1,4-Cyclohexanedicarboxylic acid, 1,4-diisononyl ester

Endpoint summary

Description of key information

One repeated dose toxicity study (28 days) according to OECD guideline 422 was conducted with DINCD and revealed no test-item related signs of toxicity. The NOAEL for systematic toxicity is above the highest administered dose 1000 mg test item/kg b.w./day, p.o..

In addition, a repeated dose toxicity study (90 days) accoding to OECD TG 408 was conducted with DINCD and revealed no test-item related effects. Based on these results, the NOAEL was established as being at least 1000 mg/kg/day for males and females.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25.09.2020-22.03.2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.3100 (90-Day Oral Toxicity in Rodents)

Deviations:

no

GLP compliance:

yes

Species:

rat

Strain:

Wistar

Details on species / strain selection:

The rat was chosen as the animal model for this study as it is an accepted rodent species for chemical toxicity testing by regulatory agencies.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: 6-7 weeks
- Weight at study initiation: 166-208g (m); 115-148g (f)
- Housing: Up to 5 animals of the same sex and same dosing group together.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 14d

DETAILS OF FOOD AND WATER QUALITY:
Municipal tap water

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22°C
- Humidity (%): 31-61%
- Air changes (per hr): ten or more
- Photoperiod (hrs dark / hrs light): 12h light and 12h dark

IN-LIFE DATES: From: 21 Oct 2020 To: 21 Jan 2021

Route of administration:

oral: gavage

Details on route of administration:

The oral route of exposure was selected because this is a possible route of human exposure,
during manufacture, handling or use of the test item.

Vehicle:

corn oil

Details on oral exposure:

The first day of dosing was designated as Day 1. The dose formulations were stirred continuously during dosing. The doses were given using a plastic feeding tube.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The concentrations analyzed in the formulations of Group 2-4 were in agreements with target concentrations and formulations of Group 2 and 4 were homogeneous.

Duration of treatment / exposure:

7 days a week for a minimum of 13 weeks.

Frequency of treatment:

Once daily

Dose / conc.:

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

Remarks:

control - Group 1

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

Group 2

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

Group 3

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

Group 4

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

The high-dose level should produce some toxic effects, but not excessive lethality that would prevent meaningful evaluation. The mid-dose level is expected to produce minimal to moderate toxic effects. The low-dose level should produce no observable indications of toxicity.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least once daily; from Day 1 at 0 to 1hour post-dose.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly; from Week 1 and throughout the study, and on the day of necropsy.

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly; from at least Day 1 and throughout the study.

FOOD CONSUMPTION:
- Food consumption quantitatively measured per cage; Weekly

FOOD EFFICIENCY: No

WATER CONSUMPTION: Yes
- Water consumption was monitored by visual inspection of the water bottles.

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: pretreatment period and during week 13
- Dose groups that were examined: Group 1 and 4

HAEMATOLOGY: Yes
- Time schedule for collection of blood: end of treatment
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: all animals
- Parameters checked:
White Blood Cell Count (WBC), Neutrophils (absolute), Lymphocytes (absolute), Monocytes (absolute), Eosinophils (absolute), Basophils (absolute), Large unstained cells (LUC) (absolute), Red Blood Cell Count, Reticulocytes (absolute), Red Blood Cell Distribution Width (RDW), Hemoglobin, Hematocrit, Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular hemoglobin concentration (MCHC), Platelets, Prothrombin time (PT), Activated partial thromboplastin time (APTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: end of treatment
- Animals fasted: Yes
- How many animals: all animals
- Parameters checked: Alanine aminotransferase (ALT), Triglycerides, Aspartate aminotransferase (AST), HDL and LDL Cholesterol, Alkaline Phosphatase (ALP), Sodium, Total protein, Potassium,
Albumin, Chloride, Total Bilirubin, Calcium, Urea, Inorganic Phosphate (Inorg. Phos), Creatinine, Triiodothyronine (T3), Glucose, Thyroxine (T4), Cholesterol, Thyroid-Stimulating Hormone (TSH)

URINALYSIS: No

FUNCTIONAL TESTS: Yes
- Time schedule for examinations: during week 12-13
- Dose groups that were examined: first 5 animals per sex per group
- Battery of functions tested: sensory activity / grip strength / motor activity

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (see table)

HISTOPATHOLOGY: Yes (see table)

Statistics:

Yes.
Parametric/non-parametric: one-way ANOVA F-test, Levene’s test or Kruskal-Wallis test
Incidence: Fisher’s exact test

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

Incidental salivation was seen after dosing among a few animals of all test item groups on one or more days, which was considered not toxicologically relevant. Any other clinical signs (e.g. thin fur cover, skin lesions, scabs) noted during the Dosing Period occurred within the range of background findings to be expected for rats of this age and strain.

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

In males of all treated groups, a general trend towards decreased mean body weights and body weight gains was observed from Day 1 onwards. Changes compared to the concurrent control group were very slight, i.e. statistical significance was reached for mean body weight gain at 300 and 1000 mg/kg/day over Days 36-43 only, and less than 10% decrease in body weight was observed at the end of the dosing period (Day 91). Furthermore, not always a dose-related trend was evident. Based on this, these changes were considered unrelated to treatment with the test item. Any changes observed in treated females when compared to the concurrent controls, including the statistically significantly increase in mean body weight gain at 300 mg/kg/day over Days 22-29, were considered not toxicologically relevant as these occurred in absence of a dose-response.

Food consumption and compound intake (if feeding study):

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

In males treated at 1000 mg/kg/day, a decrease in reticulocyte count (0.85x of control) compared to the control was observed. In absence of changes in correlating parameters, this finding was considered unrelated to treatment with the test item.
Remaining differences in hematology parameters in males and females, regardless of statistical significance, were considered not test item-related based on the absence of a dose response, general overlap of individual values with the range of control values, were of a magnitude of change commonly observed in rats under similar study conditions and/or were considered to have arisen as a result of slightly high control value (basophils in control males). The prolonged mean prothrombin time (PT) observed in females at 300 and 1000 mg/kg/day (both 1.05x of control) was considered to be unrelated to treatment with the test item as this occurred in the absence of a dose-related trend and mean values remained within the range considered normal for females of this age and strain.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

In males at 300 and 1000 mg/kg/day, increased calcium concentrations (1.07x and 1.06x of control, respectively) were observed compared to the control. In absence of changes in correlating parameters, this finding was considered unrelated to treatment with the test item.

Endocrine findings:

effects observed, non-treatment-related

Description (incidence and severity):

Triiodothyronine (T3) concentrations were decreased in all test item-treated females (0.91x, 0.90x, 0.86x of control, respectively; not statistically significant). Given the magnitude of change this finding was considered unrelated to treatment with the test item.

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

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):

No test item-related microscopic observations were observed in males at 100 mg/kg/day and females up to 1000 mg/kg/day.

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

In males treated at 300 and 1000 mg/kg/day, test item-related microscopic findings were noted in the kidney, consisting of an increased incidence of minimal to mild tubular hyaline droplet accumulation (up to a mild degree).

Histopathological findings: neoplastic:

no effects observed

Key result

Remarks on result:

not determinable due to absence of adverse toxic effects

Critical effects observed:

no

Conclusions:

In conclusion, administration of Diisononyl 1,4-cyclohexanedicarboxylate (DINCD) by once
daily oral gavage for 90 days was well tolerated in Wistar Han rats at dose levels of up to
1000 mg/kg/day. Increased minimal to mild tubular hyaline droplets accumulation was
observed in the kidneys of males at 300 and 1000 mg/kg/day, which was considered non-adverse.
No test item-related effects were seen in females up to 1000 mg/kg/day.
Based on these results, the No Observed Adverse Effect Level (NOAEL) was established as
being at least 1000 mg/kg/day for males and females.

Reference 2

Endpoint:

repeated dose toxicity: oral, other

Remarks:

COMBINED REPEATED DOS E TOXICITY STUDY WITH THE REPRODUCTION / DEVELOPMENTAL TOXICITY SCREENING TEST

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19.12.2016 to 23.11.2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rat

Strain:

other: CD(R) / Crl:CD(SD

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test species / Strain / Stock Rat / CD(R) / Crl:CD(SD)
Breeder: Charles River Laboratories, Research Models and Services, Germany GmbH
Sandhofer Weg 7, 97633 Sulzfeld, Germany
Body weight (at 1st administration on TD15): Males: 398.1 g - 476.9 g; Females: 233.9 g - 291.0 g
Age (at 1st administration on TD15): Males and Females: 80 days
Selection of species: The rat is a commonly used rodent species for such studies.

Number and sex of animals:
Pre-exposure period (TD 1 to TD 14): 65 female animals
A sufficient number of animals in order to grant at least 40 females with a normal oestrus cycle
for the main study and 10 for the toxicity study.
Main study (start on TD 15): 80 animals (40 males and 40 females)
A sufficient number in order to grant at least 8 pregnant females per group for evaluation of the FO generation.
Toxicity study with recovery period (start on TD15) 20 animals (10 male and 10 female)
Adaptation period: 5 days

Route of administration:

oral: gavage

Details on route of administration:

Route of administration: Oral, by gavage.
Frequency of administration: Once daily.
Administration volume 2 mL/kg b.w./day
Vehicle: Corn oil
Selection of route of administration: According to international guidelines.

Vehicle:

corn oil

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

Main study animals
Males: 2 weeks prior to mating (from test day 15 until test day 29), during
the mating period (from test day 30 until test day 41 at maximum)
and during the post-mating period until and including test day 48
(one day before sacrifice on test day 49) (i.e. 34 treatment days).
Females: (with litter) 2 weeks prior to mating (from test day 15 until test day 29), during
the mating period (from test day 30 until test day 41 at maximum)
and during the lactation period until and including test days 64 to 75
(1 to 3 days before sacrifice on test days 65 to 76) (i.e. 50 to 61 treatment days).

Toxicity study animals (with recovery period)
Males: From test day 15 until and including test day 48 (i.e. 34 treatment days).
Followed by a recovery period from test day 49 until test day 66.
Females: From test day 15 until and including test day 65 (i.e. 51 treatment days).
Followed by a recovery period from test day 66 until test day 80.

Frequency of treatment:

Once daily (from test day 15 (first day of administration) until one day before sacrifice of the last female animal of the main study)

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

Group 1 (Control (vehicle))

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

Group 2

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

Group 3

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

Group 4

No. of animals per sex per dose:

Pre-exposure period: 65 females
A sufficient number in order to obtain at least 40
females with normal oestrus cycles for the main
study and 10 females for the toxicity study.

Main study: 80 animals (40 males and 40 females)
A sufficient number in order to grant at least 8
females per group for evaluation of F0 generation.

Toxicity study with recovery period
20 animals (10 males and 10 females)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dose levels were selected by the Sponsor based on available toxicological data and a 14-day dose range finding study.
In the 14-day dose range finding study, Diisononyl 1,4-cyclohexanedicarboxylate (DINCD) was administered orally to male and female rats at dose levels of 100, 300 or 1000 mg/kg b.w./day for 2 weeks.
No premature deaths, no abnormalities in behaviour, the external appearance or the condition of the faeces and no test item-related influence on body weight or food consumption were noted for the animals of the treatment groups (100, 300 or 1000 mg/kg b.w./day).The macroscopic inspection of the inner organs and tissues revealed no pathological changes. Furthermore, no test item-related influence on the organ weights was noted for the animals of the treatment groups (100, 300 or 1000 mg/kg b.w./day).
Based on the data obtained in this dose range finding study, dose levels of 100, 300 and 1000 mg Diisononyl 1,4-cyclohexanedicarboxylate (DINCD)/kg b.w./day were selected for the present study.

Positive control:

No

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Additionally, once before the first exposure (to allow for within-subject comparisons) and once a week thereafter, detailed clinical observations were made in all adult animals.

BODY WEIGHT: Yes
- Time schedule for examinations:
Males and females were weighed on the first day of dosing, weekly thereafter and at termination. During gestation, females were weighed on days 0, 7, 14 and 20 and within 24 hours of parturition (day 1 post-partum) and on day 4 and 13 post-partum. Body weights were recorded individually for each adult animal.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes
Food intake per rat (g/rat/week) was calculated using the total amount of food given to and left by each rat in each group upon completion of a treatment week (pre-mating and gestation) or treatment period (lactation).

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Drinking water consumption was monitored daily by visual appraisal throughout the study.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood:
Main study animals
5 male and 5 female animals randomly selected from each group: At the end of the premating period on test day 29 (male and female animals)

Toxicity study animals (recovery)
Male animals
(all per group = 5 animals): On test day 66 (day of sacrifice).
Female animals
(all per group = 5 animals): On test day 80 (day of sacrifice).

- Anaesthetic used for blood collection: Yes isoflurane
- Animals fasted: Yes
- Parameters:
Haemoglobin content (HGB)
Erythrocytes (RBC)
Leucocytes (WBC)
Differential blood count (relative and absolute)
Reticulocytes (Reti)
Erythrocytes Platelets (PLT)
Haematocrit value (HCT)
Mean corpuscular volume
Mean corpuscular haemoglobin (MCH)
Mean corpuscular haemoglobin concentration (MCHC)
Thromboplastin time (TPT)
Activated partial thromboplastin time (aPTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: see under hematology
- Animals fasted: Yes
- How many animals: see under hematology
- Parameters checked were examined:

Albumin
Globulin
Albumin/Globulin ratio
Bile acids
Bilirubin (total)
Cholesterol (total)
Creatinine
Glucose
Protein (total)
Blood urea (BUN)

Calcium
Chloride
Potassium
Sodium
Sodium/Potassium ratio
BUN/Creatinine ratio
Lactate dehydrogenase (LDH)
Alanine
aminotransferase
(ALAT)
Alkaline
phosphatase
(aP)
Aspartate
aminotransferase
(ASAT)

URINALYSIS: Yes
- Time schedule for collection of urine:
Main study animals
5 male animals randomly selected from each group. At the end of the premating period on test day 29.

Toxicity study animals
All male animals On test day 29; At the end of the recovery period (test day 66)

- Metabolism cages used for collection of urine: Yes, The urine was collected for 16 hours in a URIMAX funnel cage.
- Animals fasted: Yes
- Parameters checked were examined:
Volume
pH
Specific gravity
Protein
Glucose
Bilirubin
Urobilinogen
Ketones
Haemoglobin (Hb) (approx. values)
Nitrite
Epithelial cells
Leucocytes
Erythrocytes
Organisms
Further constituents (i.e. sperm, casts)
Crystalluria

NEUROBEHAVIOURAL EXAMINATION: Yes
Screening of sensory reactivity to stimuli of different types (e.g. auditory, visual and proprioceptive stimuli) (based on Gad ), as well as the assessment of grip strength (Meyer et. al. ) and motor activity assessment were conducted as described on the following pages in five males and five females randomly selected from each study group.
The screening was conducted two hours after dosing and before any blood sampling for laboratory examinations:
Main study animals (shortly before scheduled sacrifice)
5 parental male animals per group
(randomly selected) On test day 48.

5 parental female animals per group
(randomly selected) Between test days 64 and 73.

Toxicity study animals with recovery period (at the end of the recovery period)
Male animals of the toxicity study
(all per group = 5 animals) On test day 63.

Female animals of the toxicity study
(all per group = 5 animals) On test day 78.


- Battery of functions tested: sensory activity / grip strength / motor activity

IMMUNOLOGY: No

OTHER:
DETERMINATION OF THYROID HORMONS:
1.) Thyroxine (T4):
-Blood samples were taken under isoflurane anaesthesia from animals fasted overnight always at the same time of day, as scheduled below:
Pups(at least 2 surplus pups per litter, all litters): On PND 4 and 13
All evaluated dams/ all adult males: At scheduled sacrifice
-measuring instrument: T4 ELISA (Total Thyroxine (T4) ELISA, IBL INTERNATIONAL cat. no. RE55261; batch nos. 304K096 and 304K096-2; Instrument: Tecan Sunrise)

2.) Thyroid-stimulating hormone (TSH) determination:
-The TSH level were determined in the serum samples (double determination) obtained from pups on PND 13 -
-measuring instrument: The Thyroid-stimulating hormone (TSH (rat)) ELISA, IBL INTERNATIONAL cat. no. RE45021; batch nos. V028 and V031; Instrument: Tecan Sunrise)

REPRODUCTIVE PERFORMANCE (see 7.8.2)

Sacrifice and pathology:

The animals were euthanized by carbon dioxide (CO2) inhalation, exsanguinated by cutting the aorta abdominalis, weighed, dissected and inspected macroscopically (gross necropsy) at the following times:
Main study animals
Males On test day 49 (including 34 treatment days)
Pups On PND 13
Females with litter Between PND 14 and 16
Toxicity study
Males On test day 66 (after a recovery period of 17 test days)
Females On test day 80 (after a recovery period of 14 test days)


GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

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

no effects observed

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

Increased cholesterol concentrations were noted for the male and the female animals in all treatment groups, more pronounced for the female animals than for the male animals. However, as all cholesterol concentrations of the individual female animals were within the LPT background range, this observation was considered as spontaneous and not as test item-related.

Further statistically significant changes that were not considered as test item-related or of toxicological relevance was Alkaline phosphatease (decreased in group 3 females on test day 29).

Urinalysis findings:

effects observed, non-treatment-related

Description (incidence and severity):

However, on test day 29 a slight reduction in the pH value was noted for the main study animals of the intermediate and the high dose group and the high dosed animals of the toxicity study, statistically significant for the main study animals at p ≤ 0.01. As fasting results in a reduction of the pH value of the urine in general, the slightly more pronounced reductions of the pH value that were noted at the high dose group and / or the intermediate dose group in comparison to the control group were considered as spontaneous and in the range of normal variability. This was demonstrated by the pH value that was noted for the animals of the control group at the end of the recovery period on test day 66. This pH value was even slightly below the pH value that was noted for the high dosed animals on test day 29 (pH 6.04 in comparison to 6.10).

Behaviour (functional findings):

no effects observed

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

For the female animals slightly reduced absolute and relative heart weights were noted at the high dose level, statistically significant for the absolute heart weight.
As the reduction of the heart weight at the high dose level in comparison to the control group was only slight and all values of the absolute female heart weight at the high dose level were within the LPT background range, this change was considered as spontaneous and not as test item-related.
Furthermore, no differences in the absolute and the relative heart weight were noted between the female animals of the control and the high dose group of the toxicity study at the end of the recovery period.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

The following changes were considered as spontaneous and not test item-related:
Male rats
- A cyst in the right kidney of animal no. 51 of the intermediate dose group.
- A reddened discoloured thymus of animal no. 78 of the high dose group.

Female rats
- A small adrenal gland from animal no. 61 of the intermediate dose group.
- The non-pregnant animal no. 88 of the high dose group revealed a partly pale liver. Furthermore, the cervix and the left uterus horn were thickened.
- An irregular shaped spleen with a rough surface was noted for animal no. 94 of the high dose group.

Neuropathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Other effects:

effects observed, non-treatment-related

Description (incidence and severity):

Thyroid hormone (T4) determination:
A slight decrease of 15.6% was detected in thyroxine serum levels of the dams after treatment with 1000 mg test item/kg b.w./day. The individual values of the dams showed certain variability. In addition, no dose-response relationship was observed. For this reason, the decrease is considered as spontaneous and not test item-related.
No test item-related differences were noted between the male and female animals of the control group and the male animals of the treatment groups (100, 300 or 1000 mg test item/kg b.w./day).

Key result

Dose descriptor:

NOAEL

Remarks:

systemic toxicity

Effect level:

> 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: highest tested dose

Dose descriptor:

NOAEL

Remarks:

reproductive parameter of the parental females

Effect level:

> 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: highest tested dose

Critical effects observed:

no

Conclusions:

In conclusion, the OECD guideline 422 with the test item Diisononyl 1,4-cyclohexanedicarboxylate (DINCD) revealed that the NOAEL for systemic and reproductive toxicity is above 1000 mg test item/kg b.w./day, p.o..

Executive summary:

The aim of the study was to obtain information on possible effects of the test item on general toxicity,

reproduction and/or development according to OECD guideline 422. The test item Diisononyl 1,4-cyclohexanedicarboxylate (DINCD) was

administered orally to rats at dose levels of 100, 300 or 1000 mg test item/kg b.w./day.

In order to investigate the reversibility of potential signs of general toxicity during

the main study, a toxicity study with a recovery period was additionally performed.

General toxicity - Main study and toxicity study

No premature deaths were noted.

No changes were noted in behaviour, the external appearence or the condition of the faeces.

The neurological screening, the body weight, the food consumption and the

examined haematological, biochemical and urinary parameters revealed no test

item-related differences between the control group and the treatment group.

The final examinations revealed no test item-related findings during the

macroscopic inspection at necropsy, the examination of the organ weights, the

examination of the T4 level and the microscopic examination.

Reproductive toxicity

Parantel - Generation

No test item-related influence was noted on the fertility and the gestation indices,

the pre-coital time and the gestation length.

Pups

No test item-related influence on prenatal development (post-implantation loss,

number of pups born, number of stillbirths, birth and live birth indices) were noted

at any of the tested dose levels.

No test item-related influence onpostnatal development(pup body weight, viability index, the sexual development (ano-genital distance, male nipples counting), gross abnormalities, T₄serum levels of pups on lactation days 4 and 13 and TSH serum levels of pups on lactation day 13) was noted at any of the tested dose levels.

The following no-observed-adverse-effect levels (NOAEL) were established:

General toxicity

NOAEL for systemic toxicity above 1000 mg test item/kg b.w./day, p.o.

Reproductive toxicity

a) for reproductive parameters of the parental females

NOAEL above 1000 mg test item/kg b.w./day, p.o.

b) for pre- and postnatal development

- b1) adverse effects on prenatal development (conceptus to birth)

NOAEL above 1000 mg test item/kg b.w./day, p.o.

- b2) adverse effects on postnatal development (pup)

NOAEL 1000 mg test item/kg b.w./day, p.o.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

No classification for specific target organ toxicity repeated exposure (STOT RE) is indicated according to the classification, labeling, and packaging (CLP) regulation (EC) No 1272/2008.