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

Toxicity to reproduction

Administrative data

Endpoint:

fertility, other

Remarks:

Combined repeated dose and carconogenicity test

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 reviewed 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 Osborne Mendel rats

GLP compliance:

not specified

Limit test:

no

Justification for study design:

not specified

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:

Osborne-Mendel

Details on species / strain selection:

No Data Available

Sex:

male/female

Details on test animals or test system and environmental conditions:

Rats were caged individually.

Administration / exposure

Route of administration:

oral: feed

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: Commercial Laboratory Chow diet

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
DIET PREPARATION
The test chemical was mixed into a commercial laboratory chow diet for each feeding study.

Details on mating procedure:

Mating was not performed

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No Data Available

Duration of treatment / exposure:

two-years

Frequency of treatment:

daily

Details on study schedule:

no data available

Doses / concentrations

Remarks:

0, 500, 1000 mg/kg bw/day

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 no treatment

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 nonneoplastic, 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 mamary glands were also observed for tumours.

Positive control:

no data available

Examinations

Parental animals: Observations and examinations:

no data available

Oestrous cyclicity (parental animals):

no data available

Sperm parameters (parental animals):

no data available

Litter observations:

no data available

Postmortem examinations (parental animals):

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 nonneoplastic, 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 mamary glands were also observed for tumours.

Statistics:

no data available

Reproductive indices:

no data available

Offspring viability indices:

no data available

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

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

Dermal irritation (if dermal study):

not specified

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

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Findings, such as liver enlargement, abscesses, distendedurinary bladders, ovarian cysts, distended uteri, enlarged parathyroids, enlargement of seminal vesicles and other miscellaneous changes, were present in approximately equal incidence and degree in test and control rats. No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Osborne Mendel strain on this two-year experiment.

Histopathological findings: neoplastic:

no effects observed

Description (incidence and severity):

No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Osborne Mendel rats on this two-year experiment. Rats of the Osborne- Mendel strain had more tumors (especially mammary).Incidence of tumor formation in mammary glands were found to be in control and treatment groups. Thus, they are considered to be incidemtal and non treatment related in nature and strain specific.

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

not specified

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

gross pathology

histopathology: non-neoplastic

histopathology: neoplastic

Remarks on result:

not determinable due to absence of adverse toxic effects

Results: P1 (second parental generation)

Effect levels (P1)

Remarks on result:

not measured/tested

Results: F1 generation

Effect levels (F1)

Remarks on result:

not measured/tested

Results: F2 generation

General toxicity (F2)

Other effects:

not specified

Developmental neurotoxicity (F2)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F2)

Developmental immunotoxicity:

not specified

Effect levels (F2)

Remarks on result:

not measured/tested

Overall reproductive toxicity

Reproductive effects observed:

no

Treatment related:

no

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	158	75	78
Number received by pathology	171	89	89
Number of males with tumors	25	9	10
Number of females with tumors	42	14	22
Total number of rats with tumors	67	23	32
Tumor type in each rat
Fibroadenoma, mammary	19	9	12
Adenocarcinoma	18	6	7
Lymphosarcoma	18	7	5
Lymphoblastoma	5	0	0
Fibrosacroma	0	0	4
Interstitial cell tumor	5	4	2
Endometrial sarcoma	3	1	0
Other tumors	10	1	5
Other pathology
Chronic pneumonia	34	18	16
Granular kidneys	49	23	23
Splenic enlargement	27	16	13
Splenic atrophy	17	16	9
Testicular atrophy	8	8	4
Adrenal enlargement	5	2	3
Chronic arteritis	2	2	0

Applicant's summary and conclusion

Conclusions:

Findings, such as liver enlargement, abscesses, distended urinary bladders, ovarian cysts, distended uteri, enlarged parathyroids, enlargement of seminal vesicles and other miscellaneous changes, were present in approximately equal incidence and degree in test and control rats. No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Osborne Mendel strain on this two-year experiment.Considering all the observations, the No Observed Adverse Effect Level was considered to be 1000 mg/kgbw/day in Osborne Mendel rats.

Executive summary:

Combined Repeated Dose and Carcinogenicity was performed to determine the toxic nature of the test chemical in Osborne Mendel rats. Two-year tests were run in which 400 male and female Osborne Mendel 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. Findings, such as liver enlargement, abscesses, distended urinary bladders, ovarian cysts, distended uteri, enlarged parathyroid, enlargement of seminal vesicles and other miscellaneous changes, were present in approximately equal incidence and degree in test and control rats. Rats of the Osborne- Mendel strain had more tumors (especially mammary).Incidence of tumor formation in mammary glands were found to be in control and treatment groups. Thus, they are considered to be incidental and non-treatment related in nature and strain specific. No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Osborne Mendel strain on this two-year experiment. Considering all the observations, the No Observed Adverse Effect Level was considered to be 1000 mg/kg bw/day in Osborne Mendel rats.