Disodium 8-hydroxynaphthalene-1,6-disulphonate

Toxicity to reproduction

Administrative data

Endpoint:

one-generation reproductive toxicity

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 publication

Data source

Reference

Reference Type:

publication

Title:

Effects of FD & C red no. 40 on rat intrauterine development

Author:

T. F. X. COLLINS and T. N. BLACK

Year:

1980

Bibliographic source:

Food and cosmatics toxicology, Vol. 18. pp 561 to 568, 1980

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Principles of method if other than guideline:

Embryotoxicity and teratogenicity study of FD & C RED NO. 40 in rats

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate

EC Number:

247-368-0

EC Name:

Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate

Cas Number:

25956-17-6

Molecular formula:

C18H16N2O8S2.2Na

IUPAC Name:

disodium 6-hydroxy-5-[(2-methoxy-3-methyl-4-sulfonatophenyl)diazenyl]naphthalene-2-sulfonate

Constituent 2

Reference substance name:

FD & C RED NO. 40

IUPAC Name:

FD & C RED NO. 40

Details on test material:

- Name of test material (as cited in study report): FD & C RED NO. 40 (disodium salt of 6-hydroxy-5-[(2-methoxy-5-methyl-4-sulph ophenyl)azo]-2-naphthalene sulphonic acid.)
- Molecular formula (if other than submission substance): C18H16N2O8S2.2Na
- Molecular weight (if other than submission substance): 496.4266 g/mole
- Substance type: Oragnic
- Physical state: No data available
- Impurities (identity and concentrations): 5.30% NaC1, 4.50% volatile matter,0.3 % Na2SO4 and 0.26% Schaeffer's salt (an intermediate).

Specific details on test material used for the study:

- Name of test material (as cited in study report): FD & C RED NO. 40 (disodium salt of 6-hydroxy-5-[(2-methoxy-5-methyl-4-sulph ophenyl)azo]-2-naphthalene sulphonic acid.)
- Molecular formula (if other than submission substance): C18H16N2O8S2.2Na
- Molecular weight (if other than submission substance): 496.4266 g/mole
- Substance type: Oragnic
- Physical state: No data available
- Impurities (identity and concentrations): 5.30% NaC1, 4.50% volatile matter,0.3 % Na2SO4 and 0.26% Schaeffer's salt (an intermediate).

Test animals

Species:

rat

Strain:

Osborne-Mendel

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: FDA breeding colony
- Age at study initiation: (P) x wks; (F1) x wks: No data available
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: P- female – 124.1 to 134.8 g
- Fasting period before study: No data available
- Housing: Animals were housed in stainless-steel hanging cages.
- Diet (e.g. ad libitum): Purina Laboratory Chow (Ralston-Purina Co., Inc., St. Louis, MO), ad libitum
- Water (e.g. ad libitum): Distilled water, ad libitum
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%):No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 12 hr darkness and 12 hr light.

Administration / exposure

Route of administration:

oral: drinking water

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: Distilled water

Details on exposure:

No data available

Details on mating procedure:

- M/F ratio per cage: 1:2 ratio
- Length of cohabitation: 4.30p.m. to following morning
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy Presence of sperm in the vaginal lavage were referred to as day 0 of pregnancy
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility: No data available

- Further matings after two unsuccessful attempts: [no / yes (explain)]: No data available

- After successful mating each pregnant female was caged (how): No data available

- Any other deviations from standard protocol: Within each group a male impregnated a maximum of two females.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The dye solutions were found to be stable for the duration of the experiment

Duration of treatment / exposure:

19 days

Frequency of treatment:

Daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day

Doses / concentrations 2

Dose / conc.:

260.2 mg/kg bw/day

No. of animals per sex per dose:

Totol: 55
0 mg/kg bw/day: 30
260.2 mg/kg bw/day: 25

Control animals:

yes, concurrent vehicle

Examinations

Parental animals: Observations and examinations:

Clinical signs , Body weight, water consumption were examihned.

Oestrous cyclicity (parental animals):

Any irregularity in Estrous cyclicity were examined.

Presence and location of resorption sites and implantation sites Corpora lutea were examined.

Sperm parameters (parental animals):

No data available

Litter observations:

Live and dead foetuses, weight and sexed were examined.

Postmortem examinations (parental animals):

No data available

Postmortem examinations (offspring):

Gross external malformationsunder magnification, and the crown-rump length and soft tissues variations was measured.

Statistics:

Statistical analysis were performed by using a least significant difference (LSD) test for numbers of corpora lutea per dam, implantations per dam, numbers of viable implants per dam, foetal body weights and crown-rump lengths. Freeman-Tukey arc-sine transformation for binomial proportions (Mosteller & Youtz, 1961) followed by an analysis of variance and an LSD test used for The number of resorptions per litter and the average number of foetuses with one or more variations per litter. Preimplantation loss data were transformed using the Freeman-Tukey arc-sine transformation followed by an analysis of variance and an LSD test. The numbers of litters with one or more resorptions, one or more skeletal or soft tissue variations and specific external, soft-tissue and skeletal variations were analysed by Fisher's exact one-tail test (Siegel, 1956). An analysis of variance was used to test maternal weight gain. An Armitage lest for linearity of proportions (Armitage, 1973) was also used to detect trends. Differences in water consumption were analysed by a paired t-test.

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):

No external signs of toxicity and animals appeared healthy and behaved normally in treated groups as compared to control.

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No effect on maternal weight gain was observed in treated groups as compared to control.

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

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

effects observed, treatment-related

Description (incidence and severity):

Significant decrease in water consumption was observed in treated rats as compare to control.

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:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

No significant changes in or treatment-related effects on the numbers of corpora lutea, implantations, early and late deaths and viable foetuses per litter or on the percentage of preimplantation loss were observed in treated group as compared to control.

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Description (incidence and severity):

No significant effect on percentage of early resorptions and percentage of females with more than one and with more than two resorptions were observed in treated rats as compared to control.

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

260.2 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

clinical signs

body weight and weight gain

reproductive function (oestrous cycle)

reproductive performance

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:

no mortality observed

Description (incidence and severity):

No effect on live or dead fetuses was observed in treated dose groups as compared to control.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No effect on mean fetal body weight was observed in treated groups as compared to control.

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:

no effects observed

Description (incidence and severity):

When treated with 260.2 mg/kg bw/day, No treatment related external variations were observed in litters of treated dams as compared to control.

Number of runts: Increase in number of runts was observed in litters of treated rats as compared to control.

Histopathological findings:

no effects observed

Description (incidence and severity):

Soft-tissue variations: slight increase in number of affected litters was observed in 260.2 mg/kg bw/day treated rats as compared to control.

Skeletal variations:
Slight increase in the number of bipartite sternebrae and reduced ossification of the parietal bone were observed in litters of treated dams as compared to control.
The observed increases did not appear to be dose-related.

Other effects:

no effects observed

Description (incidence and severity):

Crown-rump length: No effect on Crown-rump length were observed in litters of treated dams as compared to control.

Sex distribution: No effect on Sex distribution were observed in litters of treated dams as compared to control.

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not specified

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

260.2 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

mortality

body weight and weight gain

gross pathology

histopathology: non-neoplastic

Remarks on result:

other: No effect on Live and dead foetuses, weight, sex, gross pathology and histopathiology

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

Maternal and reproductive data for rats given FD & C red no 40 during gestation

			Autopsy finding (mean/dam)							Females with Resorption (%)
					Resorption					One or more	Two or more
Mode of addministration and dose level	no. Of pregnant females examined	mean maternal body weigh tgain (g)*	Corpora lutea*	implantations*	Early	Late	Viable foetuses*	Resorption (%)	Preimplantation
Intubation (%)
0	30	147.0 ± 3.6	13.4 ± 0.4	12.0 ± 0.5	0.5	0	11.5±0.5	4.4	10.4	43.3	10.0
0.2	25	147.5 ± 4.1	13.6 ± 0.3	12.7 ± 0.3	0.6	0	12.0±0.3	5.0	7.0	48.0	16.0

*Values are means ± SEM.

 Incidence of specifice external and soft tissue variations of foetuses of rats given FD & C red no 40 during gestation

		Incidence* in foetuses and litters of rats administered FD & C red no 40
		In the drinking water
Variation	Dose level†	0		0.2
		External examinations
No. Examined		345 (30)		301 (25)
Runt		1 (1)		4 (4)
Oedema		2 (1)
Cranial pimple		1 (1)
Exencephaly		1 (1)
		Soft tissue examination
No. Examined		180 (30)	161 (25)
Hydrourter: moderate		7 (5)	7 (5)
sever		1 (1)	1 (1)
Hydronephrosis: moderate		8 (4)	6 ()5
sever		1 (1)	1 (1)
Ectopic kidney		2 (2)	1 (1)
Pitted kidney		1 (1)	1 (1)
Deformed kidney		1 (1)
Haemorrhage		13 (8)	8 (7)
Hydrocephalus		2 (1)	1 (1)
Microphthalmia		1 (1)
oesophageal pouch		1 (1)

* No. Of foetuses affected and in parentheses, no of litters.

†The dose level is expressed as mg/kg/day as a percentage in the drinking water

Incidence of soft – tissue variations in foetusees of rats given FD & C red no 40 during gestation

	Soft tissue variations		Foetuses with one or more Soft tissue variations			Litters with one or more foetuses with variations
Drinking water (%)	Total	Mean/litter	No.	Mean/litter	% of total foetuses	No.	% of total litters
0	37	1.2	28	0.9	15.6	13	43.3
0.2	25	1.0	17	0.7	10.6	14	56.0

 

Incidence of sternebral variations in foetusees of rats given FD & C red no 40 during gestation

	Soft tissue variations		Foetuses with one or more Soft tissue variations			Litters with one or more foetuses with variations
Drinking water (%)	Total	Mean/litter	No.	Mean/litter	% of total foetuses	No.	% of total litters
0	57	1.9	49	1.6	30.2	22	73.3
0.2	58	2.3	52	2.1	37.1	20	80.0

 

Incidence of Skeletal variations in foetusees of rats given FD & C red no 40 during gestation

	Soft tissue variations		Foetuses with one or more Soft tissue variations			Litters with one or more foetuses with variations
Drinking water (%)	Total	Mean/litter	No.	Mean/litter	% of total foetuses	No.	% of total litters
0	56	1.9	42	1.4	25.9	21	70.0
0.2	58	2.3	31	1.2	22.1	14	56.0

Applicant's summary and conclusion

Conclusions:

NOAEL was considered to be 260.2 mg/kg body weight /day for F0 and F1 generation when Osborne-Mendel female rats were treated wtih FD & C RED NO. 40.

Executive summary:

In a Embryotoxicity and teratogenicity study, Osborne-Mendel female rats were treated with FD & C RED NO. 40 in the concentration of 260.2 mg/kg body weight /day in distilled water by oral drinking water. No external signs of toxicity and animals were appeared healthy and behaved normally in treated groups as compared to control. No effects on maternal weight gain and significant decrease in water consumption were observed in treated female rats as compared to control. No significant effects on numbers of corpora lutea, implantations, early and late deaths and viable foetuses per litter or on the percentage of preimplantation loss were observed in treated group as compared to control. Similarly, no significant effect on percentage of early resorptions and percentage of females with more than one and with more than two resorptions were observed in treated rats as compared to control. In addition, No effect on live or dead fetuses and mean fetal body weight and percentage of males and females per treatment group were observed as compared to control. Increase in number of runts was observed in litters, but no external variations were observed in litters of treated dams as compared to control. No effect on Crown-rump length and Sex distribution were observed in litters of treated dams as compared to control. Slight increase in number of affected litters, number of bipartite sternebrae and reduced ossification of the parietal bone were observed in litters of treated dams as compared to control. But, the observed increases did not appear to be dose-related. Therefore, NOAEL was considered to be 260.2mg/kg body weight /day for F0 and F1 generation when Osborne-Mendel female rats were treated with FD & C RED NO. 40. orally in Drinking water for 19 days.