1,2-bis(2-methoxyethoxy)ethane

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-10-15 to 2013-01-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

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

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)

Test material

Test material information

Constituent 1

Reference substance name:

1,2-bis(2-methoxyethoxy)ethane

EC Number:

203-977-3

EC Name:

1,2-bis(2-methoxyethoxy)ethane

Cas Number:

112-49-2

Molecular formula:

C8H18O4

IUPAC Name:

2,5,8,11-tetraoxadodecane

Details on test material:

Name: Triethyleneglycoldimethylether
CAS No.: 112-49-2
Chemical Name: 1,2-bis(2-methoxyethoxy)ethane
Batch No.: DEG4158261
Physical State: volatile, colourless, liquid
Density: 0.99 g/cm2 at 20°C
Active Components: 100%
Purity: > 99%
Date of Analysis: 20.09.2012
Solubility: Miscible in water at 20°C in all proportions
Storage Conditions: room temperature
Expiry Date: 03.06.2013
Safety Precautions: The routine hygienic procedures were sufficient to assure personnel health and safety.

Test animals / tissue source

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: female
Body weight at the
beginning of the study: > 2.0kg
Age at the
beginning of the study: approximately 14 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 to autoclaved hay and to Altromin 2123 maintenance diet for rabbits (lot no. 1013), 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 rabbit cages, floor 4200 cm2
-Adequate acclimatisation period (at least 5 days) under laboratory conditions

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

A dose of 0.1 mL of the test item was applied to the test site.

Duration of treatment / exposure:

The treated eye was not rinsed 24 hours after the application.

Observation period (in vivo):

The animals were observed for 72 hours after dosing.

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 24 hours before the application the eyes were also examined with the aid of a fluorescein solution (Fluoreszein SE Thilo,
Laboratories Alcon, lot no. H 201, expiry date: 12/2013). The eyes were rinsed with physiological saline 0.9% NaCl (AlleMan Pharma, lot no. 111214, expiry date: 11/2014) 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 (Reckitt Benckiser, lot no.: 5317, expiry date: 01/2017) was administered subcutaneously in order to achieve a therapeutic level of systemic analgesia. Approximately 5 minutes prior to the application of the test item, 2-3 drops of an ocular anaesthetic (proparacaine hydrochloride ophtalmic 0.5% solution, Ursapharm, lot no.: 273851, expiry date: 05/2013) 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 not rinsed 24 hours after the application.

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 in the table below.
For the calculation only the 24, 48 and 72-hour readings were used.
At the end of the observation period 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
Individual reactions of each animal were recorded at each time of observation.
Nature, severity and duration of all lesions observed were described.
For the calculation only the 24, 48 and 72-hour readings were used.
On the basis of the test results, the test substance was classified in any of the following classes in conformity with the criteria given in Annex VI to
Commission Directive 2001/59/EC [6]:
R36 Irritating to eyes
- Substances and preparations which, when applied to the eye of the animal, cause significant ocular lesions which occur within
72 hours after exposure and which persists for at least 24 hours.
Ocular lesions are significant if the mean scores of the eye irritation test cited in Annex B.5 to Commission Regulation (EC) No. 440/2008, L 142
have any of the following values:
- corneal opacity ≥ 2 but < 3,
- iris lesion ≥ 1 but ≤ 1.5,
- redness of the conjunctiva ≥ 2.5,
- oedema of the conjunctiva (chemosis) ≥ 2,
or, in case where Annex B.5 test have been completed using three animals if the lesions, on two or more animals, are equivalent to any of the above
values except that for iris lesion the value should be ≥ 1 but < 2 and for redness of the conjunctiva the value should be ≥ 2.5.
In both cases all scores at each of the reading times (24, 48 and 72 hours) for an effect should be used in calculating the respective mean values.
- Substances or preparations which cause significant ocular lesions, based on practical experience in humans.
- Organic peroxides except where evidence to the contrary is available.
R41 Risk of serious damage to eyes
Substances and preparations which, when applied to the eye of the animal cause severe ocular lesions which occur within 72 hours after exposure
and which persist for at least 24 hours.
Ocular lesions are severe if the means of the scores of the eye irritation test cited in Annex B.5 to Commission Regulation (EC) No. 440/2008, L 142 [4] have any of the values:
- cornea opacity ≥ 3,
- iris lesion > 1.5.
The same shall be the case where the test has been completed using three animals if these lesions, on two or more animals, have any of the values:
- cornea opacity ≥ 3,
- iris lesion = 2.
In both cases all scores at each of the reading times (24, 48 and 72 hours) for an effect should be used in calculating the respective mean values.
Ocular lesions are also severe when they are still present at the end of the time of observation.
Ocular lesions are also severe if the substance or preparation causes irreversible colouration of the eyes.
- Substances and preparations which cause severe ocular lesions, based on practical experience in humans.
Note:
When a substance or preparation is classified as corrosive and assigned R34 or R35, the risk of severe damage to eyes is considered implicit and
R41 is not included in the label.
On the basis of the test results, the following risk phrases shall be assigned in conformity with the criteria given in
Annex I of Regulation (EC) 1272/2008:
Eye irritant Category 1:
Irreversible effects on the eyes / serious damage to the eyes
An eye irritant category 1 is a test material that produces:
- at least in one animal effects on the cornea, iris or conjunctiva that are not expected to reverse or that have not fully reversed within 21 days;
and/or
- at least in 2 of 3 animals, a positive response of
(i) corneal opacity ≥ 3; and/or
(ii) iritis > 1.5
calculated as the mean scores following the grading at 24, 48 and 72 hours after instillation of the test material.
Eye irritant Category 2:
Reversible effects on the eyes / Irritating to the eyes
An eye irritant category 2 is a test material that produces:
- at least in 2 of 3 animals, a positive response of
(i) corneal opacity ≥ 1; and/or
(ii) iritis ≥ 1; and/or
(iii) conjunctival redness ≥ 2; and/or
(iv) conjunctival oedema (chemosis) ≥ 2
calculated as the mean scores following the grading at 24, 48 and 72 hours after instillation of the test material, and which fully reverse within an
observation period of 21 days.
 
On the basis of the test results, the following risk phrases may be assigned in conformity with the criteria given in
GHS - Globally Harmonized System of Classification and Labelling of Chemicals, third revised edition, July 2009 :
Category 1: An eye irritant Category 1 (irreversible effects on the eye) is a test material that produces:
- at least in one animal effects on the cornea, iris or conjunctiva that are not expected to reverse or that have not fully reversed within 21 days;
and/or
- at least in 2 of 3 animals, a positive response of
(i) corneal opacity ≥ 3; and/or
(ii) iritis > 1.5
calculated as the mean scores following the grading at 24, 48 and 72 hours after instillation of the test material.
Category 2A: An eye irritant Category 2A (irritating to eyes) is a test material that produces:
- at least in 2 of 3 animals, a positive response of
(i) corneal opacity ≥ 1; and/or
(ii) iritis ≥ 1; and/or
(iii) conjunctival redness ≥ 2; and/or
(iv) conjunctival oedema (chemosis) ≥ 2
calculated as the mean scores following the grading at 24, 48 and 72 hours after instillation of the test material, and which fully reverse within
21 days.
Category 2B: Within this category an eye irritant is considered mildly irritating to eyes (Category 2B), when the effects listed above are fully reversible within 7 days of observation.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 6

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 7

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 8

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 10

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 11

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 12

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Results
After the application into the eyes of three female NZW rabbits the test item produced irritant but not corrosive effects.
Neither mortalities nor significant clinical signs of toxicity were observed.
The eyes were not rinsed after the application.
Upon fluorescein examinations at the end of the observation period of 72 hours no corneal lesions were found in any animal.
Conjunctival redness, chemosis and discharge were observed in all animals.
There were no significant body weight changes during the observation period.

Any other information on results incl. tables

Eye Irritation Scores – Animal No. 1

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	1/0	1/0	0/0	0.67
	Conjunctival Chemosis	1/0	1/0	1/0	0/0	0.67
	Iris	0/0	0/0	0/0	0/0	0
	Cornea	0/0	0/0	0/0	0/0	0

T =testitem,       C = control Eye Irritation Scores – Animal No. 2

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	0/0	0/0	0.33
	Conjunctival Chemosis	1/0	1/0	0/0	0/0	0.33
	Iris	0/0	0/0	0/0	0/0	0
	Cornea	0/0	0/0	0/0	0/0	0

T =testitem,       C = control Eye Irritation Scores – Animal No. 3

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	2/0	1/0	1/0	0/0	0.67
	Conjunctival Chemosis	2/0	1/0	0/0	0/0	0.33
	Iris	0/0	0/0	0/0	0/0	0
	Cornea	0/0	0/0	0/0	0/0	0

T =testitem,       C = control Clinical Signs of Toxicity

Animal No. 1	Time after Test Item Application	Systemic Findings	Local Findings (apart from findings in Table 3)
	1 hour	nsf	discharge grade 1
	24 hours	nsf	nsf
	48 hours	nsf	nsf
	72 hours	nsf	nsf
Animal No. 2	Time after Test Item Application	Systemic Findings	Local Findings (apart from findings in Table 4)
	1 hour	nsf	discharge grade 2
	24 hours	nsf	nsf
	48 hours	nsf	nsf
	72 hours	nsf	nsf
Animal No. 3	Time after Test Item Application	Systemic Findings	Local Findings (apart from findings in Table 5)
	1 hour	nsf	discharge grade 2
	24 hours	nsf	nsf
	48 hours	nsf	nsf
	72 hours	nsf	nsf

nsf: no specific findings Absolute Body Weights in kg

	Animal No. 1	Animal No. 2	Animal No. 3
Start of Study (weight in kg)	3.1	2.9	3.2
End of Study (weight in kg)	3.2	3.0	3.3

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 Triethyleneglycoldimethylether to rabbits at a dose of 0.1 mL produced slightly irritant effects, which were fully reversible within up to 72 hours post-application. 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 [6], the test item Triethyleneglycoldimethylether has no obligatory labelling requirement for
eye irritation.
According to Annex I of Regulation (EC) 1272/2008 [4], the test item Triethyleneglycoldimethylether has no obligatory labelling requirement
for eye irritation.
According to GHS (Globally Harmonized Classification System) [8] the test item Triethyleneglycoldimethylether has no obligatory labelling
requirement for eye irritation.

Executive summary:

 

A single ocular application of the test item Triethyleneglycoldimethylether to rabbits at a dose of 0.1 mL produced slightly irritant effects, which were fully reversible within up to 72 hours post-application. 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 toAnnex VI of Commission Directive 2001/59/EC, the test item Triethyleneglycoldimethyletherhas no obligatory labelling requirement for eye irritation.

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

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