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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from authoritative database.

Data source

Reference
Reference Type:
other: authoritative database
Title:
Repeated oral administration toxicity / reproductive developmental toxicity combined test for the test chemical in rats
Author:
J-check
Year:
2019
Bibliographic source:
J-check, 2019

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Principles of method if other than guideline:
Combined Repeat Dose and Reproductive / Developmental Toxicity study of test material by Oral Administration in Rats.
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- IUPAC name: 3-hydroxy-4-[(2-methyl-5-nitrophenyl)azo]-N-phenylnaphthalene-2-carboxamide
- Name of test material (as cited in study report): C.I. Pigment Red 22
- Molecular formula (if other than submission substance):C24H18N4O4
- Molecular weight (if other than submission substance):426.43 g/mole
- Smiles: O=C(Nc1ccccc1)c1c(O)c(\N =N\c2c (ccc([N+](=O)[O-])c2)C)c2c(ccc c2)c1
- InChI:1S/C24H18N4O4/c1-15-11-12-18(28(31)32)14-21(15)26-27-22-19-10-6-5-7-16(19)13-20(23(22)29)24(30)25-17-8-3-2-4-9-17/h2-14,29H,1H3,(H,25,30)/b27-26+
- Substance type: Organic
- Physical state: Solid powder

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
- Source: Charles River Japan Co., Ltd
- Age at study initiation: 9 weeks
- Weight at study initiation: 309 to 355 g for males and from 206 to 232 g for females
- Fasting period before study: No data available
- Housing: Rat were housed stainless steel hanging type wire mesh cage, excluded for the period of pregnancy /nursing period. During pregnancy / nursing female were housed in polycarbonate cage with a floor covering for laboratory animals.
- Diet (e.g. ad libitum): Solid feed for experimental animals irradiated with ultraviolet rays after autoclave sterilized nd filtering with a pore size of 5 μm, ad libitum
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
-Temperature (°C): 22 ± 2 ° C.
- Humidity (%):55 ± 15%
- Air changes (per hr): 12 times / hour
- Photoperiod (hrs dark / hrs light): lighting 12 hours / day (7: 00-19: 00)

Administration / exposure

Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80
Details on exposure:
VEHICLE
- Justification for use and choice of vehicle (if other than water): 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80
- Concentration in vehicle: 0 (vehicle), 100, 300, 1000 mg/kg
- Amount of vehicle (if gavage): 10 mL / kg
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: 14th day
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy Plug formation or sperm in vaginal smear referred to as day 0 of pregnancy
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
42 to 47 days
Frequency of treatment:
Daily
Details on study schedule:
not specified
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day
Dose / conc.:
100 mg/kg bw/day
Dose / conc.:
300 mg/kg bw/day
Dose / conc.:
1 000 mg/kg bw/day
No. of animals per sex per dose:
Total: 96
0 (vehicle) mg/kg: 12 male, 12 female
100 mg/kg: 12 male, 12 female
300 mg/kg: 12 male, 12 female
1000 mg/kg: 12 male, 12 female
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Test substance was repeatedly administered orally to SD rats of 3 male and 3 female at 0, 100, 300 and 1000 mg / kg doses for 14 days, and as a result, No effect on in general condition, body weight, food consumption. No clear toxicity change due to administration of the test substance was observed in both organ weight and necropsy.
Based on these findings, the high dose was set at the upper limit of 1000 mg / kg prescribed in the OECD guidelines, three doses of 300 mg / kg for the medium dose and 100 mg / kg for the low dose were set at a tolerance of about 3 .
Positive control:
not specified

Examinations

Parental animals: Observations and examinations:
Mortality, clinical sign, body weight and feed Consumption were examined.
Oestrous cyclicity (parental animals):
Estrous cyclicity were examined.
Number of corpus luteums and the number of landings were examined
Sperm parameters (parental animals):
Not specified
Litter observations:
Not specified
Postmortem examinations (parental animals):
Hematology, clinical chemistry, Gross pathology and histopathology were examined.
Postmortem examinations (offspring):
Number of births, sex and Body weight were examined.
Statistics:
Regarding the weighing data, parametric data was tested for equal variance by the Bartlett method, and one-way ANOVA was performed when variance was equal. Where the variances are not equal and nonparametric data were tested by Kruskal-Wallis. When significant difference was observed between groups, multiple comparisons of Dunnett method or Dunnett type were performed. Pathological findings in counting data were examined by χ 2 for a × b . When significant difference was observed, comparison was made between control group and each test substance administered group by Armitage χ 2 test. Other counting data were tested by Fisher's direct stochastic method. The significance level of each test was 5%. For the data on newborn babies, the average value calculated for each mother was set as a sample unit.
Reproductive indices:
Breast feeding, nesting, and the presence or absence of feeding, birth rate, implantation rate, delivery rate were examined.
Offspring viability indices:
Viability on day 0 and 4 were examined.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Red feces exhibiting the same color tone as the test substance administered in all sexes were observed daily from the day following the administration start date.
When treated wtih 1000 mg/kg bw, in 2 male loose stools were observed transiently on 22 days after the start of administration, but abnormalities in general conditions considered to be toxic changes.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
No effect on survival of treated male and female rat were observed as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No significant effect on body weight and body weight gain of treated male and female rats were observed as compared to control.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No effect on food consumption of treated male and female rats was observed as compared to control.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
When treated wtih 100 mg / kg, significant decrease in prothrombin time were observed in male rats, but no change was observed in the 300 and 1000 mg / kg group, so it was judged to be an accidental change.
In females, there was no significant difference between the test substance administered group and the control group.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
When treated wtih 1000 mg / kg, significant incrase in A / G ratio were observed in male rats. However, since the total protein and albumin were not changed and were slightly high, it was considered to be fluctuating within the physiological range and it was judged that it was not caused by administration of the test substance.
When treated wtih 100 mg / kg, significant incrase in ASAT (GOT) in female rats, but no change was observed in the 300 and 1000 mg / kg group, so it was judged to be an accidental change .
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No histopathological changes were observed in treated male and female rats as compared to control.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
No effect on sexual cycle of treated female rat were observed as compared to control.
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
No significant difference was observed in mating index, fertility index, numbers of corpora lutea or implantations, implantation index, delivery index, gestation index, gestation length, parturition or maternal behavior, numbers of offspring or live offspring of treated rats as compared to control.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
reproductive function (oestrous cycle)
reproductive performance
Remarks on result:
other: No effect observed

Target system / organ toxicity (P0)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Description (incidence and severity):
No abnormal clinical signs were observed in offspring.
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
No effect on numbers of live offspring of were observed as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No significant change in body weights of offspring were observe as compared to control.
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
No gross pathological changes were observed in offsprings.
Histopathological findings:
not specified
Other effects:
not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not specified

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
clinical signs
mortality
body weight and weight gain
gross pathology
Remarks on result:
other: No effect observed

Target system / organ toxicity (F1)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified

Overall reproductive toxicity

Reproductive effects observed:
not specified
Treatment related:
not specified

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 1000 mg/kg/day for P and F1 generation when Crj:CD (SD) male and female rats treated with test material orally by gavage for 42 to 47 days.
Executive summary:

In a Combined Repeat Dose and Reproductive / Developmental Toxicity Screening Test, Crj:CD(SD) male and female rats treated with test material in the concentration of 0 (vehicle), 100, 300, 1000 mg/kg orally by gavage for 42 to 47 days according to OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test). No effect on survival of treated male and female rat was observed as compared to control. Red feces exhibiting the same color tone as the test substance administered in all sexes were observed daily from the day following the administration start date. At 1000 mg/kg bw, in 2 male loose stools were observed transiently on 22 days after the start of administration, but abnormalities in general conditions considered to be toxic changes. No significant effect on body weight and body weight gain and food consumption of treated male and female rats were observed as compared to control. Significant decrease in prothrombin time were observed in male rats at 100 mg / kg/day, but no change was observed in the 300 and 1000 mg / kg group, so it was judged to be an accidental change. In females, there was no significant difference between the test substances administered group and the control group. Significant increase in A / G ratio were observed in male rats. At 1000 mg / kg/day However, since the total protein and albumin were not changed and were slightly high, it was considered to be fluctuating within the physiological range and it was judged that it was not caused by administration of the test substance. Significant increase in ASAT (GOT) in female rats at 100 mg / kg/day, but no change was observed in the 300 and 1000 mg / kg group, so it was judged to be an accidental change. Similarly, No effect on reproductive parameters such as sexual cycle, mating index, the fertility index, numbers of corpora lutea or implantations, the implantation index, the delivery index, the gestation index, gestation length, parturition or maternal behavior, numbers of offspring or live offspring of treated rats as compared to control. Significant increase in relative liver weight in male and absolute and relative weight in female were observed at 1000 mg / kg bw as compared to control. Significant decrease in absolute spleen weight and significant increase in relative adrenal gland weight were observed at 300 mg / kg bw in male rats. Significant increase in relative adrenal gland weight were observed in male rats at 100 mg / kg bw. But, since there was no change in the 1000 mg / kg group, it was judged to be an accidental change. Colored stomach at 1000 mg / kg and red, colored contents were observed in small intestine and large intestine at 100 mg / kg. Focal bleeding of the lung, contraction failure and white spots, stomach tar-like contents, white spots of the liver, dilation of the renal pelvic cavity, uterine bloating, vaginal distension and ventricular dilation were observed. However, since there was no certain tendency in its expression status, it was judged to be an accidental change unrelated to administration of the test substance. No histopathological changes were observed in treated male and female rats as compared to control. In addition, No effect on numbers of live offspring, clinical signs and body weight of offspring were observed as compared to control. No gross pathological changes were observed in offspring. Therefore, NOAEL was considered to be 1000 mg/kg/day for P and F1 generation when Crj:CD (SD) male and female rats treated with test material orally by gavage for 42 to 47 days.