Pullulanase

Endpoint summary

Description of key information

Pullulanase is neither a skin or an eye irritant.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May 11 - June 16, 1982

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Test procedure was according to the Code of Federal Regulations 1979, title 16, § 1500.41 and adapted to OECD TG404 (1981). Test procedure according to GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1981

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Code of Federal Regulations 1979, title 16, § 1500.41

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

other: Albino rabbits, Danish Landstrain

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Bred by Novo Industri A/S, Denmark (Renamed Novo Nordisk A/S in year 1989)
- Weight at study initiation: between 2.9 - 3.3 kg
- Age at study initiation: Young adults
- Housing: Individually in galvanized steel cages with mesh floors
- Diet (e.g. ad libitum): Standard diet
- Water (e.g. ad libitum): Tap water ad libitum
- Temperature (°C): 18 to 22 °C
- Humidity (%): 40-50%
- Acclimation period: The animals were bred in-house

IN-LIFE DATES: From: 1982-05-11 To: 1982-05-14

Type of coverage:

semiocclusive

Preparation of test site:

other: Shaved, abraded and non-abraded

Vehicle:

unchanged (no vehicle)

Remarks:

moistened with saline

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g
- Concentration: undiluted, 10% w/w total protein (100 mg total protein/g)

Duration of treatment / exposure:

4 hours

Observation period:

72 hrs

Number of animals:

6

Details on study design:

The test compound pullulanase was assessed by semiocclusive application of 0.5 g of the test material moistened with saline to an area of 1 inch x 1 inch (2.5 x 2.5 cm) on the closely-clipped intact and abraded skin on the back of three male and three female albino rabbits for 4 hrs.
The study was conducted to comply with the following guideline:
- Code of Federal Regulations 1979, title 16, § 1500.41, adapted to OECD TG404.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

No erythema, eschar formation or oedema was observed at the test sites at any of the six rabbits at the examinations 30 minutes, 24, 48 or 72 hours after termination of exposure. This result was obtained for test sites of intact as well as abraded skin.

Other effects:

No other effects.

Interpretation of results:

GHS criteria not met

Conclusions:

The test material pullulanase is not irritating to skin.

Executive summary:

The acute dermal irritant effect of pullulanase was investigated according to the guideline Code of Federal Regulations 1979, title 16, § 1500.41 and adapted to OECD TG404 (1981) “Acute Dermal Irritation/Corrosion”. The study was conducted in accordance with GLP.

The study was performed on three male and three female albino rabbits, Danish Landstrain. They were each exposed to 0.5 g of the test material, moistened with saline, applied under semiocclusive conditions to each of the closely-clipped intact and abraded test sites (2.5 x 2.5 cm) on the back.

After a 4 -hour exposure period, the test item was removed from the test site and the skin was examined after 30 minutes, 24 h, 48 h and 72 h. No erythema, eschar formation or oedema was observed at any of the test sites at any of the examinations. 

In conclusion, pullulanase was not irritating to skin and shall be classified as non-irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 08 - July 27, 1982

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was performed according to GLP and the procedures were according to the Code of Federal Regulations, title 16 § 1500.42 and adapted to OECD TG405 (1981).

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Code of Federal regulations, title 16, § 1500.42

GLP compliance:

yes

Species:

rabbit

Strain:

other: Danish Landstrain

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Bred by Novo Industri, Denmark
- Weight at study initiation: between 2.7-3.2 kg
- Housing: Individually, in animal room with control of temperature (18-22°C) and relative humidity (30-35%)
- Diet : Standard diet (Ewos Brood Stock Feed for Rabbits and Guinea Pigs)
- Water (e.g. ad libitum): tap water ad libitum

IN-LIFE DATES: From: 1982-06-08 To: 1982-06-11

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

Test batch used as is:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg into the left eye

Duration of treatment / exposure:

The test material was applied to the conjunctivel sac, no further action done, i.e. no washing.

Observation period (in vivo):

The reactions were judged 1, 24, 48 and 72 hours after instillation of the test material.

Number of animals or in vitro replicates:

3 female and 3 male rabbits

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: No
SCORING SYSTEM: The ocular reactions were recorded at 1, 24, 48 and 72 hours after instillation according to the guideline.

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal: #1, #4, #5, #6

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48h

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48h

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

No corneal opacity or iris reactions were noted at any time during the test. Diffuse redness (3 rabbits, score 2), slight redness (3 rabbits, score 1) and slight chemosis (3 rabbits, score 1) were observed after 1 hour. After 24 hours only mild redness was seen in 2 rabbits (score 1). All signs were reversible, and no reactions were seen at the 48-hour reading.

Other effects:

None

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, pullulanase can be classified as a ‘non irritant’ to the eye.

Executive summary:

In accordance with the OECD guideline 405, a test for eye irritation in rabbits was performed with Pullulanase, batch PPY1323.

The eyes of six animals were checked for defects or irritation before instillation and only animals without defects were used.

100 mg of Pullulanase, batch PPY1323, was placed in the conjunctival sac of the left eye by pulling the lower eyelid away from the eyeball. The lids were gently held together for about one second after instillation. The untreated eye served as control. The ocular reactions were recorded at 1, 24, 48 and 72 hours after instillation according to the specified guideline.

No corneal opacity or iris reactions were noted at any time during the test. Diffuse or slight redness (max score 2) and slight chemosis (max score 1) were observed after 1 hour. After 24 hours only mild redness was seen in 2 rabbits (score 1). All signs were reversible, and no reactions were seen at the 48-hour reading.

In conclusion, pullulanase can be classified as a 'non-irritant' to the eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The skin and eye irritation potential of pullulanase has been tested according to OECD guidelines, and in compliance with GLP. No reactions at all were seen in the skin irritation study. In the eye irritation study, only transient conjunctival redness with or without swelling were seen. All reactions were mild, no scores were above 1. All effects in the eye irritation study cleared before the 48h reading.

The conclusion was that pullulanase did not exert any skin and eye irritation. 

Justification for classification or non-classification

The skin and eye irritation potential of pullulanase has been tested according to OECD guidelines, and in compliance with GLP. The conclusion was that pullulanase did not exert any skin and eye irritation and therefore pullulanase should not be classifed.