Xanthylium, 9-[2-(ethoxycarbonyl)phenyl]-3,6-bis(ethylamino)-2,7-dimethyl-, molybdatetungstatephosphate

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:

data from handbook or collection of data

Justification for type of information:

Data is from peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

THE TERATOGENIC POTENTIAL IN RATS AND RABBITS OF D&C YELLOW NO. 8

Author:

C. M. BURNETI, E. I. GOLDENTHAL

Year:

1986

Bibliographic source:

Fd Chem. Toxic, Vol. 24, No. 8, pp. 819 823, 1986

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

Combined repeated dose repro-devp. Screen was performed to determine the toxic nature of D & C Yellow No. 8, sodium fluorescein upon repeated exposure by oral route

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 2-(3-oxo-6-oxidoxanthen-9-yl)benzoate

EC Number:

208-253-0

EC Name:

Disodium 2-(3-oxo-6-oxidoxanthen-9-yl)benzoate

Cas Number:

518-47-8

Molecular formula:

C20H12O5.2Na

IUPAC Name:

disodium 3-oxo-3H-spiro[2-benzofuran-1,9'-xanthene]-3',6'-diolate

Details on test material:

- Name of test material : D & C Yellow No. 8, sodium fluorescein
- Molecular formula : C20H10Na2O5
- Molecular weight : 376.274 g/mol
- Substance type: Organic
- Physical state: No data

Specific details on test material used for the study:

- Name of test material : D & C Yellow No. 8, sodium fluorescein
- Molecular formula : C20H10Na2O5
- Molecular weight : 376.274 g/mol
- Substance type: Organic
- Physical state: No data
- Imprities: 13%

Test animals

Species:

rat

Strain:

other: Charles River CD Sprague-Dawley

Details on species / strain selection:

No data

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:
Female rats: Charles River Breeding Laboratories, Portage, Michigan.
Male rats: Langshaw Farms, Augusta, Michigan
- Age at study initiation: 18 weeks
- Weight at study initiation: No data
- Fasting period before study: No data
- Housing: The animals were housed in wire bottomed cages
- Diet (e.g. ad libitum): Purina Certified Rodent Chow No. 5002 ad libitum
- Water (e.g. ad libitum): Water ad libitum
- Acclimation period: 4 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.2 ± -15.5 ˚C
- Humidity (%): 50 ± 15%
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): 12 hrs light/dark cycle

IN-LIFE DATES: From: To: No data

Administration / exposure

Route of administration:

oral: gavage

Details on route of administration:

No data

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with water as vehicle at dose levels of 0, 100, 500 or 1500 mg/kg body weight and prepared daily

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

VEHICLE
- Justification for use and choice of vehicle (if other than water): No data
- Concentration in vehicle: 0, 100, 500 or 1500 mg/kg bw
- Amount of vehicle (if gavage): 10 mL/Kg
- Lot/batch no. (if required): No data
- Purity: No data

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data

Duration of treatment / exposure:

14 days

Frequency of treatment:

Daily from 6-15 days of gestation

Doses / concentrations

Remarks:

0, 100, 500 or 1500 mg/kg bw

No. of animals per sex per dose:

Total: 100 females
0 mg/Kg bw: 25 females
100 mg/Kg bw: 25 females
500 mg/Kg bw: 25 females
1500 mg/Kg bw: 25 females

Control animals:

yes, concurrent vehicle

Details on study design:

No data

Positive control:

No data

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: During the study period
- Cage side observations checked in table [No.?] were included. Mortality

DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: No data

BODY WEIGHT: Yes
- Time schedule for examinations: body weights were recorded on days 6, 9, 12 and 16 of gestation

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

FOOD EFFICIENCY:
- 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

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No data

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

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

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

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

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

OTHER: No data

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: No data

Other examinations:

No data

Statistics:

No data

Results and discussion

Results of examinations

Clinical signs:

not specified

Description (incidence and severity):

No data

Mortality:

mortality observed, non-treatment-related

Description (incidence):

Survival was 100% in the controls and the groups receiving 100 and 500 mg/kg of dye. Six rats in the high-dose group (1500 mg/kg) died during the dosing period.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

There were slight reductions in body-weight gains in 1500 mg/Kg bw group, compared with controls, throughout the dosing period

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:

effects observed, treatment-related

Description (incidence and severity):

Orange discoloration of the urine was noted in all treated rats during the treatment period.

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

At autopsy, green discoloration of the amniotic fluid was noted in 1, 10 and 16 rats in the 100, 500 and 1500 mg/kg/day groups, respectively, and the small intestines were green in colour in many rats in the 500 mg/Kg group.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Effect levels

Dose descriptor:

NOAEL

Effect level:

1 500 mg/kg bw (total dose)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No significant effects were noted at the mentioned dose level

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Table: Mean maternal body weights for rats given D & C Yellow No. 8 by gavage on days 6-19 of gestation

Days of gestation	Mean Maternal Body Weight (g)
	0	100	500	1500
0	246 ± 145	249 ± I3.7	249 ± 14.7	241 ± 12.4
6	272 ± 16.8	271 ± 14.4	276 ± 14.5	263 ± 12.2
9	279 ± 18,1	278 ± 15.9	283 ± 16.7	269 ± 16.5
12	294 ± 20.1	292 ± 17.5	296 ± 16.1	282 ± 16.9
16	321 ± 23.4	320 ± 18.9	323 ± 2(}.4	300 ± 30.7
20	379 ± 30.0	378 ± 21.2	374 ± 32.2	359 ± 33.3

 

Table 2. Mean maternal body-weight changes in rats given D & C Yellow No. 8 by gavage on days 6-19 of gestation

Days of gestation	Mean Maternal Body Weight (g)
	0	100	500	1500
0-6	26	22	27	22
6-9	7	7	7	6
9-12	15	14	13	13
12-16	27	28	27	18
16-20	58	58	51	59
6-20	107	107	98	96
0-20	133	129	125	118

Applicant's summary and conclusion

Conclusions:

The No Observed adverse effect level (NOAEL) for D & C Yellow No. 8, sodium fluorescein is considered to be 1500 mg/kg bw

Executive summary:

Combined repeated dose repro-devp. Screen was performed to determine the toxic nature of D & C Yellow No. 8, sodium fluorescein upon repeated exposure by oral route.An aqueous solution of the dye was administered by garage to groups of 25 Charles River Sprague-Dawley rats at doses of 100, 500 and 1500 mg/kg on days 6- 19 of gestation. A control group received water on a comparable basis. Individual doses were determined on the basis of body weights recorded on days 6, 9, 12 and 16 of gestation.Survival was 100% in the controls and the groups receiving 100 and 500 mg/kg of dye. Six rats in the high-dose group (1500 mg/kg) died during the dosing period. There were slight reductions in body-weight gains in 1500 mg/Kg bw group, compared with controls, throughout the dosing period.Orange discoloration of the urine was noted in all treated rats during the treatment period.At autopsy, green discoloration of the amniotic fluid was noted in 1, 10 and 16 rats in the 100, 500 and 1500 mg/kg/day groups, respectively, and the small intestines were green in colour in many rats in the 500 mg/Kg group. Based on the above observations made, the No Observed adverse effect level (NOAEL) forD & C Yellow No. 8, sodium fluorescein is considered to be 1500 mg/kg bw