2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[2-[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4)

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

The No Observed Adverse Effect level (NOAEL) for the test chemical  2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3- [2-[4-[[2-(sulfooxy) ethyl]sulfonyl]phenyl]diazenyl] -8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (607724-37-8)for reproduction toxicity in test animals were  considered to be  in range of  750-1300mg/Kg bw/day.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data of read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: as below

Principles of method if other than guideline:

WoE derived based on the experimental data from structurally and functionally similar read across chemicals.

GLP compliance:

not specified

Limit test:

no

Species:

other: Study1;Rat,Study 2; Mouse

Strain:

other: Study1;Not specified ,Study 2; CFW

Sex:

male/female

Details on test animals or test system and environmental conditions:

Study;1TEST ANIMALS- Source: Not available- Age at study initiation: 6 and 7 weeks- Weight at study initiation: 129 g-males and 105 g-females(appx)- Fasting period before study: Not available- Housing: Individual cages- Diet (e.g. ad libitum): ad libitum- Water (e.g. ad libitum): ad libitum- Acclimation period: Not availableENVIRONMENTAL CONDITIONS- Temperature (°C): Not available- Humidity (%):Not available- Air changes (per hr): Not available- Photoperiod (hrs dark / hrs light): Not availableStudy 2;Details on test animalTEST ANIMALS- Source: No data available- Age at study initiation: No data available- Weight at study initiation: yes - Fasting period before study: No data available- Housing: They were caged in groups of 15 in a room .- Diet (e.g. ad libitum): Oxoid pasteurized Breeding diet - Water (e.g. ad libitum): water available- Acclimation period: No data availableENVIRONMENTAL CONDITIONS- Temperature (°C): No data- Humidity (%):50-60%- Air changes (per hr): No data- Photoperiod (hrs dark / hrs light): 21 ± 1°C

Route of administration:

oral: feed

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: Alphacel

Remarks:

a non-nutritive cellulose

Details on exposure:

Study1;PREPARATION OF DOSING SOLUTIONS:Alphacel (a non-nutritive ce!lulose) was added so that the level of dye plus alphacel was 1.5 per cent. The control diet contained 1.5 per cent alphacel. DIET PREPARATION- Rate of preparation of diet (frequency):Not available- Mixing appropriate amounts with (Type of food):Alphacel (a non-nutritive ce!lulose) diet - Storage temperature of food: Not availableVEHICLE- Justification for use and choice of vehicle (if other than water): Alphacel(a non nutritive cellulose)-No justification available.- Concentration in vehicle: 0, appx 15,150 and 750 mg/kg - Amount of vehicle (if gavage): Not available- Lot/batch no. (if required): Not available- Purity: Not availableStudy 2;Details on exposurePREPARATION OF DOSING SOLUTIONS:DIET PREPARATION- Rate of preparation of diet (frequency):- Mixing appropriate amounts with (Type of food): Oxoid pasteurized Breeding diet- Storage temperature of food:VEHICLE- Justification for use and choice of vehicle (if other than water):- Concentration in vehicle: 0.1, 0.25,0.5 or 1.0%- Amount of vehicle (if gavage):- Lot/batch no. (if required):- Purity:

Details on mating procedure:

Study1;Mating was not performed.Study 2;Not specified

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Study 1;64 weeksStudy 2; 80 weeks

Frequency of treatment:

Daily

Details on study schedule:

not specified

Dose / conc.:

0 mg/kg bw/day (nominal)

Dose / conc.:

15 mg/kg bw/day (nominal)

Dose / conc.:

150 mg/kg bw/day (nominal)

Dose / conc.:

750 mg/kg bw/day (nominal)

Remarks:

0, 0.1, 0.25,0.5 and 1.0%

No. of animals per sex per dose:

Study1;Total: 120Control - 15 males and 15 females15 ma/kg bw - 15 males and 15 females150 mg/kg bw - 15 males and 15 females750 mg/kg bw- 15 males and 15 femalesStudy 2;Total no. Of animals0%-60 male and 60 female0.1%-30 male and 30 female0.25% -30 male and 30 female0.5-30 male and 30 female1%-30 male and 30 female

Control animals:

yes, concurrent vehicle

other: control diet contained 1.5 per cent alphacel.

Details on study design:

After 26 weeks on test, five rats of each group receiving 1.5 per cent colourwere killed for histological examination. Haemoglobin estimations were done using a slight modification of thepyridine-haemochromogen method of Rimington. At the end of the experiment, electrocardiograms and electroencephalograms were recorded from six rats of each sex on the control diet and three rats of each sex on the 1.5 per cent level of each colour,

Positive control:

not specified

Parental animals: Observations and examinations:

Study1;Mortality, Body weight, food consumption and food efficiency were examined. Study 2;The general condition and behavior of the animals were observed frequently and any mouse that showed signs of ill-health was isolated, to be returned to its cage on recovery or to be killed If its condition deteriorated. Body weight-The mice were weighed at the start of the experiment, at wk 3 and then at intervals of 2 wk until wk 73 of the experiment. Hematology-Blood samples were taken at wk 28 and 55 from the caudal vein of ten males and ten females from the control group and from the groups given the 0.5 and 1% dietary levels. At 80 wk, blood samples were collected from the aorta of all surviving mice during the autopsy. The samples were examined for haemoglobin concentration and packed cell volume, as well as for counts of erythrocytes and leucocytes. In addition, the methaemoglobin concentrations were determined in the samples collected at 80 wk. Preparations for counting the reticuiocytes and the different types of leucocytes were made but, in the absence of consistent effects on the other measurements, these counts were not carried out.Urine analysis-During wk 28, urine samples were collected over a 6-hr period from three groups of five mice of each sex from the controls and the groups on the two highest dietary levels (0.5 and 1%) of Black PN. These samples were examined for protein, reducing substances, bile salts and blood as well as for colour, pH and microscopic constituents. Organ weight was also measured.

Oestrous cyclicity (parental animals):

No data

Sperm parameters (parental animals):

No data

Litter observations:

No data

Postmortem examinations (parental animals):

Study1;Necroscopies were performed on rats that died during the test. After 26 weeks on test, five rats of each group receiving 1.5 per cent colour were killed for histological examination. Haemoglobin estimations were done(Hb,RBC,WBC,Differential cell counts) Gross examination-After 64 weeks all surviving rats were anaesthesized with ether on all organs and tissues gross examined .Organ weights recorded.Histopathological examination-Tissues with gross pathology change was studied for histopathology.Tissues examined -Lung, liver, heart, spleen, thyroid, pancreas, stomach, small intestine, kidney, urinary bladder, adrenal, testis, prostate, coagulating gland, ovary, uterus, and thymus.Study 2;The animals were killed by exsanguinations from the aorta under sodium pentobarbitone anesthesia following an overnight period without food. At autopsy, macroscopic abnormalities were recorded and the brain, heart, liver, spleen, kidneys, adrenal glands and gonads were weighed. Samples of these organs and of salivary glands, pituitary, thyroid, thymus, various lymph nodes, pancreas, urinary bladder, lungs, stomach, duodenum, ileum, colon, caecum,rectum, striped muscle (hind limb), spinal cord, uterus, aortic arch and any other tissue that appeared abnormal were preserved in 10% buffered formalin. Paraffin-wax sections of these tissues were stained with haematoxylin and eosin. All tissues from the controlmice and from those fed diet containing 1% BlackPN were examined histologically. At the lower dose levels, the examination was confined to the liver, kidneyand any tissues seen to be abnormal at autopsy

Postmortem examinations (offspring):

No data

Statistics:

Statistics –significant differences between mean values were determined by Student’s“t”test (forgrowth, food consumption and food efficiency).

Reproductive indices:

No data

Offspring viability indices:

No data

Clinical signs:

not specified

Description (incidence and severity):

Study 2;No significant change were observed in treated group compare to control group at any dose level (0, 0.1, 0.25,0.5 and 1.0%).

Dermal irritation (if dermal study):

not specified

Mortality:

no mortality observed

Description (incidence):

Study 1;No effect on survival of treated male and female rats were observed at 15, 150 and 750 mg/kg bw as compared to control. Study 2; No statistically significant differences between the number of deaths in the control mice and those given tes chemical was observed.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Decrease in mean body weight in females at 4, 32 and 16 weeks.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

Decreased in food consumption of females were observed with no correlation with the concentration given in diet.

Food efficiency:

no effects observed

Description (incidence and severity):

No significant difference in male or female food efficiency was observed as compared to control.

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):

The only significant difference observed was lowered white cell count for females at 750 mg/kg bw as compared to controls. No adverse effect on blood cells was observed.

Clinical biochemistry findings:

not specified

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):

Middle ear and respiratory infections were observed, considered to be in older rats of the colony. Chronic otitis media was observed in nearly 50 per cent of the animals. The disease was evenly distributed in the various groups.Pathological changes in the adrenal cortex were observed in 22 animals. This particular pathology was acute in nature and was not considered to be an effect of the food colours since it was observed in two of the control animals and since there was no correlation between the incidence and the concentration of colour fed.

Histopathological findings: neoplastic:

no effects observed

Description (incidence and severity):

18 per cent tumour incidence was observed in treated rats. The differences in tumour incidence are not significant according to chi-square tests.

Other effects:

no effects observed

Description (incidence and severity):

The tracings of ECG were essentially normal. No significant deviation of electrical axis observed in treated rats.

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

not specified

Dose descriptor:

NOAEL

Effect level:

750 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

mortality

body weight and weight gain

food consumption and compound intake

food efficiency

haematology

organ weights and organ / body weight ratios

gross pathology

histopathology: non-neoplastic

histopathology: neoplastic

other: No significant effect on reproductive organ weight and histopathology of sexual organs testis,ovary and uterus.

Remarks on result:

other: No toxic effect were observed.

Dose descriptor:

NOAEL

Effect level:

1 300 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No significant effect were observed on on this dose

Remarks on result:

other: No toxic effect were 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

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

not specified

Body weight and weight changes:

not specified

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:

not specified

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not specified

Dose descriptor:

other: not specified

Based on:

not specified

Sex:

not specified

Basis for effect level:

other: not specified

Remarks on result:

other: not specified

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:

The No Observed Adverse Effect level (NOAEL) for the test chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3- [2-[4-[[2-(sulfooxy) ethyl]sulfonyl]phenyl]diazenyl] -8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (607724-37-8)for reproduction toxicity in test animals were considered to be in range of 750-1300mg/Kg bw/day.

Executive summary:

Data available for the target chemicals was reviewed to determine the toxic nature of the test chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3- [2-[4-[[2-(sulfooxy) ethyl]sulfonyl]phenyl]diazenyl] -8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (607724-37-8).The studies are as mentioned below:

Reproductive study was conducted for test chemical in male and female CFW mice for 80 weeks by oral feed. When Groups of 30 male and 30 female CFW mice were given diets containing 0.1, 0.25, 0.5 and 1.0% test chemical for 80 wk, with groups of 60 mice of each sex as controls. There were no dose-related effects on body-weight gain, haematology and organ weights. The incidence of histopathological findings was not altered by the feeding of test chemical. Therefore it is considered that test chemical to the diet of male and female CFW mice at levels up to 1% (equivalent to approximately 1300 mg/kg/day) for 80 wk does not induce any reprotoxic effect.

In a chronic toxicity study, male and female rats were treated with test chemical in the concentration of 0, 15, 150 and 750 mg/kg bw orally in feed Alphacel (a non-nutritive cellulose) for 64 weeks. No effect on survival of treated male and female rats were observed at 15, 150 and 750 mg/kg bw as compared to control. Decrease in mean body weight in females at 4, 32 and 16 weeks. Decreased in food consumption of females were observed with no correlation with the concentration given in diet. No significant difference in male or female food efficiency was observed as compared to control. The only significant difference observed was lowered white cell count for females at 750 mg/kg bw as compared to controls. No adverse effect on blood cells was observed. Similarly, Decrease in liver weight at 150 mg/kg bw and spleen weight at 750 mg/kg bw in female rat. These changes were not correlated with the level of food colour in the diet and are difficult to interpret. No effects on reproductive organs were observed in treated male and female rats as compared to control. Respiratory tract infections accounted for 28 deaths. Two animals died of starvation, one of meningitis, one of a ruptured right auricle and three as the result of neoplasms. In addition, Middle ear and respiratory infections were observed, considered to be in older rats of the colony. Chronic otitis media was observed in nearly 50 per cent of the animals. The disease was evenly distributed in the various groups. Pathological changes in the adrenal cortex were observed in 22 animals. This particular pathology was acute in nature and was not considered to be an effect of the food colours since it was observed in two of the control animals and since there was no correlation between the incidence and the concentration of colour fed. 18 per cent tumour incidence was observed in treated rats. The differences in tumour incidence are not significant according to chi-square tests. The tracings of ECG were essentially normal. No significant deviation of electrical axis observed in treated rats. Therefore, NOAEL was considered to be 750 mg/kg bw when male and female rats were treated with test chemical orally in feed Alphacel (a non-nutritive cellulose) for 64 weeks.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 300 mg/kg bw/day

Study duration:

chronic

Species:

mouse

Quality of whole database:

Weight of evidence prepared from two well qualified publication.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Data available for the target chemicals was reviewed to determine the toxic nature of the test chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3- [2-[4-[[2-(sulfooxy) ethyl]sulfonyl]phenyl]diazenyl] -8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (607724-37-8).The studies are as mentioned below:

Reproductive study was conducted for test chemical in male and female CFW mice for 80 weeks by oral feed. When Groups of 30 male and 30 female CFW mice were given diets containing 0.1, 0.25, 0.5 and 1.0% test chemical for 80 wk, with groups of 60 mice of each sex as controls. There were no dose-related effects on body-weight gain, haematology and organ weights. The incidence of histopathological findings was not altered by the feeding of test chemical. Therefore it is considered that test chemical to the diet of male and female CFW mice at levels up to 1% (equivalent to approximately 1300 mg/kg/day) for 80 wk does not induce any reprotoxic effect.

In a chronic toxicity study, male and female rats were treated with test chemical in the concentration of 0, 15, 150 and 750 mg/kg bw orally in feed Alphacel (a non-nutritive cellulose) for 64 weeks. No effect on survival of treated male and female rats were observed at 15, 150 and 750 mg/kg bw as compared to control. Decrease in mean body weight in females at 4, 32 and 16 weeks. Decreased in food consumption of females were observed with no correlation with the concentration given in diet. No significant difference in male or female food efficiency was observed as compared to control. The only significant difference observed was lowered white cell count for females at 750 mg/kg bw as compared to controls. No adverse effect on blood cells was observed. Similarly, Decrease in liver weight at 150 mg/kg bw and spleen weight at 750 mg/kg bw in female rat. These changes were not correlated with the level of food colour in the diet and are difficult to interpret. No effects on reproductive organs were observed in treated male and female rats as compared to control. Respiratory tract infections accounted for 28 deaths. Two animals died of starvation, one of meningitis, one of a ruptured right auricle and three as the result of neoplasms. In addition, Middle ear and respiratory infections were observed, considered to be in older rats of the colony. Chronic otitis media was observed in nearly 50 per cent of the animals. The disease was evenly distributed in the various groups. Pathological changes in the adrenal cortex were observed in 22 animals. This particular pathology was acute in nature and was not considered to be an effect of the food colours since it was observed in two of the control animals and since there was no correlation between the incidence and the concentration of colour fed. 18 per cent tumour incidence was observed in treated rats. The differences in tumour incidence are not significant according to chi-square tests. The tracings of ECG were essentially normal. No significant deviation of electrical axis observed in treated rats. Therefore, NOAEL was considered to be 750 mg/kg bw when male and female rats were treated with test chemical orally in feed Alphacel (a non-nutritive cellulose) for 64 weeks.

Thus based on the above annotation and CLP criteria the test chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3- [2-[4-[[2-(sulfooxy) ethyl]sulfonyl]phenyl]diazenyl] -8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (607724-37-8)is not likely to exhibit reproductive toxicant substance. Hence the substance cannot be classified as reproductive toxicant.

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

Thus based on the above annotation and CLP criteria the test chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3- [2-[4-[[2-(sulfooxy) ethyl]sulfonyl]phenyl]diazenyl] -8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:4) (607724-37-8)is not likely to exhibit reproductive toxicant substance. Hence the substance cannot be classified as reproductive toxicant.