Esterification products of fatty acids, C18 (unsaturated) alkyl and adipic acid with pentaerythritol

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

- combined repeat dose toxicity study with the reproduction/developmental screening test (OECD 422): NOAEL (reproduction) >= 1000 mg/kg bw/day (m/f)

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 Feb - 13 Oct 2022

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted in 2016

Deviations:

yes

Remarks:

Minor deviations in pup identification, access to water, dosing, clinical observations, food consumption, body weight, and organ weights. No impact to the overall integrity of the study or the interpretation of the study results and conclusions.

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 11 - 12 weeks for males; 13 - 14 weeks for females
- Weight at study initiation: 271 - 317 g for males; 190 - 240 g weeks for females
- Fasting period before study: no
- Housing: Up to 5 animals of the same dosing group together in polycarbonate cages (Makrolon, MIV type, height 18 cm)
- Diet: SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany; Pellets ad libitum, except during designated procedures.
- Water: Municipal tap water; Freely available to each animal via water bottles. One female did not have access to water for a maximum of 24 h.
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 24
- Humidity (%): 40 - 70
- Air changes (per hr): >= 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: 21 Feb - 21 Jun 2022

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Dose formulations were divided into aliquots and where dispensed on each dosing occasion. The test item was formulated (w/w) and homogenized to visually acceptable levels. It was formulated at least weekly and filled out in daily portions. The dose formulation was stored at 4 °C.

VEHICLE
- Justification for use and choice of vehicle: Stability analyses performed previously in conjunction with the method development and validation study (Test Facility Study No. 20326206) demonstrated that the test item was stable in the vehicle when prepared and stored under the same conditions at concentrations bracketing those used in the present study.
- Amount of vehicle: The dose volume for each animal was based on the most recent body weight measurement. The dose formulations were stirred continuously during dose administration and doses were given using a plastic feeding tube.

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: Up to 14 days
- Detection of mating: Evidence of sperm in the vaginal lavage or by the appearance of an intravaginal copulatory plug. This day was designated Day 0 post-coitum.
- After successful mating each pregnant female was caged: individually

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analyses were performed using a laboratory validated analytical procedure.

The concentrations analyzed in the formulations of Group 2, Group 3 and Group 4 were in agreement with target concentrations (i.e., mean sample concentration results were within or equal to 90 - 110% of target concentration). No test material was detected in the Group 1 formulation.
The formulations of Group 2 and Group 4 were homogeneous (i.e., coefficient of variation ≤ 10%). Homogeneity of Group 3 was not analyzed.

Duration of treatment / exposure:

Males: 7 days a week for a minimum of 28 days, including at least 2 weeks of treatment prior to mating and during the mating period (up to and including the day before scheduled necropsy).

Females: 7 days a week for at least 14 days prior to mating (with the objective of covering at least two complete oestrous cycles), the variable time to conception, the duration of pregnancy and at least 13 days after delivery, up to and including the day before scheduled necropsy. Females were not be dosed during littering.

Pups: Not to be treated directly but could potentially be exposed to the test item in utero, via maternal milk, or from exposure to maternal urine/feces.

Frequency of treatment:

once daily

Dose / conc.:

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

Dose / conc.:

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

Dose / conc.:

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

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on the results of the Dose Range Finder (Test Facility Reference No. 20326207).
- Fasting period before blood sampling for clinical biochemistry: All males surviving to scheduled necropsy were fasted overnight with a maximum of 24 h before necropsy. Females were not be fasted before necropsy.

Positive control:

No

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once before the first administration of the test item and weekly during the treatment period (observed for mortality at least twice daily beginning upon arrival through termination/release; except on days of receipt and necropsy where frequency was at least once daily).

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: At least once daily, up to the day prior to necropsy. No clinical observations were recorded on dosing Day 2 for two animals and dosing Day 3 for one animal.

BODY WEIGHT: Yes
- Time schedule for examinations: On Day 1 of treatment (prior to dosing) and weekly
thereafter. Mated females: On Days 0, 4, 7, 11, 14, 17, and 20 post-coitum and during lactation on PND 1, 4, 7, and 13.
The terminal body weight of one female was not recorded.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Food consumption for one female was not determined during post-coitum Day 0 - 4. One male was dosed on the day of necropsy.

WATER CONSUMPTION AND COMPOUND INTAKE: No

OTHER:
HEMATOLOGY AND COAGULATION:
- Time schedule for collection of blood: On the day of scheduled necropsy
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Only for males
- How many animals: 5/sex/group
- Parameters checked: white blood cells, neutrophils, lymphocytes, monocytes, eosinophils, basophils, large unstained cells, red blood cells, reticulocytes, red blood cell distribution, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelets, prothrombin time, activated partial thromboplastin time

CLINICAL CHEMISTRY:
- Time schedule for collection of blood: On the day of scheduled necropsy
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Only for males
- How many animals: 5/sex/group
- Parameters checked: alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total protein, albumin, total bilirubin, bile acids, urea, creatinine, glucose, cholesterol, sodium, potassium, chloride, calcium, inorganic phosphate

THYROID HORMONE:
- Time schedule for collection of blood: On the day of scheduled necropsy
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Only for males
- How many animals: 5/sex/group
- Parameters checked: thyroxine (T4), thyroid-stimulating hormone (TSH)

Oestrous cyclicity (parental animals):

Oestrous cycles was evaluated by examining the vaginal cytology of samples obtained by serial vaginal lavage procedures. Daily vaginal lavage was performed for all females beginning 14 days prior to treatment (pretest period), the first 14 days of treatment and during mating until evidence of copulation was observed. Vaginal lavage was continued for those females with no evidence of copulation until termination of the mating period. On the day of necropsy, a vaginal lavage was taken to determine the stage of oestrus. This was done for all females, except for females that had to be euthanized in extremis or died spontaneously.

Sperm parameters (parental animals):

Parameters examined in male parental generations: Testis and epididymis weight.
Testes were evaluated to assess the progression of stages of the spermatogenic cycle, cell associations, and proportions expected to be present during spermatogenesis along with assessment of interstitial and supporting cell types (Leydig cells, macrophages, vasculature, and rete testis).

Litter observations:

STANDARDISATION OF LITTERS
- Performed on PND 4: yes
- If yes, a maximum of eight pups from each litter of equal sex distribution (if possible) will be selected to reduce variability among the litters.

PARAMETERS EXAMINED
The following parameters were examined in offspring:
Mortaility, clinical observations, body weights, sex, anogenital distance, areola/nipple retention, thyroid hormone. Two pups were given the same identification from PND 1 to PND 4.

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: no

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY: no

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals after a minimum of 28 days of administration.
- Maternal animals: All surviving animals PND 14 - 16 or failure to deliver.

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations.

HISTOPATHOLOGY / ORGAN WEIGHTS
The following tissues were weighed: brain, epididymis, adrenal gland, parathyroid gland, prostate gland, seminal vesicle, thyroid gland, heart, kidney, liver, ovaries, spleen, testes, thymus, uterus/cervix
Organ weights of one male and brain weight of one male were not recorded.

The following tissues were prepared for microscopic evaluation: bone marrow (sternum), femur, sternum, brain, epididymis, eye, adrenal gland, mammary gland, pituitary gland, prostate gland, seminal vesicle, thyroid gland, gut associated lymphoid tissue, heart, kidney, cecum, colon, rectum, liver, lung, mandibular lymph node, mesenteric lymph node, skeletal muscle, sciatic nerve, ovaries, duodenum, ileum, jejunum, spinal cord, spleen, stomach, testes, thymus, trachea, urinary bladder, uterus/cervix, vagina

Postmortem examinations (offspring):

SACRIFICE
On PND 4, the surplus pups were euthanized by decapitation. All remaining pups were euthanized on PND 14-16.

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations.

Reproductive indices:

The following indices were calculated:
Mating index = (number of females mated / number of females paired) * 100
Precoital time = number of days between initiation of cohabitation and confirmation of mating
Fertility index = (number of pregnant females / number of females mated) * 100
Gestation index = (number of females with living pups on Day 1 / number of pregnant females) * 100
Duration of gestation = number of days between confirmation of mating and the beginning of parturition

Offspring viability indices:

The following indices were calculated:
Post-implantation survival index = (number of offspring born / number of uterine implantation sites) * 100
Live birth index = (number of live offspring on Day 1 after littering / number of offspring born) * 100
Percentage live males at first litter check = (number of live male pups at first litter check / number of live pups at first litter check) * 100
Percentage live females at first litter check = (number of live female pups at first litter check / number of live pups at first litter check) * 100
Viability index = (number of live offspring on Day 4 before culling / number of live offspring on Day 1 after littering) * 100
Lactation index = (number of live offspring on Day 13 after littering / number of live offspring on Day 4 after culling) * 100

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

No test material-related clinical signs were noted during daily detailed clinical observations or during weekly arena observations.
Clinical signs noted during the treatment period occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study and did not show any apparent dose-related trend. At the incidence observed, these were considered to be unrelated to treatment with the test material.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Description (incidence):

No mortality occurred during the study period.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Body weights and body weight gain of treated animals remained in the same range as controls over the treatment period.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Food consumption before or after correction for body weight of treated animals was similar to the control level over the treatment period.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

No test material-related changes were noted in hematological parameters in males up to 300 mg/kg bw/day and females up to 1000 mg/kg bw/day. In males at 1000 mg/kg bw/day, lower neutrophil count was noted. No corroborating pathological findings were observed, and therefore this change was considered non-adverse.
Remaining differences in hematology parameters, regardless of statistical significance, were considered not test material-related based on the absence of a dose response or general overlap of individual values with the range of control values.
Coagulation parameters of treated rats were considered not to have been affected by treatment with the test material.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

No test material-related changes were noted in clinical chemistry parameters in males up to 300 mg/kg bw/day.
The following changes in clinical biochemistry parameters distinguished treated animals from
control animals: Higher urea levels in males at 1000 mg/kg bw/day; Higher total bilirubin (not statistically significant) for females at 100, 300 and 1000 mg/kg bw/day. No corroborating pathological findings were observed, and therefore these changes were considered non-adverse.
Any other significant changes were considered unrelated to the treatment in the absence of a dose-related response or because individual values were within range of control values.

Thyroid hormone
Serum levels of T4 in males were considered unaffected by treatment with the test material up to 1000 mg/kg bw/day.

Endocrine findings:

not specified

Description (incidence and severity):

The following ED-related parameters were investigated in the study:
- thyroid hormones
- accessory sex organ weight and histopathology
- anogenital distance
- sex organ weight and histopathology
- oestrous cyclicity
- genital abnormalities
- nipple development
- thyroid weight and histopathology
- vaginal smears
- adrenal weight and histopathology
- brain weight
- fertility
- fetal development
- gestation length
- litter size
- litter viability
- litter / pup weight
- number of implantations, corpora lutea
- number of live births
- pituitary histopathology
- post-implantation survival
- reproduction
- sex ratio

For details, please refer to the respective result fields and the endpoint summary.

Urinalysis findings:

not examined

Behaviour (functional findings):

effects observed, non-treatment-related

Description (incidence and severity):

Hearing ability, pupillary reflex, static righting reflex and grip strength were considered not to have been affected by treatment with the test material.
Lower hind leg grip strength was noted in females at 100, 300, and 1000 mg/kg bw/day (not always statistically significant). In the absence of a dose-relationship and as mean values are within the historical control range, this was considered not related to treatment with the test material.
Motor activity was considered not to be affected by treatment with the test material. All groups showed a similar motor activity habituation profile with a decreasing trend in activity over the duration of the test period.
In females at all dose levels mean total movements and ambulations were lower (not statistically significant), which could be attributed to one or two animals per group. In the absence of clear dose relationship and as the control means were slightly high, this was considered unrelated to the treatment with the test material.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

There were no test material-related microscopic observations.
All the recorded microscopic findings were within the range of background pathology encountered in rats of this age and strain. There was no test material-related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

Length and regularity of the oestrous cycle were considered not to have been affected by treatment with the test material.
All females had a regular cycle of 4 days.

Reproductive function: sperm measures:

not examined

Reproductive performance:

effects observed, non-treatment-related

Description (incidence and severity):

There was 1/10 control couples and 1/10 couples of the of the 100 mg/kg bw/day group with no offspring. No abnormalities were seen in the reproductive organs, which could account for their lack of offspring.
There were no morphological findings in the reproductive organs of either sex which could be attributed to the test material. Stage dependent qualitative evaluation of spermatogenesis in the testis was performed. The testes revealed normal progression of the spermatogenic cycle and the expected cell associations and proportions in the various stages of spermatogenesis were present.
The length and regularity of the oestrous cycle, mating index, precoital time, and number of implantation sites were considered not to have been affected by treatment with the test material.

Dose descriptor:

NOAEL

Remarks:

general toxicity

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed up to and including the highest dose tested

Dose descriptor:

NOAEL

Remarks:

reproductive toxicity

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed up to and including the highest dose tested

Critical effects observed:

no

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

No clinical signs occurred among pups surviving until scheduled necropsy that were considered to be related to treatment with the test material.
Pale appearance was noted for one pup at the high dose (1000 mg/kg bw/day) on PND 1-4 (the pup was culled on PND 4). Laboured respiration and gasping was noted for one pup (300 mg/kg bw/day) on PND 2. The nature and incidence of these clinical signs remained within the range considered normal for pups of this age, and were therefore considered not to be test material-related.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

Live birth index (number of live offspring on PND 1 as percentage of total number of offspring born) was considered not to be affected by treatment with the test material. The live birth indices were 99, 99, 98, and 100% for the control, 100, 300 and 1000 mg/kg bw/day groups, respectively.
One pup of the control group, one pup at 100 mg/kg bw/day (with no milk in the stomach), and two pups at 300 mg/kg bw/day were found dead at first litter check. These dead pups were considered to be unrelated to treatment with the test material, since the mortality incidence did not show a dose-related trend and remained within the range considered normal for pups of this age.

Viability index (number of live offspring on PND 4 before culling as percentage of number of live offspring on PND 1) was considered not to be affected by treatment with the test material. Viability indices were 99% for all groups.
One pup of the control group, and one pup each at 100, 300 and 1000 mg/kg bw/day were missing on PND 2. Pups missing were most likely cannibalized. These missing pups were considered to be unrelated to treatment with the test material, since the mortality incidence did not show a dose-related trend and remained within the range considered normal for pups of this age.

The number of live offspring on Day 13 after littering compared to the number of live offspring on Day 4 (after culling) was considered not to be affected by treatment with the test material. No pups were found dead/missing between lactation Days 5 and 13, resulting in a lactation index of 100% for all groups.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Body weights of pups were considered not to be affected by treatment with the test material.

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

Serum T4 levels in male and female pups were considered not to be affected by treatment with the test material.

Urinalysis findings:

not examined

Sexual maturation:

not examined

Anogenital distance (AGD):

no effects observed

Description (incidence and severity):

Anogenital distance (absolute and corrected for body weight) in male and female pups was considered not to be affected by treatment with the test material.

Nipple retention in male pups:

effects observed, non-treatment-related

Description (incidence and severity):

Treatment with the test material up to 1000 mg/kg bw/day had no effect on areola/nipple retention.
Two control male pups and one male pup (1000 mg/kg bw/day) had one nipple on PND 13. Two pups (1000 mg/kg bw/day) had one or two nipples. These findings remained within the range considered normal for pups of this age, and were therefore considered not to be test material-related.

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No macroscopic findings were noted among pups surviving until scheduled necropsy that were considered to be related to treatment with the test material.
The nature and incidence of macroscopic findings remained within the range considered normal for pups of this age, and were therefore considered not to be related to treatment with the test material.

Histopathological findings:

not examined

Other effects:

effects observed, non-treatment-related

Description (incidence and severity):

Gestation index (females with living pups on Day 1 compared to the number of pregnant females) and duration of gestation were considered not to be affected by treatment with the test material.
The gestation indices were 100% for all groups.

No signs of difficult or prolonged parturition were noted among the pregnant females.
Examination of cage debris of pregnant females revealed no signs of abortion or premature birth. No deficiencies in maternal care were observed.

The total number of offspring born compared to the total number of uterine implantations was considered not to be affected by treatment with the test material.
Post-implantation survival index (total number of offspring born as percentage of total number of uterine implantation sites) was 98, 95, 93 and 94% for the control, 100, 300 and 1000 mg/kg bw/day groups, respectively.
For one female (300 mg/kg/day), the number of pups was slightly higher than the number of implantations. This phenomenon is observed from time to time and is caused by normal resorption of these areas during lactation.

Litter size was considered not affected by treatment with the test material.
Live litter sizes were 13.0, 14.0, 12.6 and 12.1 living pups/litter for the control, 100, 300 and 1000 mg/kg bw/day groups, respectively.

Sex ratio was considered not to be affected by treatment with the test material.

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

not examined

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed up to and including the highest dose tested

Critical effects observed:

no

Reproductive effects observed:

no

Conclusions:

The test substance had no effect on both, reproductive performance and pup development. The No-Observed-Adverse-Effect-Levels (NOAELs) for reproductive and developmental toxicity were, therefore, concluded to be >/= 1000 mg/kg bw/day.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.7, of Regulation (EC) No. 1907/2006.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test

The repeat dose and reproductive/developmental toxicity of esterification products of fatty acids, C18 (unsaturated) alkyl and adipic acid with pentaerythritol was investigated in a study performed according to OECD guideline 422 under GLP conditions (Charles River, 2022e). The registration substance was administered once daily, 7 days/week, via gavage to groups of 10 Wistar rats per sex at doses of 100, 300, and 1000 mg/kg bw in corn oil. A control group received the vehicle alone. Males were treated for at least 28 days, including at least 2 weeks of treatment prior to mating and during the mating period (up to and including the day before scheduled necropsy). Females were treated for at least 14 days prior to mating (with the objective of covering at least two complete oestrous cycles), the variable time to conception, the duration of pregnancy and at least 13 days after delivery, up to and including the day before scheduled necropsy. Reproduction related parameters included oestrous cycle evaluations and reproduction data to calculate mating index, precoital time, fertility index, gestation index, and duration of gestation. Parameters and results in relation to repeated dose toxicity are provided in the endpoint summary in IUCLID section 7.5.

None of the reproduction related parameters were affected by treatment with the test material. The registered substance did not demonstrate any reproductive toxicity potential.

Based on the available data, no changes that could be considered adverse were observed at any of the dose level investigated. Therefore, the No Observed Adverse Effect Level (NOAEL) for reproductive toxicity was concluded to be 1000 mg/kg bw/day, for both male and female parental animals (for information on developmental toxicity please refer to developmental toxicity section below).

Effects on developmental toxicity

Description of key information

- combined repeat dose toxicity study with the reproduction/developmental screening test (OECD 422): NOAEL (developmental) >= 1000 mg/kg bw/day (m/f)

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.7, of Regulation (EC) No. 1907/2006.

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

Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test

As already mentioned under the repeated dose toxicity endpoint summary in IUCLID section 7.5 and above in 'Effects on fertility’, a combined repeated dose toxicity study with the reproduction/developmental toxicity screening test with esterification products of fatty acids, C18 (unsaturated) alkyl and adipic acid with pentaerythritol was performed according to OECD guideline 422 under GLP conditions (Charles River, 2022e). The study design and dosing regime have already been provided elsewhere in the dossier. The specific developmental parameters included post-implantation survival index, litter size, live birth index, viability index, and lactation index and clinical signs, body weights, sex ratio, thyroid hormone, areola/nipple retention, anogenital distance, and macroscopic findings in the pups.

No treatment-related effects were seen in post-implantation survival index, litter size, live birth index, viability index, lactation index, clinical signs, body weights, sex ratio, anogenital distance, and macroscopic findings. Treatment with the test material up to 1000 mg/kg bw/day had no effect on areola/nipple retention. Serum T4 levels in male and female pups were considered not to be affected by treatment with the test material. The registered substance did not demonstrate any teratogenic potential.

Based on the available data, no changes that could be considered adverse were observed at any of the dose level investigated. Therefore, the No Observed Adverse Effect Level (NOAEL) for developmental toxicity was concluded to be 1000 mg/kg bw/day, for both male and female pups (for detailed information on reproductive toxicity please refer to the section ‘Effects on fertility’ above).

Justification for classification or non-classification

According to Regulation (EC) No. 1272/2008 the data on reproductive toxicity are conclusive but not sufficient for classification.