isliFe

Repeated dose toxicity: dermal

Administrative data

Endpoint:

short-term repeated dose toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.9 (Repeated Dose (28 Days) Toxicity (Dermal))

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPP 82-2 (Repeated Dose Dermal Toxicity -21/28 Days)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Details on test material:

- Test article: CGA65047 SG 100 (A-5787 A)
- Analytical purity: 100 % (UVCB)
- Lot/batch No.: P.201845
- Storage condition of test material: room temperature

Test animals

Species:

rat

Strain:

other: Sprague-Dawley derived; Tif:RAIf (SPF); hybrids of RII/1 x RII/2

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Laboratory Animal Breeding, Pharmaceutical Division, CIBA-GEIGY Limited, 4332 Stein, Switzerland
- Age at study initiation: young adult, based on body weight ranges
- Weight range at acclimation period: 213.9-228.1 g (males), 211.8-235.7 g (females)
- Housing: individually, in macrolon cages type 3 with wire mesh tops and granulated soft wood bedding
- Diet: pelleted, certified standard diet Nafag No. 890 (NAFAG AG, Gossau, SG, Switzerland), provided ad libitum (exception: food was withheld overnight prior to blood removal performed at the end of the treatment period)
- Water: tap water, ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 55 ± 15 %
- Air changes: approximately 15 air changes/hour
- Photoperiod: 12 hours dark / 12 hours light

Administration / exposure

Type of coverage:

occlusive

Vehicle:

water

Details on exposure:

TEST SITE
- Area of exposure: dorsal area of the trunk
- Coverage: at least 10 % of the body surface area
- Type of wrap: gauze patches, aluminium foil and adhesive but non-irritating tape
- Time intervals for shavings or clipplings: once weekly

REMOVAL OF TEST SUBSTANCE
- Washing: with lukewarm water
- Time after start of exposure: 6 hours (on each day of treatment)

TEST MATERIAL/VEHICLE
- Amount applied: 4 mL/kg bw
- Concentrations: 2.5, 25 and 250 mg/mL
- Constant volume used: yes, adjusted weekly to individual animal body weight

USE OF RESTRAINERS FOR PREVENTING INGESTION: no data

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Control analyses of the test item concentration in the vehicle was carried out at all dose levels on samples collected on experimental days 1, 7, 14 and 21. Samples were analysed by RCC Umweltchemie AG, 4452 Itingen, Switzerland (Project No. 329207).

Duration of treatment / exposure:

4 weeks

Frequency of treatment:

6 hours per day, on 5 days per week

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 10, 100 and 1000 mg/kg bw/day
Basis:
nominal per unit body weight

No. of animals per sex per dose:

5 animals

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: no rationale is given in the report.

Positive control:

none

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS:
- Time schedule: twice daily on working days and once daily on weekend days for mortality, and once daily for general observations.
- Skin irritation: Following each application, approximately 17 hours after patch removal, local skin reactions at the application site were assessed according to Draize et al. (1944).

BODY WEIGHT:
- Time schedule: on study days -7, 1, 8,15, 22 and 28

FOOD CONSUMPTION:
- Food consumption for each animal was determined weekly and mean daily diet consumption calculated as g food/kg bw/day.

HAEMATOLOGY:
- Time schedule for collection of blood: once, at the end of the treatment period
- Anaesthetic used for blood collection: ether
- Animals fasted: food was withheld overnight prior to blood sampling
- How many animals: all animals
- Parameters examined: erythrocyte count, haematocrit, mean corpuscular volume, red cell volume distribution width, haemoglobin concentration, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, haemoglobin concentration distribution width, prothrombin time and leukocyte, neutrophil, eosinophil, basophil, lymphocyte, monocyte, large unstained cells and thrombocyte counts

CLINICAL CHEMISTRY:
- Time schedule for collection of blood: once, at the end of the treatment period
- Animals fasted: food was withheld overnight prior to blood sampling
- How many animals: all animals
- Parameters examined: glucose, urea, creatinine, total bilirubin, total protein, albumin, globulin, cholesterol, triglycerides, sodium, potassium, calcium, chloride and inorganic phosphorus concentration, and A/G ratio and aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase activity

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY:
The following organs or tissues were examined microscopically:
skin application site, skin remote site, liver, spleen, kidneys, thymus, thyroid with parathyroid gland, prostate, seminal vesicle, testis, epididymis, ovary

Other examinations:

ORGAN WEIGHT
The weights of the following organs were determined at necropsy:
brain, heart, liver, kidneys, adrenals, thymus, ovaries or testes, and spleen

Statistics:

Each treated group was compared to the control group by Wilcoxon's two-sample test (non-parametric) and tested for increasing or decreasing trends from control up to the respective dose group by Jonckheere's test for ordered alternatives (parametric).

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Dermal irritation:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

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 examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

not specified

Details on results:

CLINICAL SIGNS AND MORTALITY
No treatment related clinical signs were noted and all animals survived.

BODY WEIGHT AND WEIGHT GAIN
A slight body weight loss was noted in females at 1000 mg/kg bw/day during the first week of treatment. Otherwise the body weight was comparable in all groups.

FOOD CONSUMPTION
Food consumption was not influenced by treatment.

HAEMATOLOGY AND CLINICAL CHEMISTRY
No effects on haematology and clinical chemistry parameters were noted.

ORGAN WEIGHTS
There was an increased adrenal weight in males at 1000 mg/kg bw/day.

GROSS PATHOLOGY
No treatment related macroscopical findings were noted.

HISTOPATHOLOGY
The skin application site of females treated at 1000 mg/kg bw revealed epidermal hyperkeratosis and an increased severity of acanthosis. In 2/5 males at 1000 mg/kg bw, centrilobular hypertrophy of hepatocytes was noted.

Effect levels

Key result

Dose descriptor:

NOEL

Effect level:

100 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Based on slight effects on the liver and skin and due to increased adrenal weight noted at 1000 mg/kg bw/day.

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

CHEMICAL ANALYSIS OF DOSE FORMULATIONS

The calculated overall mean contents of the test item in the dose formulations used for the low, mid and high dose groups were 102, 103 and 89.7 % of the nominal concentrations, respectively (RCC Project No. 392207). The test item was proved to be stable in the vehicle under actual conditions of administration over the period of dosing.

Applicant's summary and conclusion

Conclusions:

Under the consitions of this study, the NOEL for repeated dose dermal toxicity of FeNa-EDDHA was established at 100 mg/kg bw/day.

Executive summary:

In a repeat-dose dermal toxicity study (CIBA-GEIGY Limited, 1996b), FeNa-EDDHA in distilled water was administered to the skin (clipped fur) of 5 Sprague-Dawley derived rats/sex/dose level at dose levels of 10, 100 or 1000 mg/kg bw/day for a period of 28 days (5 days per week basis). Male and female animals of the concurrent control group were treated with the vehicle only. Dermal treatment with the test item resulted in no mortality, no relevant clinical signs, no changes in food consumption, no effects on haematology and clinical chemistry parameters and no gross findings. A transient slight body weight loss was noted in females at 1000 mg/kg bw/day during the first week of treatment. There was an increase in adrenal weight in males at 1000 mg/kg bw/day. Microscopically, the skin application sites of females at 1000 mg/kg bw/day revealed epidermal hyperkeratosis associated with an increased severity of acanthosis. In 2/5 males at 1000 mg/kg bw/day centrilobular hypertrophy of hepatocytes was noted. Based on the slight effects on the liver and skin and due to the increased adrenal weight noted at 1000 mg/kg bw/day under the conditions of this study, the NOEL was established at 100 mg/kg bw/day.

This dermal toxicity study in the rat is acceptable and satisfies the requirement for test guideline OECD 410.