Ozonized olive oil

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Remarks:

Test performed in 2010 for purpose of medical device

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 27/07/2010 to 03/08/2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: ISO 10993-10:2002

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Ozonized Tetramers and Pentamers

IUPAC Name:

Ozonized Tetramers and Pentamers

impurity 1

Reference substance name:

Unknown impurities

IUPAC Name:

Unknown impurities

Constituent 2

Reference substance name:

Free Ozonized Acids

IUPAC Name:

Free Ozonized Acids

Constituent 3

Reference substance name:

Ozonized Trimers

IUPAC Name:

Ozonized Trimers

Constituent 4

Reference substance name:

Ozonized Dimers

IUPAC Name:

Ozonized Dimers

Constituent 5

Reference substance name:

Ozonized Triglycerides

IUPAC Name:

Ozonized Triglycerides

Test material form:

other: oiy gel

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Allevamento Bettinardi - Mamo (NO) - ltaly
- Weight at study initiation: 3210-3610 g at the beginning ofthe test
- Housing: Each rabbit was caged in stainless steel cages of cm 48.2x63x37 h equipped with automatic washing cycle. The housing room was lighted with fiuorescent lamps 12 hours for day. Room temperature and humidity were regulated by a conditioning pian! and were monitored daily. Recordings of the housing conditions are being retained in Eurofins Biolab S.r.l. files.
- Diet: The animals were fed with standard pellet complete diet supplied by the authorised breeder Harlan.
- Water: Filtered tap water from locai network was supplied ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature and humidity: Room temperature and humidity
- Photoperiod (hrs dark / hrs light): The housing room was lighted with fiuorescent lamps 12 hours for day.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent negative control

Amount / concentration applied:

0,5 ml of the test product were applied with a gauze (25 mmx 25 mm) directly to the skin on cranial site of each rabbit and covered with non-occlusive dressing

Duration of treatment / exposure:

The patches were removed 4 hours alter the application

Observation period:

60min, 24h, 48h, 72h

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 6,25cm2
- Type of wrap if used: no irritant gauze

REMOVAL OF TEST SUBSTANCE
- Time after start of exposure: 4h

OBSERVATION TIME POINTS
- 60min, 24h, 48h, 72h

SCORING SYSTEM:
- Method of calculation: ISO 10993-10:2002

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

edema score

Basis:

animal: Cranial (left area) and Caudal (right area): Animal number 317 - Animal number 320 - Animal number 354

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

erythema score

Basis:

animal: Craudal (right area): Animal number 317 - Animal number 320 - Animal number 354

Time point:

72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

erythema score

Basis:

animal: Cranial (left area): Animal number 317 - Animal number 320 - Animal number 354

Time point:

72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

erythema score

Basis:

animal: Caudal (right area): Animal number 317 - Animal number 320 - Animal number 354

Time point:

48 h

Score:

1

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 5

Irritation parameter:

erythema score

Basis:

animal: Cranial (left area): Animal number 317 - Animal number 320 - Animal number 354

Time point:

48 h

Score:

2

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

other: On the basis of the results, interpreted according to GHS regulation, the test substance is considered SLIGHTLY IRRITANT for skin but, according to CLP regulation 1272/08, the subtance is not classified as skin irritant (score for CLP category 2 is from 2

Results 6

Irritation parameter:

erythema score

Basis:

animal: Caudal (right area): Animal number 320 - Animal number 354

Time point:

24 h

Score:

1

Max. score:

1

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 7

Irritation parameter:

erythema score

Basis:

animal: Caudal (right area): Animal number 317

Time point:

24 h

Score:

2

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

other: On the basis of the results, interpreted according to GHS regulation, the test substance is considered SLIGHTLY IRRITANT for skin but, according to CLP regulation 1272/08, the subtance is not classified as skin irritant (score for CLP category 2 is from 2

Results 8

Irritation parameter:

erythema score

Basis:

animal: Cranial (left area): Animal number 317 - Animal number 320 - Animal number 354

Time point:

24 h

Score:

2

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

other: On the basis of the results, interpreted according to GHS regulation, the test substance is considered SLIGHTLY IRRITANT for skin but, according to CLP regulation 1272/08, the subtance is not classified as skin irritant (score for CLP category 2 is from 2

Results 9

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

1.21

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

other: On the basis of the results, interpreted according to ISO 10993-10:2002, the test product must be considered SLIGHTLY IRRITANT for the skin, but not irritant according to CLP 1272/08.

Applicant's summary and conclusion

Interpretation of results:

Category 3 (mild irritant) based on GHS criteria

Conclusions:

On the basis of the results, interpreted according to GHS regulation, the test substance is considered SLIGHTLY IRRITANT for skin but, according to CLP regulation 1272/08, the subtance is not classified as skin irritant (score for CLP category 2 is from 2,3 to 4.0)

Executive summary:

On the test substance a toxicological study was carried out to evaluate the possible locai toxic effects, throughout the following test:

 - skin irritation test according to ISO 10993-10:2002

The skinirritation test was carriedout through a semi-occlusive application; the test product was applied on the intact skin of 3 rabbits, in the dorsal region both on the left and on the right side.

Each animal had the right caudal region and left cranial region treated with the test product.

The right cranial region and the left caudal region were treated with a non irritant humidified gauze (25mmx25mm), used as control.

Reactions were evaluated 1 hours following the remaval of the patches and were evaluated againat 24, 48 and 72h after exposure.

-After 60 minutes all treated sites shown a very slight erythema.

-After 24 hours two animals shown a well defined erythema in treated cranial site and a very slight erythema in the caudal sites, one animai shown a well defined erythema in both treated sites.

Symptoms disappear completely in all animals in the caudal sites after 72 hours, in the cranial sites after 7 days.

In control regions no oedema or erythema were observed.