiomyopathy
Minimal	4	7	8*	8*	3	3	2	5
Mild	2	0	0	0	0	0	0	0

* = p < 0.05, ** = p < 0.01

Data on reproduction performance and offsprings

Table 1. Summary of reproductive parameters assessed in the repeated dose/reproductive toxicity study of naphthenic acids.
Dose (mg/kg bw)	Corn Oil Control	100 mg/kg/day	300 mg/kg/day	900 mg/kg/day
Number of females paired	12	12	12	12
Number of females mated	12	12	10	11
Number of females pregnant^a	9	12	10	11
Number of females with litters	9	12	10	11
Precoital interval (days)	1.4 ± 0.7	2.3 ± 1.1	4.2 ± 3.3 *^b	3.8 ± 3.5
Gestation length (days)	21.4 ± 0.6	21.9 ± 0.3	22.0 ± 0.5	22.1 ± 0.5
Corpora lutea	15.6 ± 2.3	14.0 ± 1.4	15.1 ± 3.0	13.8 ± 2.1
Implantation sites	15.0 ± 2.4	13.6 ± 1.1	13.0 ± 1.2	12.2 ± 3.7
Number born	14.1 ± 1.9	12.9 ± 1.1	12.0 ± 1.6	10.8 ± 3.8 *
Post-implantation loss (%)	6.0	5.1	7.7	11.5

 

a. Pregnant = uterine implantation sites

b. A single female in the 300 mg/kg bw group had a pre-coital interval of 13 days

 

* = p < 0.05, ** = p < 0.01

Table 2. Survival, viability and growth of offspring
Dose (mg/kg bw)	Corn Oil Control	100 mg/kg/day	300 mg/kg/day	900 mg/kg/day
Number of viable litters	9	12	10	11
Number of pubs born alive/litter	13.9 ± 1.9	12.9 ± 1.1	10.1 ± 4.0 *	9.6 ± 4.0 **
Percentage of pubs surviving from birth to PND 4	98.1 ± 3.8	100 ± 0.0	88.0 ± 24.5	67.7 ± 40.6
Pubs (litters) found dead or euthanized in extremis	1(1)	0(0)	12(5)	38(8)
Sex ratio (% males / litter)	58.9 ± 9.6	53.9 ± 9.6	55.2 ± 19.1	58.1 ± 22.7
Pub weight PND 1 -males	7.0 ± 0.5	6.7 ± 0.6	6.7 ± 0.5	5.7 ± 0.8 *
Pub weight PND 1 -females	6.6 ± 0.6	6.5 ± 0.6	6.4 ± 0.4	5.6 ± 1.1
Pub weight PND 4 -males	9.7 ± 1.1	9.4 ± 1.2	9.4 ± 0.9	7.2 ± 1.5 **
Pub weight PND 4 -females	9.1 ± 1.0	9.0 ± 1.0	8.8 ± 0.7	7.3 ± 1.5 **

 

* = p < 0.05, ** = p < 0.01

Conclusions:

The commercial naphthenic acid was investigated for its reproduction toxicity according to the OECD Guideliine 422. The NOAEL was 100 mg/kg bw both for parental and offsprings. The introduced study is proposed to be used to derive the reproduction toxicity of NA-DETA by read-across.

Executive summary:

The commercial naphthenic acid was investigated for its repeated dose toxicity according to the OECD Guideline 422. The applied doses were 0, 100, 300 and 900 mg/kg bw.

All animals survived to scheduled termination except two females of 900 mg/kg bw group. The terminal body weight of males and females of 900 mg/kg bw were reduced. Reduced red blood cell count for males of 900 mg/kg bw and increased white blood cell count for females of 900 and 300 mg/kg bw were observed. Liver weight was increased for animals of 900 mg/kg bw and kidney weight was increased for males only in this group. The histopathological findings comprised hepatocellular hypertrophy, hyaline-droplet nephropathy, cortial lymphoid depletion in thymus, epithelial hypertrophy and cytoplasmic vacuolation of the thyroid gland and cytoplasmic vacuolation in the adrenal cortex. The NOAEL was 100 mg/kg bw for parental animals.

With respect to the reproductive/developmental toxicity investigation, reduced live pubs at birth and reduced survival rate from birth to PND 4 was found at doses of 300 and 900 mg/kg bw and reduced pub weights at dose of 900 mg/kg bw. The NOAEL for reproductive/developmental toxicity was 100 mg/kg bw. 

The introduced study is proposed to be used to derive the reproduction toxicity of NA-DETA by read-across.

Effect on fertility: via oral route

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

100 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

One valid OECD 422 study in rats for the purpose of read-across

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

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.

Link to relevant study records

Reference

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

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

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.

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.

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.

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

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

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

Key result

Abnormalities:

no effects observed

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

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

Key result

Abnormalities:

no effects observed

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

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.

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.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

40 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

One valid OECD 414 study in rat available

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

No classification is warranted.

The commercial naphthenic acid (read-across substance) induced reduced pup survival rate at doses that were associated with apparent parental toxicity. No other effect on the reproduction performance was found.

The registration substance NA-DETA induced reduced fetal weight at dose that was associated with apparent maternal toxicity. No other effect on the maternal reproduction performance was found and no indication of teratogenicity was found.