Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

In the combined repeated dose and reproductive/ developmental toxicity screening test according to OECD guideline 422 with the read across substance Pigment Yellow 53, no reproductive toxicity was observed and a NOAEL >= 1000 mg/kg bw/d was determined.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Qualifier:
equivalent or similar 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:
SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: 4879
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crj; CD(SD)
Details on species / strain selection:
This strain was selected because this species is usually used in this kind of test and the laboratory has a high experience with this rat strain.
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc. Atsugi 6/27 breeding center
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 10 wks
- Weight at study initiation: Males: 335 - 386 g; Females: 208 - 249 g
- Housing: individually (except for acclimatisation and mating period)
- Diet: ad libitum, γ-ray irradiation fixed feed CRF-1 (Oriental Yeast Co., Ltd)
- Water: ad libitum, tap water
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 3 (actual measured range of 20 - 24 °C)
- Humidity (%): 55 ± 10 (actual measured range of 45 - 66 %)
- Air changes (per hr): 10 to 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
other: 1% carmellose sodium aqueous solution
Remarks:
hereinafter referred to as 1% CMC-Na aqueous solution
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
After grinding the test substance in a mortar, it was weighed accurately per dose, suspended in 1% CMC-Na aqueous solution to receive a predetermined concentration, and was dispersed using an ultrasonic device.

VEHICLE
- Concentration in vehicle: 10, 100, 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Lot/batch no. : 9318
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: 1 d
- Proof of pregnancy: vaginal plug or sperm in vaginal smear
- M/F ratio per cage: 1/1
- Length of cohabitation: 14 d
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): individually
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
No details on the analytical measurements were given.
The analysis of the stability performed prior to administration for the prepared solutions of concentrations of 10 and 200 mg/mL, confirmed that the test substance is stable for 24 hours at room temperature and for 7 days at refrigeration. The prepared solutions were used within 7 days after preparation. Further, the uniformity of the prepared solutions of 10, 100 and 200 mg/mL was confirmed.
The concentration of the test substance in the prepared solutions of each concentration of the initial and final preparation was analysed. The content was 98.3 to 101.7 % of the prescribed concentration, and therefore was regarded as appropriate.
Duration of treatment / exposure:
Males: 46 days
Females: 42-46 days
Frequency of treatment:
Once daily
Dose / conc.:
250 mg/kg bw/day (nominal)
Dose / conc.:
500 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
12
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The doses of the test substance were set on the basis of the results of the preliminary test for the Combined Repeated Dose and Reproductive/Developmental Toxicity Screening Test of C.I. Pigment Yellow 53 in Rats (Test No. SR-9984P). In the preliminary test, the test substance was administered once a day for 14 days at doses of 0, 500, 1000, and 2000 mg/kg to five male and five female SD rats per group. No abnormalities beside yellow feces were observed. Therefore the high dose in the main test was set to 1000 mg/kg bw/d and the lower doses were set at 500 and 250 mg/kg by dividing by a common ratio of 2, for both males and females.
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations checked: mortality, behaviour, external appearance

BODY WEIGHT: Yes
- Time schedule for examinations:
Males: administration days 1, 2, 3, 5, 7, 10 and 14; then weekly until necropsy
Females: administrations days 1, 2, 3, 5, 7, 10 and 14; pregnancy days 0, 1, 3, 5, 7, 10, 14, 17 and 20 and lactation days 0, 1 and 4

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
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at necropsy
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes
- How many animals: all male animals; 6 lactating females per dosing group
- Parameters examined: Number of red blood cells (RBC), Hematocrit (Ht), Hemoglobin (Hb), Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular hemoglobin concentration (MCHC), Reticulocyte count (Ret), Platelet count (Plat), White blood cell count (WBC), White blood cell percentage (Hemogram of WBC), Prothrombin time (PT), Activated partial thromboplastin time (APTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at necropsy
- Animals fasted: Yes
- How many animals: all male animals; 6 lactating females per dosing group
- Parameters examined: GOT, GPT, Alkaline phosphatase (ALP), γ-GTP, Glucose (Glu), Choline esterase (ChE), Total cholesterol (T-Cho), Phospholipid (PL), Triglyceride (TG), Total bilirubin (T-Bil), Urea nitrogen (UN), Creatinine (Crea), Sodium (Na), Potassium (K), Chloride (Cl), Calcium (Ca), Inorganic phosphorus (IP), Total protein (TP), A/G ratio (A/G)

URINALYSIS: Yes
- Time schedule for collection of urine: on administration days 44 - 45
- Metabolism cages used for collection of urine: Yes
- Animals fasted: No
- Parameters examined: pH, protein, glucose, ketone bodies, urobilinogen, bilirubin, occult blood, sediment, color, volume, specific gravity
Oestrous cyclicity (parental animals):
For all females, every day from 10 days prior to dosing until successful copulation, vaginal smear samples with Giemsa staining were prepared and their estrous cycle phases (proestrus, estrus, metestrus, diestrus) were determined under an optical microscope and a cycle of repeating each phase of estrous cycle two or more times at an interval of 4 to 5 days was regarded as normal. Continuation of diestrus for 7 days or more was judged to be an abnormal estrous cycle. Estrus interval was calculated from estrous cycle of each female.
Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in F1:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, viability index, body weight and necroscopy

Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals (on the day after administration Day 46)
- Maternal animals: All surviving animals (until Day 5 post partum)

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated below were prepared for microscopic examination:
Brain (cerebrum, cerebellum), pituitary, thymus, thyroid (left and right), parathyroid (left and right), adrenal gland (left and right), spleen, heart, thoracic aorta, tongue, esophagus, stomach (forestomach and glandular stomach), liver, pancreas, duodenum, jejunum, ileum (including Peyer's patches), cecum, colon, rectum, larynx, trachea, lung (including bronchial), kidney (left and right), bladder, testes (left and right), epididymis (left and right), prostate, seminal vesicles (including coagulation glands, left and right), eye (left and right), harderian gland (left and right), skin (right abdomen), sternum (including bone marrow), femur (including bone marrow, right), spinal cord (neck), skeletal muscle (right thigh), mesenteric lymph node, mandibular lymph nodes (left and right), submandibular gland (left and right), sublingual gland (left and right), parotid gland (left and right) and the sciatic nerve (right).

The following organs were weighed:
Brain, heart, liver, kidney (left and right), spleen, adrenal glands (left and right), thymus, testis (left and right) and epididymis (left and right), ovary (left and right)
Postmortem examinations (offspring):
SACRIFICE
- The offspring was sacrificed at 4 days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of external examination (including the inside of the mouth) and internal examinations including the cervical, thoracic, and abdominal viscera
Statistics:
Please refer to "Any other information on materials and methods incl. tables"
Reproductive indices:
Estrous cycle, copulation index (number of pairs with successful copulation/number of pairs mated X 100), fertility index (number of pregnant animals/number of pairs with successful copulation X 100), gestation index (number of females with live pups/number of living pregnant females X 100), gestation length, nursing index, number of pregnant females, corpora lutea and implantation sites, implantation index (number of implantation sites/number of corpora lutea X 100), delivery index (number of pups born/number of implantation sites X 100),
Offspring viability indices:
Number of live pups on day 4/number of live pups on day 0 X 100
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Males: After administration day 2 until the necropsy day, yellowish brown feces in the 500 mg/kg group, and yellow or yellowish brown feces in the 1000 mg/kg group were observed in all of 12 rats of each group. In addition to this, fracture of incisor teeth was observed in one animal of the 250 mg/kg group on administration day 42, but this was considered accidental.

Females: After administration day 2 until the necropsy day, yellowish brown feces in the 500 mg/kg group, and yellow or yellowish brown feces in the 1000 mg/kg group were observed in all 12 rats of each group. In addition, alopecia was observed in one female in the control group after day 12 of pregnancy (5 days after delivery) until the necropsy day. The scab-formation on the back in the 250 mg/kg group and the trauma and scab-formation in the lower eyelid in the 500 mg/kg group were observed each in one female. These effects were not considered as toxicologically relevant.
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:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Males: In the 250 and 1000 mg/kg groups, a significant shortening of the activated partial thromboplastin time (APTT) was observed.
Females: In the 250 mg/kg group, significant high values of mean corpuscular hemoglobin (MCH) and platelet were observed, but these changes were not observed in the higher dosing groups.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Males: A significant low value of potassium (K) was observed in the 1000 mg/kg group, but it was considered to have no toxicological significance.
Females: Compared with the control group, a significant low value of cholinesterase (ChE) in the 250 mg/kg group, a significant low value of sodium (Na) in the 500 mg/kg group, and a significant low value of GPT in the 1000 mg/kg group were observed, but, since there were no changes found in the related organs, this was considered as non treatment related.
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
In the 250 mg/kg group, a significant increase was observed in urine specific gravity, but this was not observed in the higher dosing groups. No other urine parameter was altered compared to the control group.
Behaviour (functional findings):
not examined
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):
Males: In the control group, myocardial degeneration of the heart, hyaline cast of the kidney, eosinophilic bodies of the proximal tubule epithelium, atrophy of the seminiferous tubules of testes and sperm granuloma of the epididymis were observed each in one male. Lymphocyte infiltration of the prostate was found in six other males. In the 1000 mg/kg group, myocardial degeneration of heart in three males, eosinophilic bodies of the proximal tubule epithelium in one male, atrophy of the seminiferous tubules of testes in three males, intraluminal cell debris of the epididymis in one male, and lymphocyte infiltration of the prostate in four males were observed.
No other significant differences were observed compared to the control group.
Females: In the control group, erosion of the tongue was observed in three animals, and focal necrosis of the liver, kidney cysts, dilatation of the renal pelvis, dilatation of the renal tubules, focal fibrosis of the kidney, mineralization in the cortico-medullary junction of the kidney, granuloma of the cervix, atrophy of the thymus, atrophy of the skin hair follicles and inflammation of the mammary gland were observed each in one case. In the 1000 mg/kg group, erosion of the tongue was observed in one animal, and mineralization in the cortico-medullary junction of the kidney was observed in three females. In addition, the atresia of vagina and the inflammation of the uterine horn were observed in one animal in which the atresia of vagina was seen at necropsy (infertility example). The atresia of vagina was a congenital anomaly and it was considered as the cause of infertility.
As for all the above changes, no significant difference was observed in the frequency of appearance, as compared to the control group.
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
not examined
Reproductive performance:
effects observed, non-treatment-related
Description (incidence and severity):
As for the incidence of females with normal estrous cycle, estrus interval, copulation index, fertility index, gestation index, length of gestation, and nursing index on lactation day 4, there was no significant difference observed in each dose group, compared to the control group. Unsuccessful copulation was observed in the 250 and 500 mg/kg groups, each in one pair. In the estrous cycle inspection of females of these pairs, continuous diestrus was observed. But from the fact that it was not observed in the 1000 mg/kg group, it was not considered as substance related. One non-pregnant female was observed in the 1000 mg/kg group. For this female, the atresia of vagina was observed in the pathological examination. This was assumed to cause infertility and it was considered to be the cause of infertility.
As for the number of corpora lutea, number of implantation sites, implantation index, total number of pups born, the sex ratio of pups born, number of live pups on lactation day 0, live birth index, number of live pups on lactation day 4, newborn viability index, there was no significant difference observed in each administration group, compared with the control group. Delivery index was significantly decreased in the 250 mg/kg group, but this was considered as an incidental variation since there was no significant difference observed in the dose groups of 500 mg/kg or more.
Key result
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects observed in the parental generation.
Key result
Critical effects observed:
no
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Except for the scab observed in one male of the 1000 mg/kg group, there was no abnormality observed in any of the dead pups.
Yellowish white portion of the liver was observed in one male of 250 mg/kg group and in one male of the 1000 mg/kg group, for the pups necropsied on day 4 of lactation. Trauma was noted for one female of the 500 mg/kg group. But no other abnormalities were observed.
Histopathological findings:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
No effects observed in the offspring.
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
>= 1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effects observed in the offspring.
Key result
Critical effects observed:
no
Key result
Reproductive effects observed:
no
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Guideline compliant study report
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 a combined repeated dose and reproductive/ developmental toxicity screening test according to OECD guideline 422 (MHLW, 2002), the structural analogue C.I. Pigment Yellow 53 was administered to groups of Sprague Dawley rats (12/sex) at concentrations of 0 (vehicle), 250, 500, 1000 mg/kg bw/d. Males were treated for 46 days and females from 14 days before mating to day 4 of lactation.

No changes were found in female estrous cycle, male and female copulation index, conception index, delivery index, gestation length, and nursing index on day 4 of lactation, and no changes were observed in the weight of the genital organs (testis, epididymis and ovary) and endocrine organ (adrenal glands) as well as in the necropsy thereof, in each dosing group. In addition, abnormalities were not observed in the histopathological examination of the genital organs in the 1000 mg/kg group.

From the above, no effects of test substance administration on the male and female reproduction were observed in any of the dosing groups. Therefore, the no-observed-adverse-effect level (NOAEL) of test substance administration with repeated dose under this test conditions for the reproduction of parent animals was considered to be 1000 mg/kg/day or more.

Effects on developmental toxicity

Description of key information

The test compound was tested for its prenatal developmental toxicity in Wistar rats (OECD 414, GLP) at concentrations of 100, 300 and 1000 mg/kg bw administered by gavage. No test substance-related adverse effects on dams, gestational parameters or fetuses were seen at any dose level. The no observed adverse effect level (NOAEL) for maternal and prenatal developmental toxicology is 1000 mg/kg bw/d.

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Age at study initiation: About 11-13 weeks
- Fasting period before study: no
- Housing: single
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: From GD 0 (day of supply) to the beginning of administration (GD 6), the animals will be accustomed to the environmental
conditions and to the diet.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on exposure:
Route of administration: Orally by gavage
Reason for the selection of the route of administration: The oral administration of a test substance has been proven useful worldwide in numerous studies for discovering a potential toxicological profile.
Frequency of administration/number of administrations: Once daily (GD 6-19)/14
Volume to be administered: 10 ml/kg body weight; the body weight determined most recently will be used to calculate the administration volume.
Preparation: For the test substance preparations, the specific amount of test substance will be weighed, topped up with 0.5% Carboxymethylcellulose suspension in drinking water in a calibrated beaker and intensely mixed with a homogenizer. Before and during administration, the preparations will be kept homogeneous with a magnetic stirrer.
Preparation frequency: daily
Storage conditions of the preparations: RT
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The analytical investigations of the test substance preparations will be carried out as a
separate study at the test facility Competence Center Analytics of BASF SE, 67056
Ludwigshafen, Germany, under the responsibility of the study director of this test facility.
The study will be carried out in compliance with the Principles of Good Laboratory
Practice.
Details on mating procedure:
The animals are paired by the breeder and will be supplied at noon on the day of evidence of mating; this day is referred to as GD O and the following day as GD 1.
Duration of treatment / exposure:
The day of evidence of mating is referred to as GD O and the following day as GD 1.
The test substance will be administered to the animals orally by gavage from GD 6 through GD 19
Frequency of treatment:
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:
25
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on information of the combined reprotox / repeated dose study and the 90d study, 100, 300 and 1000 mg/kg bw were chose.

The random distribution of the animals to the individual groups will be carried out on the day of supply (= GD 0) by randomly removing the animals from the transport boxes.

On GD 20, all surviving dams will be sacrificed and examined. All evaluations except for the gross-pathological evaluation and the determination of the uterus weights will be carried out by technicians unaware of the treatment groups using coded animal numbers. The fetuses will be removed from the uterus, which has been opened before and examined.
Maternal examinations:
Mortality
A check for moribund and dead animals will be made twice daily from Mondays to Fridays and once daily on Saturdays, Sundays and public holidays (GD 0 to 20).

Clinical signs
A cageside examination will be conducted at least once daily for any signs of morbidity, pertinent behavioral changes and/or signs of overt toxicity.
During the administration period (GD 6 - 19) all animals will be checked daily for any abnormal clinically signs before the administration as well as within 2 hours and within 5 hours after the administration. Abnormalities and changes will be documented for each animal.

Food consumption
Food consumption will be recorded for GD 0-1, 1-3, 3-6, 6-8, 8-10, 10-13, 13-15, 15-17, 17-19 and 19-20.

Body weight
Body weights will be recorded on GD 0, 1, 3, 6, 8, 10, 13, 15, 17, 19 and 20.

POST-MORTEM EXAMINATIONS
On GD 20, the surviving dams will be anesthetized with isoflurane, sacrificed by cervical dislocation in a randomized sequence and examined. The fetuses will be removed from the uterus, which has been opened before and examined.
Moribund animals and animals that die intercurrently will be examined if possible (with the exception of the uterus weight).
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes / No / No data
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
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
Statistics:
Means and standard deviations will be calculated. In addition, the following statistical analyses will be carried out:

Food consumption, body weight, body weight change, corrected body weight gain, carcass weight, weight of the unopened uterus, weight of the placentas and
fetuses, corpora lutea, implantations, pre- and postimplantation losses, resorptions and live fetuses --> DUNNETT's test

Number of pregnant animals at the end of the study, mortality rate (of the dams) and number of litters with fetal findings --> FISHER's exact test

Proportion of fetuses with findings per litter --> WILCOXON test
Indices:
conception rate (in %) = (number of pregnant animals / number of fertilized animals) x 100
preimplantation loss (in %) for each individual pregnant animal which underwent scheduled sacrifice = ((number of corpora lutea – number of implantations)/number of corpora lutea) x 100
The postimplantation loss (in %) for each individual pregnant animal which underwent scheduled sacrifice = ((number of implantations – number of live fetuses) / number of implantations) x 100
Historical control data:
OECD 414 studies in wistar rats between 2012 - 2016 (n=34)
Clinical signs:
no effects observed
Description (incidence and severity):
Yellowish discolored feces were recorded for all females of the high-dose group (1000 mg/kg bw/d) from GD 14 onwards until terminal sacrifice (GD 20). This feces discoloration mirrors the presence of the test substance (or its metabolites) in the gastrointestinal tract. It is not considered as an adverse toxic effect.
No further clinical signs or changes of general behavior, which may be attributed to the test substance, were detected in any female at dose levels of 100, 300 or 1000 mg/kg bw/d during the entire study period.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
The mean body weights and average body weight gain of the low-, mid- and high-dose dams (100, 300 or 1000 mg/kg bw/d) were in general comparable to the concurrent control group throughout the entire study period. This includes the slightly higher weight gain of the mid-dose dams on GD 19-20.
Food consumption and compound intake (if feeding study):
no effects observed
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
Description (incidence and severity):
The mean gravid uterus weights of the animals of test groups 1-3 (100, 300 and 1000 mg/kg bw/d) were not influenced by the test substance. The differences between these groups and the control group revealed no dose-dependency and were assessed to be without biological relevance.
Gross pathological findings:
no effects observed
Description (incidence and severity):
A yellow discolored content of the stomach was recorded in 9 out of 25 mid-dose females (36%) and in 14 high-dose females (56%), while a yellow discolored content of the small intestine was seen in 2 mid-dose (8%) and 5 high-dose females (20%). These yellow discolorations mirror presence of the test substance (or its metabolites) in the gastrointestinal tract. They are not considered as adverse, toxic effects by themselves.
No further necropsy findings which could be attributed to the test substance were seen in any dam.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
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:
effects observed, non-treatment-related
Description (incidence and severity):
There were lower numbers of (early) resorptions than control in all treatment groups, producing higher live litter sizes in return. However, all observed differences are considered to reflect the normal range of fluctuations for animals of this strain and age. Hence, they are considered to be sporadical findings, neither related to treatment nor adverse.
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
no effects observed
Details on maternal toxic effects:
The conception rate reached 88% in the control and the mid-dose groups (0 and 300 mg/kg bw/d) and 100% in the low- and high-dose groups (100 and 1000 mg/kg bw/d). With these rates, a sufficient number of pregnant females were available for the purpose of the study. There were no test substance-related and/or biologically relevant differences between test groups 0-3 in conception rate, in the mean number of corpora lutea and implantation sites or in the values calculated for the pre- and the postimplantation losses, the number of resorptions and viable fetuses.
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: no maternal toxicity at any dose level observed
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): 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
Description (incidence and severity):
There were lower numbers of (early) resorptions than control in all treatment groups, producing higher live litter sizes in return. However, all observed differences are considered to reflect the normal range of fluctuations for animals of this strain and age for historical control data. Hence, they are considered to be sporadical findings, neither related to treatment nor adverse.
Changes in postnatal survival:
not specified
External malformations:
effects observed, non-treatment-related
Description (incidence and severity):
External malformations were detected in test groups 1 and 2 (100 and 300 mg/kg bw/d). In each case, these external malformations were associated with either soft tissue or skeletal malformations. None of these malformations are considered to be related to the treatment.
The total incidence of external malformations in treated animals did not differ significantly from the concurrent control group and was covered by the historical control data
No external variations were recorded.
One unclassified external observation, i.e. blood coagulum around placenta, was recorded in two fetuses of the mid-dose group (300 mg/kg bw/d). Since the finding was without statistical significance and not related to dose, it is not considered to be test substance-related.
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
Some skeletal malformations were detected in all test groups including the control (test groups 0-3; 0, 100, 300 and 1000 mg/kg bw/d). Two fetuses had associated external findings. The incidences of these malformations were neither statistically significantly different from control nor dose dependent and therefore not considered biologically relevant.
For all test groups, skeletal variations of different bone structures were observed, with or without effects on corresponding cartilages. The observed skeletal variations were related to several parts of fetal skeletons and appeared without a relation to dosing.
The rate of affected fetuses per litter with skeletal variations was statistically significantly higher in the low- and mid-dose groups (100 and 300 mg/kg bw/d, however, the incidences of all test groups were well within the historical control range and the variations neither showed a specific pattern nor a dose-response.
In addition to the findings tabulated above, the litter incidence for ‘misshapen sternebra; unchanged cartilage’ was statistically significantly increased in test group 2 (n=20*, 91%). However, the increase was not dose-related and the value was well within the range of the historical control data (mean 92.2%, range of 62.5 - 100.0%). Therefore, it is not assessed as being treatment-related.
designated as unclassified cartilage observations, occurred in all test groups. The observed unclassified cartilage findings were related to the skull, the ribs and the sternum. The rate of ‘bipartite processus xiphoideus’ was statistically significantly higher in all test substance-treated groups (test groups 1-3: 62.4%*/65.6%*/57.6%*[p<=0.05]). As a consequence of these increases, the incidence of total fetal skeletal unclassified cartilage observations was statistically significantly higher in test groups 1 and 2 (Tab. 4.3.4.3.1.). However, the incidences of all test substance-treated groups were well within the historical control range (mean 72.2% [52.5% - 89.4%]), while the concurrent control incidence was unusually low, below the historical control range (test group 0: 42.0%). Therefore and as there is no dose-response relationship an association to the treatment and a toxicological relevance is not assumed.
Visceral malformations:
no effects observed
Description (incidence and severity):
One fetus of test group 2 (300 mg/kg bw/d) had multiple soft tissue malformations, which were associated with an external malformation.
The total incidence of soft tissue malformations did not differ significantly from the concurrent control group. The findings were covered by the historical control data and, therefore, not assessed as treatment-related.
Three soft tissue variations were detected, i.e. short innominate, dilated renal pelvis and dilated ureter. These variations were neither significantly different from the concurrent control nor dose-dependently altered. Therefore, they were not considered biologically relevant.
No soft tissue unclassified observations were recorded.
Other effects:
no effects observed
Description (incidence and severity):
Weight of the placentae
The mean placental weights of test groups 1-3 were comparable to the concurrent control group.
Key result
Dose descriptor:
NOAEL
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 at any dose level
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no

Total external malformations

 

 

Test group 0

0 mg/kg bw/d

Test group 1

100 mg/kg bw/d

Test group 2

300 mg/kg bw/d

Test group 3

1000 mg/kg bw/d

LitterFetuses

N N

22

218

25

251

22

237

25

253

 

Fetal incidence

 

N (%)

 

0.0

 

1 (0.4)

 

2 (0.8)

 

0.0

 

Litter incidence

 

N (%)

 

0.0

 

1 (4.0)

 

2 (9.1)

 

0.0

Affectedfetuses/litter

 

Mean%

 

0.0

 

0.5

 

0.8

 

0.0

mg/kg bw/d = milligram per kilogram body weight per day; N = number; % = per cent

Total soft tissue malformations

 

 

Test group 0

0 mg/kg bw/d

Test group 1

100 mg/kg bw/d

Test group 2

300 mg/kg bw/d

Test group 3

1000 mg/kg bw/d

LitterFetuses

N N

21

105

25

119

22

114

25

118

 

Fetal incidence

 

N (%)

 

0.0

 

0.0

 

1 (0.9)

 

0.0

 

Litter incidence

 

N (%)

 

0.0

 

0.0

 

1 (4.5)

 

0.0

Affectedfetuses/litter

 

Mean%

 

0.0

 

0.0

 

0.8

 

0.0

mg/kg bw/d = milligram per kilogram body weight per day; N = number; % = per cent

Total skeletal malformations

 

 

Test group 0

0 mg/kg bw/d

Test group 1

100 mg/kg bw/d

Test group 2

300 mg/kg bw/d

Test group 3

1000 mg/kg bw/d

LitterFetuses

NN

22

113

25

132

22

123

25

135

 

Fetal incidence

 

N (%)

 

1 (0.9)

 

1 (0.8)

 

2 (1.6)

 

2 (1.5)

 

Litter incidence

 

N (%)

 

1 (4.5)

 

1 (4.0)

 

2 (9.1)

 

2 (8.0)

Affectedfetuses/litter

 

Mean%

 

0.9

 

0.8

 

1.7

 

1.4

mg/kg bw/d = milligram per kilogram body weight per day; N = number; % = per cent

Total fetal skeletal variations

 

 

Test group 0

0 mg/kg bw/d

Test group 1

100 mg/kg bw/d

Test group 2

300 mg/kg bw/d

Test group 3

1000 mg/kg bw/d

LitterFetuses

N N

22

113

25

132

22

123

25

135

 

Fetal incidence

 

N (%)

 

110 (97)

 

132 (100)

 

123 (100)

 

129 (96)

 

Litter incidence

 

N (%)

 

22 (100)

 

25 (100)

 

22 (100)

 

25 (100)

Affectedfetuses/litter

 

Mean%

 

97.6

 

100.0*

 

100.0*

 

95.7

mg/kg bw/d = milligram per kilogram body weight per day; N = number; % = per cent

* = p<=0.05 (Wilcoxon-test [one-sided])

Occurrence of statistically significantly increased skeletal variations (expressed as mean percentage of affected fetuses/litter)

 

Finding

Test group 0

0 mg/kg bw/d

Test group 1

100 mg/kg bw/d

Test group 2

300 mg/kg bw/d

Test group 3

1000 mg/kg bw/d

HCD

Mean % (range)

 

Incomplete ossification of basisphenoid

 

20.3

 

29.8*

 

24.6

 

20.0

 

17.3

(7.3 - 37.6)

Bipartite ossification of tho- racic centrum; dumbbell- shaped cartilage of cen- trum

 

0.0

 

0.0

 

2.2*

 

0.0

 

0.5

(0.0 - 2.4)

mg/kg bw/d = milligram per kilogram body weight per day; HCD = Historical control data; % = per cent

* = p<= 0.05 (Wilcoxon-test [one-sided]) ** = p<=0.01 (Wilcoxon-test [one-sided])

Total unclassified cartilage observations

 

 

Test group 0

0 mg/kg bw/d

Test group 1

100 mg/kg bw/d

Test group 2

300 mg/kg bw/d

Test group 3

1000 mg/kg bw/d

LitterFetuses

N N

22

113

25

132

22

123

25

135

 

Fetal incidence

 

N (%)

 

48 (42)

 

85 (64)

 

79 (64)

 

77 (57)

 

Litter incidence

 

N (%)

 

18 (82)

 

25 (100)*Fi

 

22 (100)

 

23 (92)

Affectedfetuses/litter

 

Mean%

 

44.7

 

64.8*Wi

 

65.6*Wi

 

58.1

mg/kg bw/d = milligram per kilogram body weight per day; N = number; % = per cent

* Fi= p ≤ 0.05 (Fisher’s exact test[one-sided])

* Wi= p ≤ 0.05 (Wilcoxon-test[one-sided])

Conclusions:
Under the conditions of this prenatal developmental toxicity study, the oral administration of the test item to pregnant Wistar rats from implantation to one day prior to the expected day of parturition (GD 6-19) at a dose of 1000 mg/kg bw/d caused neither evidence of maternal nor developmental toxicity.
In conclusion, the no observed adverse effect level (NOAEL) for maternal and prenatal developmental toxicology is 1000 mg/kg bw/d.
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
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

In a prenatal developmental toxicity study the test substance Sicotan Yellow K 2011 was administered to pregnant Wistar rats daily by gavage from implantation to one day prior to the expected day of parturition (GD 6-19) to evaluate its potential maternal and prenatal developmental toxicity.

Analyses confirmed the correctness of the prepared concentrations and their homogeneous distribution in the vehicle.

Generally, clinical observations including food consumption and body weight gain revealed no toxicologically relevant difference between the animals receiving 100, 300 or 1000 mg/kg bw/d the test item and controls.

Clinical observations, food consumption and body weights/body weight gain (gross and corrected) revealed no toxicologically relevant effects in the animals receiving 100, 300 or 1000 mg/kg bw/d of the test compound. Yellowish discolored feces were recorded for all females of the high-dose group (1000 mg/kg bw/d) from GD 14 onwards until terminal sacrifice (GD 20). This feces discoloration mirrors the presence of the test substance (or its metabolites) in

the gastrointestinal tract. It is not considered as an adverse toxic effect.

No differences of toxicological relevance between the control and the treated groups (100, 300 and 1000 mg/kg bw/d) were determined for any reproductive parameters, such as conception rate, mean number of corpora lutea, mean number of implantations, as well as pre- and postimplantation loss and live litter size. Similarly, no adverse effect of the test substance on uterine and placental weights as well as fetal weight and sex distribution of the fetuses was

noted at any dose. Fetal external, soft tissue and skeletal examinations revealed that there is no effect of the test

substance on the respective morphological structures up to a dose of 1000 mg/kg bw/d.

Under the conditions of this prenatal developmental toxicity study, the oral administration of the test item to pregnant Wistar rats from implantation to one day prior to the expected day of parturition (GD 6-19) at a dose of 1000 mg/kg bw/d caused neither evidence of maternal nor developmental toxicity.

In conclusion, the no observed adverse effect level (NOAEL) for maternal and prenatal developmental toxicology is 1000 mg/kg bw/d.

Justification for classification or non-classification

The available studies are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance does not need to be classified and labelled for reproductive or developmental toxicity under Regulation (EC) No 1272/2008, as amended for the ninth time in Regulation EC No 2016/1179.