Registration Dossier

Administrative data

Description of key information

The registration substance is not irritating to skin and not irritating to eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2015-12-15 to 2016-02-23
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
Housing and Feeding Conditions:
- Semi barrier in an air-conditioned room
- Temperature: 18 +/- 3 °C (recommendations of TVT, GV-SOLAS)
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: at least 10 x / hour
- Free access to irradiated hay briquettes and to Altromin 2123 maintenance diet for rabbits (lot no. 0614), rich in crude fibre
- Free access to tap water (drinking water, municipal residue control, microbiological controls at regular intervals)
- Certificates of food, water and bedding are filed for two years at BSL Munich and afterwards archived at Eurofins Munich
- Housed in ABS-plastic or Noryl rabbit cages, floor 4200 cm2
- Adequate acclimatisation period (at least 5 days) under laboratory conditions

Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
water
Remarks:
aqua ad injectabilia (AlleMan Pharma, lot no. 503424, expiry date: 28/02/2018)
Controls:
no
Amount / concentration applied:
Dose Level
A dose of 0.5 g of the test item was applied to each test site.
Duration of treatment / exposure:
Exposure Period
The test item was held in contact with the skin throughout a 4-hour period.
At the end of the exposure period, the residual test item was removed with physiological saline NaCl 0.9% (B. Braun Melsungen, lot no. 1406788,
expiry date: 31/05/2017).
Observation period:
Observation Period
The animals were observed for 72 hours after the patch removal.
Number of animals:
3
Details on study design:
Clinical Observation:
The animals were examined for signs of erythema and oedema 1 hour after the patch removal. For the determination of classification-relevant values, the animals were examined for signs of erythema and oedema 24, 48 and 72 hours after the patch removal. Dermal irritation was scored and
recorded according to the grades in the table below . Any other signs such as hyperplasia, scaling, discolouration, fissures and scabs or any
systemic effects were also recorded.
For the initial test in one animal, the test site was also examined immediately after the patch had been removed.

Evaluation of Results:
Individual reactions for each animal were recorded according to the grades in the table below at each time of observation.
Nature, severity and duration of clinical observations were described.
The body weight changes were summarised in a tabular form.
With few exceptions, data were captured using the validated departmental computerised system E WorkBook (version 9.4.0, ID Business Solutions Ltd.).

Irritation parameter:
erythema score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Irritation parameter:
edema score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Irritant / corrosive response data:
Neither irritant nor corrosive effects were observed on the intact skin of three male rabbits (strain NZW) after a contact time of 4 hours.
Other effects:
Neither mortalities nor significant clinical signs of toxicity were observed.
There were no significant body weight changes during the observation period.

Individual Data - Evaluation of Dermal Irritation

Animal No. Timepoint Irritation
Oedema Erythema Comment
Test Item Control Test Item Control Test Item Control
1 0h 0 0 0 0 - -
1h 0 0 0 0 - -
24h 0 0 0 0 - -
48h 0 0 0 0 - -
72h 0 0 0 0 - -
2 0h         - -
1h 0 0 0 0 - -
24h 0 0 0 0 - -
48h 0 0 0 0 - -
72h 0 0 0 0 - -
3 0h         - -
1h 0 0 0 0 - -
24h 0 0 0 0 - -
48h 0 0 0 0 - -
72h 0 0 0 0 - -

0-4 = grade

Clinical Signs

Animal No.  Timepoint Systemic Findings Local Findings Comment
Test Item Test Item Control Test Item Control
1 0h nsf nsf nsf - -
1h nsf nsf nsf - -
24h nsf nsf nsf - -
48h nsf nsf nsf - -
72h nsf nsf nsf - -
2 1h nsf nsf nsf - -
24h nsf nsf nsf - -
48h nsf nsf nsf - -
72h nsf nsf nsf - -
3 1h nsf nsf nsf - -
24h nsf nsf nsf - -
48h nsf nsf nsf - -
72h nsf nsf nsf - -

* = apart from erythema/oedema; nsf = no specific findings

Absolute Body Weights (kg)

Animal No. Bodyweight (kg)
Start of Study 72 Hours Post Application
1 3.6 3.6
2 4 4
3 4.5 4.6
Interpretation of results:
GHS criteria not met
Conclusions:
The skin irritation property of the registration substance was investigated according to the Guideline OECD 404. The registration substance did not induce any effect in this study. No classification is justified.
Executive summary:

The skin irritation property or the registration substance was investigated according to the Gudieline OECD 404. Rabbits were dermally treated with 0.5g of the registration substance semi-occusively for 4 hours. No effect was found.

No classification is justified.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2015-12-15 to 2016-05-13
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Test System
Species/strain: healthy New Zealand White Rabbits, Crl: KBL (NZW)
Source: Charles River Deutschland, 97633 Sulzfeld, Germany
Sex: male
Body weight at the beginning of the study: > 2 kg
Age at the beginning of the study:
animal no. 1: 17 - 18 weeks old;
animal no. 2: 18 - 19 weeks old;
animal no. 3: 19 - 20 weeks old
Number of animals: 3
The animals were derived from a controlled full-barrier maintained breeding system (SPF). According to Art. 9.2, No. 7 of the
German Act on Animal Welfare the animals were bred for experimental purposes.


Housing and Feeding Conditions
- Semi barrier in an air-conditioned room
- Temperature: 18 +/- 3 °C (recommendations of TVT, GV-SOLAS)
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: at least 10 x / hour
- Free access autoclaved hay and to Altromin 2123 maintenance diet for rabbits (lot no. 0614), rich in crude fibre
- Free access to tap water (drinking water, municipal residue control, microbiological controls at regular intervals)
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Housed in ABS-plastic or Noryl rabbit cages, floor 4200 cm2
- Adequate acclimatisation period (at least 5 days) under laboratory conditions

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
A dose of 0.1 g of the test item was applied to the test site.
Duration of treatment / exposure:
The test item was applied at a single dose in the conjunctival sac of one eye of each test animal after pulling the lower lid away from the eyeball.
The lids were then gently held together for about 1 second in order to prevent loss of the material. The untreated contralateral eye served as control.
Observation period (in vivo):
TThe animals were observed for 72 hours after dosing.
To determine the reversibility of the observed effects, the observation period was extended to 14 days after dosing in one out of three animals.
Number of animals or in vitro replicates:
The in vivo test was performed initially using one animal.
The results of the initial test did not indicate the test item to be corrosive or a severe irritant to the eye using the procedure described.
In order to confirm the response, two additional animals were treated in the same manner.
Details on study design:
Preparation of the Animals
Approximately 24 hours before the test and immediately prior to the application both eyes of each animal were examined.
A health inspection was performed to ensure the good state of health of the animals.
Approximately 20 hours (animal no. 1) or 18 hours (animals no. 2 and 3) before the application the eyes were also examined with the aid of
a fluorescein solution (Fluoreszein SE Thilo, Alcon Pharma, lot no. H404, expiry date: 31/10/2016). The eyes were rinsed with physiological
saline 0.9% NaCl (B. Braun Melsungen, lot no. 1406788, expiry date: 31/05/2017) after the examination.
None of the animals showed eye irritation, ocular defects, or pre-existing corneal injury.


Application
One hour before the application of the test item, 0.01 mg/kg of buprenorphine was administered subcutaneously in order to achieve a
therapeutic level of systemic analgesia. Approximately 5 minutes prior to the application of the test item, 1-2 drops of an ocular anaesthetic
(proparacaine hydrochloride ophtalmic 0.5% solution) were administered in both the treated and the control eye of each animal.
The test item was applied at a single dose in the conjunctival sac of one eye of each test animal after pulling the lower lid away from the eyeball.
The lids were then gently held together for about 1 second in order to prevent loss of the material. The untreated contralateral eye served as control.
The treated eye was either not rinsed (animals no. 1 and no. 2) or rinsed with physiological saline 0.9% NaCl (see 10.5) 2.5 hour after the application.
To prevent animal no. 3 showing pain and distress after the application of the test item and was treated with the following doses of buprenorphine
and meloxicam.

Clinical Observation
The eyes were examined for signs of irritation throughout the observation period. The eye irritation was scored and recorded according
to the grades. For the calculation only the 24, 48 and 72-hour readings were used.
At all time points starting with and after 24 h, the treated eyes were examined with the aid of a fluorescein solution.
The eyes were rinsed with physiological saline 0.9% NaCl after the examination.

Evaluation of Results
IIndividual reactions for each animal were recorded according to the scoring system described at each time of observation.
For the calculation only the 24, 48 and 72-hour readings were used.
Nature, severity and duration of clinical observations were described.
The body weight changes were summarised in a tabular form.
With few exceptions, data were captured, using the validated departmental computerized system
E-Workbook (version 9.4.0, ID Business Solutions Ltd).

Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
three animals
Time point:
24/48/72 h
Score:
0.4
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
iris score
Basis:
mean
Remarks:
three animals
Time point:
24/48/72 h
Score:
0
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
three animals
Time point:
24/48/72 h
Score:
0
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
three animals
Time point:
24/48/72 h
Score:
0.1
Max. score:
1
Reversibility:
fully reversible
Irritant / corrosive response data:
After the application into the eyes of three male NZW rabbits the test item produced irritant but not corrosive ocular effects in all animals.
Neither mortalities nor significant clinical signs of systemic toxicity were observed
Upon fluorescein examinations at all time points starting with and after 24 h, no corneal lesions were found in any animal.
Conjunctival redness, chemosis and hypersecretion were observed in all animals. Iris lesion was observed in one out of three animals.
Other effects:
The body weight development of all animals was within the expected range. A slight body weight loss was observed in one of three animals
(animal no. 3). It could not clearly be concluded that this finding is test item related.
Interpretation of results:
GHS criteria not met
Conclusions:
The eye irritation property of the registration substance was investigated according to the Guideline OECD 405. The registration substance induced mild transient effects in one animal out of three treated animals. No classification is justified.
Executive summary:

The eye irritation property of the registration substance was investigated according to the Guideline OECD 405. 0.1g of the registration substance was applied to eyes of rabbits. Three rabbits were treated and there was no effect in two rabbits. One rabbit exhibited transient redness and chemosis. No classification is warranted.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Additional information

Justification for classification or non-classification

The skin irritation property or the registration substance was investigated according to the Gudieline OECD 404. Rabbits were dermally treated with 0.5g of the registration substance semi-occusively for 4 hours. No effect was found.

No classification is justified.

The eye irritation property of the registration substance was investigated according to the Guideline OECD 405. 0.1g of the registration substance was applied to eyes of rabbits. Three rabbits were treated and there was no effect in two rabbits. One rabbit exhibited transient redness and chemosis. No classification is justified.