Peroxidase

Endpoint summary

Description of key information

Peroxidase was tested in vivo for skin and eye irritation. Peroxidase was not irritating to skin and shall be classified as non-irritant. In addition, peroxidase can be classified as non-irritant to the eye.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1996-04-19 to 2000-03-30

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Bred by Novozymes A/S (formerly Novo Nordisk A/S), Denmark.
- Age at study initiation: 17-19 weeks.
- Weight at study initiation: Between 2.6 - 2.7 kg.
- Housing: Individually in plastic cages with mesh floor and automatic watering.
- Diet: 130 g standard diet daily (Altromin 2110).
- Water: Ad libitum tap water.
- Acclimation period: 7 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3 °C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

physiological saline

Remarks:

0.5 g of test substance moistened with 0.9 w/v % NaCl.

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g (equivalent to 0.49 g enzyme concentrated dry matter)

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.5 g of test substance moistened with 0.9 w/v % NaCl.

Adjacent areas of untreated skin served as control.

Duration of treatment / exposure:

4 hours

Observation period:

0.5-1, 24, 48 and 72 h after patch removal using the scoring system.
If skin reactions were observed, further assessments were conducted up to 21 days after test material application to determine the reversibility of the reactions.

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: The test compound peroxidase was assessed by semiocclusive application of 0.5 g of the test material, moisted with saline, to an area of 2.5 x 2.5 cm on the shaved skin on the back of three rabbits for 4 hrs.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): After 4 hours of exposure the patches were removed gently and the remaining test substance washed off with water.

OBSERVATION TIME POINTS
(indicate if minutes, hours or days): 0.5-1, 24, 48 and 72 h after patch removal using the scoring system. In addition, the animals were observed daily for systemic signs of toxicity.

SCORING SYSTEM:

Erythema and eschar fonnation Grade
No erythema .................................................................................................................................0
Very slight erythema (barely perceptible) ....................................................................................1
Well defined erythema ..................................................................................................................2
Moderate to severe erythema ......................................................................................................3
Severe erythema (beet redness) to slight eschar formations (injuries in depth) ........................4

Oedema formation
No oedema ....................................................................................................................................0
Very slight oedema (barely perceptible) ..................................................................................... 1
Slight oedema (edges of area well defined by definite raising) ...................................................2
Moderate oedema (raised approximately 1 mm) ...........................................................................3
Severe oedema (raised more than 1 mm and extending beyond area of exposure) .................. 4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 6 days

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 9 days

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Dermal reaction was observed at the test site of all animals at the readings 1, 24 and 48 hours after test substance removal. The scores never exceeded 2. At 72 hours after test substance removal and the following two days two animals showed redness at the site of application. From day 6 until day 8 skin reaction (redness) was seen in only one animal. The reaction never exceeded a score of one. Nine days after removal of the test substance no reaction was observed.

Other effects:

No other effects.

Interpretation of results:

not irritating

Conclusions:

Under the conditions of this test and criteria of the EEC Guideline, peroxidase batch PPX 5156, was classified as 'non-irritant to skin' as the reaction never exceeded a score higher than 1 and as the skin reaction had disappeared within the first 9 days of the 21-day observation period.

Executive summary:

The acute dermal irritant effect of peroxidase was investigated according to OECD TG 404 (1992) “Acute Dermal Irritation/Corrosion”. The study was conducted in accordance with GLP.

 

The study was performed on three New Zealand White rabbits. 0.5 g test substance (equivalent to 0.49 g enzyme concentrate dry matter) was applied once under a semi-occlusive bandage to the shaved back skin of each of 3 rabbits. The length of exposure was 4 hours. The primary skin irritation was evaluated 0.5-1, 24, 48 and 72 hours after removal of the test substance. Irritation was observed in two animals at the reading 72 hours after test substance removal. No irritation was observed nine days after test substance removal. No systemic effects were observed.

 

Under the conditions of this test and criteria of the EEC Guideline, peroxidase batch PPX 5156, was classified as 'non-irritant to skin' as the reaction never exceeded a score higher than 1 and as the skin reaction had disappeared within the 21 day observation period.

Reference 2

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

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:

From 1996-04-19 to 1996-07-12

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

OECD 405, (1987)

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

1987

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Bred by Novo Nordisk A/S
- Age at study initiation: 19 weeks
- Weight at study initiation: 2.6-2.7 kg
- Housing: Individually in plastic cages with mesh floor
- Diet: 130 g standard diet per day
- Water: Tap water ad libitum
- Acclimation period: 6-7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3°C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12 hrs/12 hrs

IN-LIFE DATES: From: 1996-04-29 To: 1996-05-03

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 g enzyme concentrate dry matter into the left eye

Duration of treatment / exposure:

The test material was applied to the conjunctival sac.

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 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.

Grading of Ocular Lesions:
CORNEA
Opacity : Degree of density (area most dense taken for reading)
No ulceration or opacity ...................................................................................................... 0
Scattered or diffuse areas of opacity (other than slight dulling of normal luster,
details of iris clearly visible) ................................................................................................ 1
Easily discernible translucent areas, details of iris slightly obscured ..................................2
Nacreous areas, no details of iris visible, size of pupil barely discernible ...........................3
Opaque cornea, iris not discernible through the opacity ......................................................4
IRIS
Normal ...................................................................................................................................0
Markedly deepened rugae, congestion, swelling, moderate circum corneal hyperaemia,
or injection, any of these or any combination thereof, iris still reacting to light (sluggish
reaction is positive) ...............................................................................................................1
No reaction to light, hemorrhage, gross destruction (any or all of these) .............................2
CONJUNCTIVAE
Redness : (Refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Blood vessels normal .............................................................................................................0
Some blood vessels definitely hyperemic (injected) ..............................................................1
Diffuse, crimson colour, individual vessels not easily discernible .........................................2
Diffuse beefy red ...................................................................................................................3
Chemosis : Lids and/or nictating membranes
No swelling .............................................................................................................................0
Any swelling above normal (includes nictating membranes) .................................................1
Obvious swelling with partial eversion of lids ........................................................................2
Swelling with lids about half closed .......................................................................................3
Swelling with lids more than half closed ................................................................................4
Flourescein 2% w/w was used, if there were signs of corneal opacity/ulcerations.

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.6

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No corneal reactions, no iris reactions were observed in any of the animals at the 1-, 24-, 48- and 72-hour reading.
Animal no. 4 showed conjunctival redness at the readings 1, 24 and 48 hours after test substance application. Rabbits no. 4 and 7 showed similar reactions at 1 and 24 hours after application. The scores were 1 in all situations. However, this was fully reversible after 72 hours.
The rabbits showed no sign of systemic reaction to the treatment. The weight of the animals was not influenced by the treatment

Other effects:

None

Interpretation of results:

GHS criteria not met

Conclusions:

The instillation of 0.1 g enzyme concentrate dry matter to the conjunctival sac of 3 rabbits caused no signs of eye irritation, and under the test conditions peroxidase is not to be classified and can be considered as 'non irritant'.

Executive summary:

In accordance with the OECD guideline 405, a test for eye irritation in rabbits was performed with Peroxidase, batch PPX 5156. The eyes of three animals were checked for defects or irritation before instillation and only animals without defects were used. 

 

0.1 g enzyme concentrate dry matter 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, iris reactions or tear secretion were noted at any time during the test. Conjunctival redness was observed after 24 hours in all three rabbits, and after 48 hours in 1 rabbit (maximum score of 1). All signs were fully reversible, and no reactions were seen after 72 hours.

 

In conclusion, peroxidase can be 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)

Additional information

Under the conditions of this test and criteria of the EEC Guideline, peroxidase batch PPX 5156, was classified as 'non-irritant to skin' as the reaction never exceeded a score higher than 1 and as the skin reaction had disappeared within the first 9 days of the 21-day observation period.

The instillation of 0.1 enzyme concentrate dry matter to the conjunctival sac of 3 rabbits caused no signs of eye irritation, and under the test conditions peroxidase is not to be classified and can be considered as 'non irritant'.

Justification for classification or non-classification

Peroxidase is not classified as a skin or eye irritant.