Disodium 2-[[5-carbamoyl-1-ethyl-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridyl]azo]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]benzenesulphonate

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

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 peer-reviewed journal.

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:

no guideline available

Principles of method if other than guideline:

Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test 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

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): C18-H16-N2-O8-S2.2Na- 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: No data available
- Weight at study initiation: 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.

IN-LIFE DATES: From: To: No data available

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: Distilled water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: Solutions of FD & C Red No. 40 were prepared with distilled water at 7.5, 15, 30, 100 and 200 mg FD & C Red No. 40/kg body weight daily on days 0-19 of gestation.

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE- Justification for use and choice of vehicle (if other than water): distilled water
- Concentration in vehicle: 0, 7.5, 15, 30, 100 and 200 mg/kg body weight/day
- Amount of vehicle (if gavage): 1 ml/100 g body weight
- Lot/batch no. (if required): No data available
- Purity: No data available

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

stability were tested

Duration of treatment / exposure:

19 days

Frequency of treatment:

Daily

Doses / concentrations

Remarks:

Doses / Concentrations:0, 7.5, 15, 30, 100 and 200 mg/kg body weight /day Basis:nominal in water

No. of animals per sex per dose:

25

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Doses were selected on the basis of teratology study of FD & C Red No. 2 (Collins & McLaughlin, 1972).
- Rationale for animal assignment (if not random): No data available
- Rationale for selecting satellite groups: No data available
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): No data available

Positive control:

No data available

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: No data available
- Time schedule: No data available
- Cage side observations checked in table [No.?] were included. No data available

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: Daily

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data available
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available

FOOD EFFICIENCY: No data available
- 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: No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Daily

OPHTHALMOSCOPIC EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available

HAEMATOLOGY: No data available
- Time schedule for collection of blood: No data available
- Anaesthetic used for blood collection: No data available
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table [No.?] were examined. No data available

CLINICAL CHEMISTRY: No data available
- Time schedule for collection of blood: No data available
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table [No.?] were examined. No data available

URINALYSIS: No data available
- Time schedule for collection of urine: No data available
- Metabolism cages used for collection of urine: No data available
- Animals fasted: No data available
- Parameters checked in table [No.?] were examined. No data available

NEUROBEHAVIOURAL EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data available

OTHER: No data available

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Other examinations:

Presence and location of resorption sites and implantation sites, Corpora lutea, Live and dead foetuses, weight and sexed of foetuses, Gross external malformationsunder magnification, and the crown-rump length and soft tissues variations of foetuses were examined.

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.

Results and discussion

Results of examinations

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.

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):

no effects observed

Description (incidence and severity):

No effect on water comsumption 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

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

no effects observed

Description (incidence and severity):

When treated wtih 7.5 and 15 mg/kg bw/day , slightly increase in percentage of early resorptions were observed but this response were appe ared to be the result of sporadic occurrences. When treated wtih 100 mg/kg bw/day, One litter was totally resorbed. The percentage of females with more than one and with more than two resorptions showed no dose-related correlation as comapred to control.

Effect levels

Dose descriptor:

NOAEL

Effect level:

200 other: mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No effect on clinical sign, water consumption, body weight and reproductive performance was observed.

Target system / organ toxicity

Critical effects observed:

no

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 (mg/kg/day)
0	24	124.1 ± 4.5	13.0 ± 0.3	11.5 ± 0.5	1.1	0.04	10.3 ±0.5	10.1	11.2	66.7	33.3
7.5	27	127.2 ± 5.6	13.4 ± 0.4	12.5 ± 0.4	1.4	0	11.0 ±0.6	11.6	6.9	48.2	29.6
15	28	134.8 ± 3.9	13.8 ± 0.4	12.7 ± 0.2	1.6	0.04	11.1 ± 0.3	12.9	7.8	71.4	35.7
30	28	128.3 ± 5.1	13.4±0.6	12.1 ± 0.5	1.2	0.04	10.9 ±0.6	10.0	10.2	64.3	21.4
100	26	123.1 ±4.4	13.6 ± 0.5	11.6 ± 0.5	0.7	0	11.0 ± 0.6	5.9	14.6	34.6	23.1
200	25	123.3 ± 4.1	13.0 ± 0.4	11.6 ± 0.4	1.0	0	10.6 ±0.5	8.3	11.4	56.0	20.

*Values are means ± SEM.

Applicant's summary and conclusion

Conclusions:

NOAEL was considered to be 200 mg/kg body weight /day when Osborne-Mendel female rats were treated wtih FD & C RED NO. 40.

Executive summary:

In a Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening, Osborne-Mendelfemale rats were treated wtih FD & C RED NO. 40 in the concentration of 0, 7.5, 15, 30, 100 and 200 mg/kg body weight /day in distilled water by oral gavage. No external signs of toxicity and animals were appeared healthy and behaved normally in treated groups as compared to control.No effects on weight gain and water consumption were observed in treated female rats as compared to control. No significant effect 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. Slightly increas in percentage of early resorptions were observed in 7.5 and 15 mg/kg bw/day but this response were appeared to be the result of sporadic occurrences. One litter was totally resorbed at 100 mg/kg bw/day. But, he percentage of females with more than one and with more than two resorptions showed no dose-related correlation as comapred to control. In addition, no effect on live or dead foetuses and mean foetal body weight and percentage of males and females per treatment group were observed as compared to control. Increased in number of runts were observed at 7.5, 15, 100 and 200 mg/kg bw/day treated dams, No compound-related external variations were observed in litters of treated dams as compared to control. Slightly decrease in males and females Crown-rump length were observed but were not significantly significant with increasing dose level. No effect on percentage of males and females per treatment group were observed as compared to control. Therefore, NOAEL was considered to be 200 mg/kg body weight /day when Osborne-Mendel female rats were treated wtih FD & C RED NO. 40 orally by gavage for 19 days.