Trisodium bis[2-[[2,4-dihydroxy-3-[(2-methyl-4-sulphophenyl)azo]phenyl]azo]benzoato(3-)]chromate(3-)

Repeated dose toxicity: dermal

Administrative data

Endpoint:

short-term repeated dose toxicity: dermal

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

1983

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: read across, no GLP, outdated design, limited number of parameters examined

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1983

Report date:

1984

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)

GLP compliance:

no

Remarks:

pre.dates GLP regulation

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

see chapter 13

IUPAC Name:

see chapter 13

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Test animals

Species:

rat

Strain:

other: Crl:CD(SD) BR

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River UK Ltd. Manston Road, Margate, UK
- Age at study initiation: 5 weeks
- Weight at study initiation: 148 -245 g
- Fasting period before study: none
- Housing: single stainless teel wire mesh
- Diet (e.g. ad libitum): ad lib
- Water (e.g. ad libitum): ad lib.
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 24
- Humidity (%): 55 - 88
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Type of coverage:

occlusive

Vehicle:

water

Details on exposure:

TEST SITE
- Area of exposure: 24 cm² in the dorsolumbar region

- Type of wrap if used: occlusive
- Time intervals for shavings or clipplings: when needed

REMOVAL OF TEST SUBSTANCE
- Washing (if done): none, but the treated area was wiped aftr 6 hours of exposure
- Time after start of exposure: 6h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 1000 mg/kg
- Concentration (if solution): 250mg/mL
- Constant volume and concentration used: yes: 4 mL/kg bw / day
- For solids, paste formed: yes/no


USE OF RESTRAINERS FOR PREVENTING INGESTION: no

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

4 weeks

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
1000 mg/kg
Basis:
nominal per unit body weight

Doses / concentrations 2

Remarks:

Doses / Concentrations:
0 mg/kg
Basis:
nominal per unit body weight

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: No sign of toxicity was een in an oral acute study in the same laboratory at a dose of 2000 mg/kg

Positive control:

no required

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: included in clinical examination

BODY WEIGHT: Yes
- Time schedule for examinations: before start, then weekly

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes, weekly

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

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: week 3
- Anaesthetic used for blood collection: Yes (diethyl ether)
- Animals fasted: Yes (16 hrs)
- How many animals: all
- Parameters examined: haemoglobin (Hb), red blood cell count (RBC) and derived fndices:
packed cell volume (PCV), mean cell haemoglobin (MCH), mean cell haemoglobin concentration (MCHC), mean cell volume (MCV)
total and differential white blood cell count (WBC)
In addition, samples were withdrawn into 3.13% trisodium citrate anticoagulant for determination of prothrombin time (PT).

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: week 3
- Animals fasted: Yes (16 hrs)
- How many animals: all
- Parameters examined:
glutamate oxaloacetate transaminase (GOT)
alkaline phosphatase (Alk.P)
blood urea nitrogen (BUN)
bilirubin
glucose
calcium
sodium
potass;um
chloride
inorganic phosphate
creatinine
total protein
albumin
albumin/globulin ratio

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (full external and internal macroscopic examination)
Organ weights of: adrenal, kidneys, liver, testes
HISTOPATHOLOGY: Yes (treated & untreated skin, adrenals, kidneys, liver, gross leasions)

Other examinations:

none

Statistics:

yes, but no detail given

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

One control male was found dead on day 22. All animals treated with Brown HH 469 survived the 4 week study period. There were no treatment-related clinical signs.

Dermal irritation:

effects observed, treatment-related

Description (incidence and severity):

Slight temporary signs of irritation (redness, very slight edema) were seen in some treated animals. No irritation was detected in controls.

Mortality:

no mortality observed

Description (incidence):

One control male was found dead on day 22. All animals treated with Brown HH 469 survived the 4 week study period. There were no treatment-related clinical signs.

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
One control male was found dead on day 22. All animals treated with the test material survived the 4 week study period. There were no treatment-related clinical signs
Slight redness of the treated skin site was noted in some females exposed to the test material on days 2-4 and in 1 male on days 23 -28.
Slightly increased redness (grade 2) was noted on the treated skin site on female 16 on days 22-29 and in female 20 on days 11-26.
Very slight oedema was also noted on female 20 on days 11-12. One female showed skin sores on its back probably due to the occlusive dressing.
No adverse skin redness was noted in control animals.

BODY WEIGHT AND WEIGHT GAIN
Overall body weight gains during the treatment period were considered to be similar in all animals.

FOOD CONSUMPTION
There were no adverse effects on food intake.

HAEMATOLOGY
There were no changes in the haematological parameters of treated animals that could be attributed to treatment with Brown HH 469.

CLINICAL CHEMISTRY
There were no changes in the clinical chemistry parameters that could be attributed to treatment with Brown HH 469.

ORGAN WEIGHTS

GROSS PATHOLOGY
The majority of treated and control skin sites were unremarkable at necropsy. A minor sore was noted in 1 female treated with the test material but this was an isolated finding. The main skin reactions were sores and fur loss (alopecia) on the abdomen due to the adhesive dressing, but this was not related to treatment. The only consistent treatment-related finding was a brown discolouration of the tail, probably due to contamination by the test article. There were no unusual findings in the internal organs examined at necropsy except in one control male which died from urinary tract obstruction.

HISTOPATHOLOGY: NON-NEOPLASTIC
Skin reactions in the treated skin site of control rats consisted primarily of low grade acanthosis. This was more noticeable in males than females. The skin sites exposed to the test material were generally similar to those of the controls.
One male and 1 female had erosions, but these were minor focal lesions, probably of traumatic origin, and the rest of the treated skin area was similar to controls. Overall, there was no evidence of any significant skin irritation due to repeated percutaneous application of the test article.
Histopathology findings in the selected tissues examined were generally infrequent and of a minor nature such as hyaline droplets in the kidney of males and leucocyte foci in the livers of both sexes. There were no findings of any unusual nature or incidence to suggest any systemic toxic effect due to test article administration.

Effect levels

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: No adverse effects observed at all.

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Dermal application of Brown HH 469 at adose level of 1000 mg/kg/day for 4 weeks produced no evidence of systemic toxicity.

Executive summary:

1.1 The study was performed to evaluate the toxicity of the test article, Brown HH 469, when administered by dermal application to intact skin of the rat for a period of 4 weeks.

1.2 Groups of Sprague Dawley rats, comprising 5 animals of each sex, were treated at dose levels of 0 and 1000 mg/kg/day.

1.3 One control animal was found dead on day 22. All animals treated with 1000 mg/kg/day Brown HH 469 survived the 4 week study period.

1.4 No treatment-related changes in clinical condition or behaviour occurred.

1.5 Similar overall body weight gains were recorded in both test and control groups.

1.6 Food consumption was similar in both groups.

1.7 There were no changes in the haematological and clinical chemistry parameters measured.

1.8 All organ weights and organ/body weight ratios were considered similar for treated and control animals.

1.9 The gross and histological appearance of the skin site treated with Brown HH 469 was similar to that of control animals.

1.10 There was no evidence of any systemic toxicity in organs examined at necropsy or in tissues selected for histological evaluation. 1.11 In conclusion, dermal application of Brown HH 469 at a dose level of 1000 mg/kg/day produced no evidence of systemic toxicity.