Mucorpepsin

Endpoint summary

Description of key information

Renilase SP 252 was tested in rabbits for skin and eye irritation.

- The skin irritation study was conducted according to the Code of Federal Regulations 1979, title 16 § 1500.41. Two of the six female animals (No. 5 and 6) revealed on the abraded area a very slight erythema evaluated 30-60 minutes after patch removal, which gave a primary irritation score of 0.16. However, the reaction was fully reversible, and no irritation signs were seen at the reading at 24 hours. The irritation score for the mean of 24, 48 and 72 hours was 0. Therefore, according to the current classification, rennilase SP 252 is classified as a 'non irritant' to the skin.

- In the eye irritation study, no corneal opacity or iris reactions were noted in any of the animals at any time during the test. Mild redness and slight chemosis were seen in all of the rabbits estimated at 1 hour. At 24 hours mild redness was seen in 1 rabbit. At 48 and 72 hours no conjunctival reactions were seen in any of the rabbits. In conclusion, rennilase SP 252 can be classified as a 'non-irritant' to the rabbit eye.

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:

1982-05-11 to 1982-06-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to GLP and according to the Code of Federal Regulations 1979, title 16, 1500.41 and adapted to OECD Guidelines for Testing Chemicals

Qualifier:

according to guideline

Guideline:

other: Study conducted according to GLP and according to the Code of Federal Regulations 1979, title 16, 1500.41 and adapted to OECD Guidelines for Testing Chemicals.

Version / remarks:

1979

Deviations:

no

GLP compliance:

yes

Remarks:

GLP equivalent at the time. The study was subject to appropriate inspection by the quality assurance unit.

Species:

rabbit

Strain:

other: Danish Landstrain

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Bred by Novozymes A/S (previously Novo Industri A/S)
- Weight at study initiation: 2.9-3.3 kg
- Housing: Individually in galvanized steel cages with mesh floors and automatic watering
- Diet (e.g. ad libitum): Standard diet

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 22°C
- Humidity (%): 40 - 50%

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

Type of coverage:

occlusive

Preparation of test site:

abraded

Remarks:

Hair was clipped and skin abraded.

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL

Duration of treatment / exposure:

4 hours

Observation period:

30-60 minutes, 24 hours, 48 hours and 72 hours.

Number of animals:

6 (3 males and 3 females)

Details on study design:

TEST SITE
- Area of exposure: 1 inch by 1 inch
- Type of wrap if used: A coat of PVC

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Not specified
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
30-60 minutes, 24 hours, 48 hours and 72 hours

SCORING SYSTEM:

Erythema and eschar formation
No erythema ....................................................................................................................................0
Very slight erythema (bar ely perceptible) ......................................................................................1
Well-defined erythema .....................................................................................................................2
Moderate to severe erythema ..........................................................................................................3
Severe erythema (beet redness) to slight eschar formations (injuries in the depth) ......................4

Edema formation
No edema ..........................................................................................................................................0
Very slight edema (bar ely perceptible) ............................................................................................1
Slight edema (edges of area well-defined by definite raising) .........................................................2
Moderate edema (raised approximately 1 millimeter) ........................................................................3
Severe edema (raised more than 1 millimeter and extending beyond the area of exposure) ......... 4

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Remarks:

Abraded skin

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Remarks:

Abraded skin

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

Two of the female animals (No. 5 and 6) revealed on the abraded area a very slight erythema evaluated 30-60 minutes after patch removal. The reaction was reversible, and no signs were seen at the reading at 24 hours.
None of the other animals showed any signs of reaction to the test substance during the observation period.

Interpretation of results:

not irritating

Conclusions:

The study was conducted according to the Code of Federal Regulations 1979, title 16 § 1500.41. Two of the six female animals (No. 5 and 6) revealed on the abraded area a very slight erythema evaluated 30-60 minutes after patch removal, which gave a primary irritation score of 0.16. However, the reaction was fully reversible, and no signs were observed at the reading at 24 hours. The irritation score for the mean of 24, 48 and 72 hours was 0. Therefore, according to the current classification, rennilase SP 252 was classified as a "non irritant" to the skin.

Executive summary:

The skin irritation effect of rennilase SP 252, batch PPR1289 was investigated according to the Code of Federal Regulations 1979, title 16 § 1500.41 and adapted to the OECD guidelines for testing of chemicals.

The study was performed on six rabbits. They were each exposed to 0.5 mL of the test material, applied under occlusive conditions to clipped intact and abraded skin (1 x 1 inch) on the back.

After a 4-hour exposure period, the test item was removed from the test site and the skin was examined after 30-60 minutes, 24 h, 48 h and 72 h.

Two of the six female animals (No. 5 and 6) revealed on the abraded area a very slight erythema evaluated 30-60 minutes after patch removal, which gave a primary irritation score of 0.16. However, the reaction was fully reversible, and no irritation signs were seen at the reading at 24 hours. The irritation score for the mean of 24, 48 and 72 hours was 0.

Therefore, according to the current classification, rennilase SP 252 was classified as a "non irritant" to the skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1982-04-20 to 1982-06-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test method was proposed by OECD Guidelines for Testing Chemicals and evaluation made according to the Code of Federal Regulations, title 16, §1500.42.

Qualifier:

according to guideline

Guideline:

other: OECD Guidelines for Testing Chemicals and evaluation made according to the Code of Federal Regulations, title 16, §1500.42.

Deviations:

no

GLP compliance:

yes

Remarks:

GLP equivalent at the time where the conduct of the study was subject to appropriate inspection by the quality assurance unit.

Species:

rabbit

Strain:

other: Danish Landstrain

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Bred by Novozymes A/S (previously Novo Industri A/S)
- Weight at study initiation: 2.9-3.2 kg
- Housing: Individually in galvanized steel cages with mesh floors and automatic watering
- Diet (e.g. ad libitum): Standard diet (Ewos Brood Stock Feed for Rabbits and Guinea Pigs).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 22 °C
- Humidity (%): 30 - 35%

IN-LIFE DATES: From: 1982-04-20 To: 1982-04-23

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): 0.1 mL into the left eye

Duration of treatment / exposure:

0.1 mL of the test substance was installed into the left eye of each rabbit by gently pulling the lower lid away from the eye ball, and the test substance was carefully introduced into the resulting cup. The lids were then gently held together for one second.
The right eye, remaining untreated, served as a control.

Observation period (in vivo):

The eyes were examined at 1, 24, 48 and 72 hours after treatment.

Number of animals or in vitro replicates:

6 rabbits (3 female + 3 male)

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 Code of Federal Regulations, title 16 §1500.42

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

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal: #1, #3, #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

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:

conjunctivae score

Remarks:

Ulceration

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. Mild redness and slight chemosis were seen in all of the rabbits estimated at 1 hour.
At 24 hours mild redness was seen in 1 rabbit.
Both observed reactions were fully reversible and at 48 and 72 hours no conjunctival reactions were seen in any of the rabbits.

Interpretation of results:

not irritating

Remarks:

Not classified, Criteria used for interpretation of results: OECD GHS

Conclusions:

In conclusion, Rennilase SP 252 was regarded as negative to potential for eye injury.

Executive summary:

In accordance with the proposed test method by OECD Guidelines for Testing Chemicals and with the evaluation conducted according to the Code of Fereral Regulation, title 1500.42, a test for eye irritation in rabbits was performed with rennilase SP 252, batch PPR 1289.

0.1 mL of rennilase SP 252, batch PPR 1289 was placed in the conjunctival sac of the left eye of six rabbits 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. Mild redness and slight chemosis were seen in all of the rabbits estimated at 1 hour. At 24 hours mild redness was seen in 1 rabbit. At 48 and 72 hours no conjunctival reactions were seen in any of the rabbits.

In conclusion, rennilase SP 252 was classified as a 'non-irritant' to the rabbit eye.

Reference 2

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

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Not classified.