3-hydroxy-4-[(2-methoxy-5-nitrophenyl)azo]-N-(3-nitrophenyl)naphthalene-2-carboxamide

Toxicity to reproduction

Administrative data

Endpoint:

reproductive toxicity, other

Remarks:

Toxicology and Carcinogenesis Study

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from NTP

Data source

Reference

Reference Type:

publication

Title:

Toxicology and Carcinogenesis Studies of C.I. Pigment Red 23 (CAS No. 6471-49-4) in F344 Rats and B6C3F1 Mice (Feed Studies)

Author:

U.S. DEPARTMENT OFCOMMERCE National Technical Information Service

Year:

1992

Bibliographic source:

NATIONAL TOXICOLOGY PROGRAM. Technical Report Series. No.411, PB93-228435

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: as below

Principles of method if other than guideline:

Toxicology and Carcinogenesis Study of C.1. Pigment Red 23 (CAS No. 6471-49-4) in F344 Rats

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

3-hydroxy-4-[(2-methoxy-5-nitrophenyl)azo]-N-(3-nitrophenyl)naphthalene-2-carboxamide

EC Number:

229-313-2

EC Name:

3-hydroxy-4-[(2-methoxy-5-nitrophenyl)azo]-N-(3-nitrophenyl)naphthalene-2-carboxamide

Cas Number:

6471-49-4

Molecular formula:

C24H17N5O7

IUPAC Name:

3-hydroxy-4-[(2-methoxy-5-nitrophenyl)azo]-N-(3-nitrophenyl)naphthalene-2-carboxamide

Details on test material:

Name: 3-hydroxy-4-[(2-methoxy-5-nitrophenyl)azo]-N-(3-nitrophenyl)naphthalene-2-carboxamide
SMILES:COc1ccc(N(=O)=O)cc1N=Nc1c2ccccc2cc(C(=O)Nc2cccc(N(=O)=O)c2)c1O
InChI:1S/C24H17N5O7/c1-36-21-10-9-17(29(34)35)13-20(21)26-27-22-18-8-3-2-5-14(18)11-19(23(22)30)24(31)25-15-6-4-7-16(12-15)28(32)33/h2-13,30H,1H3,(H,25,31)/b27-26+
Molecular Formula: C24H17N5O7
Molecular Weight: 487.4263 g/mole

Specific details on test material used for the study:

Name: 3-hydroxy-4-[(2-methoxy-5-nitrophenyl)azo]-N-(3-nitrophenyl)naphthalene-2-carboxamide
SMILES:COc1ccc(N(=O)=O)cc1N=Nc1c2ccccc2cc(C(=O)Nc2cccc(N(=O)=O)c2)c1O
InChI:1S/C24H17N5O7/c1-36-21-10-9-17(29(34)35)13-20(21)26-27-22-18-8-3-2-5-14(18)11-19(23(22)30)24(31)25-15-6-4-7-16(12-15)28(32)33/h2-13,30H,1H3,(H,25,31)/b27-26+
Molecular Formula: C24H17N5O7
Molecular Weight: 487.4263 g/mole

Test animals

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Source: Frederick Cancer Research Center (Frederick, MD)
- Age at study initiation: 56 days old
- Weight at study initiation: No data available
- Fasting period before study: No data available
- Housing: Rats were housed five per cage. Cages were rotated vertically once every two weeks.
- Diet (e.g. ad libitum):NIH-07 Rat, meal (Zeigler Bros., Inc., Gardners, PA), ad libitum
- Water (e.g. ad libitum):Tap water (Birmingham Water Works) in glass water bottles with stainless steel sippers (Edstrom Automatic Watering Systems, Waterford, WI), ad libitum
- Acclimation period:20 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):17.8° C-25.6° C
- Humidity (%):15%-85%
- Air changes (per hr): minimum 15 changes/hour
- Photoperiod (hrs dark / hrs light):12 hours/day

Administration / exposure

Route of administration:

oral: feed

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: NIH-07 Rat, meal

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: of NIH-07 Rat as meal premix with the appropriate amount of C.I. Pigment Red 23.


DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): NIH-07 Rat as meal, - Storage temperature of food:2 weeks for 45 °C

VEHICLE
- Justification for use and choice of vehicle (if other than water):No data
- Concentration in vehicle: No data
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data

Details on mating procedure:

Not specified

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

Not specified

Duration of treatment / exposure:

2 years

Frequency of treatment:

Daily

Details on study schedule:

Not specified

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day

Doses / concentrations 2

Dose / conc.:

425 mg/kg bw/day

Remarks:

for male

Doses / concentrations 3

Dose / conc.:

1 100 mg/kg bw/day

Remarks:

for male

Doses / concentrations 4

Dose / conc.:

2 100 mg/kg bw/day

Remarks:

for male

Doses / concentrations 5

Dose / conc.:

500 mg/kg bw/day

Remarks:

for female

Doses / concentrations 6

Dose / conc.:

1 300 mg/kg bw/day

Remarks:

for female

Doses / concentrations 7

Dose / conc.:

2 600 mg/kg bw/day

Remarks:

for female

No. of animals per sex per dose:

Total: 480
0 mg/kg/day: 60 male
425 mg/kg/day: 60 male
1100 mg/kg/day: 60 male
2100 mg/kg/day: 60 male

500 mg/kg/day: 60 female
1300 mg/kg/day: 60 female
2600 mg/kg/day: 60 female

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:Because levels of C.I. Pigment Red 23 as high as 2500 or 50,000 mg/kg in the feed did not adversely affect survival and mean body weights in the 17-day and 13-week studies, nor cause any chemical-related lesions, doses of 0, 425, 1100 and 2100 mg/kg bw for male and 500, 1300 and 2600 mg/kg/day for female were selected for the 2-year studies.

Positive control:

Not specified

Examinations

Parental animals: Observations and examinations:

Mortality, clinical sign, body weight and feed Consumption were examined.

Oestrous cyclicity (parental animals):

Not specified

Sperm parameters (parental animals):

Not specified

Litter observations:

Not specified

Postmortem examinations (offspring):

Not specified

Statistics:

Probability of survival was estimated by the product-limit procedure of Kaplan and Meier; Neoplasm Incidences were analyzed by adjusting for intercurrent mortality is the prevalence Dinse and Lagakos. alternative methods of statistical analysis were used for rapidly lethal neoplasms, and the Fisher exact test and the Cochran-Armitage trend test. Nonneoplastic Lesion Incidences were analyzed by the Fisher exact test was used, a procedure based on the overall proportion of affected animals. Organ and body weight data analyzed using parametric multiple comparison procedures of Dunnett and Williams. Hematology and clinical chemistry data were analyzed using nonparametric multiple comparison methods of Dunn and Shirley. Jonckheere's test was used to assess the significance of dose-response trends and to determine whether a trend-sensitive test (Williams' or Shirley's test) was more appropriate for pairwise comparisons than a test that does not assume a monotonic dose-response trend (Dunnett's or Dunn's test). Average nephropathy severity values were analyzed for significance using the Mann-Whitney U test.

Reproductive indices:

Not specified

Offspring viability indices:

Not specified

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Description (incidence and severity):

No clinical sign were considered to be treatment related.

Dermal irritation (if dermal study):

not specified

Mortality:

no mortality observed

Description (incidence):

When treated with 2100 and 2600 mg/kg bw, Significant increase in survival of treated male and female rats were observed as compared to control.

When treated with 1100 mg/kg bw, Significant increase in survival of treated male rats were observed as compared to control.
The greater survival of the exposed groups was due principally to the chemically related decreased incidence of mononuclear cell leukemia.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

When treated with 1300 and 2600 mg/kg bw, Significant decrase in body weights of treated female rats were observed as compared to control.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No effect on food consumption of treated male and female rats was observed as compared to control.

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Description (incidence and severity):

When treated with 2600 mg/kg bw in female, significant decrase in Hematocrit values, hemoglobin concentration, and erythrocyte counts were observed as compared to control. Which indicat mild anemia.
No effect in mlae rats were observed as compared to control.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

When treated with 2600 mg/kg bw in female, Significant increase in Serum total bilirubin were observed as compared to control.
This finding coupled with the mild anemia suggests a mild hemolytic process.

No effect in mlae rats were observed as compared to control.

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

When treated with 2100 mg/kg bw in male rats, Four renal tubule cell adenomas or carcinomas, renal tubule focal hyperplasia and renal tubule
adenomas were observed.

When treated with 1100 and 2100 mg/kg bw in male rats, Four renal tubule cell adenomas or carcinomas were observed.

When treated with 2500 mg/kg bw in female rats, one renal tubule adenoma was observed.
Although the trend for these renal neoplasms is significant, the incidences are low and do not exceed the historical control range of 0% to 6% in male rats.
When treated with 2100 and 2600 mg/kg bw in male and 1300 and 2600 mg/kg bw in female, significant dose-related decrease in the incidence of mononuclear cell leukemia was observed as compared to control.

No significant histopathological changes were observed in reproductive organ such as testes and mammory gland of treated male and female rats as compared to control.

Histopathological findings: neoplastic:

not specified

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:

2 100 mg/kg bw/day

Based on:

test mat.

Sex:

male

Basis for effect level:

clinical signs

mortality

body weight and weight gain

food consumption and compound intake

haematology

clinical biochemistry

organ weights and organ / body weight ratios

histopathology: non-neoplastic

other: No effect observed

Remarks on result:

other: No effect observed

Target system / organ toxicity (P0)

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

Results: F1 generation

General toxicity (F1)

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

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not specified

Effect levels (F1)

Dose descriptor:

other: not specified

Generation:

other: not specified

Based on:

not specified

Sex:

not specified

Basis for effect level:

other: not specified

Remarks on result:

other: not specified

Target system / organ toxicity (F1)

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

Overall reproductive toxicity

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

Any other information on results incl. tables

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Feed Study of C.I. Pigment Red 23

	0 ppm	10000 ppm	25000 ppm	50000 ppm
Genital System
Epididymis	(50)	(2)		(48)
Fibrous histiocytoma	1 (2%)
Preputial gland	(49)	(9)	(8)	(49)
Adenoma	3 (6%)	5 (56%)	8 (100%)	7 (14%)
Carcinoma	2 (4%)			1 (2%)
Prostate	(50)	(3)	(2)	(49)
Carcinoma, metastatic, stomach				1 (2%)
Fibrous histiocytoma	1 (2%)
Seminal vesicle	(50)	(2)		(49)
Carcinoma, metastatic, stomach				1 (2%)
Fibrous histiocytoma	1 (2%)
Testes	(50)	(44)	(44)	(49)
Bilateral, interstitial cell, adenoma	42 (84%)	43 (98%)	41 (93%)	44 (90%)
Interstitial cell, adenoma	6 (12%)	1 (2%)	3 (7%)	2 (4%)
Integumentary System
Mammary gland	(49)	(46)	(44)	(43)
Fibroadenoma	2 (4%)	2 (4%)	2 (5%)	3 (7%)
Fibroma				2 (5%)

 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2·Year Feed Study of C.I. Pigment Red 23

	0 ppm	10000 ppm	25000 ppm	50000 ppm
Genital System
Testes	(50)	(44)	(44)	(49)
Atrophy	1 (2%)		2 (5%)
Hemorrhage	1 (2%)
Bilateral, interstitial cell, hyperplasia	2 (4%)			1 (2%)
Interstitial cell, hyperplasia		1 (2%)		1 (2%)
Integumentary System
Mammary gland	(49)	(46)	(44)	(43)
Cyst		1 (2%)
Galactocele	1 (2%)	1 (2%)
Hemorrhage			1 (2%)
Hyperplasia	4 (8%)	7 (15%)	9 (20%)	6 (14%)
Inflammation, chronic	1 (2%)	1 (2%)
Duct, ectasia	3 (6%)	12 (26%)	7 (16%)	5 (12%)

Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Feed Study of C.I. Pigment Red 23

	0 ppm	10000 ppm	25000 ppm	50000 ppm
Genital System
Clitoral gland	(47)	(48)	(47)	(49)
Adenoma   Carcinoma	5 (11%) 3 (6%)	4 (8%)	3 (6%) 1 (2%)	2 (4%)
Ovary	(50)	(4)	(1)	(50)
Carcinoma			1 (100%)
Granulosa cell tumor NOS		1 (25%)
Granulosa cell tumor benign				1 (2%)
Thecoma NOS		2 (50%)
Uterus	(50)	(50)	(50)	(50)
Carcinoma, metastatic, ovary			1 (2%)
Fibrosarcoma			1 (2%)
Leiomyoma			1 (2%)
Cervix, sarcoma stromal		1 (2%)
Endometrium, polyp stromal	7 (14%)	4 (8%)	7 (14%)	13 (26%)
Endometrium, sarcoma stromal	1 (2%)	1 (2%)	1 (2%)
Vagina	(8)		(1)	(4)
Fibrosarcoma	1 (13%)
Leiomyosarcoma	1 (13%)
Sarcoma				1 (100%)
Schwannoma malignant	1 (13%)
Integumentary System
Mammary gland	(50)	(38)	(32)	(50)
Adenoma		2 (5%)	2 (6%)	1 (2%)
Carcinoma	1 (2%)	2 (5%)
Fibroadenoma	23 (46%)	24 (63%)	14 (44%)	19 (38%)

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2·Year Feed Study of C.I. Pigment Red 23^a

	0 ppm	10000 ppm	25000 ppm	50000 ppm
Genital System
Clitoral gland	(47)	(48)	(47)	(49)
Atrophy			1 (2%)
Hyperplasia	5 (11%)	3 (6%)	1 (2%)	1 (2%)
Inflammation, acute	5 (11%)	1 (2%)	5 (11%)	3 (6%)
Inflammation, chronic	3 (6%)	3 (6%)	2 (4%)	7 (14%)
Duct, cyst				1 (2%)
Duct, dilatation		1 (2%)
Duct, ectasia	1 (2%)		1 (2%)
Ovary	(50)	(4)	(1)	(50)
Follicle, cyst	2 (4%)	1 (25%)
Uterus	(50)	(50)	(50)	(50)
Cyst		2 (4%)	1 (2%)
Dilatation	4 (8%)	3 (6%)	2 (4%)	4 (8%)
Hemorrhage		1 (2%)
Inflammation, acute	1 (2%)
Necrosis	1 (2%)
Celvix, cyst		2 (4%)
Celvix, inflammation, acute	1 (2%)
Endometrium, hyperplasia, cystic	1 (2%)
Vagina	(8)		(1)	(4)
Inflammation, acute	1 (13%)
Inflammation, chronic				1 (25%)
Integumentary System Mammary gland	(50)	(38)	(32)	(50)
Galactocele	1 (2%)
Hyperplasia	4 (8%)	4 (11%)	5 (16%)	4 (8%)
Inflammation, acute		1 (3%)
Duct, ectasia	30 (60%)	20 (53%)	22 (69%)	18 (36%)
Duct, hyperplasia		1 (3%)
Skin	(50)	(8)	(2)	(49)
Acanthosis		2 (25%)
Hyperkeratosis		2 (25%)

Applicant's summary and conclusion

Conclusions:

NOAEL was considered to be 2100 mg/kg/day for male and 2600 mg/kg bw for female P generation when F344 male and female rats treated with C.1. Pigment Red (3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide) orally in diet for 2 years.

Executive summary:

In a Toxicology and Carcinogenesis Study, F344 male and female rat were treated with C.1. Pigment Red (3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide) in the concentration of 0,425, 1100 and 2100 mg/kg bw for male and 0, 500, 1300 and 2600 mg/kg/day for female orally in diet for 2 years. The average daily ingestion of C.1. Pigment Red 23 was approximately 425, 1,100, or 2,100 mg/kg body weight per day for male rats and 500, 1,300, or 2,600 mg/kg for females. Significant increase in survival of treated male and female rats were observed at 2100 and 2600 mg/kg bw as compared to control. Significant increase in survival of treated male rats were observed at 1100 mg/kg bw as compared to control. The greater survival of the exposed groups was due principally to the chemically related decreased incidence of mononuclear cell leukemia. No clinical sign were considered to be treatment related. Significant decrease in body weights of female rats were observed at 1300 and 2600 mg/kg bw as compared to control.No effect on food consumption of treated male and female rats was observed as compared to control.In female, significant decrease in Hematocrit values, hemoglobin concentration, and erythrocyte counts at 2600 mg/kg bw as compared to control. Which indicate mild anemia. No effect in male rats was observed as compared to control. Significant increase in Serum total bilirubin were observed in female rats at 2600 mg/kg bw as compared to control. This finding coupled with the mild anemia suggests a mild hemolytic process. No effect in male rats was observed as compared to control. Similarly, at 1300 and 2600 mg/kg bw in female rats, significant increase in relative kidney and brain weight were observed. In addition, at 2100 mg/kg bw in male rats, Four renal tubule cell adenomas or carcinomas, renal tubule focal hyperplasia and renal tubule adenomas were observed. At 1100 and 2100 mg/kg bw in male rats, Four renal tubule cell adenomas or carcinomas were observed. At 2500 mg/kg bw in female rats, one renal tubule adenoma was observed. Although the trend for these renal neoplasms is significant, the incidences are low and do not exceed the historical control range of 0% to 6% in male rats. At 2100 and 2600 mg/kg bw in male and 1300 and 2600 mg/kg bw in female,significant dose-related decrease in the incidence of mononuclear cell leukemia was observed as compared to control.No significant histopathological changes were observed in reproductive organ such as testes and mammary gland of treated male and female rats as compared to control.Therefore, NOAEL was considered to be 500 mg/kg/day for P and F1 generation whenWistarmale and female rats treated withIndigo Carmineorally in dietfor 2 years. Therefore,NOAEL was considered to be 2100 mg/kg/day for male and 2600 mg/kg bw for female P generation whenF344male and female rats treated withC.1. Pigment Red (3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide) orally in diet for 2 years.