Ethyldimethyl[2-[(2-methyl-1-oxoallyl)oxy]ethyl]ammonium ethyl sulphate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Start of the experimental phase: June 10, 2014 ; Termination of the in-life phase: June 25, 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study on a well characterized test material.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

other: liquid

Details on test material:

- Name of test material (as cited in study report): FlocrylTM MADAM/MECL 75%
- Molecular formula (if other than submission substance):
- Molecular weight (if other than submission substance):
- Smiles notation (if other than submission substance):
- InChl (if other than submission substance):
- Structural formula attached as image file (if other than submission substance): see Fig.
- Substance type:
- Physical state: Clear to slightly yellow liquid
- Analytical purity: Monomer concentration 74.9%
- Impurities (identity and concentrations):
- Composition of test material, percentage of components:
- Isomers composition:
- Purity test date: 04.12.2013
- Lot/batch No.: MADC 1-43-13-03-E
- Expiration date of the lot/batch:
- Radiochemical purity (if radiolabelling):
- Specific activity (if radiolabelling):
- Locations of the label (if radiolabelling):
- Expiration date of radiochemical substance (if radiolabelling):
- Stability under test conditions:
- Storage condition of test material: At room temperature, not stored together with oxidizing and self-igniting products.
- Other:

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Isoquimen, SL, Carretera de Sant Miquel del Fai, Km 3.5, 08182 Sant Feliu de Codines, Spain.
- Age at study initiation: Approx. 7.5 - 8.5 months
- Weight at study initiation: 3.8 - 4.8 kg
- Housing: For 8 hours following test item application, the animals were kept singly in restrainers which allowed free movement of the head but prevented a complete body turn, wiping of the eyes with the paws and excluded irritation of the eye by excrements and urine.
During the acclimatisation period and after the 8-hour period in restrainers, the animals were kept singly in cages with dimensions of 380 mm x 425 mm x 600 mm (manufacturer: Dipl.Ing. W. EHRET GmbH, 16352 Schönwalde, Germany).
- Diet (e.g. ad libitum): The food was available ad libitum before and after the exposure period.
- Water (e.g. ad libitum): Tap water was offered daily ad libitum.
- Acclimation period:


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20°C ± 3°C (maxi¬mum range).
- Humidity (%): The relative humidity amounted to 30% - 70% (maximum range).
- Air changes (per hr): 15 to 20 times
- Photoperiod (hrs dark / hrs light): Rooms were lit (150 lux at approx 1.5 m room height) and darkened on a 12-hour light/12-hour dark cycle.


IN-LIFE DATES: From: June 01, 2014 To: Termination of the in-life phase June 25, 2014

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The left eyes served as control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL of the test item were administered into one eye. The test item was placed into the conjunctival sac of the right eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second in order to prevent loss of the material. The left eye, which remained untreated, served as a control.

- Concentration (if solution):


VEHICLE
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:

Duration of treatment / exposure:

The test was performed initially using one animal. As no corrosive or severe irritant effects were observed in this animal, 2 further animals were employed 24 hours after start of the initial test.
Sixty minutes prior to test item administration, 0.01 mg Buprenovet®/kg b.w. were administered by subcutaneous injection to all animals to provide a therapeutic level of systemic analgesia to avoid or minimize pain and distress.
Five minutes prior to the test item administration, two drops of Ophtocain®, a topical anaesthetic, was applied to each eye of all animals, to the right eye, in which the test item was to be applied, and to the left eye, which served as control.
Eight hours after administration, Buprenovet 0.01 mg/kg, s.c. and Metacam 0.5 mg/kg, s.c. were administered to provide a continued therapeutic level of systemic analgesia.
No washout after 24 hours was performed.

Observation period (in vivo):

The eyes were examined ophthalmoscopically with a slit lamp prior to the administration and 1, 24, 48, 72 hours and 4 to 6 days after the administration. The eye reactions were observed and registered.

Number of animals or in vitro replicates:

3 female rabbits

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: Eight hours after administration, Buprenovet 0.01 mg/kg, s.c. and Metacam 0.5 mg/kg, s.c. were administered to provide a continued therapeutic level of systemic analgesia. No washout after 24 hours was performed.

- Time after start of exposure: Eight hours after administration


SCORING SYSTEM:
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 lustre), details of iris clearly visible 1
easily discernible translucent area, 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 circumcorneal hyperaemia, or injection, iris reactive to light (a sluggish reaction is considered to be an effect) 1
haemorrhage, gross destruction, or no reaction to light 2


CONJUNCTIVAE
Redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris)
normal 0
some blood vessels hyperaemic (injected) 1
diffuse, crimson colour; individual vessels not easily discernible 2
diffuse beefy red 3

CHEMOSIS
Swelling: refers to lids and/or nictitating membranes
normal 0
some swelling above normal 1
obvious swelling with partial eversion of lid 2
swelling with lids about half-closed 3
swelling with lids more than half-closed 4


TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein
One day before and 24 hours after administration, fluorescein was applied to the eyes before being examined to aid evaluation of the cornea for possible lesions. The fluorescein test was repeated on each day of observation. (Fluorescein SE Thilo drops (ALCON PHARMA GmbH, 79108 Freiburg, Germany))

Fluorescein-Test:

DEGREE OF STAINING
no staining 0
scattered or diffuse colouration, details of iris clearly visible 1
easily discernible translucent area, details of iris slightly obscured 2
opalescent areas, details of iris not discernible, extent of pupil difficultly determinable 3
opaque cornea, iris not discernible through the opacity
INVOLVED AREA OF CORNEA
none 0
up to 1/4 of the surface 1
1/4 to 1/2 of the surface 2
1/2 to 3/4 of the surface 3
3/4 to whole surface. 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: 24h, 48h, 72h

Score:

0

Max. score:

0

Reversibility:

other: No effect

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: 24h, 48h, 72h

Score:

0

Max. score:

0

Reversibility:

other: No effect

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

other: 24h, 48h, 72h

Score:

0

Max. score:

0

Reversibility:

other: No effect

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 24h, 48h, 72h

Score:

0

Max. score:

0

Reversibility:

other: No effect

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: 24h, 48h, 72h

Score:

0

Max. score:

0

Reversibility:

other: No effect

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: 24h, 48h, 72h

Score:

0

Max. score:

0

Reversibility:

other: No effect

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: 24h, 48h, 72h

Score:

1

Max. score:

1

Reversibility:

not fully reversible within: 4 days

Remarks on result:

other: Mean score

Results 8

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: 24h, 48h, 72h

Score:

1.6

Max. score:

2

Reversibility:

not fully reversible within: 6 days

Remarks on result:

other: Mean score

Results 9

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

other: 24h, 48h, 72h

Score:

1

Max. score:

1

Reversibility:

not fully reversible within: 3 days

Remarks on result:

other: Mean score

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 24h, 48h, 72h

Score:

0

Max. score:

0

Reversibility:

other: No effect

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 24h, 48h, 72h

Score:

1

Max. score:

1

Reversibility:

not fully reversible within: 4 days

Remarks on result:

other: Mean score

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: 24h, 48h, 72h

Score:

0

Max. score:

0

Reversibility:

other: No effect

Irritant / corrosive response data:

The irises were not affected by instillation of the test item.
The fluorescein tests performed 24 and 48 hours after instillation revealed corneal staining in animal no. 2 (up to 1/4 of the surface).

Other effects:

There were no systemic intolerance reactions.

Any other information on results incl. tables

Acute eye irritation/corrosion test of Flocryl™ MADAM/MECL 75% in rabbits
Examination of the treated eye
Time after administration	CORNEA	IRIS	CONJUNCTIVAE		Fluorescein Test
	Opacity		Redness*	Chemosis**	Grade	Area
	Animal number: 1/2/3
Right eye: 0.1 mL Test item/animal
Before dosing	0/0/0	0/0/0	0/0/0	0/0/0	-/-/-	-/-/-
60 minutes	0/0/0	0/0/0	1/1/1	0/0/0	-/-/-	-/-/-
24 hours	0/0/0	0/0/0	1/2/1	0/1/0	0/1/0	0/1/0
48 hours	0/0/0	0/0/0	1/2/1	0/1/0	0/1/0	0/1/0
72 hours	0/0/0	0/0/0	1/1/0	0/1/0	0/0/0	0/0/0
4 days	0/0/-	0/0/-	0/1/-	0/0/-	0/0/-	0/0/-
5 days	-/0/-	-/0/-	-/1/-	-/0/-	-/0/-	-/0/-
6 days	-/0/-	-/0/-	-/0/-	-/0/-	-/0/-	-/0/-
* Refers to palpebral and bulbar conjunctivae; excluding cornea and iris
** Swelling: refers to lids and/or nictitating membrane
- No examination

 

 

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

FlocrylTM MADAM/MECL 75% was not irritating to eyes

Executive summary:

According to the EC-Commission directive 67/548/EEC and its subsequent amendments on the approximation of the laws, regulations and administrative provision relating to the classification, packaging and labelling of dangerous substances and the results obtained under the present test conditions MADAMQUAT MC was not irritating to eyes, hence, no labelling is required.

Also, according to the Globally Harmonized System (GHS) and EC-Regulation 1272/2008 and subsequent regulations, the test item is non-irritating to eyes; classification and labelling of the substance is not necessary.