Insulin Aspart Ethyl Ester

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000.04.04 - 2000.07.04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

Storage temperature of samples, the humidity in the animal holding room, and the time of recording the dermal reactions deviated from what stated in the protocol. None of the deviation is considered to have affected the study outcome.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

Storage temperature of samples, the humidity in the animal holding room, and the time of recording the dermal reactions deviated from what stated in the protocol. None of the deviation is considered to have affected the study outcome.

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

yes

Remarks:

Storage temperature of samples, the humidity in the animal holding room, and the time of recording the dermal reactions deviated from what stated in the protocol. None of the deviation is considered to have affected the study outcome.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

other: Cream powder (84% w/w)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL (MI3 dried)
- Source and lot/batch No.of test material: C202483
- Expiration date of the lot/batch: 2001.05.31
- Purity: 98.4%
- Purity test date: 2001.05.31

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: < -10 degrees Celcius
- Stability under test conditions: NA
- Solubility and stability of the test substance in the solvent/vehicle: Soluble in vehicle (water)
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: NA

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Purified water (Elgastat purifier used)
- Final preparation of a solid: 100mg/ml MI3 in water, pH of 8.0.

FORM AS APPLIED IN THE TEST (if different from that of starting material): solid dissolved in purified water.

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Rabbit (New Zealand White rabbit)
- Sex: male and female
- Age at study initiation: 10 - 14 weeks.
- Weight at study initiation: 2.35 - 2.41 kg
- Housing: Single housed in floor-pens with wood chips bedding
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: NA

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-22
- Humidity (%): 40 – 80 %
- Air changes (per hr): at least 10 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours/12 hours

IN-LIFE DATES: From: To: at least 10 - 13 days

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.5 mL of MI3 moistened with water (paste)

Duration of treatment / exposure:

4 hours

Observation period:

one hour, 24, 48 and 72 hours after removal of patch and dressing.

Number of animals:

3 ( one male and two females)

Details on study design:

TEST SITE
- Area of exposure: 30 x 20 mm
- % coverage: NA
- Type of wrap if used: Dense elastic gauze bandage (semi-occlusive)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Light brushing and swabbing with moistened cotton wool
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) one hour, 24, 48 and 72 hours after application.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No indication of irritation or corrosive response. Individual and group mean values for dermal changes ( Erythema and Oedema) in the tested animals were zero at 0, 24, 48 and 72 hours.

Other effects:

No other effects observed

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The Study was conducted to determine the irritation caused to intact rabbit skin following a single (4 hour), semi-occluded, topical application of MI3. The study was designed in accordance with OECD guideline method 404. One male and two female rabbits were exposed to a single dose of 0.5mL MI3 applied to 30 x 20 mm area.All animals were killed by an intravenous injection of an overdose of barbiturate at day 4. No reactions were observed following a single semi-occluded, topical application of MI3 to intact rabbit skin for four hours.

Executive summary:

The Study was conducted to determine the irritation caused to intact rabbit skin following a single (4 hour), semi-occluded, topical application of MI3. The study was designed in accordance with OECD guideline method 404, the commission directive 92/69/EEC, Method B4, and the US EPA Health effects test Guidelines OPPTS 870.2500. A single dose of 0.5 mL MI3 that was applied to a 30 x 20 mm area on the clipped dorsum of each of three rabbits, one male and two females. The treated area of skin was covered by a semi- occlusive bandage for 4 hours and assessed for dermal reaction the following three days. All animals were killed by an intravenous injection of an overdose of barbiturate at day 4.

In conclusion, no dermal reactions were observed following a single semi-occluded, topical application of MI3 to intact rabbit skin for four hours.