C18 unsaturated fatty acids, reaction products with 1-aminopropan-2-ol, maleic anhydride and sodium bisulfite

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

A key study for reproductive toxicity was performed in rats by means of an OECD 422 study with a read-across liquid formulation containing 41.5% active ingredient (CAS 68784 -08 -7) given by oral gavage at dose levels of 100, 300 and 1000 mg act. ingr./kg bw/day. No reproductive toxicity effects were observed up to 1000 mg/kg bw. At the dose of 1000 mg/kg bw, decrease body weight and other systemic effects were observed, therefore NOAEL for systemic toxicity was 300 mg/kg bw/day, whereas NOAEL for reproductive toxicity was 1000 mg/kg bw/day.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
For details please refer to Read Across Justification Document, Section 13.2

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
For details please refer to Read Across Justification Document, Section 13.2

3. ANALOGUE APPROACH JUSTIFICATION
For details please refer to Read Across Justification Document, Section 13.2

4. DATA MATRIX
For details please refer to Read Across Justification Document, Section 13.2

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

read-across source

Limit test:

no

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Slightly increased salivationwas noted in one male rat dosed at 1000 mg/kg bw/day.
No signs of clinical toxicity were noted in all female treatment groups.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Description (incidence):

No premature deaths were noted in the male and female rats treated with 100, 300 or 1000 mg/kg bw/ day.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Slight reduction of body weight and body weight gain was noted in male and female rats dosed at 1000 mg/kg bw/day.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

oral gavage study
Slight reduction in food consumption was noted in male and female rats dosed at 1000 mg/kg bw/day

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Increased number of eosinophils in intermediate dose females on test day 15 lacking dose dependence (p≤0.05)

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

In the high dose group (1000 mg/kg bw/day) a statistically significant (p≤0.01) increase (males: +66%; females: +46%)) was noted for the plasma activity of ALAT.
In the male high dose group (1000 mg/kg bw/day) a decrease in albumin on test day 15 (p≤0.01) was noted but not considered test-item related (slight alteration in comparison to control animals without biological relevance).
In the female intermediate dose group (300 mg/kg bw/day) an increase in bile acids on test day 15 (p≤0.05) was noted but not considered test-item related (lacking dose dependence).

Urinalysis findings:

not examined

Behaviour (functional findings):

effects observed, non-treatment-related

Description (incidence and severity):

No test item-related influence was noted for the fore- and hindlimb grip strength in any male/female treatment group (100, 300 or 1000 mg/kg bw/day).
No test item-related influence was noted on the spontaneous motility of the rats in any of the treatment groups.
In the male low dose group (100 mg/kg bw/day) a decrease in hindlimb grip strength on test day 36 (p≤0.05) was noted but not considered test-item related (lacking dose dependence).

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

No test item-related influence in observational screening was noted for any treatment group (100, 300 or 1000 mg/kg bw/day).
In the high dose males a statistically significant decreased mean body temperature was noticed in comparison to the control group. This was considered to be not test item related.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

No test item-related microscopic changes were seen in the reproductive organs for both males and females.
A test item-related squamous cell hyperplasia was noted in the forestomachs from 5/5 male and 5/5 female animals of the high dose group (1000 mg/kg bw/day). These test item-related stomach changes were localized in the zone adjoining the glandular stomach mucosa (forestomach or
non-glandular mucosa) and attained statistical significance for both sexes (p≤0.01). Occasionally the squamous cell hyperplasia with subsequent hyperkeratinization was associated with acute inflammation of the submucosa in the non-glandular stomach (for 2/5 males and 1/5 females).
Examination of the stomachs from the animals (5 per group) of the low- and intermediate dose groups (100 and 300 mg/kg bw/day) did not reveal any test item-related changes. As humans lack a forestomach, the relevance of these changes for humans is questionable.

Histopathological findings: neoplastic:

not examined

Other effects:

no effects observed

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

effects observed, non-treatment-related

Reproductive performance:

effects observed, non-treatment-related

Description (incidence and severity):

No statistically significant differences were noted in the length of the pre-coital time between the control group and the treatment groups.
There were no statistically significant differences for the female fertility rates between the control and the treatment groups.
No test item-related influence was noted on the gestation length of the females in any of the treatment groups compared to the control group.
No statistically significant differences were noted in the number of corpora lutea between the control and the treatment groups of P0 dams.
No statistically significant differences were noted in the number of implantation sites between the control and the treatment groups of P0 dams.
No statistically significant differences were noted in the total number of born pups (alive and dead) or in the number of pups born alive between the control group and the treatment groups of P0 dams.
No test item-related differences were noted between the control group and the treatment groups of P0 dams with respect to the reproduction indices which were calculated from the above mentioned parameters (Fertility index, Gestation index, Birth index, Live birth index, Pre-implantaion losss and Post-implantation loss).
Parameters with statistically significant differences in comparison to the control group which were considered as to be not test item related:
-Increased Birth Index in low and intermediate dose females
-Increased Pre-implantation loss in low dose females
-Decreased Post-implantation loss in low and intermediate dose females

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No premature deaths were noted in the male and female rats treated with 100, 300 and 1000 mg/kg bw/day.
No signs of clinical toxicity were noted for the male and female rats of the low and intermediate dose groups (100 and 300 mg/kg bw/day).
A slightly increased salivation was noted in one male rat, no further signs of clinical toxicity were noted for the male and female rats of the high dose group (1000 mg/kg bw/day). No test item related influence was noted for the male and female rats of all treatment groups (100, 300 and 1000 mg/kg bw/day) during the observational and functional (grip strength and spontaneous motility) neurological
screenings.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
A slight reduction in body weight was noted for the male and female rats of the high dose group (1000 mg/kg bw/day). For the male rats the reduction in body weight was noted from test day 8 (4.4%) until test day 36 (5.5%) , statistically significant (p≤0.05) in comparison to the control group on test day 8. Body weight gain was accordingly reduced in the high dose group during the whole study, mostly pronounced and statistically significant at the end of the first test week on test day 8 (p≤0.01), with a reduction in body weight gain by 35.4% in comparison to the control group. During the further course of the study the differences in body weight gain between the high dose group and the control group declined, leading to a body weight gain at the end of the study of 42.1%, the control group revealed a body weight gain of 50.5%.
For the high dose female rats a slight, not statistically significant, reduction in body weight was noted of 3.7% and 3.4% was noted on test days 8 and 15 (pre-mating). Body weight gain was 6.6% on test day 8 and 10.6% on test day 15 in the high dose group in comparison to 10.7% (test day 8) and 14.6% (test day 15) in the control group. Body weight was decreased during the whole gestation period between 7.6% on gestation day 0 and 6.5% on gestation day 20, statistically significant (p≤0.05) on gestation days 7 and 14 with 9.7% and 7.9% decrease, respectively. A small reduction in body weight gain was noted in the high dose group on gestation day 0 (15% in comparison to 18%. On gestation day 20 body weight gain was nearly identical in the high dose group (59.7%) and the control group (59.4%). During lactation a slight but not significant reduction in body weight was noted in the intermediate dose female rats by 4.4 % on lactation day 0 and 5.6% on lactation day 4. In the high dose group the body weight was statistically significantly (p≤0.05) reduced on lactation days 1 and 4 by 9.1% and 9.5%. No noticeable differences were noted in body weight gain during the lactation period with 5.6% in the high dose group and 6.1% in the control group.

A slightly statistically significant (p ≤ 0.01) increase in relative food consumption by 10.3% was noted in the high dose males during the 2nd test week. This was caused by the reduced body weight of the rats of the high dose group.
A slightly statistically significant (p≤0.05) decrease in relative food consumption by 7.4% was noted in the high dose females during the first test week.

HAEMATOLOGICAL FINDINGS
No test item-related influences were noted between the control group and the treatment groups for the haematological parameters, i.e. the haemoglobin content, the number of erythrocytes, leucocytes, reticulocytes and platelets, the haematocrit value, the thromboplastin time (TPT, aPTT), the mean corpuscular volume (MCV), the mean corpuscular haemoglobin (MCH) and the mean corpuscular haemoglobin concentration (MCHC). No test item-related changes were noted in the relative and absolute differential blood counts.
Statistically significant increased number of eosinophils (p≤0.05) was noted in the intermediate dose females on test day 15. This was considered to be not test item related ( lacking dose dependence).

CLINICAL BIOCHEMISTRY
In the high dose male and female group (1000 mg/kg bw/day) a statistically significant (p≤0.01) increase was noted for the plasma activity of ALAT.
In males no test item related influence was noted for the plasma levels of globulin, the albumin/globulin ratio, bilirubin (total), cholesterol (total), creatinine, glucose, urea in blood, sodium, potassium,calcium, chloride, the activity of the alkaline phosphatise (aP), the activity of ASAT and the serum levels of the bile acids. Decreased albumin in the high dose males (p≤0.01) was considered to be not test item related (slight alteration in comparison to control animals without biological relevance and 4/5 individual values within range of LPT Background Data))
In females no test item related influence was noted for the plasma levels of albumin, of globulin, the albumin/globulin ratio, bilirubin (total), cholesterol (total), creatinine, glucose, protein (total), urera in blood, sodium, potassium, calcium, chloride, the activity of the alkaline phosphatase (aP) and the
serum levels of the bile acids. Increased bile acids in the intermediate dose females (p≤0.05) was considered to be not test item related (lacking dose dependence).

BEHAVIOUR (FUNCTIONAL FINDINGS)
The functional neurological screenings were performed on test day 36 for the male rats and between test days 40-50 for the female rats, 2 hours after dosing.
No test item-related influence was noted for the fore- and hindlimb grip strength in any male/female treatment group (100, 300 or 1000 mg/kg bw/day).
Hinglimb grip strength in low dose males was statistically significant decreased on test day 36 (p≤0.05) but lacking dose dependence and was considered to be not test item related.
No test item-related influence was noted on the spontaneous motility of the rats in any of the treatment groups. No test item-related influence on relative and absolute organ weights was noted for the rats treated with 100, 300 or 1000 mg/kg bw/day in comparison to the control group.
Statistically significant differences in the relative organ weights compared to the control, which are not considered to be test item-related were:
- increased relative weight of left gonads in low and high dose males (p≤0.05)
- increased relative weight of right kidney in high dose females (p≤0.05)
- increased relative weight of right adrenal in high dose males (p≤0.05)
Statistically significant differences in the absolute organ weights compared to the control, which are not considered to be test item-related were:
-decreased absolute brain weight in high dose males (p≤0.05)
-decreased absolute heart weight by 16.6% in high dose females (p≤0.05)

GROSS PATHOLOGY (PARENTAL ANIMALS)
Macroscopic inspection at autopsy for the males was performed on test day 37.
No test-item related findings were noted in the low and intermediate dose group (100 and 300 mg/kg bw/day).
However, one animal (no. 3) with a reduction in the size of the testes was noted in the control group.
At the low dose group (100 mg/kg bw/day) animal no. 21 showed a partly reddened thymus and animal no. 29 a thickened right prostate.
In the intermediate dose group (300 mg/kg bw/day) one animal (no. 49) with macroscopic changes in the stomach (yellowish contents, detachment of mucosa) was found.
All changes are considered to be not test item-related but spontaneous due to the low number of occurrence.
In the high dose group (1000 mg/kg bw./day) 2 animals with macroscopic changes in the stomach were noted: Animal no. 68 showed a whitish thickening in the cardia part of the stomach and in the stomach of animal no. 69 a yellowish content was noted.
These findings were considered to be test item-related, because they were associated with microscopic changes in the forestomach from the animals of the high dose group.
Macroscopic inspection at autopsy for the females was performed between test days 43 and 54.
No test-item related macroscopic changes were noted in any treatment group (100, 300 and 1000 mg/kg bw/day) in females.
In 2 of the 3 non-pregnant rats (nos 16 and 54) from the control and the intermediate dose group (300mg/kg bw/day) a thickened uterus was noted.
These findings were considered to be not test item-related but spontaneous due to the low number ofoccurrence.

NEUROPATHOLOGY (OBSERVATIONAL FINDINGS)
The observational screenings were performed on test day 36 for the male rats and between test days 40-50 for the female rats, 2 hours after dosing.
No test item-related influence was noted for any treatment group (100, 300 or 1000 mg/kg bw/day).
A decrease in mean body temperature (p≤0.05) in the high dose males on test day 36 was considered to be not test item related. The slight alteration in comparison to control animals was without biological relevance (only 0.4°C difference to controls)

HISTOPATHOLOGY (PARENTAL ANIMALS)
Histopathological examination performed on one testicle and one epididymis with special emphasis on the qualitative stages of spermatogenesis (proliferative, meiotic and spermiogenic phases) and histopathology of the interstitial testicular structure of all adult male animals of the highest dose group and the control group following H & E and PAS staining, did not reveal any test item-related effects.
No test item-related microscopic changes were seen in the reproductive organs for both males and females.
A pulmonary congestion was noted in the lungs from 4/5 male (control 0/5) and 5/5 female rats (control 4/5) of the high dose group (1000 mg/kg bw/day). As pulmonary congestion is occasionally seen in rats as a background finding, this change was not considered to be test item-related.
A test item-related squamous cell hyperplasia was noted in the fore-stomachs from 5/5 male and 5/5 female animals of the high dose group (1000 mg/kg bw/day). These test item-related stomach changes were localized in the zone adjoining the glandular stomach mucosa (forestomach or
13 the squamous cell hyperplasia with subsequent hyperkeratinization was associated with acute inflammation of the submucosa in the non-glandular stomach (for 2/5 males and 1/5 females).
Examination of the stomachs from the animals (5 per group) of the low- and intermediate dose groups (100 and 300 mg/kg bw/day) did not reveal any test item-related changes. As humans lack a fore-stomach, the relevance of these changes for humans is questionable.
All other microscopic changes observed were either coincidental, or lie within the normal background alterations which may be seen in untreated rats of this age and strain.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
All animals were successfully mated as determinated by positive sperm detection. No statistically significant differences were noted in the length of the pre-coital time between the control group and the treatment groups.
There were no statistically significant differences for the female fertility rates between the control and the treatment groups.
No test item-related influence was noted on the gestation length of the females in any of the treatment groups compared to the control group.
No statistically significant differences were noted in the number of corpora lutea between the control and the treatment groups of P0 dams.
No statistically significant differences were noted in the number of implantation sites between the control and the treatment groups of P0 dams.
No statistically significant differences were noted in the total number of born pups (alive and dead) or in the number of pups born alive between the control group and the treatment groups of P0 dams.
No test item-related differences were noted between the control group and the treatment groups of P0 dams with respect to the reproduction indices which were calculated from the above mentioned parameters (Fertility index, Gestation index, Birth index, Live birth index, Pre-implantation loss and Post-implantation loss).
Parameters with statistically significant differences in comparison to the control group which were considered as to be not test item related:
-Increased Birth Index in low and intermediate dose females (p≤0.05)
-Increased Pre-implantation loss in low dose females (p≤0.05)
-Decreased Post-implantation loss in low and intermediate dose females (p≤0.05).

Key result

Dose descriptor:

NOAEL

Remarks:

reproductive toxicity

Effect level:

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

Based on:

act. ingr.

Sex:

male/female

Remarks on result:

other: highest dose tested

Key result

Dose descriptor:

NOAEL

Remarks:

general toxicity

Effect level:

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

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

clinical signs

body weight and weight gain

food consumption and compound intake

clinical biochemistry

gross pathology

histopathology: non-neoplastic

Key result

Critical effects observed:

no

Details on results:

No P1 generation: screening study

Clinical signs:

not examined

Dermal irritation (if dermal study):

not examined

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

No test item-related differences were noted between the survival index of the control group and the treatment groups.
No live born pup from the control group and the low and intermediate dose groups died between lactation day 1 and 4, leading to a survival index of 100% for these groups.
In the high dose group (1000 mg/kg bw/day) 9 of 10 dams revealed a survival index of 100%, whereas all 3 live born pups from dam no. 77 were found dead without milk on lactation day 2, leading to a survival index of 0.0% for dam no. 77 and a survival index of 97.6% for the pups of the high dose group. This small decrease in the survival index of the pups of the high dose group was regarded as to be spontaneous.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No test item-related influence was noted on the mean litter weight of the pups in all treatment groups.
No test item-related influence was noted on the total litter weight of the pups in all treatment groups.

Food consumption and compound intake (if feeding study):

not examined

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

Sexual maturation:

not examined

Description (incidence and severity):

Pups were sacrificed on day 4 of lactation.

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Description (incidence and severity):

The external examinations of the pups at sacrifice on day 4 of lactation revealed no test item-related external visible changes or gross abnormalities after treatment of the parental animals with 100, 300 or 1000 mg/kg bw/day. No milk was noted in the stomachs of the 3 dead pups from dam no. 77. This observation was spontaneous and considered as not test-item related.

Histopathological findings:

not examined

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

not examined

VIABILITY (OFFSPRING)
No test item-related influence was noted on the survival rate of the pups.
No live born pup from the control group and the low and intermediate dose groups died between lactation day 1 and 4, leading to a survival index of 100% for these groups.
In the high dose group (1000 mg/kg bw/day) 9 of 10 dams revealed a survival index of 100%, whereas all 3 live born pups from dam no. 77 were found dead without milk on lactation day 2, leading to a survival index of 0.0% for dam no. 77 and a survival index of 97.6% for the pups of the high dose group. This small decrease in the survival index of the pups of the high dose group was regarded as to be spontaneous.

BODY WEIGHT (OFFSPRING)
No test item-related influence was noted on the mean litter weight of the pups in all treatment groups.
No test item-related influence was noted on the total litter weight of the pups in all treatment groups.

GROSS PATHOLOGY (OFFSPRING)
The external examinations of the pups at sacrifice on day 4 of lactation revealed no test item-related external visible changes or gross abnormalities after treatment of the parental animals with 100, 300 or 1000 mg/kg bw/day. No milk was noted in the stomachs of the 3 dead pups from dam no. 77. This observation was spontaneous and considered as not test-item related.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

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

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: development of F1 offspring

Remarks on result:

other: highest dose tested

Key result

Critical effects observed:

no

No F2 generation: screening study

Key result

Reproductive effects observed:

no

Table 1. Fertility and Reproductive parameters Parental generation

Parameter	Group 1 Control	Group 2 100 mg/kg	Group 3 300 mg/kg	Group 4 1000 mg/kg
No. of females evaluated for pre-coital time	10	10	10	10
Mean precoital interval (days)	4.5	3.4	3.9	2.6
No. of females evaluated for fertility	10	10	10	10
Number of pregnant dams	8	9	9	10
Fertility index (%)	80	90	90	100
No. of females evaluated for gestation length	8	9	9	10
Gestation length (days)	22.0	22.2	22.2	22.1
Number of dams with live pups	8	9	9	10
Gestation Index (%)	100	100	100	100
Corpora lutea(total)	110	137	129	142
Corpora lutea(mean)	13.8	15.2	14.3	14.2
Implantation sites (total)	110	131	128	139
Implantation sites (mean)	13.8	14.6	14.2	13.9
Number of pups at birth (total)	96	125	123	127
Number of pups at birth (mean)	12.0	13.9	13.7	12.7
Birth Index (mean %)	90.4	95.5	95.9	90.1
Birth Index (total^# %)	87	95^1	96^1	90.1
Number of stillbirths	0	0	1	0
No. of dams with stillborn pups	0	0	1	0
Number of live born pups (total)	96	125	123	126
Number of live born pups (mean)	12.0	13.9	13.7	12.6
Live birth index (mean %)	100.0	100.0	100.0	97.5
Live birth index (total^#1 %)	100	100	100	99
Pre-implantation loss (mean %)	0.0	4.4	0.7	1.9
Pre-implantation loss (total^#2 %)	0.0	4.4^2	0.8	2.1
Post-implantation loss (mean %)	9.6	4.5	4.1	11.2
Post-implantation loss (total^#3 %)	12.7	4.6^2	3.9^2	9.4
Number of runts	0	0	0	0
Number of malformed pups	0	0	0	0

^# based on the total No. of implantation sites and total No. of pups at birth (alive and dead)

^#1 based on the total No. live born pups and total No. of pups at birth (alive and dead)

^#2 based on the total No. corpora lutea and total No. of implantation sites

^#3 based on the total No. implantation sites and toal number of live born pups

¹ p≤0.05 Chi²-test

² p<0.05 Chi²-test

Conclusions:

CAS 68784-08-7: NOAEL (no-observed-adverse-effect level) for reproductive toxicity: >= 1000 mg/kg bw/day, p.o.

Executive summary:

The aim of the study was to obtain information on possible effects of the read-across test item on general toxicity, reproduction and/or development according to OECD guideline 422. The read-across test item was administered orally by gavage to rats at dose levels of 100, 300 and 1000 mg active ingredient/kg bw/day. The application started two weeks before mating on test day one and ended on the day or one day before sacrifice. Day of sacrifice was on test day 37 for the male rats and between lactation day 4 and 7 for the female rats.

Effects on the parental generation (general toxicity)
No test item-related premature death was noted in any treatment group (100, 300 and 1000 mg test item/kg bw/day).

No signs of clinical toxicity were noted for the male and female rats of the low and intermediate dose groups (100 and 300 mg test item/kg bw/day). A slightly increased salivation was noted in one male rat, no further signs of clinical toxicity were noted for the male and female rats of the high dose group (1000 mg test item/kg bw/day). A slight reduction in body weight was noted for the male and female rats of the high dose group (1000 mg test item/kg bw/day). For the male rats the reduction in body weight was noted from test day 8 (4.4%) until test day 36 (5.5%) and for the female rats from gestation day 0 (7.6%) until lactation day 4 (9.5%). The body weight at autopsy was reduced accordingly.

Effects on reproduction parameters and organs

No test item-related influence was noted on the reproduction parameters in any treatment group (100, 300 and1000 mg/kg bw/day).

Microscopic examination revealed no changes in the reproductive organs from the male and female rats of the high dose group (1000 mg/kg bw/day).

Effects on the F₀-generation

NOAEL (no-observed-adverse-effect level): 300 mg/kg bw/day, p.o.

Effects on reproductive toxicity

NOAEL (no-observed-adverse-effect level): >=1000 mg/kg bw/day, p.o.

 

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

High quality (Klimisch 1)

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Reproductive screening

A key study with read-across substance Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1-oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts (CAS 68784-08-7) was performed by means of an oral combined repeated dose and reproduction/development screening study according to OECD guideline 422 (Hansen, 2013d). The test item (liquid formulation) containing 41.5% active ingredient was administered orally by gavage to rats at dose levels of 100, 300 and 1000 mg act. ingr./kg bw/day for for at least 28 days in male rats and at least 39 days in females. No test item-related premature death was noted in any treatment group. No signs of clinical toxicity were noted for the male and female rats of the low and intermediate dose groups (100 and 300 mg/kg bw/day), whereas slightly increased salivation was noted in one male rat as the only finding at 1000 mg/kg bw/day. A slight reduction in body weight was noted for the male and female rats dosed at 1000 mg/kg bw/day. Other parameters such as neurological observations, haematology and serum chemistry, as well as gross and histopathology are discussed in the repeated dose toxicity section 7.5. No test item-related influence was noted on the reproductive parameters in any treatment group (100, 300 and1000 mg/kg bw/day). Microscopic examination revealed no changes in the reproductive organs from the male and female rats of the high dose group (1000 mg/kg bw/day). NOAEL for paternal/maternal toxicity was 300 mg/kg bw/day, whereas NOAEL for reproductive toxicity was >= 1000 mg/kg bw.

  

Conclusion

An oral gavage reproductive screening study with read-across substance Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1-oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts (CAS 68784-08-7) showed a NOAEL of 300 mg/kg bw for paternal/maternal systemic toxicity, whereas 1000 mg/kg bw was NOAEL for reproductive and developmental toxicity as well as for neurotoxicity. Based on the absence of reproductive findings in the screening study, no further testing is needed.

Effects on developmental toxicity

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
For details please refer to Read Across Justification Document, Section 13.2

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
For details please refer to Read Across Justification Document, Section 13.2

3. ANALOGUE APPROACH JUSTIFICATION
For details please refer to Read Across Justification Document, Section 13.2

4. DATA MATRIX
For details please refer to Read Across Justification Document, Section 13.2

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

read-across source

Clinical signs:

no effects observed

Description (incidence and severity):

No signs of clinical toxicity were noted in the investigated female treatment groups (100, 300 or 1000 mg/kg bw/day) during the whole study.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

A slight reduction in body weight was noted for the female rats of the high dose group (1000 mg/kg bw/day). The reduction in body weight was noted from gestation day 0 (7.6%) until lactation day 4 (9.5%). The body weight at autopsy was reduced accordingly.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

oral gavage study
A slightly statistically significant (p≤0.05) decrease in relative food consumption by 7.4% was noted in the high dose group (1000 mg/kg bw/day) during the first test week.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Increased number of eosinophils in intermediate dose females on test day 15 lacking dose de
pendence (p≤0.05).
No test item-related influence was noted.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

In the female high dose group (1000 mg/kg bw/day) a statistically significant (p≤0.01) increase (+46%)) was noted for the plasma activity of ALAT.
In the female intermediate dose group (300 mg/kg bw/day) an increase in bile acids on test day 15 (p≤0.05) was noted but not considered test-item related (lacking dose dependence).

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

No test item-related influence was noted for the fore- and hindlimb grip strength in any female treatment group (100, 300 or 1000 mg/kg bw/day).
No test item-related influence was noted on the spontaneous motility of the rats in any of the treatment groups.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

No test item-related influence on relative and absolute organ weights was noted for the rats treated with 100, 300 or 1000 mg/kg bw/day in comparison to the control group.
Statistically significant differences in the relative organ weights compared to the control, which are not considered to be test item-related were:
- increased relative weight of right kidney in high dose females (slight alteration in comparison to controls without biological relevance and not accompanied by an increase in absolute organ weight)
-decreased absolute heart weight by 16.6% in high dose females (p≤0.05).

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No test-item related macroscopic changes were noted in any treatment group (100, 300 and 1000 mg/kg bw/day) in females.
In 2 of the 3 non-pregnant rats (nos 16 and 54) from the control and the intermediate dose group (300 mg/kg bw/day) a thickened uterus was noted.
These findings were considered to be not test item-related but spontaneous due to the low number of occurrence.

Neuropathological findings:

no effects observed

Description (incidence and severity):

No test item-related influence in observational screening was noted for any treatment group (100, 300 or 1000 mg/kg bw/day).

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

No test item-related microscopic changes were seen in the reproductive organs for females.
A test item-related squamous cell hyperplasia was noted in the forestomachs from 5/5 female animals of the high dose group (1000 mg/kg bw/day). These test item-related stomach changes were localized in the zone adjoining the glandular stomach mucosa (forestomach or non-glandular mucosa) and attained statistical significance (p≤0.01). Occasionally the squamous cell hyperplasia with subsequent hyperkeratinization was associated with acute inflammation of the submucosa in the non-glandular stomach (1/5 females). Examination of the stomachs from the animals (5 per group) of the low- and intermediate dose groups (100 and 300 mg/kg bw/day) did not reveal any test
item-related changes. As humans lack a fore-stomach, the relevance of these changes for humans is questionable.

Histopathological findings: neoplastic:

not examined

Other effects:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
No premature deaths were noted in the female rats treated with 100, 300 and 1000 mg/kg bw/day.
No signs of clinical toxicity were noted for the female rats treated with 100, 300 and 1000 mg/kg bw/day.

BODY WEIGHT AND FOOD CONSUMPTION
A slight reduction in body weight was noted for the female rats of the high dose group (1000 mg/kg bw/day).
For the high dose female rats a slight, not statistically significant, reduction in body weight was noted of 3.7% and 3.4% was noted on test days 8 and 15 (pre-mating). Body weight gain was 6.6% on test day 8 and 10.6% on test day 15 in the high dose group in comparison to 10.7% (test day 8) and 14.6% (test day 15) in the control group. Body weight was decreased during the whole gestation period between 7.6% on gestation day 0 and 6.5% on gestation day 20, statistically significant (p≤0.05) on gestation days 7 and 14 with 9.7% and 7.9% decrease, respectively. A small reduction in body weight gain was noted in the high dose group on gestation day 0 (15% in comparison to 18%. On gestation day 20 body weight gain was nearly identical in the high dose group (59.7%) and the control group (59.4%). During lactation a slight but not significant reduction in body weight was noted in the intermediate dose female rats by 4.4 % on lactation day 0 and 5.6% on lactation day 4. In the high dose group the body weight was statistically significantly (p≤0.05) reduced on lactation days 1 and 4 by 9.1% and 9.5%. No noticeable differences were noted in body weight gain during the lactation period with 5.6% in the high dose group and 6.1% in the control group.
A slightly statistically significant (p≤0.05) decrease in relative food consumption by 7.4% was noted in the high dose females during the first test week.

HAEMATOLOGICAL FINDINGS
No test item-related influences were noted between the control group and the treatment groups for the haematological parameters, i.e. the haemoglobin content, the number of erythrocytes, leucocytes, reticulocytes and platelets, the haematocrit value, the thromboplastin time (TPT, aPTT), the mean corpuscular volume (MCV), the mean corpuscular haemoglobin (MCH) and the mean corpuscular haemoglobin concentration (MCHC). No test item-related changes were noted in the relative and absolute differential blood counts.
Statistically significant increased number of eosinophils (p≤0.05) was noted in the intermediate dose females on test day 15. This was considered to be not test item related ( lacking dose dependence).

CLINICAL BIOCHEMISTRY
In the high dose group (1000 mg/kg bw/day) a statistically significant (p≤0.01) increase was noted for the plasma activity of ALAT.
In females no test item related influence was noted for the plasma levels of albumin, of globulin, the albumin/globulin ratio, bilirubin (total), cholesterol (total), creatinine, glucose, protein (total), urera in blood, sodium, potassium, calcium, chloride, the activity of the alkaline phosphatase (aP) and the serum levels of the bile acids. Increased bile acids in the intermediate dose females (p≤0.05) was considered to be not test item related (lacking dose dependence).
BEHAVIOUR (FUNCTIONAL FINDINGS)
The functional neurological screenings were performed between test days 40-50 for the female rats, 2 hours after dosing.
No test item-related influence was noted for the fore- and hindlimb grip strength in any treatment group (100, 300 or 1000 mg/kg bw/day).
No test item-related influence was noted on the spontaneous motility of the rats in any of the treatment groups.

ORGAN WEIGHTS(MATERNAL)
No test item-related influence on relative and absolute organ weights was noted for the rats treated with 100, 300 or 1000 mg/kg bw/day in comparison to the control group.
Statistically significant differences in the relative organ weights compared to the control, which are not considered to be test item-related were:
- increased relative weight of right kidney in high dose females (p≤0.05)
Statistically significant differences in the absolute organ weights compared to the control, which are not considered to be test item-related were:
-decreased absolute heart weight by 16.6% in high dose females (p≤0.05)

GROSS PATHOLOGY (MATERNAL)
Macroscopic inspection at autopsy for the females was performed between test days 43 and 54.
No test-item related macroscopic changes were noted in any treatment group (100, 300 and 1000 mg/kg bw/day) in females.
In 2 of the 3 non-pregnant rats (nos 16 and 54) from the control and the intermediate dose group (300 mg/kg bw/day) a thickened uterus was noted.
These findings were considered to be not test item-related but spontaneous due to the low number of occurrence.

NEUROPATHOLOGY (OBSERVATIONAL FINDINGS)
The observational screenings were performed between test days 40-50 for the female rats, 2 hours after dosing.
No test item-related influence was noted for any treatment group (100, 300 or 1000 mg/kg bw/day).

HISTOPATHOLOGY (MATERNAL)
No test item-related microscopic changes were seen in the reproductive organs for females.
A pulmonary congestion was noted in the lungs from 4/5 male (control 0/5) and 5/5 female rats (control 4/5) of the high dose group (1000 mg/kg bw/day). As pulmonary congestion is occasionally seen in rats as a background finding, this change was not considered to be test item-related.
A test item-related squamous cell hyperplasia was noted in the fore-stomachs from 5/5 male and 5/5 female animals of the high dose group (1000 mg/kg bw/day). These test item-related stomach changes were localized in the zone adjoining the glandular stomach mucosa (forestomach or
non-glandular mucosa) and attained statistical significance for both sexes (p≤0.01). Occasionally the squamous cell hyperplasia with subsequent hyperkeratinization was associated with acute inflammation of the submucosa in the non-glandular stomach (for 2/5 males and 1/5 females).
Examination of the stomachs from the animals (5 per group) of the low- and intermediate dose groups (100 and 300 mg/kg bw/day) did not reveal any test item-related changes. As humans lack a fore-stomach, the relevance of these changes for humans is questionable.
All other microscopic changes observed were either coincidental, or lie within the normal background alterations which may be seen in untreated rats of this age and strain.

Number of abortions:

no effects observed

Description (incidence and severity):

No statistically significant differences were noted in the total number of born pups (alive and dead) or in the number of pups born alive between the control group and the treatment groups of P0 dams.No statistically significant differences were noted in the number of implantation sites between the control and the treatment groups of P0 dams.

Pre- and post-implantation loss:

effects observed, non-treatment-related

Description (incidence and severity):

Parameters with statistically significant differences in comparison to the control group which were considered as to be not test item related:
-Increased Pre-implantation loss in low dose females (p≤0.05)
-Decreased Post-implantation loss in low and intermediate dose females (p≤0.05)

Total litter losses by resorption:

no effects observed

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

Changes in pregnancy duration:

no effects observed

Description (incidence and severity):

No test item-related influence was noted on the gestation length of the females in any of the treatment groups compared to the control group.
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.DescriptionIncidenceAndSeverityEffectsOnPregnancyDuration): No test item-related influence was noted on the gestation length of the females in any of the treatment groups (100, 300 and 1000 mg/kg bw/day) compared to the control group.

Changes in number of pregnant:

no effects observed

Description (incidence and severity):

All animals were successfully mated as determinated by positive sperm detection. No statistically significant differences were noted in the length of the pre-coital time between the control group and the treatment groups.
No test item-related differences were noted between the control group and the treatment groups (100, 300 and 1000 mg act. ingr./kg bw/day) with respect to the fertility index.

Details on maternal toxic effects:

Maternal toxic effects:yes. Remark: Slight reduction in body weight in male and female rats dosed at 1000 mg/kg bw/day; other changes were noted as well for clinical biochemistry, macroscopic and microscopic examination

Details on maternal toxic effects:
A slight reduction in body weight was noted for female rats of the high dose group (1000 mg test item/kg bw/day). For the female rats the reduction in body weight was noted from gestation day 0 (7.6%) until lactation day 4 (9.5%). The body weight at autopsy was reduced accordingly.

The laboratory examinations revealed an increased ALAT activity for female rats of the high dose group (1000 mg test item/kg bw/day).

Microscopic examination revealed the occurrence of squamous cell hyperplasia in the non-glandular mucosa of the forestomach from female rats of the high dose group (1000 mg test item/kg bw/day).

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

act. ingr.

Basis for effect level:

changes in number of pregnant

changes in pregnancy duration

early or late resorptions

effects on pregnancy duration

number of abortions

pre and post implantation loss

total litter losses by resorption

Remarks on result:

other: developmental toxicity

Remarks:

highest dose tested

Key result

Abnormalities:

no effects observed

Description (incidence and severity):

No test item related influence was noted on the mean and the total litter weight of the pups on lactation day 1 and 4.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): Pups were sacrificed on day 4 of lactation.

Reduction in number of live offspring:

no effects observed

Description (incidence and severity):

No statistically significant differences were noted in the total number of born pups (alive and dead) or in the number of pups born alive between the control group and the treatment groups of P0 dams.

Changes in sex ratio:

not examined

Changes in litter size and weights:

no effects observed

Description (incidence and severity):

No test item related influence was noted on the mean and the total litter weight of the pups on lactation day 1 and 4.

Changes in postnatal survival:

no effects observed

Description (incidence and severity):

No test item-related differences were noted between the survival index of the control group and the treatment groups (100, 300 and 1000 mg act. ingr./kg bw/day).

External malformations:

no effects observed

Description (incidence and severity):

No visible gross abnormalities were noted between the control and the treatment groups.

Skeletal malformations:

not examined

Visceral malformations:

not examined

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects. Remark: No test item related influence was noted on the survival rate and the mean and total body weights of the pups.

Details on embryotoxic / teratogenic effects:
No test item related influence was noted on the survival rate and the mean and total body weights of the pups.
Effects on the development of the F1 offsprings (pups) NOAEL (no-observed-adverse-effect level): above 1000 mg/kg bw/day, p.o.

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

act. ingr.

Sex:

male/female

Remarks on result:

other: highest dose tested

Key result

Abnormalities:

no effects observed

Key result

Developmental effects observed:

no

Table 1.Developmental parameters

Parameter	Group 1 Control	Group 2 100 mg/kg	Group 3 300 mg/kg	Group 4 1000 mg/kg
Number of pregnant dams	8/10	9/10	9/10	10/10
Number of pups at birth (total)	96	125	123	127
Number of pups at birth (mean)	12.0	13.9	13.7	12.7
No. of live pups at birth (total)	96	125	123	126
No. of live pups at birth (mean)	12.0	13.9	13.7	12.6
No. of live pups on day 4 (total)	96	125	123	123
No. of live pups on day 4 (mean)	12.0	13.9	13.7	13.7
Viability index day 1-4 (mean %)	100.0	100.0	100.0	90.0
Viability index day 1-4 (total^# %)	100.0	100.0	100.0	97.6
Mean body weight day 0/1 male and female pups (g)	6.23	6.34	6.29	6.11
Mean body weight day 4 male and female pups (g)	8.51	9.07	9.11	8.73
Mean total litter weight day 0/1, male and female pups (g)	75.00	87.38	85.91	77.09
Mean total litter weight day 4, male and female pups (g)	102.54	124.08	123.28	118.94
External examinations of pups (No. of litters/pregnant dams) - no externally visible  abnormalities      - stomach without milk	8/8 0/8	9/9 0/9	9/9 0/9	10/10 1/10 (3/3)*

#: total number of live pups on the respective lactation day

* 3 live born pups from dam no. 77 were found dead without milk on lactation day 2.

Conclusions:

CAS 68784-08-7: NOAEL (no-observed-adverse-effect level) of the F1 offspring: above 1000 mg/kg bw/day, p.o.

Executive summary:

The aim of the study was to obtain information on possible effects of the read-across test item on general toxicity, reproduction and/or development according to OECD guideline 422. The read-across test item was administered orally by gavage to rats at dose levels of 100, 300 and 1000 mg active ingredient/kg bw/day. The application started two weeks before mating on test day one and ended on the day or one day before sacrifice. Day of sacrifice was on test day 37 for the male rats and between lactation day 4 and 7 for the female rats.

Effects on the parental generation (general toxicity)
No test item-related premature death was noted in any treatment group (100, 300 and 1000 mg/kg bw/day).

No signs of clinical toxicity were noted for the male and female rats of the low and intermediate dose groups (100 and 300 mg/kg bw/day).

A slightly increased salivation was noted in one male rat, no further signs of clinical toxicity were noted for the male and female rats of the high dose group (1000 mg/kg bw/day).

No test item related influence was noted for the male and female rats of all treatment groups (100, 300 and 1000 mg/kg bw/day) during the observational and functional (grip strength and spontaneous motility) neurological screenings.

A slight reduction in body weight was noted for the male and female rats of the high dose group (1000 mg/kg bw/day). For the male rats the reduction in body weight was noted from test day 8 (4.4%) until test day 36 (5.5%) and for the female rats from gestation day 0 (7.6%) until lactation day 4 (9.5%). The body weight at autopsy was reduced accordingly.

The laboratory examinations revealed an increased ALAT activity for the male and female rats of the high dose group (1000 mg/kg bw/day) and a decrease in the albumin concentration for the male rats of the high dose group.

No test item-related changes were noted during the macroscopic inspection at autopsy with the exception of changes in the stomach from 2 male animals from the high dose group (1000 mg/kg bw/day).

Microscopic examination revealed the occurrence of squamous cell hyperplasia in the non-glandular mucosa of the forestomach from the male and female rats of the high dose group (1000 mg/kg bw/day). Further microscopic findings occurred in form of pulmonary congestion in the male rats from the high dose group.

Effects on reproduction parameters and organs

No test item-related influence was noted on the reproduction parameters in any treatment group (100, 300 and 1000 mg/kg bw/day).

Microscopic examination revealed no changes in the reproductive organs from the male and female rats of the high dose group (1000 mg/kg bw/day).

Effects on the development of the F₁offsprings (pups)

No test item related influence was noted on the survival rate and the mean and total body weights of the pups.

External examination of the pups revealed no visible changes related to the test item.

The following no-observed-effectlevelswere established:

Effects on the F₀-generation

NOAEL (no-observed-adverse-effect level): 300 mg/kg bw/day, p.o.

Effects on reproductive toxicity

NOAEL (no-observed-adverse-effect level): above 1000 mg/kg bw/day, p.o.

Reproductive indices:

Effects on the development of the F₁offsprings (pups)

NOAEL (no-observed-adverse-effect level): above 1000 mg/kg bw/day, p.o.

 

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

High quality (Klimisch 1)

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

Developmental screening

Supporting data for absence of developmental toxicity were available from an oral combined repeated dose and reproduction/development screening study according to OECD guideline 422 (Hansen, 2013d) with read-across test item Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1-oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts (CAS 68784-08-7). The test item was administered orally by gavage to rats with a formulation containing 41.5% active ingredient at dose levels of 100, 300 and 1000 mg act. ingr./kg bw/day for for at least 28 days in male rats and at least 39 days in females. No test item-related premature death was noted in any treatment group. No signs of clinical toxicity were noted for the male and female rats of the low and intermediate dose groups (100 and 300 mg/kg bw/day), whereas slightly increased salivation was noted in one male rat as the only finding at 1000 mg/kg bw/day. A slight reduction in body weight was noted for the male and female rats dosed at 1000 mg/kg bw/day. Other parameters such as neurological observations, haematology and serum chemistry are discussed in the repeated dose toxicity section.

No test item-related influence was noted on the developmental toxicity parameters in any treatment group (100, 300 and 1000 mg/kg bw/day). Microscopic examination revealed no changes in the reproductive organs from the male and female rats of the high dose group (1000 mg/kg bw/day). No test item related influence was noted on the survival rate and the mean and total body weights of the pups. External examination of the pups revealed no visible changes related to the test item. NOAEL for paternal/maternal toxicity was 300 mg/kg bw/day, whereas NOAEL for developmental toxicity was >= 1000 mg/kg bw.

  

Conclusion

An oral gavage reproductive screening study with registered substance showed NOAEL of 300 mg/kg bw for paternal/maternal systemic toxicity, whereas 1000 mg/kg bw was NOAEL for reproductive and developmental toxicity.

Based on the absence of developmental findings in the screening study, no further testing is needed.  

Mode of Action Analysis / Human Relevance Framework

There is no evidence for species specific effects of the substance. Therefore, the results of the studies are regarded as relevant for humans.

Justification for classification or non-classification

Based on the results and according to the CLP Regulation (No. 1272/2008 of 16 December 2008), the test substance does not have to be classified and has no obligatory labelling requirement for reproductive and developmental toxicity.