Registration Dossier

Administrative data

Description of key information

Repeated dose toxicity: via oral route;

NOAEL was considered to be in a dose range of 1000-1300 mg/kg bw when the test animals were  treated with test chemical.

Repeated inhalation study:

According to Annex IX of the REACH regulation, testing by the inhalation route is appropriate only if exposure of humans via inhalation is likely. Taking into account the low vapour pressure of the substance 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt (503155-49-5) which is reported as 6.788058e-45 mmHg at 25 C. Also considering the particle size distribution of the substance the majority of the particles was found to be in the size of 150 micron to 106 micron which is much larger size range compared to the inhalable particulate matter .Thus, exposure to inhalable dust, mist and vapour of the test chemical is highly unlikely. Therefore this study is considered for waiver.

Repeated dermal study;

The acute toxicity value for 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt (503155-49-5)  (as provided in section 7.2.3) is >2000 mg/kg body weight. Also, given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt   shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
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 structurally similar chemicals
Justification for type of information:
Data for the target chemical is summarized based on the structurally similar chemicals
Reason / purpose:
read-across source
Related information:
Composition 1
Reason / purpose:
read-across source
Related information:
Composition 1
Reference:
Composition 0
Qualifier:
according to
Guideline:
other: as below
Principles of method if other than guideline:
WoE report is based on two repeated dose toxicity studies on rats
1.To evaluate the toxicity of test chemical in male and female mice by chronic study.
2.Chronic toxicity study was conducted to evaluate the toxic potential of test chemical in CD male and female rats by oral feed.
GLP compliance:
not specified
Limit test:
no
Test material information:
Composition 1
Species:
other: 1. Mice, 2. Rat
Strain:
other: 1.CFW strain, 2. Charies river CD®
Sex:
male/female
Details on test animals and environmental conditions:
1.Details on test animal
TEST ANIMALS
- Source: Specified-pathogen-free colony
- Age at study initiation: Male : 21-22, female: 18-19
- Weight at study initiation: Male: 21 – 22 g, Female: 18 – 19 g
- Fasting period before study: No data available
- Housing: They were caged in groups of 15 in a room maintained at 21±1°C with a relative humidity of 50-60%.
- Diet (e.g. ad libitum): Oxoid pasteurized breeding diet , ad libitum
- Water (e.g. ad libitum): water available , ad libitum
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 1°C
- Humidity (%):50-60%
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 21 ± 1°C No data available


2.- Source: Charies river Breeding Laboratories. Iac. Wilningeon
- Age at study initiation: (P) x wks; (F1) x wks: P: 36 days
- Weight at study initiation: (P) Males: 75-155 g, Female: 75-114 g
- Fasting period before study: No data available
- Housing: Animals were housed individually in hanging wire mash cages.
- Diet (e.g. ad libitum): Purina Laboratory diet, ad libitum. During post weaning segment Purina La
boratory diet, Rodent Laboratory Chow@#5001 and Certified Rodent Chow were used respectively.
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: 15 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 71 °
- Humidity (%): 53 %
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 12 hr light/12hr dark
Route of administration:
oral: feed
Vehicle:
other: 1 & 2. Purina Laboratory diet
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
1.80 weeks, 2. 126 week
Frequency of treatment:
Daily
Remarks:
0.1, 0.25,0.5 or 1.0% (130,325,650,1300 mg/kg bw/d)
Remarks:
0, 25, 150 and 1000 mg/kg bw
No. of animals per sex per dose:
1. 30 male and 30 female mice


2. Total : 1300
F0 generation
0 mg/kg body weight/day: 60 male, 60 Female
0 mg/kg body weight/day: 60 male, 60 Female
25 mg/kg body weight/day: 60 male, 60 Female
150 mg/kg body weight/day: 60 male, 60 Female
1000 mg/kg body weight/day: 60 male, 60 Female
F1 generation
0 mg/kg body weight/day: 70 male, 70 Female
0 mg/kg body weight/day: 70 male, 70 Female
25 mg/kg body weight/day: 70 male, 70 Female
150 mg/kg body weight/day: 70 male, 70 Female
1000 mg/kg body weight/day: 70 male, 70 Female
Control animals:
yes, concurrent vehicle
Details on study design:
Not specified.
Positive control:
Not specified.
Observations and examinations performed and frequency:
1.Observations and examinations performed & frequency
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Frequently
- Cage side observations checked in table [No.?] were included.: ill-health and mortality were obsaerved.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Frequently

BODY WEIGHT: Yes
- Time schedule for examinations: At week 0, 15, 39, 57 and 73 of study

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available
- Time schedule for examinations: No data available

OPHTHALMOSCOPIC EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available

HAEMATOLOGY: Yes
- Time schedule for collection of blood:
At week 28 and 55 from 10 males and 10 females from the control group and from the groups of 0.5 and 1.0% dietary levels.
At 80 wk, blood samples were collected from the aorta of all surviving mice during the autopsy.
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 20 animals (10 male and 10 female)
- Parameters were examined: counting the reticulocyte and leucocytes.

CLINICAL CHEMISTRY: No data
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters were examined: No data

URINALYSIS: Yes
- Time schedule for collection of urine: At 28 wks at 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 1.0%) of Black PN.
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters were examined: reducing substances, bile salts and blood as well as for colour, pH and microscopic constituents


2. CAGE SIDE OBSERVATIONS: Yes
DETAILED CLINICAL OBSERVATIONS: Yes
F0; the clinical sign and mortality was observed twice daily
Pups: the clinical sign and mortality was observed once daily
F1: the clinical sign and mortality was observed twice daily
BODY WEIGHT: Yes
- Time schedule for examinations:
F0; the body weight was observed weekly. For females body weight was observed individually on 0,6,15 and 21 of gestation
Pups: The pups were weighed as a litter on 1 and 4 day of lactation. Even on 21 day they were observed for lactation.
F1: the body weight was observed weekly for first 14 weeks and biweekly for next 12 weeks and once mortality thereafter.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg
body weight/day: Yes ,
F0; the food consumption was observed weekly for 1st 10 weeks.
F1: the food consumption was observed weekly for 1st 14 weeks. Biweekly (the second 7 days of every
2 week)the next 12 weeks and once monthly (7 days during the last week of each month)
- Compound intake calculated as time-weighted averages from the consumption and body weight gain
data: Yes
F0; Compound consumption was calculated from the diet consumption and body weight measurement.
F1: Compound consumption was calculated from the diet consumption and body weight measurement.
FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted
averages from the consumption and body weight gain data: No data available
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations:
F0; the ophthalmoscopic examination was observed for all rats till 15.
F1: the ophthalmoscopic examination was observed for all male rats on 3, 6,12, 18 and 21 month while 2
4 month for females..
HAEMATOLOGY: - Yes, Blood was obtained by puncture of orbital sinus pleura.
- Time schedule for collection of blood: During the3, 6,12, 18 and 21 of month for male while on 24
month for females
- Anaesthetic used for blood collection: No data available
- Animals fasted: Yes
- How many animals: 10 rats /sex were selected .While only 9 rats were selected from 1000 mg/kg/day at
21 months.
- Parameters checked in table [No.?] were examined. Hemoglobin, Haematocrits, Differential leukocyte
count, Erythrocyte reticulocytes count
CLINICAL CHEMISTRY: Yes, Blood was obtained by puncture of orbital sinus pleura.
- Time schedule for collection of blood: During the3, 6,12, 18 and 21 of month for male while on 24 mont
h for females
- Anaesthetic used for blood collection: No data available
- Animals fasted: Yes
- How many animals: 10 rats /sex were selected .While only 9 rats were selected from 1000 mg/kg/day at
21 months.
- Parameters checked in table [No.?] were examined. Glucose ,BUN,SGOT,SGPT Creatinine and Serum
total protein
URINALYSIS: Yes
- Time schedule for collection of urine: 3, 6,12, 18 and 21 of month for female
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked in table [No.?] were examined. pH ,specific gravity, Quantative test for protein,
glucose ,bilirubin, Ketones occult blood colour and appearance.
NEUROBEHAVIOURAL EXAMINATION: No data available
OTHER: AT 20 months of study, blood was collected from 2 fasted rats /sex/group and send for virology
and microbiology test. The blood was taken from rat who was showing the sign of pulmonary distress.
Sacrifice and pathology:
1. GROSS PATHOLOGY: Yes ,
After 24 month, all surviving mice were sacrificed by carbon dioxide asphyxiation. Complete external examination was performed, including all orifices, each carcasses and content of abdomen, thoracic and cranial cavities were examined.
HISTOPATHOLOGY: Yes, The following tissue were weighed and fixed in buffered neutral formalin. Brain ,liver kidney and spleen , abdominal aorta ,adrenal (2)bone and bone marrow, blood amear2,eye, gall bladder,ovary,testis (2)epididymides, heart ,intestine, liver ,lung, lymph node, mammary gland,mandibular,salivary gland nerve pancreas, pituitary ,seminal vesicle, skeleton muscle, skin, spinal cord ,spleen,stomach,thymus,trachea, thyroid ,urinary bladder, uterus and prostrate.
Other examinations
OTHER: Stool analysis was also performed

2.
GROSS PATHOLOGY: Yes,
F1: After 12 months of the dietary consumption 10 rats / sex /group was scarified by carbon di oxide asphyxiation. Rats were observed for macroscopic changes in brain ,kidney ,liver, spleen, testes, thyroid, heart, uterus and ovaries.
HISTOPATHOLOGY: Yes, Animals were observed for microscopic changes. The following tissue were s
tained with Hematoxylin and eosin abdominal aorta ,adrenal (2)bone and bone marrow, blood amear2,eye, gall bladder,ovary,testis (2)epididymides, heart ,intestine, liver ,lung, lymph node, mammary gland,mandibular,salivary gland nerve pancreas, pituitary ,seminal vesicle, skeleton muscle, skin, spinal cord ,spleen,stomach,thymus,trachea,thyroid ,urinary bladder, uterus and prostrate.
Statistics:
1. chi-square test, Student's t test
2. Statistically analysis were performed by using fertility indices, gestation 4, 14 and 21 day survival indices were compared by using Chi- square test and /or Fishur’s extract probability. The mean number of liveborn pups per litter and the litter mean per weight at day 0,4,14 and 21 were compared by analysis of
variance (one way classification), Barletts test for homogenicty of variances and the appropriate t-test (for equal and unequal variances).
For F1 rat: all pair wise statistical comparison were two tailed with the probability level for significance at 0.01. Body weight, food consumption and absolute and relative body weight were compared by analysis of variances and appropriate t-test. Data for animals with malignant tumors, with beings and all tumors
combined were analyzed separately by sex.
Homogenicity was analysed by cor’s test and cehan-Breslow generalized Krunkal-Wallis test. For the
histopathologically
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
1. No significant changes were observed in treated group dose level compare to control.
2. F0; No statically significant change were observed in the treated rat at dose level of 0, 25, 150, or 1000 mg/kg bw/day compare to control.
Pups: No statically significant change was observed in the treated rat at dose level of 0, 25, 150, or 1000 mg/kg bw/day compare to control.
F1: No statically significant change were observed in the treated rat at dose level of 0, 25, 150, or 1000 mg/kg bw/day compare to control.
Description (incidence):
1. : There were no statistically significant differences between the number of deaths in the control mice and those given
2. Mortality
F0; When treated with 150 mg/kg bw, 3 rats were died.
When treated with 1000 mg/kg bw one rat died and in control 3 rats were died.
Pups: No statically significant change were observed in the treated rat at dose level of 0, 25, 150, or 1000 mg/kg bw/day compare to control.
F1: No statically significant change were observed in the treated rat at dose level of 0, 25, 150, or 1000 mg/kg bw/day compare to control.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
1. No significant changes were observed in treated group compare to control.
2. F0; No effect on body weight of treated rat was observed as comparable to control.
F1: When treated with 1000 mg/kg bw, significant decrease in body weight was observed in male rat as
comparable to control.
When treated with 25 and 150 mg/kg bw, slightly decreased body weight was observed as comparable to control.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):

2. F0; No effect on food consumption of treated male and female rats was observed as comparable to control.
F1: No effect on food consumption of treated male and female offspring’s was observed as comparable to control.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
no effects observed
Description (incidence and severity):
2. F0; No effect on Opthalmoscopic examination of treated male and female rats was observed as comparable to control.
F1: No effect on Opthalmoscopic examination of treated male and female offspring’s was observed as comparable to control.
Haematological findings:
no effects observed
Description (incidence and severity):

2. F0; No effect on Hematology of treated male and female rats was observed as comparable to control.
F1: No effect on Hematology of treated male and female offspring’s was observed as comparable to control.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
2. F0; No effect on clinical chemistry of treated male and female rats was observed as comparable to control.
Urinalysis findings:
no effects observed
Description (incidence and severity):
2. F0; No effect on urine analysis of treated male and female rats was observed as comparable to control.
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):

2. F0; No effect on organ weight of treated male and female rats was observed as compared to control
F1: No effect on organ weight of treated male and female offspring was observed as compared to control.
Gross pathological findings:
no effects observed
Description (incidence and severity):

2. F0; No gross pathological changes were observed in treated male and female rats as compared to control.
F1: No gross pathological changes were observed in treated male and female offspring as compared to control.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
1. Most of the tumours in the study occurred with either a comparable or a greater incidence in the control groups than in the treated mice .Several isolated tumours were identified in mice given the lower levels of Black PN, without comparable findings in the controls or in the highest dose group. Mammary fibroadenoma, a uterine fibromyoma, squamous-cell carcinoma were seen at different concentration.
The frequency of histopathological findings in the treated animals did not differ significantly from those in the controls. Hence, no relationship was obvious between these findings and treatment with Black PN. Only in the case of adenomas of the lung and of the mammary tissue did more than one tumour of a given type occur among the treated animals of any group.

2. F0; An acceleration of testicular changes (degeneration of the testicular tubules) was observed in treated male rat, a common effect in ageing rats. However, at the termination of the study, the increased incidence was not of statistical significance.
F1: An acceleration of testicular changes (degeneration of the testicular tubules) was observed in treated male rat, a common effect in ageing rats. However, at the termination of the study, the increased incidence was not of statistical significance.
Histopathological findings: neoplastic:
not specified
Other effects:
no effects observed
Description (incidence and severity):
2. Virology:
No effect on virology of treated male and female rats was observed as compared to control.
Microbiology:
No effect on Microbiology of treated male and female rats was observed as compared to control
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
ophthalmological examination
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
other: Virology and Microbiology
Remarks on result:
other: No effect observed
Dose descriptor:
NOAEL
Effect level:
1 300 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No significant effect were observed at this dose
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
Conclusions:
NOAEL was considered to be in a dose range of 1000-1300 mg/kg bw when the test animals were treated with test chemical.
Executive summary:

Repeated dose toxicity: via oral route;

The data available for the test chemical was reviewed to determine the toxic nature of 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt (503155-49-5) repeated exposure by oral route. The study is as mentioned below:

 As the read across substances share high similarity in structure and functional group .Therefore, it is acceptable to derive information on toxicity from the analogue substance for target test substance. Repeated dose toxicity test were performed on mice with different concentrations from0.1, 0.25,0.5 or 1.0% (130,325,650,1300 mg/kg bw/d)for 80 wk. 30 males and 30 females was used for the treatment and group of 60 mice of each sex as control.The general condition and behaviour 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. 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. Blood sample were taken at wk 28 and 55.At 80 wk, blood samples were collected from the aorta of all surviving mice during the autopsy. For haematology blood samples were collected and for urine sample from 6-hr period from three groups of five mice of each sex from the controls and the groups on the two highest dietary levels of test chemical. Histopathology was also conducted.There were no dose-related effects on body-weight gain, haematology or organ weights. The incidence of histopathological findings, including tumours, was not altered by the feeding of test chemical.Therefore, the endpoint for the repeated dose toxicity was considered to be NOAEL at 1 % (1300 mg/kg/day) concentration of test substance to mice.

 

In a chronic toxicity and carcinogenicity study, Charies river CD® male and female rat were exposed to test chemical in the concentration 0, 0, 25, 150 and 1000 mg/kg/day orally in feed. In the parental generation, 3 rats were died at 150 mg/kg bw, one rat died at 1000 mg/kg bw and 3 rats were died in in control. No effect on body weight and food consumption, ophthalmoscopic examination, haematology, clinical chemistry, urinalysis, virology and microbiology of treated male and female rats was observed as comparable to control. Similarly, No effect on reproductive performance such as Fertility index, gestation index and effect on parturition and lactation were observed as compared to control. No effect onorgan weight and gross pathology were observed as compared to control in F0 and F1 generation. In addition, anacceleration of testicular changes (degeneration of the testicular tubules) was observed in treated male rat, a common effect in ageing rats. However, at the termination of the study, the increased incidence was not of statistical significance. Therefore, NOAEL was considered to be 1000 mg/kg/day for F0 generation and F1 generation when Charies river CD® male and female rat were exposed to test chemical orally in feed.

 

Based on the data available from the test chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt (503155-49-5) does not exhibit repeated dose oral toxicity in the range of 1000-1300 mg/kg bw. Hence the test chemical is not likely to classify as a repeated dose oral toxicity as per the criteria mentioned in CLP regulation.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
chronic
Species:
rat
Quality of whole database:
Woe is prepared from Klimisch 2 publication.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
Wavier

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
Waiver

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Mode of Action Analysis / Human Relevance Framework

Additional information

 Repeated dose toxicity: via oral route;

The data available for the test chemical was reviewed to determine the toxic nature of 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt (503155-49-5) repeated exposure by oral route. The study is as mentioned below:

 As the read across substances share high similarity in structure and functional group .Therefore, it is acceptable to derive information on toxicity from the analogue substance for target test substance. Repeated dose toxicity test were performed on mice with different concentrations from0.1, 0.25,0.5 or 1.0% (130,325,650,1300 mg/kg bw/d)for 80 wk. 30 males and 30 females was used for the treatment and group of 60 mice of each sex as control.The general condition and behaviour 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. 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. Blood sample were taken at wk 28 and 55.At 80 wk, blood samples were collected from the aorta of all surviving mice during the autopsy. For haematology blood samples were collected and for urine sample from 6-hr period from three groups of five mice of each sex from the controls and the groups on the two highest dietary levels of test chemical. Histopathology was also conducted.There were no dose-related effects on body-weight gain, haematology or organ weights. The incidence of histopathological findings, including tumours, was not altered by the feeding of test chemical.Therefore, the endpoint for the repeated dose toxicity was considered to be NOAEL at 1 % (1300 mg/kg/day) concentration of test substance to mice.

 

In a chronic toxicity and carcinogenicity study, Charies river CD® male and female rat were exposed to test chemical in the concentration 0, 0, 25, 150 and 1000 mg/kg/day orally in feed. In the parental generation, 3 rats were died at 150 mg/kg bw, one rat died at 1000 mg/kg bw and 3 rats were died in in control. No effect on body weight and food consumption, ophthalmoscopic examination, haematology, clinical chemistry, urinalysis, virology and microbiology of treated male and female rats was observed as comparable to control. Similarly, No effect on reproductive performance such as Fertility index, gestation index and effect on parturition and lactation were observed as compared to control. No effect onorgan weight and gross pathology were observed as compared to control in F0 and F1 generation. In addition, anacceleration of testicular changes (degeneration of the testicular tubules) was observed in treated male rat, a common effect in ageing rats. However, at the termination of the study, the increased incidence was not of statistical significance. Therefore, NOAEL was considered to be 1000 mg/kg/day for F0 generation and F1 generation when Charies river CD® male and female rat were exposed to test chemical orally in feed.

 

Based on the data available from the test chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt (503155-49-5) does not exhibit repeated dose oral toxicity in the range of 1000-1300 mg/kg bw. Hence the test chemical is not likely to classify as a repeated dose oral toxicity as per the criteria mentioned in CLP regulation.

Repeated inhalation study:

According to Annex IX of the REACH regulation, testing by the inhalation route is appropriate only if exposure of humans via inhalation is likely. Taking into account the low vapour pressure of the substance 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt (503155-49-5) which is reported as 6.788058e-45 mmHg at 25 C. Also considering the particle size distribution of the substance the majority of the particles was found to be in the size of 150 micron to 106 micron which is much larger size range compared to the inhalable particulate matter .Thus, exposure to inhalable dust, mist and vapour of the test chemical is highly unlikely. Therefore this study is considered for waiver.

Repeated dermal study;

The acute toxicity value for 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt (503155-49-5)  (as provided in section 7.2.3) is >2000 mg/kg body weight. Also, given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt   shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

Based on the data available for the target chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt (503155-49-5) does not exhibit toxic nature upon repeated exposure by oral, inhalation and dermal route of exposure and hence is not likely to classify as per the criteria mentioned in CLP regulation.

 

Justification for classification or non-classification

Based on the data available for the target chemical 2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt (503155-49-5) does not exhibit toxic nature upon repeated exposure by oral, inhalation and dermal route of exposure and hence is not likely to classify as per the criteria mentioned in CLP regulation.