Registration Dossier

Administrative data

Description of key information

Skin irritation (OECD 404): not irritating

Eye irritation (OECD 437): not irritating

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:
12 Nov - 24 Nov 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
adopted 17 Jul 1992
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Version / remarks:
1992
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
THE DEPARTMENT OF HEALTH OF THE GOVERNMENT OF THE UNITED KINGDOM
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 12-16 weeks
- Weight at study initiation: 2.73-3.02 kg
- Housing: animals were housed individually in suspended metal cages
- Diet: STANRAB SQC Rabbit Diet (Special Diets Services LTD, Witham, UK), ad libitum
- Water: mains drinking water, ad libitum
- Acclimation period: minimum 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23
- Humidity (%): 30-70
- Air changes (per hr): approx. 15
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 0.5 mL
Duration of treatment / exposure:
3 min and 1 and 4 h (one animal)
4 h (two animals)
Observation period:
7 days
Reading time points: 1, 24, 48 and 72 h and 7 days
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: area on the back
- Type of wrap: The test substance was applied to the skin and covered with gauze patch (2.5 cm x 2.5 cm). The patch was secured in position with a strip of surgical adhesive tape (BLENDERM, 2.5 cm x 4.0 cm). Additionally, the trunk was wrapped in an elasticated corset (TUBIGRIP).

REMOVAL OF TEST SUBSTANCE
- Washing: The skin was cleaned of residual test substance by gentle swabbing with cotton wool soaked in 74% Industrial Methylated Spirits.
- Time after start of exposure: 3 min and 1 and 4 h (one animal); 4 h (two animals)

SCORING SYSTEM: Draize scoring system
Irritation parameter:
erythema score
Basis:
animal: #1 and #3
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: loss of skin elasticity after 72 h, slight desquamation after 7 days
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: mean after 24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 h
Irritant / corrosive response data:
Very slight erythema was noted at all treated skin sites at the 1 h observation and at two treated skin sites at the 24, 48 and 72 h observations. Very slight oedema was noted at all treated skin sites at the 1 h observation, at two treated skin sites at the 24 h observation and at one treated skin site at the 48 and 72 h observations. Loss of skin elasticity was noted at two treated skin sites at the 72 h observation. Slight desquamation was noted at two treated skin sites after 7 days.
One treated skin site appeared normal at the 24-hour observation.

3 min exposure period: No evidence of skin irritation was noted during the study.

1 h exposure period: Very slight erythema was noted at the treated skin site at the 24, 48 and 72 h observations. Loss of skin elasticity was noted at the treated skin site at the 72 h observation. Slight desquamation was noted at the treated skin site after 7 days..

Table 1. Results of the skin irritation study following 4 h exposure.

Observation time

Rabbit no.

1

2

3

Erythema

Edema

Erythema

Edema

Erythema

Edema

1 h

1

1

1

1

1

1

24 h

1

1

0

0

1

1

48 h

1

1

0

0

1

0

72 h

1LE

1

0

0

1LE

0

7 days

0D

 

0

0

0D

0

Mean value
24 + 48 + 72 h

1.00

1.00

0

0

1.00

0.33

D: Desquamation (slight)

LE: Loss of skin elasticity

Table 2. Results of the skin irritation study following 3 min and 1 h exposure.

Observation time

Exposure period

3 min

1 h

Erythema

Edema

Erythema

Edema

1 h

0

0

0

0

24 h

0

0

1

0

48 h

0

0

1

0

72 h

0

0

1LE

0

7 days

0

0

0D

0

D: Desquamation (slight)

LE: Loss of skin elasticity

Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
Under the conditions of this skin irritation study in rabbits the test substance was not irritating to the skin.
Executive summary:

A single 4-hour, semi-occluded application of the test material to the intact skin of three rabbits produced very slight erythema and very slight edema. Incidents of loss of skin elasticity were also noted. Slight desquamation was noted at two treated skin sites at the 7-day observation. One treated skin site appeared normal at the 24-hour observation. No corrosive effects were noted. Under the conditions of this skin irritation study in rabbits the test substance was not irritating to the skin.

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

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
19 June 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Version / remarks:
July 2013
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Hess. Ministerium für Umwelt, Klimaschutz, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany
Species:
other: cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Source: Schlachthof Bensheim, Bensheim, Germany
- Characteristics of donor animals: at least 9 month old
- Storage, temperature and transport conditions of ocular tissue: The isolated eyes were transported in Hank's Buffered Salt Solution (HBSS) at ambient temperature.
- Time interval prior to initiating testing: The corneae were isolated on the same day after delivery of the eyes and directly used in the BCOP test.
- Indication of any existing defects or lesions in ocular tissue samples: All eyes were carefully examined macroscopically for defects. Those presenting defects such as vascularization, pigmentation, opacity and scratches were discarded.


Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount applied: 0.75 mL

POSITIVE SUBSTANCE
- Amount applied in the test: 0.75 mL
- Purity: 99%

NEGATIVE CONTROL
- Amount applied in the test: 0.75 mL
Duration of treatment / exposure:
10 min at 32 ± 1 °C
Duration of post- treatment incubation (in vitro):
2 h
Number of animals or in vitro replicates:
triplicates for each treatment and control group
Details on study design:
SELECTION AND PREPARATION OF CORNEAS:
The cornea was carefully removed from the eye using scalpel and rounded scissors. A rim of about 2 mm of tissue (sclera) was left for stability and handling of the isolated cornea. Each cornea was mounted in a specially designed cornea holder.

QUALITY CHECK OF THE ISOLATED CORNEAS:
At the end of the equilibration period, the basal opacity was determined (t0). Each cornea with a value of the basal opacity >7 was discarded.

TREATMENT METHOD:
The cornea holder consists of anterior and posterior compartments, which interface with the epithelial and endothelial sides of the cornea, respectively. The endothelial side of the cornea was positioned against the sealing ring (O-ring) of the posterior part of the holder. The cornea was gently flattened over the O-ring but stretching was avoided. The anterior part of the holder was positioned on the top of the cornea and fixed in place with screws. Both compartments of the holder were filled with incubation medium. The posterior compartment was filled first to return the cornea to its natural convex position. The anterior compartment received the test substance or the controls on the surface of the corneae. The corneae were incubated in a horizontal position at 32 ± 1°C in the water-bath for 10 minutes.

REMOVAL OF TEST SUBSTANCE:
The test substance was rinsed off from the application side with saline.

- POST-EXPOSURE INCUBATION: 2 h in a vertical position

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Corneal opacity was determined by the amount of light transmission through the cornea via an opacitometer (OP_KiT opacitometer, Electro Design, France).
- Corneal permeability: The passage of sodium fluorescein dye was measured with the aid of a microplate reader (Versamax Molecular Devices) at 490 nm.

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA:
Test substance with an IVIS > 55 was regarded as severe irritant/corrosive and labelled Category 1.
Test substance with an IVIS ≤ 3 was regarded as non-irritant and labelled in no category.
Test substance with an IVIS > 3; ≤ 55 no prediction can be made.
Irritation parameter:
in vitro irritation score
Run / experiment:
mean value of 3 corneae
Value:
0.81
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
With the negative control (0.9% (w/v) NaCl solution in deionised water) neither an increase of opacity nor permeability of the corneae could be observed.
The positive control (2-ethoxyethanol) showed clearly increased opacity and distinctive permeability of the corneae fullfilling the criteria as severe irritating/corrosive.
Relative to the negative control, the test substance did not cause an increase of the corneal opacity or permeability.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The negative control resulted in an IVIS which was within two standard deviations of the current historical mean.
- Acceptance criteria met for positive control: The positive control resulted in opacity and permeability values that were less than the established upper limits for background opacity and permeability values for bovine corneae treated with the respective negative control.

Table 1: Results after 10 min incubation time.

Test group

Opacity value =
Difference (t130-t0) of Opacity

Permeability at 490 nm (OD490)

IVIS

Mean IVIS

 

Mean

 

Mean

 

 

Negative

control

0

0.33

0.051

0.053

0.77

1.13

1

0.054

1.81

0

0.054

0.81

Positive

control

61.67*

0.994*

76.58

70.00

53.67*

0.768*

65.19

57.67*

0.705*

68.24

Test substance

-0.33*

0.013*

-0.14

0.81

0.67*

0.017*

0.92

1.67*

-0.001*

1.65

* corrected values

Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
Under the conditions of the BCOP assay the test substance was not irritating to the eye. Application of the test substance to bovine corneae resulted in a calculated mean IVIS of 0.81.
Executive summary:

Relative to the negative control, the test item JACINTHAFLOR® did not cause any increase of the corneal opacity or permeability. The calculated mean in vitro irritancy score was 0.81. According to OECD 437 the test item is not categorized (GHS).

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin

The skin irritation potential of the test substance was determined by an in vivo skin irritation test in rabbits according to OECD Guideline 404 and in compliance with GLP (2000). In this study the test substance was not irritating to skin.

 

Eye

The eye irritation potential of the test substance was determined in a bovine corneal opacity and permeability (BCOP) test according to OECD Guideline 437 and in compliance with GLP (2015). In this study the test substance was not irritating to the eye.

Justification for classification or non-classification

The available data on skin and eye irritation do not meet the criteria for classification according to Regulation (EC) 1272/2008