Mannanase, endo-1,4-β-

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 5, 2002 – November 8, 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Active enzyme protein of mannan endo-1,4-beta-mannosidase (EC no. 253-446-5, CAS no. 37288-54-3, EC name: Mannanase, endo-1,4-.beta.-, Enzyme Class No.: 3.2.1.78)

Molecular formula:

Not applicable, see remarks

IUPAC Name:

Active enzyme protein of mannan endo-1,4-beta-mannosidase (EC no. 253-446-5, CAS no. 37288-54-3, EC name: Mannanase, endo-1,4-.beta.-, Enzyme Class No.: 3.2.1.78)

Constituent 2

Reference substance name:

Protein as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available

IUPAC Name:

Protein as a constituent of enzyme deriving from the fermentation or extraction process

Constituent 3

Reference substance name:

Inorganic salts as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available. See remarks.

IUPAC Name:

Inorganic salts as a constituent of enzyme deriving from the fermentation or extraction process

Constituent 4

Reference substance name:

Carbohydrates constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available. See remarks.

IUPAC Name:

Carbohydrates constituent of enzyme deriving from the fermentation or extraction process

Constituent 5

Reference substance name:

Lipids as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available. See remarks.

IUPAC Name:

Lipids as a constituent of enzyme deriving from the fermentation or extraction process

Test material form:

liquid

Details on test material:

-Substance type: UVCB
-Physical state: liqiud
-Storage: The test article was stored in the freezer and the refrigerator

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Animals were procured from Millbrook Breeding Labs, Amherst MA 10/23/02. Three healthy animals were selected after an equilibration period of at least five days. The test animals were born the week of August 4th, 2002 and pre-test body weights ranged from 2.3 to 2.5 kg.

Animals were identified by cage notation and metal ear tags. Animals were each housed in a separate suspended wire cage. Bedding was placed beneath each cage and changed at least three times per week. Fresh food (PMI Rabbit Chow Diet #5321) was provided daily. Water was freely available at all times. The animal room was used only for actue testing on rabbits. The room was temperature and humidity controlled, had a 12 hour light/dark cycle, and was kept clean and vermin free.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Remarks:

The test article was used as received.

Controls:

no

Amount / concentration applied:

The test article was dosed by volume at 0.5 mL per rabbit.

Duration of treatment / exposure:

The test area was covered with a 4 ply surgical gauze patch and the torso was wrapped with plastic in a semi-occlusive manner and secured with non-irritating tape. The test article was kept in contact with the skin for 4 hours, at which time the wrappings were removed.

Observation period:

Observation of any dermal reactions were recorded at 1, 24, 48 and 72 hours after patch removal. The skin was also evaluated for ulceration and necrosis or any evidence of tissue destruction.

Number of animals:

Three (two males and one female)

Details on study design:

The dorsal area of the trunk of each animal was clipped free of hair on the day prior to the application of the test article. The site of administration was approximately 2.5 cm x 2.5 cm. The test article was dosed by volume at 0.5 mL per rabbit. The test area was covered with a 4 ply surgical gauze patch and the torso was wrapped with plastic in a semi-occlusive manner and secured with non-irritating tape. The test article was kept in contact with the skin for 4 hours, at which time the wrappings were removed. Observation of any dermal reactions were recorded at 1, 24, 48 and 72 hours after patch removal. Residual test article was removed from the test site by gentle washing with distilled water prior to scoring observations.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Reversibility:

other: Not applicable. No erythema observed.

Results 2

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Reversibility:

other: Not applicable. No erythema observed.

Results 3

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Reversibility:

other: Not applicable. No erythema observed.

Results 4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Reversibility:

other: Not applicable. No edema observed.

Results 5

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Reversibility:

other: Not applicable. No edema observed.

Results 6

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Reversibility:

other: Not applicable. No edema observed.

Irritant / corrosive response data:

There were no erythema or edema noted. There were no overt signs of systemic toxicity. The Primary Irritation Score was determined to be 0.

Other effects:

None.

Any other information on results incl. tables

Systemic observations were also made at 1, 24, 48, and 72 hours after patch removal. All observations were normal at all time points in all test animals. There were no abnormal clinical signs observed and all body weight changes were normal.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test article is not irritating to skin. Mannanase shall be classified as 'Non Irritant'.

Executive summary:

Three New Zealand White rabbits (2 males : 1 female) were used. The test article (0.5 mL) was applied to one intact dermal site/rabbit. The test article was kept in contact with the skin for 4 hours, at which time the wrappings were removed. Dermal reactions were scored at 1, 24, 48 and 72 hours after patch removal. There were no erythema or edema noted at any time during the study period. There were no overt signs of systemic toxicity. The Primary Irritation Score was determined to be 0 and mannanase shall be classified as 'Non Irritant'.