Disodium 3-hydroxy-4-[(4-methyl-2-sulphonatophenyl)azo]-2-naphthoate

Endpoint summary

Description of key information

Chronic oral toxicity study was published by Environmental Protection Agency (National Technical Reports Library, 1992) to find the oral toxic nature of disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate /(5858-81-1) Other name: D&C Red 6 upon repeated exposure in mice. In a chronic toxicity study, CD male and female mice were exposed to disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate in the concentration 0 ,0, 75, 1500 and 7500 mg/kg bw/day oral feed. Slight compound related increase in mortality was observed in male mice during last 6 months of study at 7500 mg/kg bw. Hair and exposed skin area appeared red, a reddish brown ventral abdomen and/or anogenital region was observed reddish brown in colour at 1500 and 7500 mg/kg bw and anogenital region was observed orange in colour at 75 mg/kg bw. No effect on body weight, food consumption, Organ weight and haematological value of treated mice was observed as compare to control. Similarly, No effect on gross pathology of treated male and female mice was observed as compared to control. Revealed degenerative renal changes were observed in 7500 mg/kg bw treated male mice but it was not compound related . Therefore, NOAEL was considered to be 1500 mg/kg/day for CD male and female mice, when they were exposed to disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate by oral feed.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data is from secondary source.

Qualifier:

according to guideline

Guideline:

other: As mention below

Principles of method if other than guideline:

To evaluate the toxicity of Pigment Red 57 in male and female mice by chronic study.

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

Name of the test chemical: disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylateCommon Name: D & C Red No. 6Molecular Formula: C18H12N2Na2O6SMolecular Weight: 430.3468 g/molInChI: 1S/C18H14N2O6S.2Na/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*+1/p-2/b20-19+;;Substance Type: OrganicPhysical State: SolidSMILES:Cc1ccc(N=Nc2c3ccccc3cc(C(=O)O{-}.[Na]{+})c2O)c(S(=O)(=O)O{-}.[Na]{+})c1

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

Details on test animalTEST ANIMALS- 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: 24-30 g, Female: 20-24 g- Fasting period before study: No data available - Housing: Animals were housed individually in suspended wire mesh cages and identified ear tagged. - Diet (e.g. ad libitum): Basal laboratory diet, ad libitum. Change from Rodent Laboratory Chow@#5001 to Certified Rodent Chow. - Water (e.g. ad libitum): water, ad libitum- Acclimation period: 2 weeksENVIRONMENTAL 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: Purina Laboratory diet

Details on oral exposure:

VEHICLE- Concentration in vehicle: 0, 75, 1500 and 7500 mg/kg bw/day

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

2 years

Frequency of treatment:

Daily

Dose / conc.:

0 other: mg/kg bw/day

Dose / conc.:

75 other: mg/kg bw/day

Dose / conc.:

1 500 other: mg/kg bw/day

Dose / conc.:

7 500 other: mg/kg bw/day

No. of animals per sex per dose:

Total : 6000 mg/kg body weight/day: 60 male, 60 Female 0 mg/kg body weight/day: 60 male, 60 Female 75 mg/kg body weight/day: 60 male, 60 Female 1500 mg/kg body weight/day: 60 male, 60 Female 7500 mg/kg body weight/day: 60 male, 60 Female

Control animals:

yes, concurrent vehicle

Details on study design:

Not specified.

Positive control:

Not specified.

Observations and examinations performed and frequency:

Observations and examinations performed & frequencyCAGE SIDE OBSERVATIONS: Yes - Time schedule: The mice were observed thrice daily (Monday to Friday) and twice daily (weekend and holiday) for toxicity, morbidity and mortality.DETAILED CLINICAL OBSERVATIONS: Yes - Time schedule: Detail observation recorded weekly.BODY WEIGHT: Yes - Time schedule for examinations: Individual body weight were recorded weekly during the first 14 weeks of study, biweekly (the second seven days of every 2 weeks) during the next 12 weeks and monthly (seven days during the third week of each month.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 , Individual food consumption were recorded weekly during the first 14 weeks of study, biweekly (the second seven days of every 2 weeks) during the next 12 weeks and monthly (seven days during the third week of each month.- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No dataFOOD 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: Yes ,but available on request ,but not in this paper. WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes, but available on request, but not in this paper. OPHTHALMOSCOPIC EXAMINATION: No data availableHAEMATOLOGY: Yes - Time schedule for collection of blood: On 3, 6, 12 and 18 month.- Anaesthetic used for blood collection: Yes (identity) / No / No data- Animals fasted: No data available- How many animals: 10 /mice /sex/group- Parameters checked in table [No.?] were examined.CLINICAL CHEMISTRY: Yes / No / No data- Time schedule for collection of blood:- Animals fasted: Yes / No / No data- How many animals:- Parameters checked in table [No.?] were examined.URINALYSIS: Yes NEUROBEHAVIOURAL EXAMINATION: No data available.

Sacrifice and pathology:

Sacrifice and pathologyGROSS 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

Statistics:

Statistically analysis was performed for two tailed pair wise statistical comparisons. Body weight, clinical pathology parameters, organ weight, compared by analysis of one way variances homoginicity of variances by Barrletts test and survival indices compared using Chi- square test were used.

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

No significant changes were observed in treated group dose level 75, 1500 and 7500 mg/kg bw/day compare to control.

Mortality:

mortality observed, treatment-related

Description (incidence):

Mortality; Slight compound related increase in mortality was observed in male mice during last 6 months of study at 7500 mg/kg bw.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

No significant changes were observed in treated group 75, 1500 and 7500 mg/kg bw/day compare to control.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

No significant changes were observed in treated group 75, 1500 and 7500 mg/kg bw/day compare 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):

No significant changes were observed in treated group dose level 75, 1500 and 7500 mg/kg bw/day compare to control.

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

Description (incidence and severity):

No significant changes were observed in treated group dose level 75, 1500 and 7500 mg/kg bw/day compare to control.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No significant changes were observed in treated group dose level 75, 1500 and 7500 mg/kg bw/day compare to control.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

Statically significant renal changes were observed in the dose level of 7500 mg/kg bw/day of treated male but it was not related to the compound consumed.No changes were observed on the treated female at dose level of 75, 1500 and 7500 mg/kg bw/day compare to control.

Histopathological findings: neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

Statically significant renal changes were observed in the dose level of 7500 mg/kg bw/day of treated male but it was not related to the compound consumed.No changes were observed on the treated female at dose level of 75, 1500 and 7500 mg/kg bw/day compare to control.

Other effects:

not specified

Key result

Dose descriptor:

NOAEL

Effect level:

1 500 other: mg/kg body weight/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

histopathology: non-neoplastic

mortality

Remarks on result:

other: No significant effect were observed on mortality and histopathology.So as the substance can be considered to be toxic

Critical effects observed:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Survival after 104 weeks

		No. of survivor/No of initiated
Sr no .	Treatment level	Male	Female
1	Control1	30/60	24/60
2	Control 2	33/60	26/60
3	75 mg/kg bw/day	26/60	26/60
4	1500 mg/kg bw/day	23/60	23/60
5	7500 mg/kg bw/day	17/60	24/60

Conclusions:

NOAEL was considered to be 1500 mg/kg/day for male and female CD mice ,when they were exposed to disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate byoral feed.

Executive summary:

In a chronic toxicity study, CD male and female mice were exposed to disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate in the concentration 0 ,0, 75, 1500 and 7500 mg/kg bw/day oral feed. Slight compound related increase in mortality was observed in male mice during last 6 months of study at 7500 mg/kg bw. Hair and exposed skin area appeared red, a reddish brown ventral abdomen and/or anogenital region was observed reddish brown in colour at 1500 and 7500 mg/kg bw and anogenital region was observed orange in colored at 75 mg/kg bw. No effect on body weight, food consumption, Organ weight and hematological value of treated mice was observed as compare to control. Similarly, No effect on gross pathology of treated male and female mice was observed as compared to control. Revealed degenerative renal changes were observed in 7500 mg/kg bw treated male mice but it was not compound related . Therefore, NOAEL was considered to be 1500 mg/kg/day for CD male and female mice, when they were exposed to disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate by oral feed.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 500 mg/kg bw/day

Study duration:

chronic

Species:

mouse

Quality of whole database:

Data is taken from K4 National Technical Reports Library

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

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

Additional information

Repeated dose oral toxicity:

Various experimental studies were reviewed to determine the toxic nature of disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate /(5858-81-1) Other name: D&C Red 6 and Lithol rubine B(R201)upon repeated exposure by oral route. The studies are as mentioned below:

Chronic oral toxicity study was published by Environmental Protection Agency (National Technical Reports Library, 1992) to find the oral toxic nature of disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate /(5858-81-1) Other name: D&C Red 6 upon repeated exposure in mice. In a chronic toxicity study, CD male and female mice were exposed to disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate in the concentration 0 ,0, 75, 1500 and 7500 mg/kg bw/day oral feed. Slight compound related increase in mortality was observed in male mice during last 6 months of study at 7500 mg/kg bw. Hair and exposed skin area appeared red, a reddish brown ventral abdomen and/or anogenital region was observed reddish brown in colour at 1500 and 7500 mg/kg bw and anogenital region was observed orange in colour at 75 mg/kg bw. No effect on body weight, food consumption, Organ weight and haematological value of treated mice was observed as compare to control. Similarly, No effect on gross pathology of treated male and female mice was observed as compared to control. Revealed degenerative renal changes were observed in 7500 mg/kg bw treated male mice but it was not compound related . Therefore, NOAEL was considered to be 1500 mg/kg/day for CD male and female mice, when they were exposed to disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate by oral feed.

Supported by another experimental study which was published by Environmental Protection Agency (National Technical Reports Library, 1992) to find the oral toxic nature of disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate /(5858-81-1) Other name: D&C Red 6 upon repeated exposure in rat . In a chronic toxicity and carcinogenicitystudy,Charies river CD®male and female rat were exposed to disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3- oxidonaphthalene -2-carboxylate 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 andfood consumption,ophthalmoscopic examination, haematology, clinical chemistry, urinalysis, virology and microbiologyof tAreated male and female rats was observed as comparable to control. Similarly, No effect on reproductive performance such as Fertilityindex, gestation index and effect on parturition and lactation were observed as compared to control. No effect organ weight and gross pathology were observed as compared to control in F0 and F1 generation. In addition, 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. 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 todisodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate orally in feed.

 

The data available for the target chemical disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate /(5858-81-1) Other name: D&C Red 6 and Lithol rubine B(R201) is insufficient to classify the chemical as toxic. Also the NOAEL value range can be close to1500mg/kg bw/day . Based on the observations made, D&C Red 6 does not exhibit toxicity upon repeated exposure by oral route. Hence the test chemical is not likely to classify as a toxicant upon repeated exposure by oral route. Further testing is required to clearly judge the test chemical as toxic upon repeated exposure.

Justification for classification or non-classification

Thus based on the above annotation for the target chemical disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate /(5858-81-1) Other name: D&C Red 6 and Lithol rubine B(R201) is insufficient to classify the chemical as toxic. Also the NOAEL value range can be close to1500mg/kg bw/day . Based on the observations made, D&C Red 6 does not exhibit toxicity upon repeated exposure by oral route. Hence the test chemical is not likely to classify as a toxicant upon repeated exposure by oral route. Further testing is required to clearly judge the test chemical as toxic upon repeated exposure.