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

Repeated dose toxicity: oral

Administrative data

Endpoint:

repeated dose toxicity: oral

Remarks:

combined repeated dose and carcinogenicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from peer reviwed journal

Data source

Reference

Reference Type:

publication

Title:

Chronic Toxicity of the given test chemical to Rats

Author:

K. J. DAVIS ET AL.

Year:

1966

Bibliographic source:

TOXICOLOGY AND APPLIED PHARAMACOLOGY

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: Combined Repeated Dose and Carcinogenicity

Principles of method if other than guideline:

Combined Repeated Dose and Carcinogenicity was performed to determine the toxic nature of the test chemical in Sprague Dawley rats

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 3-[(2,4-dimethyl-5-sulphonatophenyl)azo]-4-hydroxynaphthalene-1-sulphonate

EC Number:

224-909-9

EC Name:

Disodium 3-[(2,4-dimethyl-5-sulphonatophenyl)azo]-4-hydroxynaphthalene-1-sulphonate

Cas Number:

4548-53-2

Molecular formula:

C18H16N2O7S2.2Na

IUPAC Name:

Disodium 3-[(2,4-dimethyl-5-sulphonatophenyl)azo]-4-hydroxynaphthalene-1-sulphonate

Test material form:

solid

Details on test material:

- Name of test material (as cited in study report): Ponceau SX
- Molecular formula : C18H16N2O7S2.2Na
- Molecular weight : 480.4276 g/mol
- Smiles notation: [Na+].[Na+].Cc1cc(C)c(cc1N=Nc2cc(c3ccccc3c2O)S(=O)(=O)[O-])S(=O)(=O)[O-]
- InChl :1S/C18H16N2O7S2.2Na/c1-10-7-11(2)16(28(22,23)24)8-14(10)19-20-15-9-17(29(25,26)27)12-5-3-4-6-13(12)18(15)21;;/h3-9,21H,1-2H3,(H,22,23,24)(H,25,26,27);;/q;2*+1/p-2/b20-19+;;
- Substance type: Organic
- Physical state: Solid

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Rats were caged individually.

Administration / exposure

Route of administration:

oral: feed

Details on route of administration:

The test chemical was mixed into a commercial laboratory chow diet for each feeding study.

Vehicle:

other: commercial laboratory chow diet

Details on oral exposure:

The test chemical was mixed into a commercial laboratory chow diet for each feeding study.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

2yrs

Frequency of treatment:

Daily

Doses / concentrations

Remarks:

0, 500, 1000 mg/kg bw

No. of animals per sex per dose:

control - 200(100/ sex)
500 mg/kgbw/day - 100 (50/sex)
1000 mg/kgbw/day - 100 (50/sex)

Control animals:

yes, concurrent vehicle

Details on study design:

Two-year tests were run in which 400 rats were distributed into various treatment groups. The test chemical was mixed into a commercial laboratory chow diet for each feeding study.Rats were individually caged. Each animal received in the Pathology Laboratory was given a careful gross examination, and all lesions were recorded. Of those not received, most were discarded because of advanced postmortem autolysis. So far as can be determined, none of the discarded animals had tumors. Lesions which could not clearly be identified as neoplastic or non-neoplastic, and tumors, the type of which was uncertain or uncommon, were also sectioned and examined microscopically. Tissues sectioned include the following: liver, lung, heart, spleen, pancreas, stomach, small in testine, colon, kidney, adrenal, thyroid, testis (or ovary and uterus), leg muscles, leg bones with included marrow, and any tumor or other unusual condition.Sections of the mammary glands were also observed for tumours.

Examinations

Observations and examinations performed and frequency:

The animals were observed for survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Other examinations:

No data available

Statistics:

No data available

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

The ingestion of the test chemical had no effect on survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes in the test animals

Mortality:

no mortality observed

Description (incidence):

The ingestion of the test chemical had no effect on survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes in the test animals

Body weight and weight changes:

no effects observed

Description (incidence and severity):

The ingestion of the test chemical had no effect on survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes in the test animals

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

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

The ingestion of the test chemical had no effect on survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes in the test animals

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Sprague Dawley strain on this two-year experiment.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Sprague Dawley strain on this two-year experiment.

Histopathological findings: neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Sprague Dawley strain on this two-year experiment.

Other effects:

not specified

Effect levels

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical signs

histopathology: neoplastic

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Lesions observed in the 2 year feeding study

Parameter	Control	1%	2%
Number of rats started	200	100	100
Number of survivors at 80 weeks	122	66	54
Number received by pathology	147	83	74
Number of males with tumors	14	5	4
Number of females with tumors	24	11	10
Total number of rats with tumors	38	16	14
Tumor type in each rat
Fibroadenoma, mammary	21	9	9
Adenocarcinoma	1	1	0
Lymphosarcoma	5	4	4
Lymphoblastoma	4	0	0
Fibrosacroma	3	0	0
Interstitial cell tumor	0	0	0
Endometrial sarcoma	0	0	0
Other tumors	5	2	4
Other pathology
Chronic pneumonia	30	13	13
Granular kidneys	31	9	18
Splenic enlargement	12	9	8
Splenic atrophy	12	3	4
Testicular atrophy	13	4	4
Adrenal enlargement	7	2	7
Chronic arteritis	9	2	5

Applicant's summary and conclusion

Conclusions:

The ingestion of the test chemical had no effect on survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes in the test animals. No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Sprague Dawley strain on this two-year experiment. Hence, the No Observed Adverse Effect level was considered to be 1000 mg/kgbw/day in both male and female Sprague Dawley rats.

Executive summary:

Combined Repeated Dose and Carcinogenicity was performed to determine the toxic nature of the test chemical in Sprague Dawley rats. Two-year tests were run in which 400 male and female Sprague Dawley rats were distributed into various treatment groups. The treatment groups were - 100 rats/dose (50/sex) and 100 rats/sex for controls. The test chemical (500 mg/kg bw/day, 1000 mg/kg bw/day) was mixed into a commercial laboratory chow diet for each feeding study. Rats were individually caged. Each animal received in the Pathology Laboratory was given a careful gross examination, and all lesions were recorded. Of those not received, most were discarded because of advanced postmortem autolysis. So far as can be determined, none of the discarded animals had tumors. Lesions which could not clearly be identified as neoplastic or non-neoplastic, and tumors, the type of which was uncertain or uncommon, were also sectioned and examined microscopically. Tissues sectioned include the following: liver, lung, heart, spleen, pancreas, stomach, small intestine, colon, kidney, adrenal, thyroid, testis (or ovary and uterus), leg muscles, leg bones with included marrow, and any tumor or other unusual condition. Sections of the mammary glands were also observed for tumors. The ingestion of the test chemical had no effect on survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes in the test animals. No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Sprague Dawley strain on this two-year experiment. Hence, the No Observed Adverse Effect level was considered to be 1000 mg/kg bw/day in both male and female Sprague Dawley rats.