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Key value for chemical safety assessment

Effects on fertility

Description of key information

Fertility

A repeated-dose toxicity combined with a reproductive/developmental toxicity screen, conducted according (OECD Test Guideline No 422), was undertaken on the registered substance, an ester of 1,2,4 -benzenetricarboxylic acid with C8 linear side chains. No adverse effects on oestrous cycle, copulation, fertility, delivery or lactation and no changes related to gestation index, gestation length, numbers of corpora lutea, implantation sites or implantation index were found. There were no changes in sex ratio, body weight, viability or morphology of pups. The No Observed Adverse Effect Level (NOAEL) for reproductive toxicity was 500 mg/kg/day for both parent animals and offspring, this being the highest dose level investigated.

Reproductive toxicity - rat: NO(A)EL 500 mg/kg/day

Gene expression associated with reproductive toxicity: No repression of genes in the testicular mal-development pathway

This substance is manufactured by the lead registrant in quantities >1000 tpa. As such, a two-generation reproductive toxicity study (OECD Test Guideline No 443) is required in accordance with REACH Regulation 1907/2006, Annex X, Section 8.7.3 of Column 1. It is proposed to conduct the OECD 443 with the structural analogue TM 08-10 and then read across to the results of this study for TM08, once ECHA provides the final compliance check for TM08-10.

TM8-10 is a structural analogue of TM 8, as it is 1,2,4 -benzenetricarboxylic acid with linear C8- and C10 -alcohols, produced by esterification of trimellitic anhydride with a mixture of linear C8-C10-alcohols (40 - 60 % C8 and 40 - 60 % C10) rather than C8 alcohol alone, as in TM8.

Link to relevant study records

Referenceopen allclose all

Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
- Lot/batch No.: C-120
Species:
rat
Strain:
other: Crj:CD:SD
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Japan
- Age at study initiation: (P) x 10 weeks;
- Weight at study initiation: (P) Males: 388-420 g; Females: 242-277 g
- Fasting period before study: No
- Housing: wire mesh cages, in pairs for mating
- Use of restrainers for preventing ingestion (if dermal): Not applicable
- Diet (e.g. ad libitum): yes, pelleted diet
- Water (e.g. ad libitum): yes, tap water
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-25
- Humidity (%): 50-65
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12:12

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Prepared weekly by dissolving required weight of the substance in corn oil and stored refrigerated in airtight containers in the dark until use.


VEHICLE
- Justification for use and choice of vehicle (if other than water): Substance is poorly soluble in water
- Concentration in vehicle: As required to achieve nominal dose, up to 25% w/v
- Amount of vehicle (if gavage): 2mL/kg
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: up to 14 days (until sperm detected in vagina).
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged (how): singly
- Any other deviations from standard protocol: None reported
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
No data except that concentration of preparations were analysed and the results were in the range 98-102% of nominal. Formulated substance stable for at least 8 days.
Duration of treatment / exposure:
Males: from 14 days before pairing for 42days
Females: from 14 days before pairing to day 4 of lactation
Frequency of treatment:
Daily during treatment period
Remarks:
Doses / Concentrations:
0 (control), 30, 125 and 500 mg/kg/day
Basis:
nominal conc.
No. of animals per sex per dose:
13 males & 13 females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on results of 14 day preliminary study at dose levels of 500 and 1000 mg/kg/day. Effects on body weight, increased liver weight and oedema of gastric mucosa noted in animals given 1000 mg/kg/day
- Rationale for animal assignment (if not random): Random, stratified body weight
Positive control:
No
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily, parents & foetuses

DETAILED CLINICAL OBSERVATIONS: No data


BODY WEIGHT: Yes
- Time schedule for examinations: Males: Weekly; Females: days 0, 7, 14, 20, gestation days 1, 7, 14 and parturition, lactation days 0 and 4.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes - Males: days 1-2, 7-8, 14-15, 29-30, 35-36 and 41-42; Females: days 1-2, 7-8, 14-15, days 7-8, 14-15 and 20-21 of gestation, 3-4 of lactation

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
Oestrous cyclicity (parental animals):
Examined pre-dose and during dosing period
Sperm parameters (parental animals):
Parameters examined in P males/group : yes
- testis weight and microscopic pathology, epididymis weight and microscopic pathology

Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities, :

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was/was not determined for pups born or found dead
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals following 42 days of treatment
- Maternal animals: All surviving animals day 4 of lactation

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations of all organs & including the cervical, thoracic, and abdominal viscera. Pups: all organs

HISTOPATHOLOGY / ORGAN WEIGHTS
The following tissues were weighed: Brain, heart, thymus, liver, kidneys, spleen, adrenals, testes, epididymides
The following tissues were prepared for microscopic examination: Brain, heart, thymus, liver, kidneys, spleen, adrenals, testes, epididymides, stomach, prostate, urinary bladder, lungs (and bronchi), ileum, trachea, mandibular lymph node, seminal vesicles and coagulating gland, duodenum, jejunum, caecum, colon, rectum, mesenteric lymph nodes, pituitary gland, thyroid gland, sciatic nerve, spinal cord, bone marrow (from femur), ovary, uterus, vagina
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed at 4 days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination). Pups: found dead & abnormalities

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.
Statistics:
Sexual cycle, copulation rate and conception rate were examined using Fisher's exact test
Clinical signs and histopathological findings were graded by Mann-Whitney U test with the sum of the positive findings. Significant difference tests were performed between the control group by one-sided test of direct probability of Fischer.
Other data, as a sample value or the average value of each litter, were obtained for each individual and tested for uniformity of the variance of each group by the method of Bartlett. If the distribution was uniform, ANOVA and significance was examined using the multiple comparison method of Dunnett. If the distribution was not uniform, a Kruskal-Wallis rank test was performed with significance between the groups examined by Dunnett test. The significance level was 5%.
Reproductive indices:
Copulation Index: No. of pairs with successful copulation/no. of pairs mated X 100
Fertility Index: No of pregnant females/no. of pairs with successful copulation X 100
Implantation index: No. of implantation sites/no. of corporea lutea X 100
Delivery index: No. of pups born/no. of implantation sites X 100
Gestation index: No. of females with live pups delivered/no. of pregnant females X 100
Nursing index: No. of females nursing live pups/no. of females with normal delivery X 100
Offspring viability indices:
Live birth index: No. of live pups at birth/no. of pups at birth X 100
Viability index: No. of live pups on d4/no.of live pups at birth
Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
hepatocellular hypertrophy of liver of males at 500 mg/kg/day
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS): No deaths occurred except for one female treated at 500 mg/kg/day which died on day 23 of gestation. Temporally increase in salivation after dosing was observed in animals treated at 500 mg/kg/day.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS): Decreased body weight gain from day 7 to 14 of gestation observed in females treated at 500 mg/kg/day. No adverse effects on body weights in males. No effects on food consumption in either sex.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS): No data

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS): No effects

ORGAN WEIGHTS (PARENTAL ANIMALS): No effects

GROSS PATHOLOGY (PARENTAL ANIMALS): No effects

HISTOPATHOLOGY (PARENTAL ANIMALS): Hypertrophy of hepatocytes in the centrilobular zone of the liver in males treated at 500 mg/kg/day. No adverse changes in other organs.
Dose descriptor:
LOEL
Effect level:
500 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
organ weights and organ / body weight ratios
histopathology: non-neoplastic
Critical effects observed:
no
Lowest effective dose / conc.:
500 mg/kg bw/day (nominal)
System:
hepatobiliary
Organ:
liver
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed
VIABILITY (OFFSPRING): No effects

CLINICAL SIGNS (OFFSPRING): No effects

BODY WEIGHT (OFFSPRING): No effects

GROSS PATHOLOGY (OFFSPRING): No effects

Dose descriptor:
NOEL
Generation:
F1
Effect level:
500 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Absence of effects on pup weight; sex ratio; survival index; viability index
Critical effects observed:
no
Reproductive effects observed:
not specified

Oestrus cycle

 

Dose level (mg/kg body weight/day

0

30

125

500

4 day cycle

12

13

13

13

Irregular cycle

1

0

0

0

Length of oestrus cycle (days

4.1 ± 0.2 (13)

4.0± 0.0 (13)

4.0 ± 0.0 (13)

4.0 ± 0.0 (13)

 

 

 

Reproductive performance

 

Dose level (mg/kg body weight/day

0

30

125

500

Number of mated pairs

13

13

13

13

Number of copulated pairs

13

13

13

13

Copulation index

100

100

100

100

Number of pregnant animals

12

13

13

13

Fertility index

92.3

100

100

100

Number of pairing days

2.3 ± 1.3 (13)

2.5 ± 1.1 (13)

1.8 ± 1.0 (13)

2.2 ± 1.1 (13)

Frequency of oestrus

1.0 ± 0.0 (13)

1.0 ± 0.0 (13)

1.0 ± 0.0 (13)

1.0 ± 0.0 (13)

 

 

 

Developmental toxicity parameters

 

Dose level (mg/kg body weight/day

0

30

125

500

Number of pregnant animals

12

13

13

13

Number of pregnant animals with live young

12

13

13

12 *

Gestation index

100

100

100

92.3

Gestation length (days)

22.6 ± 0.5

22.2 ± 0.4

22.5 ± 0.5

22.6 ± 0.5

Number of corpora lutea

17.3 ± 1.5

17.9 ± 2.5

17.0 ± 1.8

16.9 ± 2.3

Number of implantation sites

15.8 ± 1.5

16.3 ± 1.4

15.8 ± 1.3

15.2 ± 2.6

Implantation index

91.6 ± 7.8

91.6 ± 6.5

93.6 ± 6.1

90.3 ± 12.8

 

 

 

 

 

Lactation day 0:

 

 

 

 

Number of pups born

14.7 ± 2.3

15.2 ± 1.9

14.7 ± 1.4

13.3 ± 4.0

Delivery index

92.8 ± 7.4

93.3 ± 7.1

92.9 ± 6.5

88.1 ± 20.7

Number of pups alive

14.1 ± 1.8

15.1 ± 1.8

14.5 ± 1.4

13.2 ± 4.0

Birth index

89.5 ± 7.8

92.4 ± 7.3

91.9 ± 6.8

86.9 ± 20.1

Live birth index

96.6 ± 6.3

99.1 ± 2.3

99.0 ± 2.5

98.8 ± 2.8

Pup weight (g)   - Males

7.1 ± 0.8

6.8 ± 0.5

7.1 ± 0.8

7.2 ± 1.0

                          - Females

6.7 ± 0.8

6.4 ± 0.5

6.5 ± 0.7

6.9 ± 1.0

Sex ratio (% males)

45.3 ± 12.9

53.5 ± 12.3

46.6 ± 11.5

53.0 ± 18.2

 

 

 

 

 

Lactation day 4:

 

 

 

 

Number of live pups

13.8 ± 1.7

14.8 ± 1.6

14.1 ± 1.1

12.9 ± 3.9

Viability index

98.3 ± 4.1

98.6 ± 2.7

98.6 ± 3.6

98.4 ± 5.4

Pup weight (g)   - Males

10.6 ± 2.6

10.5 ± 1.5

11.1 ± 1.6

11.5 ± 2.3

                          - Females

10.2 ± 2.7

10.0 ± 1.3

10.4 ± 1.4

11.1 ± 2.3

Sex ratio (% males)

44.8 ± 12.4

53.2 ± 12.4

46.3 ± 11.4

53.5 ± 17.4

 

* One female found dead on gestation day 23

Conclusions:
A repeated-dose toxicity combined with a reproductive/developmental toxicity screen conducted according to OECD Test Guideline No 422 found no adverse effects on oestrous cycle, copulation, fertility, delivery or lactation and no changes related to gestation index, gestation length, numbers of corpora lutea, implantation sites or implantation index. There were no changes in sex ratio, body weight, viability or morphology of pups. The No Observed Adverse Effect Level (NOEL) for reproductive and developmental toxicity was considered to be 500 mg/kg/day for both parent animals and offspring, this being the highest dose level investigated.
Executive summary:

A repeated-dose toxicity combined with a reproductive/developmental toxicity screen conducted according to OECD Test Guideline No 422 found no adverse effects on oestrous cycle, copulation, fertility, delivery or lactation and no changes related to gestation index, gestation length, numbers of corpora lutea, implantation sites or implantation index. There were no changes in sex ratio, body weight, viability or morphology of pups. The No Observed Adverse Effect Level (NOEL) for reproductive and developmental toxicity was considered to be 500 mg/kg/day for both parent animals and offspring, this being the highest dose level investigated.

Endpoint:
extended one-generation reproductive toxicity - with F2 generation (Cohorts 1A, and 1B with extension)
Type of information:
experimental study planned (based on read-across)
Justification for type of information:
This information will be submitted later based on ECHA decision number CCH-D-2114440644-51-0
The complex mixture TM 8-10 is a structural analogue of TM 8, as it is 1,2,4 -benzenetricarboxylic acid with linear C8- and C10 -alcohols, produced by esterification of trimellitic anhydride with a mixture of linear C8-C10-alcohols (40 - 60 % C8 and 40 - 60 % C10) rather than C8 alcohol alone, as in TM8.
This is also subject to an ECHA compliance check and a testing proposal for an extended one generation reproduction toxicology test (OECD 443): Draft CCH-XXXXXXXXXX-XX-XX/F) sent to all registrant of JS-C810trimellitate, 19th August 2020.
The existing developmental toxicology information (OECD 414) for two species with TM08 and the rat with TM 08-10, indicates the NOAEL for developmental effects on both substances are at or above 1000 mg/kg/bw.
In order to prevent the excessive use of animals, it is proposed that only one EOGRT is conducted, for TM 08-10, and that this is used as read across for TM 08.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 443 (Extended One-Generation Reproductive Toxicity Study)
GLP compliance:
yes
Justification for study design:
Guideline study planned for structural analogue
Reproductive effects observed:
not specified
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
500 mg/kg bw/day
Study duration:
subacute
Species:
rat
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

In addition, by analogy with the phthalate esters, the principle concern for benzenetricarboxylic acids with linear carbon chains alcohols might be expected to be for potential effects for the developing male testes. This aspect has been studied on a structural analogue of the substance (TM 8 -10 - see below) using RNA transcriptional profiling in an assay that subscribes to the principle of the 3R’s and reported in 7.9.4: Specific investigations; other studies.

TM8-10 is a structural analogue of TM 8, as it is 1,2,4 -benzenetricarboxylic acid with linear C8- and C10 -alcohols, produced by esterification of trimellitic anhydride with a mixture of linear C8-C10-alcohols (40 - 60 % C8 and 40 - 60 % C10) rather than C8 alcohol alone, as in TM8.

Short description of key information:
Fertility - Rat: NO(A)EL - 500 mg/kg/day

No expected effect on male reproductive organs.

Effects on developmental toxicity

Description of key information

Developmental toxicity / teratogencity

The results of two high quality prenatal developmental toxicity studies (OECD Test Guideline No 414) are available for both the rat and the 2nd species, rabbit, conducted with the registered substance: an ester of 1,2,4 -benzenetricarboxylic acid with C8 linear side chains (TM 8).

Both studies provided a NOAEL for maternal toxicity of 1000 mg/kg/day, the highest concentration tested. For developmental toxicity the NOAEL was also 1000 mg/kg/day in both species.

In addition, a prenatal developmental toxicity studies (OECD Test Guideline No 414), as weight of evidence, is provided for the multi-constituent substance 1,2,4 -Benzenetricarboxylic acid, decyl octyl ester (TM 8-10), as a structural analogue of TM8. The results of this study the maternal toxicity was 300 mg/kg, although the embryo foetal developmental NOAEL was 1000 mg/kg/day, comparable to TM8.

As supporting information, an in-silico QSAR analysis utilising the Danish QSAR assessment models (Battery; CASE Ultra; Leadscope: SciQSAR) for the teratogenic potential is included. This also predicted that the teratogenic potential in humans of trioctyl benzene-1,2,4-tricarboxylate was negative.

This substance is manufactured by the lead registrant in quantities >1000 tpa. As such, a two-generation reproductive toxicity study (OECD Test Guideline No 443) is required in accordance with REACH Regulation 1907/2006, Annex X, Section 8.7.3 of Column 1. It is proposed to conduct the OECD 443 with the structural analogue TM 08-10 and then read across to the results of this study for TM08, once ECHA provides the final compliance check for TM08-10.

Developmental toxicity - rat: NO(A)EL 1000 mg/kg/day

Developmental toxicity study - rabbit: NO(A)EL 1000 mg/kg/day

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 2017-05-09 to 2017-06-11
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
Adopted 22nd January 2001
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Polynt Lot No. 3606217009
- Expiration date of the lot/batch: 09 Jan 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient conditions
- Stability under test conditions: Stable
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: None

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None
Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS srl, San Pietro al Natisone, Italy
- Age at study initiation: circa 13 weeks
- Weight at study initiation: 203 - 259 g
- Fasting period before study: None
- Housing: Group caged except during mating
- Diet (e.g. ad libitum): Commercial laboratory rodent dient, ad-libitum
- Water (e.g. ad libitum): Municipal drinking water, ad-libitum
- Acclimation period: circa 4 weeks


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24 deg C
- Humidity (%): 40 - 70 %
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2017-05-09 To: 2017-06-11
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:

- VEHICLE
- Justification for use and choice of vehicle (if other than water): Commonly used vehicle for non-water miscible materials
- Concentration in vehicle: 0, 25, 75 and 250 mg/mL
- Amount of vehicle (if gavage): 4 mL/kg bw
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The formulation procedure was checked in the range from 1 to 250 mg/mL by chemical analysis for concentration and homogeneity to confirmthat the method was suitable.
Samples of the formulations prepared during Weeks 1 and 3 of the dosing phase were analysed to check the homogeneity and concentration.
Details on mating procedure:
- Impregnation procedure: Cohoused
- M/F ratio per cage: 1/1
- Length of cohabitation: Until mating verified
- Verification of same strain and source of both sexes: Yes
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as Day 0 of pregnancy
- Any other deviations from standard protocol: No
Duration of treatment / exposure:
From Day to Day 19 post coitum (13 days)
Frequency of treatment:
Daily
Duration of test:
20 days post coitum
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Vehicle control
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
24 mated females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Based on data from screening studies
- Rationale for animal assignment: Random
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

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

FOOD CONSUMPTION: Yes
- Food consumption determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes - At intervals over Days 0-6, 6-9, 9-12, 12-15 and 15-20

WATER CONSUMPTION: No

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation Day 20
- Organs examined: Detailed examination with particular attention to ovaries and uterus
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
Statistics:
For continuous variables the significance of the differences amongst group means was assessed by Dunnett's test or a modified t test, depending on the homogeneity of data.
Statistical analysis of non-continuous variables was carried out by means of the Kruskal-Wallis test and intergroup differences between the control and treated groups assessed by a non-parametric version of the Williams test.
Indices:
Pre-implantation loss
Post-implantation loss
Total implantation loss
Sex ratio
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Significantly reduced body weight and body weight gain in animals dosed at 1000 mg/kg bw/day
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Reduced food consumption in animals dosed at 1000 mg/kg bw/day
Gross pathological findings:
no effects observed
Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed
Dose descriptor:
LOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
food consumption and compound intake
Dose descriptor:
NOEL
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: Lack of treatment related effects
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
other: Lack of treatment related effects
Abnormalities:
no effects observed
Developmental effects observed:
no

Body weight (g) of dams - Group mean data

 

 

 

Gestation day:

Treatment

 

0

6

9

12

15

20

 

(n)

23

23

23

23

23

23

Control

Mean

233.12

257.83

266.89

278.76

297.89

373.46

 

SD

10.52

10.80

11.61

10.89

14.68

25.64

 

(n)

23

23

23

23

23

23

100 mg/kg

Mean

235.71

258.33

268.14

279.27

295.16

372.80

 

SD

12.70

11.49

11.68

11.01

11.88

12.55

 

(n)

22

22

22

22

22

22

300 mg/kg

Mean

234.61

257.08

266.14

276.12

294.56

371.29

 

SD

11.62

11.97

110.8

12.01

12.87

19.92

 

(n)

23

23

23

23

23

23

1000 mg/kg

Mean

231.91

255.48

262.70

272.62

288.40*

355.69*($)

 

SD

11.81

11.68

11.07

11.73

13.63

23.18

  

 

Food consumption (g/animal/day) of dams - Group mean data

 

 

 

Gestation day:

Treatment

 

6

9

12

15

20

 

(n)

9

9

9

9

9

Control

Mean

20.21

18.90

20.13

20.79

22.99

 

SD

1.72

1.10

2.12

2.59

1.47

 

(n)

8

8

8

8

8

100 mg/kg

Mean

19.82

19.22

18.55

18.52

22.02

 

SD

0.87

2.05

0.72

1.37

2.55

 

(n)

9

9

9

9

9

300 mg/kg

Mean

20.59

19.51

19.20

19.33

23.39

 

SD

1.80

2.05

1.52

1.46

1.11

 

(n)

10

10

10

10

10

1000 mg/kg

Mean

20.35

17.28

17.37*($)

18.37*

22.35

 

SD

0.97

2.17

2.50

1.97

3.11

 

 

Uterus weight (g) of dams - Group mean data

 

 

 

Terminal body weight

Absolute weight gain

Gravid uterus weight

Treatment

 

(g)

(g)

(g)

 

(n)

23

23

23

Control

Mean

371.23

62.94

75.15

 

SD

25.84

10.06

18.48

 

(n)

23

23

23

100 mg/kg

Mean

370.42

55.76

78.96

 

SD

11.78

8.31

7.97

 

(n)

22

22

22

300 mg/kg

Mean

370.59

57.96

78.01

 

SD

15.73

11.70

11.76

 

(n)

23

23

23

1000 mg/kg

Mean

352.09*

44.83*

75.33

 

SD

22.26

11.47

13.47

 

 

Macroscopic findings in dams - Group incidence

 

 

Treatment:

Control

100 mg/kg

300 mg/kg

1000 mg/kg

 

Number in group:

24

24

24

24

Finding

Number examined:

10

10

10

10

Uterus

Not pregnant

1

1

2

1

 

Unilateral implantation

1

0

0

0

 

 

 

 

 

 

No abnormalities

 

22

23

22

23

 

 

Litter data - Group mean data

 

 

 

Corpora

Implantations

Uterine deaths

Viable foetuses

%

Implantation loss

Litter weight

Treatment

 

Lutea

 

Early

Late

Total

Male

Female

males

Pre-

Post-

Total

(g)

 

(n)

23

23

23

23

23

22

23

22

23

23

23

23

Control

Mean

13.78

13.70

0.52

0.00

13.17

6.00

7.17

43.43

0.48

3.69

4.17

48.05

 

SD

3.67

3.59

0.73

0.00

3.52

2.54

2.37

15.61

2.31

5.06

5.22

13.47

 

(n)

23

23

23

23

23

23

23

23

23

23

23

23

100 mg/kg

Mean

15.04

14.70

0.74

0.00

13.96

7.13

6.83

51.74

2.07

4.83

6.84

51.55

 

SD

2.01

1.89

1.48

0.00

2.06

2.24

2.44

14.64

5.92

9.01

10.32

8.05

 

(n)

22

22

22

22

22

22

22

22

22

22

22

22

300 mg/kg

Mean

14.86

14.45

1.18

0.00

13.27

6.64

6.64

49.03

2.14

8.05

10.15

50.47

 

SD

2.62

2.06

1.62

0.00

2.39

2.15

1.59

13.53

6.01

11.29

11.59

8.88

 

(n)

23

23

23

23

23

23

23

23

23

23

23

23

1000 mg/kg

Mean

14.26

13.83

0.65

0.00

13.17

6.91

6.26

51.67

2.58

4.87

7.38

49.46

 

SD

2.75

2.50

1.11

0.00

2.81

2.79

2.05

14.03

6.66

8.70

10.33

9.71

 

 

 

External examination of foetuses - Group incidence

 

 

 

 

 

Number of foetuses

 

Number of litters

Treatment

Organ

Category

Observation

Observed

Affected

%

 

Observed

Affected

%

 

Whole foetus

 

No abnormalities observed

303

298

98.35

 

23

-

-

 

 

AN

Small

303

5

1.65

 

23

4

17.39

100 mg/kg

Whole foetus

 

No abnormalities observed

321

321

100.00

 

23

23

100.00

300 mg/kg

Whole foetus

 

No abnormalities observed

292

291

99.66

 

22

-

-

 

 

AN

Small

292

1.00

0.34

 

22

1

4.55

1000 mg/kg

Whole foetus

 

No abnormalities observed

303

303

100.00

 

23

23

100.00

 


 

Skeletal examination of foetuses - Group incidence

 

 

 

 

 

Number of foetuses

 

Number of dams

Treatment

Organ

Category

Observation

Observed

Affected

%

 

Observed

Affected

%

Control

Forepaw(s)

AN

Metacarpal(s) no ossification 4th

157

16

10.19

 

23

9

39.13

 

Hindpaw(s)

AN

Metatarsal(s) no ossification 4th

157

1

0.64

 

23

1

4.35

 

Lumbar vertebrae

AN

Centrum dumbbell shaped

157

1

0.64

 

23

1

4.35

 

Lumbar vertebrae

VA

Centrum incomplete ossification

157

1

0.64

 

23

1

4.35

 

Pelvic girdle

AN

Pubis incomplete ossification

157

2

1.27

 

23

2

8.70

 

Ribs

VA

Rudimentary 14th

157

26

16.56

 

23

15

65.22

 

Ribs

VA

Short 14th

157

1

0.64

 

23

1

4.35

 

Sacral vertebrae

AN

Arch(es) incomplete ossification

157

1

0.64

 

23

1

4.35

 

Sacral vertebrae

AN

Arch(es) no ossification

157

1

0.64

 

23

1

4.35

 

Skull

AN

General incomplete ossification

157

1

0.64

 

23

1

4.35

 

Skull

AN

Temporal incomplete ossification

157

27

17.20

 

23

15

65.22

 

Skull

AN

Palatine incomplete ossification

157

1

0.64

 

23

1

4.35

 

Skull

VA

Interparietal incomplete ossification

157

1

0.64

 

23

1

4.35

 

Skull

VA

Supraoccipital incomplete ossification

157

1

0.64

 

23

1

4.35

 

Sternebrae

AN

No ossification 6th

157

1

0.64

 

23

1

4.35

 

Sternebrae

AN

Bipartite 5th

157

1

0.64

 

23

1

4.35

 

Sternebrae

AN

Asymmetrical ossification

157

3

1.91

 

23

3

13.04

 

Sternebrae

AN

Rudimentary 5th

157

1

0.64

 

23

1

4.35

 

Sternebrae

AN

Bipartite

157

2

1.27

 

23

1

4.35

 

Sternebrae

AN

Asymmetrical ossification 5th

157

2

1.27

 

23

2

8.70

 

Sternebrae

VA

Incomplete ossification 6th

157

17

10.83

 

23

10

43.48

 

Sternebrae

VA

No ossification 5th

157

4

2.55

 

23

3

13.04

 

Sternebrae

VA

Incomplete ossification 5th

157

15

9.55

 

23

10

43.48

 

Sternebrae

VA

Incomplete ossification

157

6

3.82

 

23

4

17.39

 

Thoracic vertebrae

AN

Centrum no ossification

157

1

0.64

 

23

1

4.35

 

Thoracic vertebrae

AN

Centrum bipartite

157

1

0.64

 

23

1

4.35

 

Thoracic vertebrae

AN

Centrum asymmetrical ossification

157

1

0.64

 

23

1

4.35

 

Thoracic vertebrae

VA

Centrum incomplete ossification

157

10

6.37

 

23

7

30.43

 

Whole foetus

 

No abnormalities detected

157

86

54.78

 

-

-

-

100 mg/kg

Forepaw(s)

AN

Metacarpal(s) no ossification 4th

156

12

7.69

 

23

7

30.43

 

Hindpaw(s)

AN

Metatarsal(s) no ossification 4th

156

2

1.28

 

23

2

8.70

 

Lumbar vertebrae

VA

Centrum incomplete ossification

156

1

0.64

 

23

1

4.35

 

Pelvic girdle

AN

Ischium incomplete ossification

156

1

0.64

 

23

1

4.35

 

Pelvic girdle

MA

Pubis no ossification

156

1

0.64

 

23

1

4.35

 

Ribs

VA

14 ribs

156

2

1.28

 

23

1

4.35

 

Ribs

VA

Rudimentary 14th

156

26

16.67

 

23

14

60.87

 

Skull

AN

Temporal incomplete ossification

156

18

11.54

 

23

10

43.48

 

Skull

AN

Palatine incomplete ossification

156

1

0.64

 

23

1

4.35

 

Skull

VA

Supraoccipital incomplete ossification

156

2

1.28

 

23

2

8.70

 

Sternebrae

AN

Asymmetrical ossification 5th

156

4

2.56

 

23

4

17.39

 

Sternebrae

AN

Asymmetrical ossification

156

3

1.92

 

23

2

8.70

 

Sternebrae

AN

No ossification

156

3

1.92

 

23

3

13.04

 

Sternebrae

AN

No ossification 6th

156

1

0.64

 

23

1

4.35

 

Sternebrae

AN

Rudimentary 5th

156

3

1.92

 

23

2

8.70

 

Sternebrae

VA

Incomplete ossification 6th

156

26

16.67

 

23

11

47.83

 

Sternebrae

VA

No ossification 5th

156

5

3.21

 

23

5

21.74

 

Sternebrae

VA

Incomplete ossification

156

4

2.56

 

23

4

17.39

 

Sternebrae

VA

Incomplete ossification 5th

156

12

7.69

 

23

7

30.43

 

Thoracic vertebrae

AN

Centrum no ossification

156

1

0.64

 

23

1

4.35

 

Thoracic vertebrae

VA

Centrum incomplete ossification

156

4

2.56

 

23

4

17.39

 

Thoracic vertebrae

VA

Centrum asymmetrical dumb-bell shaped

156

1

0.64

 

23

1

4.35

 

Thoracic vertebrae

VA

Centrum dumb-bell shaped

156

4

2.56

 

23

4

17.39

 

Whole foetus

 

No abnormalities detected

156

50

32.05

 

-

-

-

300 mg/kg

Forepaw(s)

AN

Metacarpal(s) no ossification 4th

151

9

5.96

 

22

4

18.18

 

Ribs

VA

14 ribs

151

6

3.97

 

22

1

4.55

 

Ribs

VA

Short 14th

151

1

0.66

 

22

1

4.55

 

Ribs

VA

Rudimentary 14th

151

33

21.85

 

22

15

68.18

 

Skull

AN

Temporal incomplete ossification

151

13

8.61

 

22

10

45.45

 

Skull

VA

Supraoccipital incomplete ossification

151

1

0.66

 

22

1

4.55

 

Sternebrae

AN

Bipartite 5th

151

1

0.66

 

22

1

4.55

 

Sternebrae

AN

Asymmetrical ossification

151

2

1.32

 

22

1

4.55

 

Sternebrae

AN

No ossification

151

1

0.66

 

22

1

4.55

 

Sternebrae

AN

Asymmetrical ossification 5th

151

2

1.32

 

22

2

9.09

 

Sternebrae

VA

No ossification 5th

151

4

2.65

 

22

3

13.64

 

Sternebrae

VA

Incomplete ossification

151

3

1.99

 

22

2

9.09

 

Sternebrae

VA

Incomplete ossification 5th

151

15

9.93

 

22

8

36.36

 

Sternebrae

VA

Incomplete ossification 6th

151

12

7.95

 

22

7

31.82

 

Thoracic vertebrae

VA

Centrum dumb-bell shaped

151

6

3.97

 

22

4

18.18

 

Thoracic vertebrae

VA

Centrum incomplete ossification

151

5

3.31

 

22

3

13.64

 

Thoracic vertebrae

VA

Centrum asymmetrical dumb-bell shaped

151

2

1.32

 

22

2

0.09

 

Whole foetus

 

No abnormalities detected

151

86

56.95

 

-

-

 

1000 mg/kg

Forepaw(s)

AN

Metacarpal(s) no ossification 4th

154

4

2.60

 

23

3

13.04

 

Pelvic girdle

AN

Pubis incomplete ossification

154

1

0.65

 

23

1

4.35

 

Ribs

VA

Rudimentary 14th

154

18

11.69

 

23

10

43.48

 

Ribs

VA

14 ribs

154

1

0.65

 

23

1

4.35

 

Ribs

VA

Short 14th

154

3

1.95

 

23

3

13.04

 

Skull

AN

Temporal incomplete ossification

154

17

11.04

 

23

11

47.83

 

Sternebrae

AN

Asymmetrical ossification

154

2

1.30

 

23

2

8.70

 

Sternebrae

AN

Asymmetrical ossification 5th

154

8

5.19

 

23

6

26.09

 

Sternebrae

VA

Incomplete ossification 5th

154

8

5.19

 

23

6

26.09

 

Sternebrae

VA

Incomplete ossification 6th

154

8

5.19

 

23

4

17.39

 

Thoracic vertebrae

AN

Centrum bipartite

154

1

0.65

 

23

1

4.35

 

Thoracic vertebrae

VA

Centrum incomplete ossification

154

2

1.30

 

23

2

8.70

 

Thoracic vertebrae

VA

Centrum dumb-bell shaped

154

3

1.95

 

23

3

13.04

 

Whole foetus

 

No abnormalities detected

154

99

64.29

 

-

-

-

 

 

Visceral examination of foetuses - Group incidence

 

 

 

 

 

Number of foetuses

 

Number of dams

Treatment

Organ

Category

Observation

Observed

Affected

%

 

Observed

Affected

%

Control

Abdomen

VA

Haemorrhagic

146

2

1.37

 

23

2

8.70

 

Brain

AN

Ventricles enlarged moderate

146

1

0.68

 

23

1

4.35

 

Brain

VA

Ventricles enlarged slight

146

2

1.37

 

23

2

8.70

 

Great vessels

VA

Innominate artery longer

146

3

2.05

 

23

2

8.70

 

Heart

AN

Atrium enlarged

146

8

5.48

 

23

5

21.74

 

Heart

AN

Ventricle enlarged

146

1

0.68

 

23

2

8.70

 

Heart

AN

Pericardial fluid

146

6

4.11

 

23

4

17.39

 

Heart

VA

Ventricle enlarged

146

1

0.68

 

23

2

8.70

 

Kidneys

AN

Ectopic

146

13

8.90

 

23

8

34.78

 

Kidneys

AN

Pelvic dilatation moderate

146

1

0.68

 

23

1

4.35

 

Kidneys

VA

Pelvic dilatation slight

146

5

3.42

 

23

4

17.39

 

Testis

AN

Displaced

146

9

6.16

 

23

5

21.74

 

Ureter

AN

Enlarged moderate

146

11

7.53

 

23

10

43.48

 

Ureter

AN

Kinked moderate

146

2

1.37

 

23

2

8.70

 

Ureter

VA

Kinked slight

146

9

6.16

 

23

8

34.78

 

Ureter

VA

Enlarged slight

146

27

18.49

 

23

16

69.57

 

Whole foetus

 

No abnormalities detected

146

85

58.22

 

-

-

-

100 mg/kg

Abdomen

VA

Haemorrhagic

156

6

3.85

 

23

3

13.04

 

Great vessels

VA

Innominate artery longer

156

1

0.64

 

23

1

4.35

 

Heart

AN

Atrium enlarged

156

19

12.18

 

23

9

39.13

 

Heart

AN

Pericardial fluid

156

18

11.54

 

23

7

30.43

 

Heart

VA

Ventricle enlarged

156

2

1.28

 

23

2

8.70

 

Kidneys

AN

Ectopic

156

22

14.10

 

23

10

43.48

 

Kidneys

AN

Pelvic dilatation moderate

156

1

0.64

 

23

1

4.35

 

Kidneys

VA

Pelvic dilatation slight

156

2

1.28

 

23

2

8.70

 

Testis

AN

Displaced

156

19

12.18

 

23

10

43.48

 

Ureter

AN

Kinked moderate

156

2

1.28

 

23

2

8.70

 

Ureter

AN

Enlarged moderate

156

10

6.41

 

23

6

26.09

 

Ureter

VA

Kinked slight

156

19

12.18

 

23

12

52.17

 

Ureter

VA

Enlarged slight

156

26

16.67

 

23

17

73.91

 

Whole foetus

AN

Generalised oedema slight

156

1

0.64

 

23

1

4.35

 

Whole foetus

 

No abnormalities detected

156

69

44.23

 

-

-

-

300 mg/kg

Abdomen

VA

Haemorrhagic

140

2

1.43

 

22

2

9.09

 

Great vessels

VA

Innominate artery short

140

1

0.71

 

22

1

4.55

 

Heart

AN

Ventricle enlarged

140

1

0.71

 

22

5

22.73

 

Heart

AN

Pericardial fluid

140

10

7.14

 

22

7

31.82

 

Heart

AN

Atrium enlarged

140

12

8.57

 

22

8

36.36

 

Heart

VA

Ventricle enlarged

140

4

2.86

 

22

5

22.73

 

Kidneys

AN

Pelvic dilatation moderate

140

2

1.43

 

22

2

9.09

 

Kidneys

AN

Ectopic

140

14

10.00

 

22

9

40.91

 

Kidneys

VA

Pelvic dilatation slight

140

2

1.43

 

22

2

9.09

 

Testis

AN

Displaced

140

5

3.57

 

22

4

18.18

 

Thoracic cavity

AN

Haemorrhage

140

2

1.43

 

22

2

9.09

 

Ureter

AN

Enlarged moderate

140

11

7.86

 

22

9

40.91

 

Ureter

AN

Kinked moderate

140

4

2.86

 

22

3

13.64

 

Ureter

MA

Kinked extreme

140

1

0.71

 

22

1

4.55

 

Ureter

VA

Enlarged slight

140

32

22.86

 

22

14

63.64

 

Ureter

VA

Kinked slight

140

15

10.71

 

22

10

45.45

 

Whole foetus

AN

Generalised oedema slight

140

1

0.71

 

22

1

4.55

 

Whole foetus

 

No abnormalities detected

140

68

48.57

 

-

-

-

1000 mg/kg

Abdomen

VA

Haemorrhagic

147

11

7.48

 

23

6

26.09

 

Great vessels

AN

Innominate artery absent

147

1

0.68

 

23

1

4.35

 

Great vessels

MA

Absence of aortic arch

147

1

0.68

 

23

1

4.35

 

Great vessels

VA

Innominate artery short

147

2

1.36

 

23

2

8.70

 

Heart

AN

Pericardial fluid

147

11

7.48

 

23

8

34.78

 

Heart

AN

Atrium enlarged

147

7

4.76

 

23

5

21.74

 

Kidneys

AN

Ectopic

147

12

8.16

 

23

8

34.78

 

Kidneys

VA

Pelvic dilatation slight

147

5

3.40

 

23

5

21.74

 

Testis

AN

Displaced

147

4

2.72

 

23

4

17.39

 

Thoracic cavity

AN

Haemorrhage

147

1

0.68

 

23

1

4.35

 

Ureter

AN

Kinked moderate

147

1

0.68

 

23

1

4.35

 

Ureter

AN

Enlarged moderate

147

10

6.80

 

23

7

30.43

 

Ureter

MA

Kinked extreme

147

1

0.68

 

23

1

4.35

 

Ureter

MA

Enlarged extreme

147

1

0.68

 

23

1

4.35

 

Ureter

VA

Kinked slight

147

8

5.44

 

23

7

30.43

 

Ureter

VA

Enlarged slight

147

16

10.88

 

23

12

52.17

 

Whole foetus

 

No abnormalities detected

147

96

65.31

 

-

-

-

 

Key to tables

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

** = mean value of group is significantly different from control at p < 0.01

($) = Modified t test if group variances are inhomogeneous ($)

AN = Anomoly

VA = Variation

MA = Malformation

Conclusions:
Developmental toxicity has been examined in rats at dose levels of 100, 300 or 1000 mg/kg bw/day administered during teh period of organogenesis.
No exposure related developmental toxic effects were observed and, as a result, the highest dose investigated, 1000 mg/kg bw/day, was considered to be the No Observed Adverse Effect Level (NOAEL) for embryo-foetal effects.
Executive summary:

Developmental toxicity has been examined in rats using methods described in OECD test guideline No. 414. The substance was administered during the period of organogenesis at levels of 100, 300 or 1000 mg/kg bw/day.

No exposure related developmental toxic effects were observed and, as a result, the highest dose investigated, 1000 mg/kg bw/day, was considered to be the No Observed Adverse Effect Level (NOAEL) for embryo-foetal effects.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental Start Date: 23/07/2019 (dosing); Experimental Completion Date:14/08/2019 (necropsy)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
25 June 2018
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
Batch 3606218211
Retest 15 May 2020
Species:
rabbit
Strain:
New Zealand White
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
0.5% w/w
Details on exposure:
Dose volumes (5 mL/kg) calculated on indvidual bodyweights on Days 6, 9, 12, 15, 18,21,24 and 27 pc
Analytical verification of doses or concentrations:
yes
Remarks:
Liquid chromatography with UV detection; method validated at test lab
Details on analytical verification of doses or concentrations:
Dosing solutions prepared daily; samples from first and last week of dosing were analysed and were within acceptable range
Details on mating procedure:
Single females introduced to single sexually mature male from same supplier, and retained for at least 1 hour following successful mating observed.
Duration of treatment / exposure:
Day 6 to day 28 post coitum
Frequency of treatment:
Daily
Duration of test:
Day six to euthanasia at day 29 post coitum
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Control plus carrier
Dose / conc.:
100 mg/kg bw/day
Dose / conc.:
300 mg/kg bw/day
Dose / conc.:
1 000 mg/kg bw/day
No. of animals per sex per dose:
20 mated females per dose
Control animals:
yes, concurrent vehicle
Maternal examinations:
Measurements and observations: Mortality, abortions, clincal signs, bodyweights, food consumption.
Thyroid was dissected free of fat, weighed and examined histopathologically for all controls, high dose and euthanised animals.
Ovaries and uterine content:
The following were recorded:
Gravid uterine weights
Number of corpora lutea for pregnant animals
Number of implantations for pregnanat animals
Uteri and individual uterine horms without impanatations immersed in 20% ammonium sulphide for evidence of early embryonic death
Fetal examinations:
The following observations were made:
Number, sex and weight of live and dead foetus
Number of interuterine deaths; early or late resorptions
Gross placentae evaluation
The thoracic and abdominal cavities were opened and examined
Head sections on each foetus, skeletal exam and brain ventricles
Head from half of all foetus removed/preserved for examination of internal structures; eye, brain, nasal passage and tongue
Skeletal examination for all groups for structural deviations (malformations, anomalies, variants)
Statistics:
Continuous variables: group means asessed by Dunnets test or modified t test.
Non continuos variables Kruskal Wallis test and intergroup difference assessed with non-parametric version of the Williams test.
Statistical significance; p < 0.05
Indices:
Pre-implantation loss
Post-implantation loss
Total implantation loss
Sex ratio
Historical control data:
Yes
Clinical signs:
no effects observed
Description (incidence and severity):
No treatment-related clinical signs were recorded.
Mortality:
no mortality observed
Description (incidence):
No treatment related effects; 1 mid dose (300 mg/kg/day) and 1 high dose (1000 mg/kg/day) animal was terminated early.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
No treatment related effects. Average body weights did not differ although the highest dose group had significantly lower body weight gain at gestation day 9.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
300 mg/kg bw; significantly reduced food consumption on days 9 and 21
1000 mg/kg bw; reduced food consumption on days 9, 12 and 21
Recovered by end of treatment phase
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Description (incidence and severity):
Except for the two sacrificed females (1 at 300 mg/kg; 1 at 1000 mg/kg), in which dark red staining in the urogenital region was observed, there were no treatment related findings in dams compared to control animals.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Other effects:
no effects observed
Number of abortions:
effects observed, non-treatment-related
Description (incidence and severity):
Abortions occurred in one female of the mid-dose (300 mg/kg/day) and in one of the high
dose level (1000 mg/kg/day).
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
Two dams were found to be not pregnant at the end of the study; one in the low dose and one in the high dose. Given the lack of a dose response this was regarded as incidental.
Other effects:
no effects observed
Details on maternal toxic effects:
None oberved
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: Signs of maternal toxicity
Abnormalities:
no effects observed
Fetal body weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
At 300 mg/kg and 1000 mg/kg statistically significantly reduced foetal weight compared to control, but these did not follow a dose related pattern. Assumed to be delayed development effects of early stage maternal toxicity, linked with reduced food consumption.
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
effects observed, treatment-related
Description (incidence and severity):
Only observed external abnormality was the smaller size of foetuses; 36.7% of foetus in treated groups cf to 4.7% in the control group were less than 35 g.
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
Severe skeletal findings were recorded in a litter of the low dose group. One foetus in low dose showed malformation; absence of tail and absence of all caudal vertebrae.
In the same litter skeletal examination identified one other foetus with fused causdal vetrabrae.

Skeletal examination recorded a number of unossified bones. This was related to the small size of fetus (<35g). It was suggested this was related to some retardion of fetal growth.
Visceral malformations:
effects observed, non-treatment-related
Description (incidence and severity):
One foetus with skeletal malformations from a litter in the low dose group showed agenesis of left kideny plus hyploplasia azygous lobe of lung and dark area in liver. In the high dose group two foetus from one litter showed enlarged bilteral brain ventricle.

One foetus (Dam no. X1050057) showed malformations like the absence of the tail and the agenesis of the left kidney. In the same foetus, additional abnormalities included hypoplasia of the azygous lobe of the lung and the presence of dark area in the liver. In the high dose group, two foetuses (Dam no. X1050159) showed enlarged bilateral brain ventricles.

In the absence of a dose relationship, the low occurrence and the different nature of malformation, these major abnormalities were considered to be incidental.
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
Folded retina
Details on embryotoxic / teratogenic effects:
Some retardation of foetal growth observed in mid and high doses with reduced foetal weights and delayed ossification anomalies. This was related to maternal toxicity.
Observed malformations were limited to single foetus in litters at low and high dose and were not observed to be dose dependent
Key result
Dose descriptor:
NOAEL
Remarks:
foetal
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no direct dose related teratological effects.
Key result
Abnormalities:
effects observed, treatment-related
Localisation:
external: paw
skeletal: pelvic girdle
skeletal: hindlimb
Description (incidence and severity):
Regarded as effect of reduced delayed growth in higher number of small feotus as a result of reduced food consumption and body weight gain in dams.
Key result
Developmental effects observed:
yes
Lowest effective dose / conc.:
1 000 mg/kg bw/day (nominal)
Treatment related:
no
Relation to maternal toxicity:
developmental effects as a secondary non-specific consequence of maternal toxicity effects
Dose response relationship:
no

Body weight (kg)

Group

Day

Post-coitum

0 (mating)

6

9

12

15

18

21

24

27

29

Control

No

20

20

20

20

20

20

20

20

20

20

Mean

3.898

4.074

4.084

4.084

4.162

4.221

4.247

4.276

4.307

4.325

SD

0.3196

0.3175

0.3395

0.3329

0.3086

0.2833

0.3187

0.3077

0.3062

0.3077

100 mg/kg

No

19

19

19

19

19

19

19

19

19

19

Mean

3.994

4.143

4.169

4.191

4.218

4.258

4.282

4.307

4.339

4.361

SD

0.3413

0.3275

0.3380

0.3457

0.3530

0.3313

0.3271

0.3077

0.2923

0.2946

300 mg/kg

No

20

20

20

20

20

20

20

20

20

20

Mean

3.871

3.992

3.978

4.011

4.019

4.071

4.060

4.061

4.076

4.098

SD

0.2753

0.2753

0.2553

0.2746

0.2744

0.2896

0.3097

0.3148

0.3012

0.2973

1000 mg/kg

No

19

19

19

19

19

19

19

19

19

19

Mean

3.897

4.057

4.013

4.053

4.085

4.093

4.113

4.141

4.159

4.232

SD

0.2779

0.2463

0.2319

0.2150

0.1992

0.2483

0.2599

0.2847

0.3095

0.2775

Uterus weights; corrected body weights and absolute weight gain; females with live foetus

Group

Day

Post-coitum

Gravid uterus weight (g)

Corrected body weight (g)

Absolute body weight gain (g)

Total body weight (kg)

Thyroid gland (g)

Throid gland to terminal body weight (%)

Control

No

20

20

20

20

20

20

Mean

461.34

3858.15

-215.85

4.320

0.2492

0.0058

SD

108.97

302.09

138.46

0.3035

0.06016

0.00147

100 mg/kg

No

19

19

19

19

19

19

Mean

446.33

3901.58

-241.11

4.297

0.2615

0.0061

SD

66.05

302.63

207.00

0.3656

0.05887

0.00144

300 mg/kg

No

19

19

19

19

20

20

Mean

399.31

3689.16

-303.53

4.088

0.2479

0.0061

SD

108.10

302.63

207.57

0.2932

0.05776

0.00132

1000 mg/kg

No

18

18

18

18

18

19

Mean

410.16

3810.94

242.39

4.196

0.2331

0.0056

SD

100.69

252.20

256.64

0.2931

0.03775

0.00093

Macroscopic findings

Findings

Control

100 mg/kg

300 mg/kg

1000 mg/kg

Number of animals

20

20

19

19

Number examined

20

20

19

19

Abnormal caecum contents

0

1

0

0

Hairloss: forelimbs

0

1

0

1

Hairloss; hindlimbs

0

1

0

1

Hairloss; forepaws

1

0

0

0

Hairloss; head

1

0

0

0

Hair loss; skin

0

0

3

3

Abnormal sized gall bladder

0

0

1

0

Uterus; not pregnant

0

1

0

1

No abnormalities observed

18

15

14

14

Litter data and sex ratios (group data)

Group

 

Corpora lutea

Implantation

Pre imp loss (%)

Early resorptn.

Late resorptn.

Total resorptn.

Foetus sex

Males

Foetal sex

Females

Male (%)

Control

No

20

20

20

20

20

20

20

20

20

Mean

8.15

7.95

2.38

0.15

0.4

0.55

3.75

3.65

49.2

SD

2.28

2.24

5.09

0.49

0.68

1

1.74

4.26

18.64

100 mg/kg

No

19

19

19

19

19

19

19

19

19

Mean

8.63

8.32

3.58

0.11

0.47

0.58

3.47

3.74

56.05

SD

1.74

1.73

5.51

0.32

0.69

0.84

1.74

1.33

15.88

300 mg/kg

No

19

19

19

19

19

19

19

19

19

Mean

8.42

8.21

2.51

0.32

0.58

0.9

3.58

3.74

52.32

SD

2.12

2.15

5.03

0.75

1.26

1.33

1.74

1.85

22.05

1000 mg/kg

No

18

18

18

18

18

18

18

18

18

Mean

8.78

8.22

7.39

0.61

0.17

0.78

3.56

3.89

51.00

SD

2.02

2.42

12.17

1.38

0.38

1.52

1.62

1.94

24.27

Litter data; viable foetus and foetal weights

Group

 

Dead Foetus

Live foetus

Post implant loss (%)

Litter weight (g)

Foetal weight (g)

Male foetal wt (g)

Female foetal wt (g)

Control

No

20

20

20

20

20

20

20

Mean

0.05

7.35

7.38

313.21

43.44

43.24

43.6

SD

0.22

2.11

11.96

78.62

4.45

5.77

4.27

100 mg/kg

No

19

19

19

19

19

19

19

Mean

0.05

7.68

8.89

302.17

40.13

40.62

39.31*

SD

0.23

1.77

13.47

46.17

5.29

5.53

5.52

300 mg/kg

No

19

19

19

19

19

18

18

Mean

0.05

7.26

13.15

257.77

36.38*

36.82*

34.73*

SD

0.23

2.56

20.47

89.46

6.72

7.14

5.68

1000 mg/kg

No

18

18

18

18

18

18

18

Mean

0.00

7.44

7.44

282.71

38.43*

37.45*

37.45*

SD

0.00

2.01

14.10

75.93

4.95

4.89

4.89

*significant p <0.05

Foetal macroscopic observations; internal/external

Finding

Control

100 mg/kg

300 mg/kg

1000 mg/kg

Number of dams/foetus

20/148

20/147

20/139

20/134

Number litters examined

20

19

19

18

Litters(l)/foetus(f)

l/f

l/f

l/f

lf

Abnormalities

tail; short

0/0

1/1

0/0

0/0

liver: area

0/0

1/1

0/0

0/0

lung hypoplasia

0/0

1/1

0/0

1

foetus; small

4/7

7/31

10/51

8/33

Malformation

 

tail absent

0/0

1/1

0/0

0/0

enlarged brain Ventricle

0/0

0/0

0/0

1/2

Kidney (left) agenesis

0/0

1/1

0/0

0/0

Dead foetus

1/1

1/1

1/1

0/0

No abnormalities detected (% foetal)

20/140 (94.59)

19/114 (77.55)

17/87

(62.59)

17/101 (75.37)

Foetal visceral observations

Finding

Control

100 mg/kg

300 mg/kg

1000 mg/kg

Number of examined litter(l)/foetus(f)

20/69

19/67

18/65

18/64

No abnormalities detected

20/64

19/65

18/65

17/58

% normal foetus

92.75

97.01

100

90.63

Variants

l/f

l/f

l/f

l/f

Brain; slight enlarged ventricle

4/4

1/1

0/0

1/5

Folded retina, bilateral

1/1

0/0

0/0

0/0

Folded retina unilateral

0/0

1/1

0/0

1/1

 

Foetal skeletal observations

Finding

Control

100 mg/kg

300 mg/kg

1000 mg/kg

Number of examined litters (l)/foetus (f):

20/147

19/146

19/138

18/134

Anomalies

l/f

l/f

l/f

l/f

Caudal vertebra(e); fused

0/0

1/1

0/0

0/0

Caudal vert.; malposition

0/0

1/1

0/0

1/2

One or more incomplete ossification

0/0

0/0

0/0

2/2

Cervical vertebral centrum(s);

no ossification(s)

0/0

0/0

0/0

1/1

Forelimbs: throclea no ossification

1/1

3/6

4/11

6/20

Forelimbs: head no ossification

12/20

18/48

17/71

14/54

Forepaw: Distal phalanx no ossification 1st

0/0

0/0

0/0

1/1

One/more distal phalanx no ossification

3/6

4/5

2/2

1/2

One/more distal phalanges no ossification

5/8

8/13

6/16

3/8

Proximal phalanx no ossification 1st

2/3

2/2

6/15

3/5

Hindlimb: Femur head no ossification

11/18

11/22

13/54

15/53

Hind paw; astralagus incomplete ossification

0/0

0/0

0/0

1/1

Hind paw; astralagus no ossification

0/0

0/0

3/7

1/1

Lumbar vertebrae arch; additional vertebra

0/0

1/1

0/0

1/1

Pelvic girdle; articulation point absent

0/0

0/0

1/2

0/0

Pelvic girdle; additional articulation point

11/16

4/5

11/17

8/9

Pubis; no ossification

0/0

0/0

1/5

1/1

Ribs; floating 13th

12/21

10/14

12/14

8/18

Ribs; one of more bifurcated

0/0

0/0

0/0

1/1

Skull; nasal incomplete ossification

0/0

0/0

0/0

1/1

Skull; sutural bone

0/0

1/1

0/0

1/1

Sternebra(e); additional ossification site

2/2

0/0

0/0

0/0

Sternebra(e); asymmetrical ossification

2/2

2/2

4/5

3/3

Sternebra(e): bipartite

0/0

1/1

1/1

3/3

Sternebra(e): fused

1/1

0/0

1/3

1/1

Sternebra(e); no ossification

2/4

7/11

2/5

5/10

Sternebra(e); abnormal shape

0/0

0/0

1/1

0/0

Sternebra(e); incomplete ossification

0/0

1/1

1/1

0/0

Sternebra(e); rudimentary 6th

1/1

1/1

0/0

1/1

Hyoid; incomplete ossification

1/1

2/2

1/1

5/7

Hyoid; no ossification

0/0

0/0

1/2

3/3

Malformation

l/f

l/f

l/f

l/f

All vertebrae; no ossification

0/0

1/1

0/0

0/0

Variant

l/f

l/f

l/f

l/f

Caudal vertebra(e); less than 12th

0/0

0/0

3/3

1/1

Forepaw; prox. phalange(s) no ossification

0/0

0/0

0/0

1/1

Hindlimbs; tibia head no ossification

16/57

15/64

17/96

16/89

Hind paw; distal phalange(s) no ossification

0/0

0/0

1/2

1/1

Hind paw; distal phalange(s) incomplete

0/0

0/0

1/1

0/0

Pelvic girdle; insertion 2ndsacral vertebrae

8/18

8/20

11/33

11/40

Ribs; 13 ribs

19/98

18/73

18/77

18/93

Ribs; rudimentary 13th

2/3

0/0

0/0

1/1

Rib; short 13th

18/67

18/54

18/69

17/57

Skull; anterior fontanelle enlarged

1/2

1/2

4/5

3/6

Skull; posterior fontanelle enlarged

0/0

0/0

1/1

2/2

Sternebra(e); no ossification 5th

12/23

13/36

11/41

11/49

Sternebra(e); incomplete ossification

12/26

13/25

11/21

14/21

Sternebra(e); rudimentary 5th

4/5

7/8

9/10

4/7

Conclusions:
Slight maternal toxicity was observed at 300 and 1000 mg/kg/day based on reduced food consumption and reduced body weight gain in the early stages of gestation. These effects were not observed later in the study. The reduced foetal weight observed at these dosages may be a secondary effect of obeseved early maternal toxicity. However, no adverse effects were seen on foetuses at termination including the skeletal and fixed head examinations.

Therefore, it can be concluded that 1000 mg/kg was considered the NOAEL (No Observed Adverse Effect Level) for maternal toxicity and 1000 mg/kg for the developmental toxicity of rabbits, orally administered with Trioctyl benzene-1,2,4-tricarboxylate (DIPLAST TM8).
Executive summary:

The effects of Trioctyl benzene-1,2,4-tricarboxylate (DIPLAST TM8) were investigated in New Zealand White rabbits during pregnancy and embryo-foetal development.  Females were in-house mated with sexually mature males of the same strain, held as ERBC stock animals, and then assigned to 4 groups of 20 animals each at dose levels of 0, 100, 300 & 1000 mg/kg/day.

All femaleswere administered by oral gavage during the gestation period from Day 6 through Day 28 post coitum . The dose volume was set at 5 mL/kg body weight. Females from the control group (Group 1) received 0.5% carboxymethylcellulose (0.5% CMC) as vehicle at the same dose volume during the same treatment period. Mortality check, clinical signs, body weight and food consumption were recorded during the in-life phase. At completion of the study period, females were caesarean-sectioned on Day 29 post coitum and subjected to detailed post mortem examination. Gravid uterus was weighed and the uterine content examined for the number of implantations, intra-uterine deaths and live foetuses. Live foetuses were weighed, sexed and observed for external and internal abnormalities. The ovaries were also examined and the corpora lutea counted.

Maternal Findings

Two rabbits were sacrificed for abortion, one each in the mid-dose (300 mg/kg/day) and high dose (1000 mg/kg/day) groups on gestation Day 25 and 28, respectively.  All the remaining females were sacrificed at termination and only two were found not pregnant, one each in the low and high dose groups (100 and 1000 mg/kg/day).  The final number of females with live foetuses on gestation Day 29 was: 20 in the control group, 19 in the low and mid-dose groups and 18 in the high dose group.

No treatment-related signs were recorded in the study.

The body weight gain was statistically significantly lower in the high dose group when compared to the control group on gestation Day 9.

Food consumption was statistically significantly reduced in the high dose group on gestation Days 9, 12 and 21 and in the mid-dose group on gestation Days 9 and 21.

These effects on food consumption were not apparent by the end of the gestation period.

No differences were seen in the uterus weight, corrected body weight (terminal body weight minus the uterus weight) and the absolute weight gain, calculated subtracting the uterus weight and the body weight at gestation Day 6 from the terminal body weight, between control and treated groups.

There was no effect on terminal body weight or on the thyroid gland weight, absolute or relative to body weight, in treated females, when compared to controls.

A single female in Groups 3 & 4 was sacrificed for humane reasons on Day 25 and 28 post coitum, respectively. At post mortem observation, both females showed dark red staining in the urogenital region.  No treatment-related changes were noted in all other (scheduled) terminations.

Upon histopathological examination, no treatment related changes were noted in thyroid gland of females receiving Trioctyl benzene-1,2,4-tricarboxylate at 1000 mg/kg.

Foetal Findings

Statistically significantly lower mean foetal weight was observed in the mid- and high dose groups compared to controls. This reduction was seen both in male and female foetuses. In addition, the mean female foetal weight was found statistically significantly lower also in the low dose group.

Upon external examination, the most recurrent abnormality was the small size of the foetuses (b.w. less than 35 g) which was observed up to 36.7% in treated groups versus 4.7% in the control group. No treatment related findings were observed at internal examination.

Upon visceral examination, no treatment related findings were recorded between groups when compared to controls.

Upon skeletal examination, no treatment related findings were recorded between groups when compared to controls.

Conclusion

In conclusion, slight maternal toxicity was observed at 300 and 1000 mg/kg/day based on the reduced body weight gain and food consumption in the early stages of gestation. The reduced foetal weight observed at these dosages may be a secondary effect of the early stage maternal toxicity. However, no adverse effects were seen on foetuses at termination including the skeletal and fixed head examinations.

Therefore, it can be concluded that 1000 mg/kg was considered the NOAEL for maternal toxicity and 1000 mg/kg was the NOAEL for the developmental toxicity in rabbits, orally administered with Trioctyl benzene-1,2,4-tricarboxylate (DIPLAST TM8).

Endpoint:
developmental toxicity
Type of information:
(Q)SAR
Adequacy of study:
supporting study
Study period:
August 10, 2016
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification
Justification for type of information:
The model is been assessed according to the OECD principles for the validation of QSAR, to generate a transparent, understandable, reproducible and verifiable result.
Qualifier:
according to
Guideline:
other: ECHA Guidance on information requirements and chemical safety assessment - Chapter R.06: QSARs and grouping of chemicals
Principles of method if other than guideline:
Battery algorithm
The models are made in three independent systems: CASE Ultra (CU), Leadscope Predictive Data Miner (LS) and SciQSAR (SQ). Based on predictions from each of the applied systems, a battery prediction is made using a so-called battery algorithm. The battery approach can give more reliable predictions and can also expand the applicability domain, which was shown in a previous pilot project including 32 different models and the three systems mentioned above (not published).

For the teratogenic potential, QSAR predictions are made in each of the independent QSAR model systems and combined into a battery prediction by using the criteria shown in the following table. The first column shows the total number of predictions (positive/negative) in domain. The next two columns show the number of positive and negative predictions, respectively. The final battery prediction based on the individual predictions is shown in the fourth column.

Total POS/NEG POS NEG Battery prediction (a) Remarks
in domain in domain in domain
3 3 0 POS_IN
3 0 3 NEG_IN
3 2 1 POS_IN
3 1 2 INC_OUT EXCEPT when CU and LS are both NEG_IN,
or (see remark) in this case the battery call is NEG_IN
NEG_IN

(a) POS, positive; NEG, negative; INC, inconclusive; IN, inside applicability domain; OUT, outside applicability domain.
(b) Less weight is put on an SQ POS compared to LS or CU POS in cases where LS and CU agree on a NEG in AD prediction, because SQ in many cases has lower specificity than LS and CU.
GLP compliance:
no
Limit test:
no
Species:
other: Humans
Dose descriptor:
other: (Q)SAR prediction
Based on:
other: (Q)SAR prediction
Basis for effect level:
other: (Q)SAR prediction
Remarks on result:
other: no teratogenic potential based on (Q)SAR prediction
Dose descriptor:
other: (Q)SAR prediction
Based on:
other: (Q)SAR prediction
Basis for effect level:
other: (Q)SAR prediction
Remarks on result:
other: no teratogenic potential based on (Q)SAR prediction
Developmental effects observed:
no

 Developmental Toxicity

Exp

Battery

CASE Ultra

Leadscope

SciQSAR

Teratogenic Potential in Humans

 

NEG_IN

NEG_IN

POS_OUT

NEG_IN

Conclusions:
As result of the in-silico assessment, the teratogenic potential in humans of trioctyl benzene-1,2,4-tricarboxylate was estimated to be negative.
Executive summary:

As result of the in-silico assessment, the teratogenic potential in humans of trioctyl benzene-1,2,4-tricarboxylate was estimated to be:negative.

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
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

Both the registered substance and a structurally related substance, an ester of 1,2,4 -benzenetricarboxylic acid with mixed C8 and C10 linear side chains, have been tested in rats to detect the effects on pregnant animals when the materials were administered during the period of organogenesis at dose levels of 0, 100, 300 and 1000 mg/kg/day. Study design was based on OECD Test Guideline No. 414. No exposure related developmental toxic effects were observed in the study. In both cases a dose level of 300 mg/kg/day was considered the NOAEL for maternal toxicity and a dose level of 1000 mg/kg/day the NOAEL for embryo-foetal effects.

Toxicity to reproduction: other studies

Additional information

A study with a structurally related substance, an ester of 1,2,4 -benzenetricarboxylic acid mixed C8 and C10 linear side chains, of effects on gene expression associated with developmental toxicity in male rat foetal testes by transcriptional profiling indicate that the substance does not cause repression of genes in the testicular mal-development pathway indicating that the substance is unlikely to cause testicular dysgenesis in rats as is seen with some phthalates (7.9.4).

Justification for classification or non-classification

An OECD screening study of reproductive toxicity revealed no functional changes in fertility or reproductive performance.

A sub-chronic toxicity study on a structurally related substance showed no indications of possible reproductive effects, examinations including spermatogenic staging, oestrus cycle, weights of ovaries and testes, microscopic and macroscopic observations.

A study on a structurally related substance of effects on gene expression associated with developmental toxicity in male rat foetal testes by transcriptional profiling indicate that the substance does not cause repression of genes in the testicular mal-development pathway indicating that the substance is unlikely to cause testicular dysgenesis in rats.

A developmental toxicity study on the same structurally related substance revealed no effects on developmental toxicity at a dose level of 1000 mg/kg/day in rats. A second developmental study, in rabbits, on the same structurally related substance als show no maternal or developmental toxicity at a dose level of 1000 mg/kg/day.

In accordance with Regulation (EC) No. 1272/2008 no toxicologically significant effects are evident sufficient to warrant classification.