4',5'-dibromo-3',6'-dihydroxyspiro[isobenzofuran-1(3H),9'-[9H]xanthene]-3-one

Repeated dose toxicity: dermal

Administrative data

Endpoint:

repeated dose toxicity: dermal

Remarks:

combined repeated dose and carcinogenicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from peer reviewed journal

Data source

Reference

Reference Type:

publication

Title:

SICIN PAINTING STUDIES IN MICE WITH 14 FD & C AND D & C COLORS: FD & C Blue No. 1 , Red No. 3, and Yellow No. 5, D & C Red No. 7, Red No. 9, Red No. 10, Red No. 19, Red No. 21 , Red No. 27, Red No. 31, Red No. 36, Orange No. 5, Orange No. 10, & Orange 17

Author:

STEVEN CARSON

Year:

1984

Bibliographic source:

Journal of Toxicology. Cutaneous and Ocular Toxicology 3(4), 357-370 (1984)

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: as below

Principles of method if other than guideline:

Repeated dose toxicity/ carcinogenicity study was performed to determine the toxicity of the test compound D & C Orange no. 5.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

4',5'-dibromo-3',6'-dihydroxyspiro[isobenzofuran-1(3H),9'-[9H]xanthene]-3-one

EC Number:

209-876-0

EC Name:

4',5'-dibromo-3',6'-dihydroxyspiro[isobenzofuran-1(3H),9'-[9H]xanthene]-3-one

Cas Number:

596-03-2

Molecular formula:

C20H10Br2O5

IUPAC Name:

4',5'-dibromo-3',6'-dihydroxy-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one

Details on test material:

- Name of test material: D & C Orange no. 5- Molecular formula: C20H10Br2O5- Molecular weight: 490.10 g/mol- Substance type: Organic- Purity: 96%

Test animals

Species:

mouse

Strain:

ICR

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Strain details: 100 ICR (Swiss Webster derived)- 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 availableENVIRONMENTAL CONDITIONS- Temperature (°C): No data available- Humidity (%):No data available- Air changes (per hr): No data available- Photoperiod (hrs dark / hrs light): No data availableIN-LIFE DATES: From: To: No data available

Administration / exposure

Type of coverage:

open

Vehicle:

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: according to the rate of hair growthREMOVAL OF TEST SUBSTANCE- Washing (if done): No data available- Time after start of exposure: No data availableTEST MATERIAL- Amount(s) applied (volume or weight with unit): 0.1 ml of the vehicle containing 133.4 mg test material- Concentration (if solution): 0.1ml- Constant volume or concentration used: yes- For solids, paste formed: no data availableVEHICLE- Justification for use and choice of vehicle (if other than water): No data available- Amount(s) applied (volume or weight with unit): no data- Concentration (if solution):0.1 ml- Lot/batch no. (if required): Not data- Purity: no data availableUSE 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

Duration of treatment / exposure:

492 days

Frequency of treatment:

Twice weekly

Doses / concentrations

Remarks:

Doses / Concentrations:1 mg (mean dose-136.5 mg)Basis:no data

No. of animals per sex per dose:

50

Control animals:

yes

Details on study design:

No data available

Positive control:

The positive control (3, 4-benzpyrene), the observed effects in rodents were as expected for this known dermal carcinogen. Ninety-four percent of the mice in the positive control group developed typical gross skin lesions associated with the area of treatment in an average latent period of 245 days.

Examinations

Observations and examinations performed and frequency:

Observations and examinations performed & frequencyCAGE SIDE OBSERVATIONS: Yes - Time schedule: daily- Cage side observations checked in table [No.?] were included.DETAILED CLINICAL OBSERVATIONS: no data- Time schedule: no dataBODY WEIGHT: no data- Time schedule for examinations: no dataFOOD 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 dataFOOD 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 dataWATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data- Time schedule for examinations: No dataOPHTHALMOSCOPIC EXAMINATION: No data - Time schedule for examinations:- Dose groups that were examined: No dataHAEMATOLOGY: 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 dataCLINICAL 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 dataOTHER:

Sacrifice and pathology:

GROSS PATHOLOGY: Yes HISTOPATHOLOGY: Yes

Other examinations:

No data

Statistics:

No data

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Dermal irritation:

not specified

Mortality:

no mortality observed

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

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Details on results:

Effect- Spleen: Extramedullary HematopoiesisVehicle control: 5/50 female mice, 8/50 male miceTest material: 6/50 female mice, 4/50 male mice

Effect levels

Dose descriptor:

NOAEL

Effect level:

136.5 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No observed incidence of neoplasia

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The dermal application of 1 mg (136.5 mg/Kg bw) of D & C Orange no. 5 failed to increase the incidence of neoplasia. Hence the No Observed Adverse Effect Level (NOAEL) was assessed to be 1mg (136.5 mg/Kg bw).

Executive summary:

Repeated dose toxicity/ carcinogenicity study is carried out to find toxicity of D & C Orange no. 5. 100 ICR (Swiss Webster derived) mice, 50 males and 50 females treated with 0.1 ml dose containing 1mg of D & C Orange no. 5. An additional three groups of 100 mice were treated with the vehicle (distilled water), and another group of 100 mice was treated with 3, 4-benzpyrene (positive control) dissolved in acetone. Initially, the hair on the dorsal area of each animal was clipped with an animal clipper free of lubricating oil. Subsequent periodic clipping was performed according to the rate of hair growth. This area of approximately 6 cm² was treated twice weekly. Observation were made daily for mortality and gross toxicity. No adverse effect were observed related to D & C Orange no. 5. The dermal application of 1 mg (136.5 mg/Kg bw) of D & C Orange no. 5 failed to increase the incidence of neoplasia. Hence the No Observed Adverse Effect Level (NOAEL) was assessed to be 1mg (136.5 mg/Kg bw).