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Administrative data

Description of key information

Trypsin was tested for irritation in vivo in rabbit.

Skin irritation: Under the test conditions employed Porcine and Bovine concentrate of Pancreas Trypsin Novo had primary irritation scores of 2.8 and 3.6, respectivly. In the current GHS classification criteria it corresponds to Category 2 (irritant).

Eye irritation: Under the test conditions employed, Pancreas Trypsin Novo 3.3 special was regarded as Irritating to eyes, Category 2A. Besides the eye redness that carried a score of 1 up to 7 days after treatment, all effects were fully reversible within 72 hours.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

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 the available in vitro test methods are not applicable for the substance and therefore an in vivo skin irritation study was conducted
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
November 12, 1981 - January 15, 1982
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test procedure was according to the Code of Federal Regulations 1979, title 16, § 1500.41
Reference:
Composition 0
Qualifier:
according to
Guideline:
other: Code of Federal Regulations 1979, title 16, § 1500.41
Deviations:
no
GLP compliance:
no
Test material information:
Composition 1
Specific details on test material used for the study:
Porcine concentrate of Pancrease Trypsin NOVO
- Physical state: Brown powder
- Name of test material (as cited in study report): Porcine concentrate of Pancras Trypsin NOVO
- Lot/batch No.: FTTE 193
- Storage condition of test material: Stability was kept by placing the test substance in refrigerator

Bovine concentrate of Pancrease Trypsin NOVO
- Physical state: Brown powder
- Name of test material (as cited in study report): Bovine concentrate of Pancras Trypsin NOVO
- Lot/batch No.: FTTE 056
- Storage condition of test material: Stability was kept by placing the test substance in refrigerator
Species:
rabbit
Strain:
other: Albino rabbits, Danish Landstrain
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Bred by Novo Industry A/S
- Weight at study initiation: 2 - 3 kg
- Housing: Individually in galvanized steel cages with mesh floors.
- Diet (e.g. ad libitum): Standard diet (Ewos Brood Stock Feed for Rabbits and Guinea Pigs)
- Water (e.g. ad libitum): Automatic, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-22°C
- Humidity (%): 40-60%

IN-LIFE DATES: From: 1981-11-17 To: 1981-11-20
Type of coverage:
occlusive
Preparation of test site:
other: Abraded and non-abraded
Vehicle:
physiological saline
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g of the test material (Porcine and Bovine Trypsin concentrate) moistened with 0.4 mL physiological saline.
Duration of treatment / exposure:
24 hours
Observation period:
72 hours
Number of animals:
6
Details on study design:
The test compounds, Porcine and Bovine Trypsin concentrate were assessed by occlusive application of 0.5 g of the test material moistened with 0.4 mL physiological saline to an areal 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 24 hrs.

Irritation parameter:
overall irritation score
Remarks:
bovine trypsin
Basis:
mean
Time point:
24/48/72 h
Score:
3.6
Reversibility:
not specified
Irritation parameter:
overall irritation score
Remarks:
porcine trypsin
Basis:
mean
Time point:
24/48/72 h
Score:
2.8
Reversibility:
not specified
Irritation parameter:
erythema score
Remarks:
porcine trypsin
Basis:
mean
Remarks:
intact skin
Time point:
24/48/72 h
Score:
0
Irritation parameter:
erythema score
Remarks:
porcine trypsin
Basis:
mean
Remarks:
abraded skin
Time point:
24 h
Score:
2.67
Reversibility:
not reversible
Irritation parameter:
erythema score
Remarks:
porcine trypsin
Basis:
mean
Remarks:
abraded skin
Time point:
72 h
Score:
3.67
Reversibility:
not specified
Irritation parameter:
edema score
Remarks:
porcine trypsin
Basis:
mean
Remarks:
Intact skin
Time point:
24 h
Score:
0
Irritation parameter:
edema score
Remarks:
porcine trypsin
Basis:
mean
Remarks:
intact skin
Time point:
48 h
Score:
0
Irritation parameter:
edema score
Remarks:
porcine trypsin
Basis:
mean
Remarks:
abraded skin
Time point:
24 h
Score:
3.83
Reversibility:
fully reversible within: 72 h
Irritation parameter:
edema score
Remarks:
porcine trypsin
Basis:
mean
Remarks:
abraded skin
Time point:
72 h
Score:
0.83
Reversibility:
not specified
Irritation parameter:
erythema score
Remarks:
bovine trypsin
Basis:
mean
Remarks:
intact skin
Time point:
24 h
Score:
1.33
Reversibility:
not reversible
Irritation parameter:
erythema score
Remarks:
bovine trypsin
Basis:
mean
Remarks:
intact skin
Time point:
72 h
Score:
1.67
Reversibility:
not specified
Irritation parameter:
erythema score
Remarks:
bovine trypsin
Basis:
mean
Remarks:
abraded skin
Time point:
24 h
Score:
2.17
Reversibility:
not reversible
Irritation parameter:
erythema score
Remarks:
bovine trypsin
Basis:
mean
Remarks:
abraded skin
Time point:
72 h
Score:
3.67
Reversibility:
not specified
Irritation parameter:
edema score
Remarks:
bovine trypsin
Basis:
mean
Remarks:
intact skin
Time point:
24 h
Score:
0.83
Reversibility:
fully reversible
Irritation parameter:
edema score
Remarks:
bovine trypsin
Basis:
mean
Remarks:
intact skin
Time point:
72 h
Score:
0
Irritation parameter:
edema score
Remarks:
bovine trypsin
Basis:
mean
Remarks:
abraded skin
Time point:
24 h
Score:
4
Reversibility:
fully reversible
Irritation parameter:
edema score
Remarks:
bovine trypsin
Basis:
mean
Remarks:
adraded skin
Time point:
72 h
Score:
0.67
Reversibility:
not specified
Irritant / corrosive response data:
Porcine Trypsin 24 hours reading:
In 6/6 rabbits, intact skin sites applicated Porcine Typsin concentrate showed no reaction, while abraded skin sites showed well-defined to moderate erythema (score 2-3) and moderate to severe edema (score 3-4).

Porcine Trypsin 72 hours reading:
In 6/6 rabbits, intact skin sites applicated Porcine Typsin concentrate showed no reaction, while the main part of abraded skin sites showed a slight superficial eschar formation (score 4) but no chemosis.

Bovine Trypsin 24 hours reading:
In 4/6 rabbits, intact skin sites applicated Bovine Trypsin concentrate showed very slight to moderate erythema and edema (score 1-3), while intact skin sites in 2/6 rabbits showed no reactions. Abraded skin sites showed almost the same reaction as described for Procine Trypsin.

Bovine Trypsin 72 hours reading:
In 3/6 rabbits, intact skin applicated Bovine Trypsin concentrate showed well-defined erythema to slight superficial eschar formation (score 2-4) while intact skin sites in 3/6 rabbits showed no reactions. Abraded skin sites showed almost the same reaction as described for Porcine Trypsin.

Calculation of a primary irritation score for Porcine and Bovine Trypsin concentrate yielded 2.8 and 3.6, respectively
Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
Under the test conditions employed Porcine and Bovine concentrate of Pancreas Trypsin Novo had primary irritation scores of 2.8 and 3.6, respectivly. In the current GHS classification criteria it corresponds to Category 2 (irritant).
Executive summary:

An in vivo skin irritation test was conducted to assess the potential of the test substances, Porcine (batch FTTE 193) and Bovine (batch FTTE 056) Trypsin concentrate to cause skin irritation. The Code of Federal regulations 1979, title 16, § 1500.41 was followed.

0.5 g of each test substance moistened with 0.4 mL physiological saline was applied under occlusive conditions on clipped intact and abraded skin on 6 rabbits (Danish Landstrain) for 24 hours. After 24 hours the patches were removed from the rabbits and resulting reactions evaluated. Readings were again made 72 hours after exposure.

The primary irritation score for Porcine Trypsin concentrate (24 and 72 hours reading on intact and abraded skin) was 2.8.

The primary irritation score for Bovine Trypsin concentrate (24 and 72 hours reading on intact and abraded skin) was 3.6.

Both test substances were found to be significant more irritating on abraded skin then on intact skin. The test substances were both classified as "moderate irritant" to skin.

Under the test conditions employed Porcine and Bovine concentrate of Pancreas Trypsin Novo had primary irritation scores of 2 .8 and 3.6, respectivly, and were both classified as "moderate irritant" to skin. In the current GHS classification criteria it corresponds to Category 2.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

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 the available in vitro test methods are not applicable for the substance and therefore an in vivo eye irritation study was conducted
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
November 9, 1981 - January 15, 1982
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test procedure was according to the Code of Federal Regulations 1979, title 16, § 1500.42
Reference:
Composition 0
Qualifier:
according to
Guideline:
other: Code of Federal Regulations 1979, title 16, § 1500.42
Deviations:
no
GLP compliance:
no
Test material information:
Composition 1
Species:
rabbit
Strain:
other: Albino rabbits, Danish Landstrain
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Bred by Novo Industry A/S
- Weight at study initiation: 2 - 3 kg
- Housing: Individually in galvanized steel cages with mesh floors.
- Diet (e.g. ad libitum): Standard diet (Ewos Brood Stock Feed for rabbits and guinea pigs)
- Water (e.g. ad libitum): Automatic, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-22°C
- Humidity (%): 40-60%

IN-LIFE DATES: From: 1981-11-09 To: 1981-11-16
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g
- Concentration (if solution): Undiluted test material
Observation period (in vivo):
The reactions were judged 1, 24, 48 and 72 hours and 7 days after treatment.
Number of animals or in vitro replicates:
6
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): No washing

TOOL USED TO ASSESS SCORE:
Except for the examination at 1 hour, one drop of 1% fluorescein sodium was used for the eye examinations. The excess of fluorescein was flushed out with 0.9% sodium chloride solution.
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Time point:
other: 1 h
Score:
1.67
Max. score:
2
Reversibility:
not fully reversible within: 72 hours
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 1 h
Score:
0
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 1 h
Score:
0.33
Max. score:
1
Reversibility:
fully reversible within: 48 h
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: 1 h
Score:
3.33
Max. score:
4
Reversibility:
fully reversible within: 48 h
Irritation parameter:
conjunctivae score
Remarks:
ulceration
Basis:
mean
Time point:
other: 1 h
Score:
0
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
24 h
Score:
0.67
Max. score:
1
Reversibility:
fully reversible within: 48 h
Irritation parameter:
iris score
Basis:
mean
Time point:
24 h
Score:
0.67
Max. score:
1
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Time point:
24 h
Score:
2.83
Max. score:
3
Reversibility:
not fully reversible within: 72 h
Irritation parameter:
chemosis score
Remarks:
chemosis
Basis:
mean
Time point:
24 h
Score:
0.16
Max. score:
1
Reversibility:
fully reversible within: 48 h
Irritation parameter:
conjunctivae score
Remarks:
ulceration
Basis:
mean
Time point:
24 h
Score:
0
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
72 h
Score:
0
Irritation parameter:
iris score
Basis:
mean
Time point:
72 h
Score:
0
Irritation parameter:
conjunctivae score
Remarks:
reddness
Basis:
mean
Time point:
other: 48/72 h
Score:
1.83
Max. score:
3
Reversibility:
not fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
mean
Time point:
72 h
Score:
0
Irritation parameter:
conjunctivae score
Remarks:
ulceration
Basis:
mean
Time point:
72 h
Score:
0
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
7 d
Score:
0
Irritation parameter:
iris score
Basis:
mean
Time point:
7 d
Score:
0
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Time point:
7 d
Score:
0.83
Max. score:
1
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
48 h
Score:
0
Max. score:
0
Irritation parameter:
iris score
Basis:
mean
Time point:
48 h
Score:
0
Max. score:
0
Irritation parameter:
chemosis score
Basis:
mean
Time point:
48 h
Score:
0
Max. score:
0
Irritation parameter:
conjunctivae score
Remarks:
ulceration
Basis:
mean
Time point:
48 h
Score:
0
Max. score:
0
Irritant / corrosive response data:
Corneal reactions: Scattered areas of corneal opacity were seen at 1 and 24 hours in 2 and 4 rabbits respectively. No corneal opacity was noted at 48 hours.

Iris reactions: Congestion was seen at 24 hours in 2 rabbits. No iris reactions were noted at 48 hours.

Conjunctival reactions: Some hyperemic blood vessels or diffuse crimson color and swelling with lids about or more than half closed were seen at 1 hour in all the rabbits. At 24 hours the conjunctiva was diffuse crimson or beefy red in all the rabbits but no chemosis was seen. At day 7 only conjunctival reactions of score 1 were observed.

Besides the eye redness that carried a score of 1 for up to 7 days after treatment, all effects were fully reversible within 72 hours.
Interpretation of results:
Category 2A (irritating to eyes) based on GHS criteria
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the test conditions employed, Pancreas Trypsin Novo 3.3 special was regarded as Irritating to eyes, Category 2A. Besides the eye redness that carried a score of 1 for up to 7 days after treatment, all effects were fully reversible within 72 hours.
Executive summary:

A test for eye irritation in rabbits was performed with Pancreas Trypsin Novo 3.3 special, batch FTI 1164 in accordance with the Code of Federal Regulations 1979, title 16, § 1500.42.

The eyes of the six rabbits (three male and three females) were examined for defects 24 hours prior to the test. Just before instillation of the test substance the left eye of each rabbit was anaesthetized with two drops of 1% tetracain chloride. 0.1 g of Pancreas Trypsin Novo 3.3 special, batch FTI 1164 was placed in the left eye of each rabbit by gently pulling the lower lid away from the eye ball, and then introducing the test substance into the conjunctival sac. The lids were gently held together for one second. The right eye served as control.

The ocular reactions were recorded at 1, 24, 48, 72 hours and 7 days after instillation according to the specified guideline. Except for the examination at 1 hour, 1 drop of 1% fluorescein sodium was used.

Corneal reactions: Scattered areas of corneal opacity were seen at 1 and 24 hours in 2 and 4 rabbits respectively. No corneal opacity was noted at 48 hours.

Iris reactions: Congestion was seen at 24 hours in 2 rabbits. No iris reactions were noted at 48 hours.

Conjunctival reactions: Some hyperemic blood vessels or diffuse crimson color and swelling with lids about or more than half closed were seen at 1 hour in all the rabbits. At 24 hours the conjunctiva was diffuse crimson or beefy red in all the rabbits but no chemosis was seen. At day 7 only conjunctival reactions of score 1 were observed.

Besides the eye redness that carried a score of 1 for up to 7 days after treatment, all effects were fully reversible within 72 hours.

In conclusion, under the test conditions employed, Pancreas Trypsin Novo 3.3 special was regarded as Irritating to eyes, Category 2A.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Additional information

Justification for classification or non-classification

Category 2, Irritant.