Nitrile hydratase recombinant from E.coli

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 September 2003 to 29 September 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Nitrile hydratase recombinant from E.coli

EC Number:

617-328-0

Cas Number:

82391-37-5

Molecular formula:

Not available for a UVCB substance

IUPAC Name:

Nitrile hydratase recombinant from E.coli

Test material form:

liquid

Details on test material:

- Appearance: Grey liquid (in dissolved state)
- Storage conditions: Frozen conditions

Test animals

Species:

rabbit

Strain:

other: Japanese white

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 17 weeks
- Weight at study initiation: 3.06-3.20 kg
- Housing: Rabbits were housed in stainless steel or aluminium cages. (63 cm (D) x 50 cm (W) x 36 cm (H)). There were one rabbit per cage.
- Diet: approx. 120 g of solid fodder was given once a day
- Water: ad libitum
- Acclimation period: 8 Days

ENVIRONMENTAL CONDITIONS
- Temperature: 21.5 – 22.6°C
- Humidity: 43 – 66%
- Air changes: 15 times/hour
- Photoperiod: 12 hours/day artificial lighting (06:00 – 18:00 lamp-lighting)

Test system

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.5 mL per site

Duration of treatment / exposure:

3 minutes, 1, 4 hours

Observation period:

1, 24, 48 and 72 hours

Number of animals:

3 animals

Details on study design:

TEST SITE
- Area of exposure: Test material was coated uniformly on 2.5 x 2.5 cm lint cloth
- Type of wrap if used: Lint cloth was covered with paraffin film and fixed with gauze and an adhesive plaster, covered with bandage.

REMOVAL OF TEST SUBSTANCE
- Washing: Applied area was wiped with gauze dipped in warm water
- Time after start of exposure: 3 minutes initially on animal No.1. Since no formation of crust, the test material was applied for 4 hours to the remaining sections on animals No.1 to 3.

OBSERVATION TIME POINTS
- Pasted sections of all the three animals with the test material were judged and evaluated after 1, 24, 48 and 72 hours from the removal of the pastings.

SCORING SYSTEM:
- Method of calculation: Draize method

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 2

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 3

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 5

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 6

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

The individual scores for erythema/eschar and edema are given in Table 'Average irritation score on skin - part 1' and Table 'Average irritation score on skin - part 2'. The skin to which the test material was pasted for 3 minutes, 1 hour and 4 hours showed no abnormality in all of the animals during any of the observation periods.

Any other information on results incl. tables

Average irritation score on skin - part 1

Animal no.	Treatment	Item	Administration time (minute)
			3	60
1	Test Material	Erythema and eschar	0	0
		Edema	0	0
		Total	0	0

 

Average irritation score on skin - part 2

Animal No.	Treatment	Patched sites	Item	Time (hour) after removed of patch				P.I.I.	Irritation grade
				1	24	48	78
1	Test Material	left	Erythema and eschar	0	0	0	0	0.0	-
			Edema	0	0	0	0
			Total	0	0	0	0
2	Test Material	left	Erythema and eschar	0	0	0	0
			Edema	0	0	0	0
			Total	0	0	0	0
3	Test Material	left	Erythema and eschar	0	0	0	0
			Edema	0	0	0	0
			Total	0	0	0	0
Mean				0.0	0.0	0.0	0.0
1	Non-treatment	right	Erythema and eschar	0	0	0	0	0.0	-
			Edema	0	0	0	0
			Total	0	0	0	0
2	Non-treatment	right	Erythema and eschar	0	0	0	0
			Edema	0	0	0	0
			Total	0	0	0	0
3	Non-treatment	right	Erythema and eschar	0	0	0	0
			Edema	0	0	0	0
			Total	0	0	0	0
Mean				0.0	0.0	0.0	0.0

Applicant's summary and conclusion

Interpretation of results:

other: Not classified according to EU criteria

Conclusions:

Under the conditions of this study, the test material caused no skin irritation.

Executive summary:

The potential of the test material to cause skin irritation was investigated in accordance with the standardised guideline OECD 404, under GLP conditions.

The test material was pasted under sealed condition to the skin of Japanese white rabbits for 3 minutes, 1 hour and 4 hours, and its skin irritation was investigated. All of the animals showed no abnormality on their skin to which the test material was pasted under sealed condition for 3 minutes, 1 hour and 4 hours during each of the observation periods. The primary irritation index P.I.I. as calculated from the average of the marks for 24, 48 and 72 hours after the removal of KNH-4 was 0.0. During the observation periods, none of the animals showed any abnormality in their general conditions.

Under the conditions of this study, the test material caused no skin irritation.