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Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

NOAEL was considered to be 1000 mg/kg/day for male and female for P and F1 generation when Crj:CD (Sprague-Dawley) male and female rats were treated with Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid orally by gavage for male 42 days and female from 14 days prior to mating to Day 3 of lactation.

Thus, comparing this effect with the criteria of CLP regulation, Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid can be not classified for reproductive toxicity. 

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from J-check
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 screening Test of D & C Red No. 7 in Rats
GLP compliance:
not specified
Limit test:
no
Specific details on test material used for the study:
- Name of test material (as cited in study report): 2-Naphthalenecarboxylic acid, 3-hydroxy-4-[(4-methyl-2-sulfophenyl)azo]-, calcium salt- Molecular formula (if other than submission substance): C18H14N2O6S.Ca- Molecular weight (if other than submission substance): 424.445 g/mole- Smiles notation (if other than submission substance): c12c(c(c(C(=O)[O-])cc1cccc2)O)\N=N\c1c(cc(C)cc1)S(=O)(=O)[O-].[Ca+2]- InChl (if other than submission substance): 1S/C18H14N2O6S.Ca/c1-10-6-7-14(15(8-10)27(24,25)26)19-20-16-12-5-3-2-4-11(12)9-13(17(16)21)18(22)23;/h2-9,21H,1H3, (H,22,23)(H,24,25,26);/q;+2/p-2/b20-19+;- Substance type: Organic - Physical state: Solid
Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
- Age at study initiation: (P) x wks; (F1) x wks: 8 weeks - Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: female 196.5 to 227.4 g, males 253.5 ~ 299.6 g- Fasting period before study:- Housing: Animals were individually housed in a wire mesh floor cage (Nihon Cage) and raised. For the mother animals after 18th gestation, metal floorboards were laid on the floor of breeding cages, and wood chips (White Flake (R), Japan Charles River) were appropriately supplied as bedding.- Diet (e.g. ad libitum): Solid feed (CA - 1, Japan Clea), ad libitum- Water (e.g. ad libitum): Tap water, ad libitum- Acclimation period: 1 week ENVIRONMENTAL CONDITIONS- Temperature (°C): 24 ± 1 °C.- Humidity (%): 55 ± 5%- Air changes (per hr): Ventilation frequency of about 15 times / hour - Photoperiod (hrs dark / hrs light): Lighting room of a barrier system adjusted for 12 hours of lighting (7 am to 7 pm).
Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: 5% Gum Arabic solution
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Test substance was suspended in an aqueous solution of 5% gum arabic, and the content was adjusted so that the dosage volume at one time would be 10 ml / kg at any dose. VEHICLE- Justification for use and choice of vehicle (if other than water): Test substance was almost insoluble in water and acetone and hence 5% Gum Arabic solution were used. - Concentration in vehicle: 0, 100, 300, 1000 mg/kg/day - Amount of vehicle (if gavage): 10 ml / kg bw - Lot/batch no. (if required):- Purity:
Details on mating procedure:
- M/F ratio per cage:1: 1. - Length of cohabitation:14 days - Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancyConfirmation of copulation was made every morning by examining the presence of sperm in the vaginal plug and vaginal plaque in the vagina and females confirmed to be mated were separated from the male starting from that day as the 0th day of gestation. - After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.Not specified - Further matings after two unsuccessful attempts: [no / yes (explain)]Not specified- After successful mating each pregnant female was caged (how): Individually
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
Males, 42 daysFemales, from 14 days prior to mating to Day 3 of lactation
Frequency of treatment:
Daily
Dose / conc.:
0 mg/kg bw/day (actual dose received)
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:
Total: 1040 (vehicle) mg/kg/day: 13 male, 13 female 100 mg/kg/day: 13 male, 13 female 300 mg/kg/day: 13 male, 13 female 1000 mg/kg/day: 13 male, 13 female
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose was selected based on Preliminary dose setting test. 0 (solvent control) and 1000 mg / kg of D & C Red No. 7 were orally administered repeatedly to 5 male and 5 female Sprague-Dawley (Crj: CD) rats once a day for 14 days and the administration end date, and repeated administration toxicity of D & C Red No. 7 to male and female rats was examined. Thus for the purpose of searching for toxicity, the maximum dose was 1000 mg / kg / day, and the following were divided by the common ratio 3 to set the intermediate dose to 300 mg / kg / day and the lowest dose to 100 mg / kg / day.
Parental animals: Observations and examinations:
Survival, Clinical sign, Body weight and weight gain and food consumption were examined.
Oestrous cyclicity (parental animals):
Not specified
Sperm parameters (parental animals):
Not specified
Litter observations:
Birth rate, litter weight and Sex ratio were examined.
Postmortem examinations (parental animals):
Organ weight, gross pathology and histopathology was examined.
Postmortem examinations (offspring):
Grossly visible abnormalities was examined.
Statistics:
The mating rate and conception rate were subjected to χ ^ 2 test. All other data were tested for uniformity of variance of each group by the Bartlett method, with the value obtained for each individual or the average value for each litter as one sample. As a result, when the variance is made uniform, a one-way analysis of variance is carried out, and when significance is observed between the groups, the control group and each test substance administered group and the control group are administered by the Dunnett method or the Scheff method The difference between the mean values was tested between. When the variance was not uniform, Kruskal-Wallis rank test was conducted, and when significantness was observed between the groups, a Dunnett type or Scheff type test for the difference between the control group and each test substance administered group went. The level of significance was set to 5% and 1%.
Reproductive indices:
Female fertility index, copulation index, Gestation index, viability index, Live birth index, implantation index, delivery index, Number of females with stillborn pups were examined.
Offspring viability indices:
Yes
Clinical signs:
no effects observed
Description (incidence and severity):
Red feces exhibiting the same color tone as the test substance administered was observed daily during the administration period. In addition, the formation of crust was observed in a very small number of animals, but abnormalities in general condition regarded as toxicity change were not observed in either administration group in both males and females.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
No effect on survival of treated male and female rats were observed.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No significant differences were observed between body weight and increasing amount between each test substance administered group and control group in any period before commencement, during pregnancy, and after parturition.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No significant change was observed in any of the test substance administered groups as compared with the control group.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Description (incidence and severity):
When treated with 300 and 1000 mg/kg bw, significant decrease in mean red blood cell hemoglobin level and dose-dependent decrease in white blood cell count were observed but no significant difference was observed between the group and the control group. No change suggesting the effect of the test substance was observed for the results of the leukocyte classification.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
When treated with 1000 mg/kg bw, significant decrease in total cholesterol, potassium concentration and significantly increased chlorine concentration and GOT activity and Significant decreases in inorganic phosphorus and calcium concentrations were observed in treated rats. No other presumed changes due to the administration of the test substance were observed in the other test items.
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
When treated with 1000 mg/kg bw, significant increase in kidney weight were observed in treated rats as compred to control. No effect on absolute or relative weight of testis and epididymis were observed between the control and the treated groups.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No treatment related macroscopic change was observed in treated rats as compared to control.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Increased incidence of renal tubule regeneration in males at 300 and 1000 mg/kg bw, and the increased incidence of renal tubule regeneration and necrotic or foamy tubular epithelial cells in females at 100, 300 and 1000 mg/kg bw were observed. These lesions increased in severity and incidence in a dose-dependent manner.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
No effect on Number pregnant per dose level, Female fecundicity index, Female fertility index, Male fertility index, Number aborting, Number of resorptions, Gestation index, Live birth index, Number of females with live born pups, Number of females with all stillborn pups, Post implantation loss, Number of corpus luteum, number of implantation and implantation and duration of Pregnancy was observed in treated rats as compared to control.
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:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
organ weights and organ / body weight ratios
gross pathology
reproductive performance
Remarks on result:
other: No effect on reproductive performance and organ
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
No effect on viability on nursing day 0 and nursing 4th day of pups were observed as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No significant effect on birth weight at any measurement time were observed 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:
not specified
Histopathological findings:
not specified
Other effects:
no effects observed
Description (incidence and severity):
Sex ratio: No difference was observed in the sex ratio between the groups.Birth rate: No effect on birth rat of pups were observed as compared to control.
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
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:
viability
clinical signs
mortality
body weight and weight gain
other: Birth rate and Sex ratio
Remarks on result:
other: No effect observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Reproductive effects observed:
not specified
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
NOAEL was considered to be 1000 mg/kg/day for male and female for P and F1 generation when Crj:CD (Sprague-Dawley) male and female rats were treated with Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid orally by gavage for male 42 days and female from 14 days prior to mating to Day 3 of lactation.
Executive summary:

In a reproduction/developmental screening Test,  Crj:CD (Sprague-Dawley) male and female rats were treated with Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid in the concentration of0,100, 300, 1000 mg/kg/day orally by gavage for Males 42 days and female from 14 days prior to mating to Day 3 of lactation. Red feces exhibiting the same color tone as the test substance administered was observed daily during the administration period. In addition, the formation of crust was observed in a very small number of animals, but abnormalities in general condition regarded as toxicity change were not observed in either administration group in both males and females. No effect on survival of treated male and female rats was observed. No significant differences were observed between body weight and increasing amount between each test substance administered group and control group in any period before commencement, during pregnancy, and after parturition. No significant change was observed in food consumption any of the test substance administered groups as compared with the control group. Similarly, significant decrease in mean red blood cell hemoglobin level and dose-dependent decrease in white blood cell count were observed at 300 and 1000 mg/kg bw, but no significant difference was observed between the group and the control group. No change suggesting the effect of the test substance was observed for the results of the leukocyte classification. Significant decrease in total cholesterol, potassium concentration and significantly increased in chlorine concentration and GOT activity and Significant decreases in inorganic phosphorus and calcium concentrations were observed at 1000 mg/kg bw in treated rats. No other presumed changes due to the administration of the test substance were observed in the other test items. Significant increase in kidney weight was observed at 1000 mg/kg bw in treated rats as compared to control. No effect on absolute or relative weight of testis and epididymis were observed between the control and the treated groups. No treatment related macroscopic change was observed in treated rats as compared to control. Increased incidence of renal tubule regeneration in males at 300 and 1000 mg/kg bw, and the increased incidence of renal tubule regeneration and necrotic or foamy tubular epithelial cells in females at 100, 300 and 1000 mg/kg bw. These lesions increased in severity and incidence in a dose-dependent manner. In addition, no reproductive effect were observed such as on Number pregnant per dose level, Female fecundicity index, Female fertility index, Male fertility index, Number aborting, Number of resorptions, Gestation index, Live birth index, Number of females with live born pups, Number of females with all stillborn pups, Post implantation loss, Number of corpus luteum, number of implantation and implantation and duration of Pregnancy of female, and Sex ratio, viability and body weight of pups on day 0 and 4. Therefore, NOAEL was considered to be 1000 mg/kg/day for male and female for P and F1 generation when Crj:CD (Sprague-Dawley) male and female rats were treated with Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid orally by gavage for male 42 days and female from 14 days prior to mating to Day 3 of lactation.

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:
Data is Klimisch 2 and from J-check
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Reproductive toxicity:

In different studies, Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid has been investigated for reproductive toxicity to a greater or lesser extent. Study based on in vivo experiments, i.e. most commonly in rats and rabbits for Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid and summarized below 

In a experimental study conducted by J-check (Ministry of Health, Labour and Welfare", "Ministry of the Environment" and "National Institute of Technology and Evaluation, J –Check, 2018) and given by OECD SIDS (OECD HPV, Chemicals Programme. SIDS Dossier, 1993), Crj:CD (Sprague-Dawley) male and female rats were treated with Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid in the concentration of0,100, 300, 1000 mg/kg/day orally by gavage for Males 42 days and female from 14 days prior to mating to Day 3 of lactation. Red feces exhibiting the same color tone as the test substance administered was observed daily during the administration period. In addition, the formation of crust was observed in a very small number of animals, but abnormalities in general condition regarded as toxicity change were not observed in either administration group in both males and females. No effect on survival of treated male and female rats was observed. No significant differences were observed between body weight and increasing amount between each test substance administered group and control group in any period before commencement, during pregnancy, and after parturition. No significant change was observed in food consumption any of the test substance administered groups as compared with the control group. Similarly, significant decrease in mean red blood cell hemoglobin level and dose-dependent decrease in white blood cell count were observed at 300 and 1000 mg/kg bw, but no significant difference was observed between the group and the control group. No change suggesting the effect of the test substance was observed for the results of the leukocyte classification. Significant decrease in total cholesterol, potassium concentration and significantly increased in chlorine concentration and GOT activity and Significant decreases in inorganic phosphorus and calcium concentrations were observed at 1000 mg/kg bw in treated rats. No other presumed changes due to the administration of the test substance were observed in the other test items. Significant increase in kidney weight was observed at 1000 mg/kg bw in treated rats as compared to control. No effect on absolute or relative weight of testis and epididymis were observed between the control and the treated groups. No treatment related macroscopic change was observed in treated rats as compared to control. Increased incidence of renal tubule regeneration in males at 300 and 1000 mg/kg bw, and the increased incidence of renal tubule regeneration and necrotic or foamy tubular epithelial cells in females at 100, 300 and 1000 mg/kg bw. These lesions increased in severity and incidence in a dose-dependent manner. In addition, no reproductive effect were observed such as on Number pregnant per dose level, Female fecundicity index, Female fertility index, Male fertility index, Number aborting, Number of resorptions, Gestation index, Live birth index, Number of females with live born pups, Number of females with all stillborn pups, Post implantation loss, Number of corpus luteum, number of implantation and implantation and duration of Pregnancy of female, and Sex ratio, viability and body weight of pups on day 0 and 4. Therefore, NOAEL was considered to be 1000 mg/kg/day for male and female for P and F1 generation when Crj:CD (Sprague-Dawley) male and female rats were treated with Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid orally by gavage for male 42 days and female from 14 days prior to mating to Day 3 of lactation.

In another experimental study summarized by EFSA (EFSA Journal, 2010; 8 (5): 1586), female rats were treated with Litholrubine BK (Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid)in the concentration of0,5, 16 and 50 mg/kg bw/day orally by gavage for days 6-15 of pregnancy. No adverse effects were observed on maternal weight gain, number of resorptions and fetal deaths, fetal weight and viability, litter size and incidence of fetal malformations and skeletal aberrations were observed in treated female rats. Therefore, NOAEL was considered to be 50 mg/kg/day for P and F1 generation when female rats were treated with Litholrubine BK (Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid) orally by gavage for days 6-15 of pregnancy.

Further supported by experimental study summarized by EFSA (EFSA Journal, 2010; 8 (5): 1586), female rabbits were treated with Litholrubine BK (Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid) in the concentration of0,5, 16 and 50 mg/kg bw/day orally by gavage for days 6-18 of pregnancy. No adverse effects were observed on maternal weight gain, number of resorptions and fetal deaths, fetal weight and viability, litter size and incidence of fetal malformations and skeletal aberrations were observed in treated female rats. Therefore, NOAEL was considered to be 50 mg/kg/day for P and F1 generation when female rabbits were treated with Litholrubine BK (Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid) orally by gavage for days 6-18 of pregnancy.

Thus, based on the above studies on Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid, it can be concluded that Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid is non toxic to reproduction till dose of 1000 mg/kg bw. Thus, comparing this effect with the criteria of CLP regulation, Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid can be not classified for reproductive toxicity. 

Effects on developmental toxicity

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on the above studies on Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid, it can be concluded that Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid is non toxic to reproduction till dose of 1000 mg/kg bw. Thus, comparing this effect with the criteria of CLP regulation, Calcium salt of 3-hydroxy-4 - [(4-methyl-2-sulfophenyl) azo] -2-naphthalenecarboxylic acid can be not classified for reproductive toxicity.