Dialuminium tris[2-(2,4,5,7-tetrabromo-6-oxido-3-oxoxanthen-9-yl)benzoate]

Repeated dose toxicity: dermal

Administrative data

Endpoint:

repeated dose toxicity: dermal, other

Remarks:

Combined repeated dose & carcinogenicity

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:

Skin Painting Studies In Mice With 14 FD & C And D & C Colors: FD & C Blue No. 1, Red No. 3 & Yellow No. 5, D & C Red No. 7, Red No. 9, Red No. 10, Red No. 19, Red No. 2 1 , Red No. 27, Red No. 31, Red No. 36, Orange No. 5, Orange No. 10 & Orange No 17

Author:

Steven Carson

Year:

1984

Bibliographic source:

J. Toxicol. Cut. & Ocular Toxicol. 3(4), 357-370 (1984)

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

Combined repeated dose & carcinogenicity by the dermal route was performed to determine the dermal toxic nature of the test compound 2',4',5',7'-Tetrabromofluorescein upon repeated application.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

2',4',5',7'-Tetrabromofluorescein

IUPAC Name:

2',4',5',7'-Tetrabromofluorescein

Constituent 2

Reference substance name:

2-(3,6-dihydroxy-2,4,5,7-tetrabromoxanthen-9-yl)-benzoic acid

EC Number:

239-138-3

EC Name:

2-(3,6-dihydroxy-2,4,5,7-tetrabromoxanthen-9-yl)-benzoic acid

Cas Number:

15086-94-9

Molecular formula:

C20H8Br4O5

IUPAC Name:

2-(2,4,5,7-tetrabromo-3,6-dihydroxy-9H-xanthen-9-yl)benzoic acid

Test material form:

solid

Details on test material:

- Name of test material (as cited in study report): D & C Red no. 21
- Molecular formula: C20H8Br4O5
- Molecular weight: 647.8942 g/mol
- Substance type: Organic
- Physical state: Solid
- Impurities (identity and concentrations): 98% pure

Specific details on test material used for the study:

- Name of test material: D & C Red no. 21
- Molecular formula: C20H8Br4O5
- Molecular weight: 647.8942 g/mol
- Substance type: Organic
- Physical state: Solid
- Impurities (identity and concentrations): 98% pure

Test animals

Species:

mouse

Strain:

ICR

Remarks:

Swiss Webster derived

Details on species / strain selection:

No data

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: No data
- Age at study initiation: No data
- Weight at study initiation: No data
- Fasting period before study: No data available
- Housing: Each animal was assigned an identification number and individually housed in a supported wire cage.
- Diet (e.g. ad libitum): No data available
- Water (e.g. ad libitum): No data available
- 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): No data available

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

Administration / exposure

Type of coverage:

open

Vehicle:

other: Distilled water

Details on exposure:

TEST SITE
- Area of exposure: 6 cm²
- % coverage: No data available
- Type of wrap if used: No data available
- Time intervals for shavings or clipplings: Subsequent periodic clipping was performed according to the rate of hair growth

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No data available
- Time after start of exposure: No data available

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml of the vehicle containing 140.1 mg test material
- Concentration (if solution): 0.1ml
- Constant volume or concentration used: yes
- For solids, paste formed: no data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): No data available
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): 140.1 mg
- Lot/batch no. (if required): Not data
- Purity: no data available

USE OF RESTRAINERS FOR PREVENTING INGESTION: no data

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data available

Duration of treatment / exposure:

18 months/ 473 days

Frequency of treatment:

Twice weekly

Doses / concentrations

Remarks:

Doses / Concentrations: 140.1 mg

No. of animals per sex per dose:

Total: 500
140.1 mg: 50/sex
0 mg/Kg bw: 150/sex
Positive control: 150/sex

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Positive control:

3,4-benzpyrene

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations checked in table [No.?] were included. Mortality and morbundity

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Twice weekly gross toxicity was noted

BODY WEIGHT: no data
- Time schedule for examinations: no data

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: No data
- 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:
- 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.

DERMAL IRRITATION (if dermal study): No data
- Time schedule for examinations: 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.

URINALYSIS: No data
- 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.

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

OTHER:

Sacrifice and pathology:

GROSS PATHOLOGY: Yes, Animals that died, those sacrificed moribund, and those surviving the 18-month experimental period were necropsied. Tissues from following organs were collected: the brain, pituitary, thyroid, thymus, liver, spleen, kidney, adrenal, stomach, small intestines, large intestines, urinary bladder, axillary lymph nodes, testes, ovary, skin from area of treatment, any tissue masses, grossly abnormal organs, or other tissues.

HISTOPATHOLOGY: Yes, Tissues selected for histopathology were sectioned, stained with hematoxylin and eosin and was examined by a pathologist. The following tissues were selected for examination: skin and any grossly abnormal organs and tissues of all animals in the color experimental groups; skin and any grossly abnormal organs and tissues of approximately 50 vehicle control animals. complete pathology on five males and five females randomly selected vehicle control animals that survived the 18-month experimental period; and complete pathology on five males and five females randomly selected vehicle control animals that survived the 18-month experimental period; and complete pathology of five male and five female animals from the positive control group that included induced skin lesions.

Other examinations:

No data available

Statistics:

No data available

Results and discussion

Results of examinations

Clinical signs:

not specified

Dermal irritation:

not specified

Mortality:

mortality observed, treatment-related

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

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:

effects observed, treatment-related

Histopathological findings: neoplastic:

effects observed, treatment-related

Other effects:

not specified

Details on results:

Clinical signs and mortality:
Clinical signs: No data
Mortality: The percent survival of the treated mice changed with the duration of the treatment. The survival was 97% in 4 months, 93% in 10 months, 68% in 12 months and 50% in 18 months respectively.

Dermal irritation: No data

Body weight and weight gain: No data

Food consumption and compound intake: No data

Food efficiency: No data

Water consumption and compound intake: No data

Opthalmoscopic examination: No data

Haematology: No data

Clinical chemistry: No data

Urinanalysis: No data

Neurobehaviour: No data

Organ weights: No data

Gross pathology: No data

Histopathology: The incidence of extramedullary hematopoesis of the spleen (5 male and 5 females: 10%) in the test group was consistent with that found in the distilled water vehicle control groups. The incidence in the three distilled water control groups was 13 % , 18 % , and 22 % for a mean of 17.7 % . The test group did not show a greater incidence than a control group.

The incidence of ectoparasitism was greater in the dye treated group than found in the vehicle controls. This increase in skin mite infestation may have contributed to the increase in epidermal change dermatitis, acanthosis, and hyperkeratosis observed in the dye treated groups.

Although the number of neoplasias involving the mammary glands or internal organs was noted in the test group, there was, however, no apparent change in their pattern which could be attributed to the dermal application of the test dye compound.

The incidence and severity of lesions of interstitial nephritis, cystitis, amyloidosis, and bronchopneumonia though observed in the test groups but there were no significant variations that could be attributed to application of the test compound.

Effect levels

Dose descriptor:

NOAEL

Effect level:

140.1 other: mg

Based on:

test mat.

Sex:

male/female

Basis for effect level:

gross pathology

histopathology: neoplastic

histopathology: non-neoplastic

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Table: Percent survival

Months	5	10	16	18
D & C Red no 21	97	93	68	50

Table: Details of microscopic analysis

Animal Number	Experimental Days Observed	Location of Lesion	Microscopic Diagnosis
7F	532	Swelling in left in-guinal area	Mammary gland adenocarcinoma
47F	557	Swelling in left hindleg	Hemangiosarcoma
54M	558	Nodule in liver and heart	Liver-hypertrophic nodules heart-organizing thrombus in left atrium
61M	558	Tissue mass in liver	Hepatic cell adenoma
73M	558	Tissue mass in liver	Hepatic cell adenoma
84M	558	Tissue mass in liver and nodule in kidney	Liver-hepatic cell adenoma kidney-moderately severe interstinal nephritis
87M	558	Tissue mass in liver	Hepatic cell adenoma
94M	558	Tissue mass in liver	Hypertrophic nodule

Applicant's summary and conclusion

Conclusions:

The No Observed Adverse Effect Level (NOAEL) for the test compound 2',4',5',7'-Tetrabromofluorescein is considered to be 140.1 mg when ICR mice were exposed with the test compound for 473 days.

Executive summary:

Combined repeated dose & carcinogenicity by the dermal route was performed to determine the dermal toxic nature of the test compound 2',4',5',7'-Tetrabromofluorescein upon repeated application. The test chemical was applied to the clipped dorsal area mice. The animals were observed for mortality, clinical signs and gross pathology and histopathology was performed. The percent survival of the treated mice changed with the duration of the treatment. The survival was 97% in 4 months, 93% in 10 months, 68% in 12 months and 50% in 18 months respectively. The incidence of extramedullary hematopoesis of the spleen (5 male and 5 females: 10%) in the test group was consistent with that found in the distilled water vehicle control groups. The incidence in the three distilled water control groups was 13 % , 18 % , and 22 % for a mean of 17.7 % . The test group did not show a greater incidence than a control group. The incidence of ectoparasitism was greater in the dye treated group than found in the vehicle controls. This increase in skin mite infestation may have contributed to the increase in epidermal change dermatitis, acanthosis, and hyperkeratosis observed in the dye treated groups. Although the number of neoplasias involving the mammary glands or internal organs was noted in the test group, there was, however, no apparent change in their pattern which could be attributed to the dermal application of the test dye compound. The incidence and severity of lesions of interstitial nephritis, cystitis, amyloidosis, and bronchopneumonia though observed in the test groups but there were no significant variations that could be attributed to application of the test compound. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) for the test compound 2',4',5',7'-Tetrabromofluorescein is considered to be 140.1 mg when ICR mice were exposed with the test compound for 473 days.