Registration Dossier

Diss Factsheets

Administrative data

Description of key information

The test item is neither a skin irritant nor a eye irritant.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Oct 05 - Oct 19, 1993
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
None
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
Strain: Rabbit, IVA:NZW, males (m) and females (f)
Source: SAVO-Ivanovas, Kisslegg
Age: about 36 weeks
Initial Weight: 3,96 (3.36-4.67) kg
Total Number of Animals: 3 (2m, 1 f)
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
other: paraffin oil
Controls:
not required
Amount / concentration applied:
0,5 g
Duration of treatment / exposure:
4 hours
Observation period:
1, 24, 48, 72 hours after removal of patches and then daily up to day 14
Number of animals:
2m, 1f
Details on study design:
Identification and adaptation
Healthy young rabbits were allocated to the different groups.The animals were kept in the experimental room for more than 7 days to allow for acclimatization.

Assignment
2 male and 1 female rabbit were assigned for this study.


Housing and diet
All rabbits were housed in an air-conditioned room of 39 m2 in the rabbit experimental station with daylight. They were kept separately in cages type KK 017 (floor area: 75 x 54.5 = 4087 cm2 , height: 38 cm; manufacturer: Hulskamp) on metal grids. The paper underneath the cages was changed three times a week. The cages were machine­ cleaned before the start of the studies.

Temperature and humidity were measured using a thermo­ hygrograph (type 252, Wilhelm Lambrecht KG, Göttingen).
The room temperature varied from 19 to 21°C and the humidity from 33 to 57 %.

The rabbits received Altromin Standard Diet TPF(R) 2113, pellets and acidified, fully demineralized water from an automatic drinking device ad libitum.

The diet is checked periodically by an independent and German Government approved testing laboratory, according to the regulations of the manufacturer. Analysis includes both qualitative and quantitative evaluation for heavy metals, aflatoxins, pesticides, and antibiotics.


Vehicle
Name: Liquid paraffin


Observations for clinical symptoms
The rabbit was examined for skin alterations, behavior, and general condition 1 hour after removal of the patch, after 24, 48, 72 hours, and then daily up to experimental day 14.

Skin changes at the application sites were evaluated according to the DRAIZE-, OECD-and EEC recommendations.

Grading scale for evaluation:

Erythema and eschar formation

Scores
--------------------------------------------------
No erythema 0

Very slight erythema (barely perceptible) 1

Well defined erythema 2

Moderate to severe erythema 3

Severe erythema (beet redness) to eschar
formation preventing grading of erythema 4
--------------------------------------------------
Maximum possible: 4
--------------------------------------------------

Edema formation
Scores
--------------------------------------------------
No edema 0

Very slight edema (barely perceptible) 1

Slight edema (edges of area well
defined by definite raising) 2

Moderate edema (raised approx. 1 mm) 3

Severe edema (raised more than 1 mm
and extending beyond area of exposure) 4
--------------------------------------------------
Maximum possible: 4
--------------------------------------------------

Total possible irritation score (maximum): 8

Mean score = Mean grading for erythema or edema of all the rabbits per time point
Mean cumulative score = Mean grading of all readings for erythema and edema per time point
Mean score per animal = Mean grading for each animal of erythema or edema (24, 48, and 72 hours after removal of the patches)
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.7
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.67
Max. score:
1
Reversibility:
fully reversible
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.67
Max. score:
1
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.67
Max. score:
1
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
animal #2
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
Remarks on result:
other: see text for details
Irritant / corrosive response data:
The first examination of the treated skin sites followed 1 hour after removal of the patches. Then examinations of the skin sites were performed daily for further 14 days.
In the primary skin irritation test, erythema (scores 1 and 2) were observed from day 2 - 14, and edema (scores 1 and 2) from day 3 - 10 of the study. In two rabbits scales were observed from day 8 - 12 of the study.
Other effects:
No signs of systemic toxicity were detected. The body weight developent corresponded to that of the untreated rabbits of the same age. No mortality was observed during the course of the study.
Interpretation of results:
GHS criteria not met
Conclusions:
According to the result of the irritation scores, the test material has not to be classified as irritant for the skin.
Executive summary:

Purpose

The purpose of this assay was to identify the skin irritation/corrosion potential of the test material when applied to the intact skin of previously shaven rabbits for a 4 hours period under semiocclusive conditions.

This study should provide a rational basis for risk assessment to the irritating potential of the test item in man.

Study Design

To test for primary skin irritation, 0.5 g of the test material was mixed with some drops of liquid paraffin to ensure good contact with the skin. Afterwards the test material was spread onto 6 cm2 patches and applied to the intact skin of previously shaven rabbits for a 4 hours period under semiocclusive conditions. The study was started as an initial test with one animal. This animal showed signs of irritation (well defined erythema, edema, brittle skin, and scales) and no more animals were treated. The first examination of the treated skin sites followed 1 hour after removal of the patches. Thereafter, examinations were performed daily for further 14 days.

Results

No signs of pain were observed after administration. The body weight developent corresponded to that of the untreated rabbits of the same age. No mortality was observed during the course of the study.

In the primary skin irritation test, erythema (scores 1 and 2) were observed from day 2 - 14, and edema (scores 1 and 2) from day 3 - 10 of the study. In two rabbits scales were observed from day 8 - 12 of the study.

Conclusion

According to the result of the irritation scores, the test material has not to be classified as irritant for the skin.

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

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Oct 05 - Oct 19, 1993
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
None
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Strain: Rabbit, IVA:NZW, males (m) and females (f)
Source: SAVO-Ivanovas, Kisslegg
Age: about 36 weeks
Initial Weight: 4.15 (3.86 - 4.34) kg
Total Number of Animals: 3 (2m, 1 f)
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
0,1 g
Observation period (in vivo):
1 hour after treatment, after 24, 48, and 72 hours, then daily up to day 5 of the experimental part.
Number of animals or in vitro replicates:
2m, 1f
Details on study design:
-- Identification and adaptation
Healthy young rabbits were allocated to the different groups.The animals were kept in the experimental room for more than 7 days to allow for acclimatization.


-- Assignment
2 male and 1 female rabbit were assigned for this study.

-- Housing and diet
All rabbits were housed in an air-conditioned room of 39 m2 in the rabbit experimental station with daylight. They were kept separately in cages type KK 017 (floor area: 75 x 54.5 = 4087 cm2 , height: 38 cm; manufacturer: Hulskamp) on metal grids. The paper underneath the cages was changed three times a week. The cages were machine­ cleaned before the start of the studies.

Temperature and humidity were measured using a thermo­ hygrograph (type 252, Wilhelm Lambrecht KG, Göttingen).
The room temperature varied from 19 to 21°C and the humidity from 33 to 57 %.

The rabbits received Altromin Standard Diet TPF(R) 2113, pellets and acidified, fully demineralized water from an automatic drinking device ad libitum.

The diet is checked periodically by an independent and German Government approved testing laboratory, according to the regulations of the manufacturer. Analysis includes both qualitative and quantitative evaluation for heavy metals, aflatoxins, pesticides, and antibiotics.


-- Preparation
Before the application, the test material was ground in a mortar using a pestle.


–- Administration and dose level
To ensure that only rabbits with normal eyes were included in the study, approximately 24 hours before treatment ophthalmological examinations were performed after instillation of a 0.15 % fluorescein solution (Dr. Thilo & Co.) using an ophthalmoscope according to Eisenhut (Basel). Animals with eye defects, injury or irritation were excluded.
0.1 g test material was instilled into the conjunctival sac of the left eye of each animal by gently pulling the lower lid away from the eyeball to form a cup into which the test material was placed. The right eye remained untreated and served as control. After instillation, the eyelids were closed for about 30 seconds. Rinsing of the eyes was not performed.


-- Observation for clinical symptoms
The rabbits were investigated for eye irritation and for changes in behavior and general condition 1 hour after treatment, and then after 24, 48, 72 hours and so on for a period of further 4 days.
All rabbits were weighed before the start of the study and on days 5 and 8.
Local changes of the eye were evaluated according to the recommendations of the OECD (see below). The classification followed the Appendix VI of the EEC Directive 79/831.

-- Evaluation of eye reactions
- Cornea Scores
A) 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 lustre), details of iris clearly visible 1
Easily discernible translucent area, details of iris slightly obscured 2
Nacrous area, no details of iris visible, size of pupil barely
discernible 3
Opaque cornea, iris not discernible through the opacity 4

B) Area of cornea involved not evaluated (Rating according to the DRAIZE method)

- Iris
A) Normal 0
Markedly deepened rugae, congestion, swelling, moderate
circumcorneal hyperemia, or injection, any of these or combination
of any 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 Scores
A) Redness (refers to palpebral and bulbar conjunctivae,
cornea and iris)
Blood vessels normal 0
Some blood vessels definitely hyperemic (injected) 1
Diffuse, crimson color, individual vessels not easily discernible 2
Diffuse beefy red 3

B) 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

C) Discharge (Rating according to the DRAIZE method )
No discharge 0
Any amount different from normal (does not include small
amounts observed in inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs,
and of a considerable area around the eye 3

Mean score of all animals = Mean grading for irritations of cornea, iris, and conjunctivae per time point
Mean score per animal = Mean grading for each animal of irritations of cornea, iris, and conjunctivae (1, 24, 48, and 72 hours after application)
Maximum value = Maximum grading of a sign of irritation within a period
Irritation parameter:
iris score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
fully reversible
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
fully reversible
Irritation parameter:
chemosis score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
fully reversible

All animals survived the observation period. Body weight development of the treated rabbits was inconspicuous.

No signs of clinical toxicity were detected. No signs of irritation were observed at the cornea or iris. All animals showed redness (score 1) and discharge (score 1) one hour after instillation of the test material. No abnormalities were detected in the untreated eyes.

Eve irritation of all animals (mean score):

  24 h  48 h  72 h 
Cornea (mean /max) 0/0 0/0 0/0
 Iris (mean / max) 0/0 0/0 0/0
 Conjunctivae (mean / max) 0/0 0/0 0/0
Interpretation of results:
GHS criteria not met
Conclusions:
No eye irritating potential could be detected, thus the test material should not be classified as eye irritant according to EU Regulation No. 1272/2008 (CLP).
Executive summary:

Purpose

The purpose of this primary eye irritation assay was to provide information on possible health hazards in case of acute eye contact with a test material and serve as a rational basis for risk assessment to the eye irritating potential of the test item in man.

Study Design

To test for eye irritation a test according to the OECD-Guidelines for Testing of Chemicals No. 405, the annex to Directive 79/8319 EEC, and the recommendations of DRAIZE (1959) was performed.

Results

No signs of irritation were observed at the cornea or iris. In one animal the conjunctivae showed redness (score 1) at the first reading. Later on, no signs of irritation were seen. Thereafter no signs of irritation were observed. The untreated eyes were unchanged.

Conclusion

No eye irritating potential could be detected, thus the test material should not be classified as eye irritant according to EU Regulation No. 1272/2008 (CLP).

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

Additional information

Justification for selection of skin irritation / corrosion endpoint:
This study was performed according to GLP and the methods applied are fully compliant with OECD TG 404.

Justification for selection of eye irritation endpoint:
This study was performed according to GLP and the methods applied are fully compliant with OECD TG 405.

Justification for classification or non-classification

Based on the provided data, the test item is not classified skin/eye irritation/corrosion according to Regulation (EC) No 1272/2008.