Reaction products of Glycerin formal and 2-Propenoic acid, 2-methyl-, methyl ester

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21.05.2013 to 13.08.2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

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

Reference substance name:

Reaction products of Glycerin formal and 2-Propenoic acid, 2-methyl-, methyl ester

EC Number:

945-527-5

Cas Number:

1620329-57-8

Molecular formula:

C8H12O4

IUPAC Name:

Reaction products of Glycerin formal and 2-Propenoic acid, 2-methyl-, methyl ester

Test material form:

liquid

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Remarks:

New Zealand White Rabbits, Crl: KBL (NZW)

Details on test animals or tissues and environmental conditions:

Species/strain: healthy New Zealand White Rabbits, Cr!: KBL (NZW)
Source: Charles River Deutschland, 97633 Sulzfeld, Germany
Sex: female
Body weight at the beginning of the study: > 2 kg
Age at the beginning of the study: approximately 33-42 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 ofTVT [11], GV-SOLAS 0)
Relative humidity: 55 ± 10%
Artificial light, sequence being 12 hours light, 12 hours dark
Air change: at least 10 x I hour
Free access to autoclaved hay and to Altromin 2123 maintenance diet for rabbits (lot no. 105 6), rich in crude fibre
Free access to tap water ( drinking water, municipal residue control, microbiological controls at regular intervals)
Housed in ABS - plastic rabbit cages, floor 4200 cm2
Adequate acclimatisation period (at least 5 days) under laboratory conditions

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 before the application the eyes were also examined with the aid of a fluorescein solution. The eyes were rinsed with physiological saline 0.9% NaCl after the examination.
None of the animals showed eye irritation, ocular defects, or pre-existing corneal injury.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 ml

Duration of treatment / exposure:

24 h

Observation period (in vivo):

Animal no. 1 was observed for 72 hours after dosing. To determine the reversibility of the observed effects, the observation period was extended up to 15 days after dosing.
Animals no. 2 and 3 were observed for 72 hours after dosing.

Details on study design:

REMOVAL OF TEST SUBSTANCE
animal No. 1 was rinsed with physiological saline 0.9% NaCl 24 hours after the application
animal No. 2 and 3 was not rinsed 24 hours after the application

SCORING SYSTEM:
GRADING OF OCULAR LESIONS
Cornea Grade
Opacity: degree of density (readings should be taken from most dense area)*
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
Maximum possible: 4
* The area of corneal opacity should be noted
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
Hemorrhage, gross destruction, or no reaction to light ............................................................... 2
Maximum possible: 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
Maximum possible: 3
Chemosis
Swelling (refers to lids and/or nictating membranes)
Normal ......................................................................................................................................... 0
Some swelling above normal ....................................................................................................... 1
Obvious swelling, with partial eversion of lids ........................................................................... 2
Swelling, with lids about half closed ........................................................................................... 3
Swelling, with lids more than half closed.................................................................................... 4
Maximum possible:



TOOL USED TO ASSESS SCORE: fluorescein

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

fully reversible within: 15 days

Results 2

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.34

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Results 4

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3.67

Max. score:

4

Reversibility:

fully reversible within: 8 days

Results 5

Irritation parameter:

chemosis score

Basis:

animal: #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 6

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 5 days

Results 7

Irritation parameter:

iris score

Basis:

animal: #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 8

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.34

Max. score:

4

Reversibility:

fully reversible within: 11 days

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal: #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Any other information on results incl. tables

Eye irritation scores

 

Animal No. 1	Single data
		Time post application				Average score (24, 48 and 72 hours)
		1 hour	24 hours	48 hours	72 hours
		T/C	T/C	T/C	T/C
	Conjunctival Redness	2/0	3/0	3/0	3/0	3.00
	Conjunctival Chemosis	3/0	3/0	4/0	4/0	3.67
	Iris	0/0	1/0	1/0	1*/0	1.00
	Cornea	2/0	2/0	2/0	3/0	2.34

 

Animal No. 1	Single data- Prologed Observation period
		Time post application
		4 days	5 days	6 days	7 days	8 days
		T/C	T/C	T/C	T/C	T/C
	Conjunctival Redness	3/0	2/0	2/0	2/0	2/0
	Conjunctival Chemosis	3/0	1/0	1/0	1/0	0/0
	Iris	1*/0	0/0	0/0	0/0	0/0
	Cornea	3/0	1/0	1/0	1/0	1/0

 

 

Animal No. 1	Single data- Prologed Observation period
		Time post application
		9 days	10 days	11 days	12 days	13 days
		T/C	T/C	T/C	T/C	T/C
	Conjunctival Redness	2/0	1/0	1/0	1/0	1/0
	Conjunctival Chemosis	0/0	0/0	0/0	0/0	0/0
	Iris	0/0	0/0	0/0	0/0	0/0
	Cornea	1/0	1/0	0/0	0/0	0/0

 

 

Animal No. 1	Single data- Prologed Observation period
		Time post application
		14 days	15 days
		T/C	T/C
	Conjunctival Redness	1/0	0/0
	Conjunctival Chemosis	0/0	0/0
	Iris	0/0	0/0
	Cornea	0/0	0/0

 

Animal No. 2	Single data
		Time post application				Average score (24, 48 and 72 hours)
		1 hour	24 hours	48 hours	72 hours
		T/C	T/C	T/C	T/C
	Conjunctival Redness	1/0	1/0	1/0	0/0	0.67
	Conjunctival Chemosis	1/0	0/0	0/0	0/0	0.00
	Iris	0/0	0/0	0/0	0/0	0.00
	Cornea	0/0	0/0	0/0	0/0	0.00

 

Animal No. 3	Single data
		Time post application				Average score (24, 48 and 72 hours)
		1 hour	24 hours	48 hours	72 hours
		T/C	T/C	T/C	T/C
	Conjunctival Redness	1/0	1/0	0/0	0/0	0.34
	Conjunctival Chemosis	1/0	0/0	0/0	0/0	0.00
	Iris	0/0	0/0	0/0	0/0	0.00
	Cornea	0/0	0/0	0/0	0/0	0.00

 

T-test item, C=control; *=detailed observation of iris was not possible due to sanois discharge

 

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the present study, a single ocular application of the test item Glycerinformal methacrylate to rabbits at a dose of 0.1 mL produced irritant effects, which were fully reversible within 48 hours (animal no.3), 72 hours (animal no. 2) or 15
days (animal no. !). Neither mortalities nor significant clinical signs of toxicity were observed.
In conformity with the EC criteria for classification and labelling requirements for dangerous substances and preparations according to Annex VI of Commission Directive 2001/59/EC [7], the test item Glycerinformal methacrylate has no obligatory labelling
requirement for eye irritation.
According to Annex I of Regulation (EC) 1272/2008 , the test item Glycerinformal methacrylate has no obligatory labelling requirement for eye irritation.
According to GHS (Globally Harmonized Classification System)] the test item Glycerinformal methacrylate has no obligatory labelling requirement for eye irritation.

Executive summary:

In an acute in vivo eye irritation/corrosion test according OECD 405 with Glycerolformal methacrylate, a single ocular application of the test item to rabbits at a dose of 0.1 mL produced irritant effects,

which were fully reversible within 48 hours (animal no.3), 72 hours (animal no. 2) or 15 days (animal no.!).Neither mortalities nor significant clinical signs of toxicity were observed.

Inconformity with the EC criteria for classification and labelling requirements for dangerous substances and preparations according to Annex VI of Commission Directive

2001/59/EC [7], the test item Glycerinformal methacrylate has no obligatory labelling requirement for eye irritation.

According to Annex I of Regulation (EC) 1272/2008 the test item Glycerinformal methacrylate has no obligatory labelling requirement for eye irritation.

According to GHS (Globally Harmonized Classification System) the test item Glycerinformal methacrylate has no obligatory labelling requirement for eye irritation.