Bis(2-ethylhexyl) terephthalate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

72 hours

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Reliable without restriction; the study was conducted according to GLPs and OECD Guideline 405 .

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

Due to a technician error, the animals did not receive local anesthetic prior to instillation of the test material. This deviation did not impact the quality or integrity of the study.

GLP compliance:

yes

Remarks:

Compliant with two exceptions; information about the test material characterization retained by the Study Sponsor, and Study Director did not fill in proposed Experimental Start and Termination dates before signing the protocol or prior to study start.

Test material

Test material information

Constituent 1

Reference substance name:

Bis(2-ethylhexyl) terephthalate

EC Number:

229-176-9

EC Name:

Bis(2-ethylhexyl) terephthalate

Cas Number:

6422-86-2

Molecular formula:

C24H38O4

IUPAC Name:

1,4-bis(2-ethylhexyl) benzene-1,4-dicarboxylate

Constituent 2

Reference substance name:

Reference substance 001

Cas Number:

6422-86-2

Details on test material:

-Test substance (as cited in the study report): 1, 4 Diethylhexyl terephthalate
-Batch number: TD5023342
-Purity: 98.21% (as provided on Certificate of Analysis). Impurities included water (0.0179%) and the ethylhexyl-methyl ester (1.60%).

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Test animals:
-Source: Robinson Services, Inc. Clemmons, NC
-Sex: 1 male,and 2 females
-Age at study initiation: young adult
-Acclimation period: 23 days
-Housing: singly housed in suspended stainless-steel cages with mesh floor
-Diet: pelleted Purina Rabbit Chow #5326
-Water: filtered tap water ad libitum
-Method of animal identification: Uniquely numbered stainless steel ear tag

Environmental Conditions:
-Temperature: 20 - 24 °C
-Photoperiod: 12 hours light/12 hours dark

Study Dates:
-Study Initiation Date: 8 March 2006
-Experimental Start Date: 10 March 2006
-Experimental Completion Date: 13 March 2006

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The other eye of each rabbit was not treated and served as a control.

Amount / concentration applied:

0.1 mL undiluted

Duration of treatment / exposure:

The test substance (0.1 mL) was instilled into the conjunctival sac of the right eye of each rabbit by pulling the lower lid away from the eyeball. The upper and lower lids were then gently held together for approximately 1 second. Eyes were not washed.

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3

Details on study design:

Prior to the test, both eyes of a group of animals were examined for pre-existing ocular irritation. One drop of 2% ophthalmic fluorescein sodium was instilled into both eyes of each rabbit. The eyes were rinsed with physiological saline (0.9% NaCl) approximately 30 seconds later and subsequently evaluated for corneal damage using an ultraviolet light source. Three healthy animals without evidence of preexisting ocular irritation were selected for use in the study based on this screening procedure. The test substance (0.1 mL) was instilled into the conjunctival sac of the right eye of each rabbit by pulling the lower lid away from the eyeball. The upper and lower lids were then gently held together for approximately 1 second. The other eye of each rabbit was not treated and served as a control. Ocular irritation, including effects on the cornea, iris, and conjunctiva were evaluated according to the method of Draize et al. (1944), using a high intensity white light (Mag Lite) 1, 24, 48 and 72 hours after instillation. In addition, flourescein dye was used as described above to evaluate corneal damage 24 hours after treatment. Individual scores were recorded for each animal. The average score for all rabbits was determined for each evaluation time. The time interval with the highest mean score for all rabbits was used to classify the test material according to the method of Kay and Calandra (1962). The animals were observed at least once daily for signs of gross toxicity and behavioral changes during the test period. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhea and coma.

References Cited:
Kay, JH and Calandra, JC, 1962. Interpretation of eye irritation tests. J. Soc. Cos. Chem. 13:281-289.

Draize et al., 1944. Methods for the study of irritation and toxicity of substances applied topically to the skin and mucous membranes. J. Pharmacol. Exp. Ther., 82:377-390.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Time point:

other: 1 hour

Score:

0

Max. score:

10.7

Reversibility:

fully reversible within: 72 hours

Results 2

Irritation parameter:

chemosis score

Basis:

other: all animals

Time point:

other: 1 hour only

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: all other chemosis scores at 24 and 48 hours were 0

Results 3

Irritation parameter:

other: redness score

Basis:

other: all animals

Time point:

other: 1 hour

Score:

0

Max. score:

2

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: Diffuse, deep crimson red, individuals not easily discernable

Results 4

Irritation parameter:

other: redness score

Basis:

other: all animals

Time point:

other: 24 hours

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: Injected above normal

Results 5

Irritation parameter:

other: redness score

Basis:

other: all animals

Time point:

other: 48 hours

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Results 6

Irritation parameter:

other: discharge score

Basis:

animal #1

Time point:

other: 1 hour

Score:

0

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: Discharge with moistening of the lids and hair of a considerable area around the eye.

Results 7

Irritation parameter:

other: discharge score

Basis:

other: animals #2 and #3

Time point:

other: 1 hour

Score:

0

Max. score:

2

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: Discharge with moistening of the lids and hair adjacent to the lids.

Results 8

Irritation parameter:

other: discharge score

Basis:

other: all animals

Time point:

other: 24 hours

Score:

0

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: The assigned score of 1 was based on the presence of any amount of discharge different from that normally seen in the untreated rabbit eye.

Results 9

Irritation parameter:

cornea opacity score

Basis:

other: all animals

Time point:

other: all time points

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

other: no corneal opacity was observed at any time during the study

Results 10

Irritation parameter:

iris score

Basis:

other: all animals

Time point:

other: all time points

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

other: no iritis was observed at any time during the study

Results 11

Irritation parameter:

cornea opacity score

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 12

Irritation parameter:

chemosis score

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 13

Irritation parameter:

iris score

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 14

Irritation parameter:

conjunctivae score

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No corneal opacity or iritis was observed in any treated eye at any time during the study. All three treated eyes exhibited conjunctivitis 1, 24 and 48 hours after instillation but were normal at 72 hours. Redness scores in the 3 rabbits were 2 (diffuse, deep crimson red, individual vessels not easily discernable), 1 (injected above normal), and 1 at 1, 24 and 48 hours, respectively. Chemosis scores of 1 (any swelling above normal) were recorded at 1 hour for all 3 animals (all other chemosis scores were 0). Discharge scores of 3 (discharge with moistening of the lids and hairs; considerable area around the eye), 2 (discharge with moistening of the lids and hairs just adjacent to the lids), and 2 were recorded in the three animals one hour after instillation. Discharge scores of 1 were observed in all 3 animals at 24 hours. The mean irritation scores at 1, 24 and 48 hours were 10.7 (12 + 10 + 10/3), 4.0 (4 + 4 + 4/3) and 2.0 (2 + 2 + 2/3) (respectively). All animals were free of ocular irritation by 72 hours. The maximum mean total score was 10.7, which corresponded to a classification of minimally irritating using the Kay and Calandra (1962) classification system. According to their guidance, since all scores were not 0 at 48 hours, the classification was increased one level (to mildly irritating from minimally irritating).

Other effects:

Other than minimal eye irritation, there were no other signs of toxicity or abnormal behavior.

Any other information on results incl. tables

		Rabbit 1				Rabbit 2				Rabbit 3
		Hours				Hours				Hours
		1	24	48	72	1	24	48	72	1	24	48	72
I. Cornea
A. Opacity		0	0	0	0	0	0	0	0	0	0	0	0
B. Area		4	4	4	4	4	4	4	4	4	4	4	4
(AxB)x5		0	0	0	0	0	0	0	0	0	0	0	0
II. Iris
A. Values		0	0	0	0	0	0	0	0	0	0	0	0
Ax5		0	0	0	0	0	0	0	0	0	0	0	0
III. Conjunctivae
A. Redness		2	1	1	0	2	1	1	0	2	1	1	0
B. Chemosis		1	0	0	0	1	0	0	0	1	0	0	0
C. Discharge		3	1	0	0	2	1	0	0	2	1	0	0
(A+B+C)x2		12	4	2	0	10	4	2	0	10	4	2	0
Total		12	4	2	0	10	4	2	0	10	4	2	0

Applicant's summary and conclusion

Interpretation of results:

other: minimally irritating

Remarks:

Criteria used for interpretation of results: OECD GHS

Conclusions:

The maximum mean total score in a primary eye irritation study in rabbits was 10.7, which corresponded to a classification of minimally irritating according to the test scheme of Kay and Calandra (1962). According to their guidance, since all scores were not 0's at 48 hours, the classification was increased one level (to mildly irritating). There was no evidence of corneal opacity or iritis at any time during the study. Conjunctival redness was observed in all animals at 1, 24 and 48 hours but all animals were normal at the 72 hour observation period and the calculated mean score following grading at 24, 48 and 72 hours after installation of the test material was <2. Conjunctival chemosis (grade 1) was also observed in all animals at the 1 hour observation period but all animals were normal

at the 24 hour observation period. Based on these results, di (2-ethylhexyl) terephthalate is not classified as an ocular corrosive or irritant according to GHS.

Executive summary:

In an acute ocular irritation study, 0.1 mL of di (2-ethylhexyl) terephthalate was administered into the conjunctival sac of one eye of three New Zealand White Rabbits. The untreated eye of each rabbit served as a control. The eyes were examined at 1, 24, 48, and 72 hours post-instillation and graded according to the method of Draize. There was no evidence of corneal opacity or iritis at any time during the study. Signs of irritation were limited to conjunctivitis. Chemosis (grade 1) which was observed at the 1 hour observation period was absent at all later observation periods. Conjunctival redness (grade 2 at 1 hour and grade 1 at 24 and 48 hours) had cleared by 72 hours. The pattern of irritation which was observed led to a classification of the test substance as mildly irritating using the method of Kay and Calandra. However, the mean test scores did not meet the criteria for classification as corrosive or irritating to the eye under GHS. Based on the results of this study, di (2-ethylhexyl) terephthalate may cause slight transient ocular irritation and presents a low toxicity hazard.