Diallyl 2,2'-oxydiethyl dicarbonate

Endpoint summary

Description of key information

Skin and eye irritation studies conducted to evaluate the irritation potential of CR-39 were performed prior to the adoption of the OECD guidelines and the GLP regulations. The studies are acceptable, well-documented, which meet basic scientific principles (Klimish et al., 1997). The key study acute Eye Irritation Study in Albino rabbits with 100% DDC is a GLP study (report date 1981-03-24).

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:

January-June 1979

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP study, but acceptable, well-documented study report which meets basic scientific principles.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Principles of method if other than guideline:

Six animals (3 males/3 females) were exposed to 0.5 mL of test material for 6 hours (for the first test). The test material was applied undiluted on the saddle area of an animal at one intact and one abraded site, and 0.2 ml sterile physiological saline was added to the 0.5 ml of test material at the other one intact and one abraded site. A second and third trial were performed using the same animals. After 26 days following the first test, the second test was conducted and test material was applied to intact skin of the abdomen of rabbits. After 21 days following the second test, test material was applied to intact and abraded site of rabbit abdomen. For the third test another batch of sample was used.

GLP compliance:

no

Remarks:

The study was performed prior to the adoption of the GLP regulations

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Sweetwater Farm INc., Box 293, Hillsboro Ohio 45133
- Age at study initiation: data not available
- Weight at study initiation: 1.82 to 2.16 kg at the time of delivery, six days prior to study initiation (as determined by the supplier)
- Housing: data not available
- Diet (e.g. ad libitum): data not available
- Water (e.g. ad libitum):data not available
- Acclimation period: six days


ENVIRONMENTAL CONDITIONS
data not available

Type of coverage:

occlusive

Preparation of test site:

other: one site intact; one site abraded

Vehicle:

physiological saline

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): based on specific density 1.143 of test material the concentration used was 0.5715 mg/mL

0.2 mL steril physiological saline (vehicle) was added to the 0.5 mL of test material at one intact and one abraded site (in the first test).

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.2 mL
- Concentration (if solution): 0.9% sodium chloride
- Lot/batch no. (if required): data not available
- Purity:data not available

Duration of treatment / exposure:

6 hours

Observation period:

The reactions were scored immediately after removal of the patches (6-hour reading) and again at 24, 48 and 72 hours. The reactions were scored again on days 7, 9, 11 and 13.

Number of animals:

Six animals (Rabbits no. 1-3 were females; rabbits no. 4-6 were males).

Details on study design:

Applications of test substance to each animal are shown in Table 1 in "Any other information on materials and methods incl. tables"
TEST SITE
- Area of exposure: saddle area for first test and abdomens for second and third tests
- % coverage: data not available
- Type of wrap if used: a one-inch square surgical gauze patch, two layers thick on each of the patch sites.
After application of the patches, the trunk of each rabbit was wrapped with rubber dental damming, which was secured with staples. An outer layer of gauze and tape was placed around the trunk of each animal. The animals were restrained in Newmann harnesses for 6 hours

REMOVAL OF TEST SUBSTANCE
- Washing (if done): any residual sample was gently sponged from the skin with a moistened towel
- Time after start of exposure: 6 hours


SCORING SYSTEM: see scoring key Table 2 in "Any other information on materials and methods incl. tables"

Twenty-six days following the initial application of materials, the abdomens of each of the six rabbits was clipped. To one patch site in the center of the shaved area, 0.5 ml of the sample (first batch) was applied on each of the rabbits. On one rabbit (#2), 25 µl of the test material was applied to another test site anterior to the central site. The sites were left intact. The test materials was applied under one-inch gauze patches and the binding materials and Newmann harness were used as described previously. The reaction were scored as in the first test.
Twenty-one days following the second application of the test material, a third trial was performed. For this test the second batch of sample was used. The abdomens of the six rabbits were again shaved. Two application sites, lateral to the ventral longitudinal midline of the rabbit, were- prepared. Patch sites were each approximately 5 cm from the midline and at least 10 cm from each other, vOne patch site on each rabbit was left intact; one site was abraded. For each site, 0.5 ml of the test material was applied under a gauze patch as described above. The binding materials and Newmann harnesses were applied as described above.

Irritation parameter:

erythema score

Remarks:

first application

Basis:

animal #1

Time point:

48 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: Necrosis and edema noted on biopsied area on day 11 (only on intact site with undiluted test material)

Irritation parameter:

erythema score

Remarks:

first application

Basis:

animal #2

Time point:

other: 48 hours to 13 days

Max. score:

4

Reversibility:

not fully reversible within: 13 days

Remarks on result:

other: Very slight erythema at 24 hours; At 48 hours, moderate to severe erythema (purple); at 72 hours and up to to day 13 irritative effects progressed to necrosis (and sloughing) with severe erythema and very slight edema

Irritation parameter:

erythema score

Remarks:

first application

Basis:

animal #3

Time point:

other: 7 - 9 days

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: on day 11 necrosis noted at biopsied site and slight edema

Irritation parameter:

erythema score

Remarks:

first application

Basis:

animal #4

Time point:

other: 11 day

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: necrosis is noted on biopsied site on day 11

Irritation parameter:

erythema score

Remarks:

first application

Basis:

animal #5

Time point:

other: 11 day

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: necrosis noted at biopsied site

Irritation parameter:

erythema score

Remarks:

first application

Basis:

animal #6

Time point:

other: 11 day

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: necrosis noted at biopsied site

Irritation parameter:

erythema score

Remarks:

second application

Basis:

animal: 1-6

Time point:

other: 6, 24, 48 and 72 hours

Score:

0

Max. score:

0

Reversibility:

other: no effects

Remarks on result:

other: animal 2 was treated with 0.5 mL and 25 µL of test material (second application).

Irritation parameter:

erythema score

Remarks:

third application

Basis:

animal: all animals except animal 3

Time point:

other: 6, 24 48, 72 and 96 hours and 7 days

Score:

0

Max. score:

0

Reversibility:

other: no effects

Remarks on result:

other: third application

Irritation parameter:

erythema score

Remarks:

third application

Basis:

animal #3

Score:

4

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: third application

Irritation parameter:

edema score

Remarks:

first application

Basis:

animal #1

Time point:

other: 11 day

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: necrosis and edema noted on biopsied sites (only on intact site with undiluted test material)

Irritation parameter:

edema score

Remarks:

first application

Basis:

animal #2

Time point:

other: 72 hours-11 days

Max. score:

2

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: At 72 hours to day 5 - very slight edema; at days 7 to 9 irritative effects progressed to slight edema and on day 11 necrosis noted on biopsied sites

Irritation parameter:

edema score

Remarks:

first application

Basis:

animal #3

Time point:

other: 9-11 day

Max. score:

2

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: only on intact site (with test material and physiological saline)

Irritation parameter:

edema score

Remarks:

first application

Basis:

animal #4

Time point:

other: 11 day

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: necrosis noted at biopsied site and on raised patch of skin

Irritation parameter:

edema score

Remarks:

first application

Basis:

animal #5

Time point:

other: 13 day

Score:

0

Max. score:

0

Reversibility:

other: no effects

Irritation parameter:

edema score

Remarks:

first application

Basis:

animal #6

Time point:

other: 13 day

Score:

0

Max. score:

0

Reversibility:

other: no effects

Irritation parameter:

edema score

Remarks:

second application

Basis:

animal: 1-6

Time point:

other: 6, 24, 48 and 72 hours

Score:

0

Max. score:

0

Reversibility:

other: no effects

Remarks on result:

other: animal 2 was treated with 0.5 mL and 25 µL of test material (second application).

Irritation parameter:

edema score

Remarks:

third application

Basis:

animal: 1-6

Time point:

other: 6, 24, 48, 72, and 96 hours and 7 days

Score:

0

Max. score:

0

Reversibility:

other: no effects

Remarks on result:

other: third application

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Reversibility:

fully reversible

Remarks on result:

positive indication of irritation

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Reversibility:

not fully reversible within: 7 and 13 day

Remarks:

on two animals

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Reversibility:

fully reversible

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

First application: at 48 hours, moderate to severe erythema was noted at all sites on Rabbit #2, and by 72 hours irritative effects progressed to necrosis with severe erythema and very slight edema. On day 5 a veterinary dermatologist observed the rabbits. He observed no visible lesions on any rabbits except No. 2, on which reddening and blackened skin were noted at each of the -four sites.

Other effects:

dermal inflammatory response, edema, vasodilation, degenerative changes in vascular walls, increased eosinophilia

First Application

As described above in Irritation/Corrosion results only rabbit 2 exhibited severe adverse effects of treatment.

On day 5 a veterinary dermatologist observed the rabbits. He observed no visible lesions on any rabbits except No. 2, on which reddening and blackened skin were noted at each of the -four sites (see Table 1 in "Overall remarks"). The biopsied materials were evaluated by a veterinary pathologist. The biopsies-from all rabbits other than #2 and the biopsy from a non-treated site on #2 revealed few abnormalities compared to the biopsies from the treated sites of #2. The basic features of the reaction at these sites were a dermal inflammatory response characterized by rather marked hemorrhage, exudation of heterophils, edema, vasodi1ation, and a degenerative change in some of the vascular walls that was characterized by increased thickness to the wall and increased eosinophilia.

Irritation observed on the dorsal area in any rabbits other than #2 after day 5 were generally associated with the biopsied areas. The hair on the back of Rabbit #2 grew back more quickly on the treated areas than on the clipped, non-treated areas.

Second Application

No signs of irritation were noted within 72 hours following application of the material. By the time of application, rabbit #3 had developed large pus-filled abscesses on its back.

Third Application

Reactions were limited to well-defined erythema with necrosis at the intact site of Rabbit #3 noted at 72 hours and subsequently (Table 1 in "Overall remarks"). Attempts to culture the pus-like material from the back of Rabbit #3 proved unsuccessful. The veterinary dermatologist observed the rabbits at the 72-hour reading, and saw no visible lesions on any of the rabbits except No. 3. At the intact site of Rabbit No. 3 the veterinary dermatologist observed a purple to deep-red area at the center of the site, blanching to a light red color on the border. The lateral aspect of the lesion showed sharp demarcation between normal and abnormal. He saw no visible lesion at the abraded site of No. 3.

The veterinary pathologist, following histopathological evaluation of the biopsied materials, found all biopsied tissue within normal ranges except for the tissue taken from the intact site of Rabbit No. 3. At this site reactions in the dermis were found consisting of edema, hemorrhage, heterophil infiltration, vasodilation and congestion.

The results of materials sent for immunofluorescent examination included no evidence of deposits of immunoglobulin in arterial walls using fluorescein-conjugated anti-rabbit IgG. Minimal linear subepidermal deposits which appeared to be in the region of the basement membrane were noted at the treated site of Rabbit #3 at which no visible lesions were noted.

Interpretation of results:

other: sporadic irritation with necrosis

Remarks:

Criteria used for interpretation of results: expert judgment

Conclusions:

Application of CR-39 Monomer to the skin of New Zealand White rabbits resulted in sporadic irritation with necrosis. The response did not appear to be related to an immune response.

Executive summary:

The study was conducted to evaluate the irritation potential of the test material CR-39 Monomer in New Zealand White rabbits.

Two different batches of test material were used in three tests: first batch (from 1979 -01 -02) for the first and the second test and second batch (from 1979 -02 -13) for the third test. Six the same animals were used in all tests. The test material in the first test was applied to saddle areas of animals, in the second and in the third tests to abdomens. Twenty-six days following the initial application

the second application was conducted and twenty-one days following the second application a third trial was performed. The reactions were scored immediately after removal of the patches (6-hour reading) and again at 24, 48 and 72 hours and on days 7, 9, 11 and 13. Removed biopsies were sent for histopathological and immunofluorescence examinations.

The first application resulted in very slight to slight erythema and very slight edema noted in Rabbit No. 1 at the 48-hour reading. Only Rabbit No. 2 exhibited irritation within seven days following application: very slight erythema at 24 hours; moderate to severe erythema at 48 hours; and at 72 hours irritative effects progressed to necrosis with severe erythema and very slight edema. Basically only this rabbit experienced a unique response to treatment that could be characterized as primarily acute and vascular. Such a reaction could be distinctly separated from all other reactions evaluated. There was no visible effects of treatment on skin of other animals. Necrosis and edema were associated only with biopsied sites.

No effect of treatment was observed during the second test.

Reactions of third application were limited to well-defined erythema with necrosis at the intact siteof Rabbit #3 noted at 72 hours and subsequently. No visible lesions on any of the rabbits except No. 3 were found as well as all biopsied tissue was within normal ranges except for the tissue taken from the intact site of Rabbit No. 3. "There was no dose relationship between the reactions that ocurred in the first test and the third test and that the pattern of reaction is very inconsistent and not reproducible at the present time. This reaction seems not to depend upon the animal that is being tested or the amount of solution or site selection and thus, one can conclude that there certainly is a reaction taking place but being not reproducible one must consider this a spurious result." (from a conversation letter of P.T. Breen Veterinary dermatologist and Dr. Vinegar Toxicologist, 1979 -03 -12; from original report).

The test material was found to be irritative in an incidental manner.

 

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

December 1979 - March 1980

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Equivalent to guideline study; the study passed a Quality Assurance audit (dated 1981-03-24)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Vehicle:

unchanged (no vehicle)

Controls:

other: the left eye served as the untreated control

Duration of treatment / exposure:

a single exposure in one eye of albino rabbits

Observation period (in vivo):

All animals were observed closely for gross signs of systemic toxicity and mortality at frequent intervals during the day of dosing and at least twice daily thereafter for a ( total of 14 days. Room conditions as well as availability of adequate food and water were checked and any noteworthy conditions recorded.

Number of animals or in vitro replicates:

A total of nine rabbits (6 not rinsed and 3 rinsed) were used.

Irritation parameter:

overall irritation score

Basis:

mean

Remarks:

of six animals

Time point:

other: 1 h

Score:

8

Reversibility:

fully reversible within: 24 h

Remarks on result:

other: not rinsed

Irritation parameter:

overall irritation score

Basis:

mean

Remarks:

of three animals

Time point:

other: 1 h

Score:

3

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: rinsed

Irritation parameter:

chemosis score

Time point:

24/48/72 h

Remarks on result:

not determinable

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48 h

Other effects:

no

Body Weights

Body weights at the termination of the study ranged from 2.15 to 2.75 kilograms. Body weight changes appear to be normal.

Observations and Mortality

No animal exhibited signs of systemic toxicity and no deaths occurred during the investigation.

Necropsy

Evaluation of the test animals at necropsy by the pathologist revealed no significant gross pathologic findings.

Primary Eye Irritation Index

Means of individual irritation scores used to calculate the primary irritation index presented in Table 3 and 4.

Table 3: Primary Eye Irritation Index in Rabbits (not rinsed)

Tissue	Examination Intervals
	1 h	6 h	24 h	48 h	72 h	4 Days	7 Days
Cornea	0	0	0	0	0	0	0
Iris	0	0	0	0	0	0	0
Conjunctiva	48	4	0	0	0	0	0
Total	48	4	0	0	0	0	0
Mean	8.0	0.67	0	0	0	0	0

Table 4: Primary Eye Irritation Index in Rabbits (rinsed)

Tissue	Examination Intervals
	1 h	6 h	24 h	48 h	72 h	4 Days	7 Days
Cornea	0	0	0	0	0	0	0
Iris	0	0	0	0	0	0	0
Conjunctiva	18	4	2	0	0	0	0
Total	18	4	2	0	0	0	0
Mean	3.0	0.67	0.33	0	0	0	0

Positive scores only noted in the conjunctiva lasted through the 24-hour examination interval. No positive findings were noted upon further examination with fluorescein dye.

Interpretation of results:

other: EU GHS criteria not met

Conclusions:

According method of Draize, when 100% DDC (diallyl diglycol carbonate) was instilled into one eye of each of nine albino rabbits (6 not rinsed and 3 rinsed) the primary eye irritation index was determined to be 8.0 and was classified as minimally irritating. Instillation of water into the treated eyes of three rabbits did not change the classification but decreased the score to 3.0. According to the guidance to Regulation (EC) No 1272/2008 (CLP Regulation) [ECHA Guidance on the Application of the CLP Criteria, Aug 2009; p. 249], Diallyl Diglycol Carbonate does not meet criteria for classification as eye irritant.

Executive summary:

The objective of this study was to determine the irritative potential of the test material following a single exposure in one eye of albino rabbits. Nine rabbits (6 not rinsed and 3 rinsed) were used.

0.1 mL of undiluted test material was instilled in the right eye of each animal. All animals were observed for signs of systemic toxicity and mortality until sacrifice on day 14. Body weights on the day of exposure and the termination were recorded. A gross necropsy were performed on the visceral and thoracic cavities. Primary Eye Irritation Index was calculated using the standard scoring system of Draize.

No positive findings were noted in two ocular tissues: cornea and iris. Positive scores were only noted in the conjunctiva lasted through 24 -hour interval. The one-hour interval provided the highest mean score (8.0) for 100% DDC (diallyl diglycol carbonate) and was therefore used as the primary irritation index and classified the test compound as minimally irritating using the method of Draize. Instillation of water into the treated eyes of three rabbits did not change the classification but decreased the mean score to 3.0.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

In the "Primary Skin Irritation Test" (Acute Toxicity Studies IBT No. A254, 1971 -09 -10) the exposure period was 24 hours. In accordance with the OECD Guideline 404 Acute Dermal Irritation-Corrosion the exposure period should be normally 4 hours. After 24 hour exposure period in three of the four rabbits, both abraded and non-abraded sites showed severe erythema and edema at 24 and 72 hour reading. Carbonic acid was found to be highly irritating to rabbit skin. Contradictive results were obtained in the irritation tests on the skin of rabbits (data published M.Lacroix et al., 1976, endpoint 7.3.1.003). The test material provoked only minimal changes in the skin after 24 hours (slight erythema in certain animals). The exposure period of 24 hours is not realistic to humans. Therefore, the study "Skin Irritation Studies of CR-39 Monomer" (report dated 1976 -06 -04) was taken forward for risk characterization. In this study undiluted test material (0.5 mL) was applied to one intact and one abraded skin site on rabbits (3/sex) under occluded conditions for 6 hours and assessed for up to 13 days after exposure. At 48 hours, moderate to severe erythema were noted on all sites and by 72 hours, irritation progressed to necrosis with severe erythema and very slight edema. Reddened and blackened skin at all sites was noted. By 13 days, edema, but not erythema, resolved. "...there is no dose relationship between the reaction that occured in the first treatment and the third treatment and the pattern of reaction inconsisitent and not reproducible... The reaction seems not to depend upon the animal that is being tested or the amount of solution or site selection and thus, one can conclude that there certainly is a rection taking place but being not reproducible and one must consider this is a spurious result." (from the conversation letter of Dr. M. Vinegar, Toxicologist and P.T. Breen, Dermatologist, dated 1979- 03 -12). Carbonic acid, oxydiethylene diallyl ester is irritating to rabbit skin.

Eye irritation

In the earlier study "Eye Irritation Test"[(Acute Toxicity Studies IBT No. A254, 1971 -09 -10) Endpoint 7.3.2.002] undiluted test material (0.1 mL) was instilled in the conjunctival sac of the right eye of five rabbits. Test eyes were not washed. Irritation was scored at 1 minute, 1, 24 and 72 hours and 7 days after administration. Conjunctival redness and slight discharge was seen which was resolved by 24 – 72 hours. In this study the test material was slightly irritating to rabbit eyes. In the Acute Eye Irritation Study in Rabbitts with 100% DDC (report date 1981 -03 -24) the principle of test design was the same as in the previous one except the number of animals. 6 not rinsed and 3 rinsed animals were tested. The cornea and the iris were not affected, the conjunctiva was affected by one- and six-hour examination and the lesions were fully reversible within 24 hours. The one-hour interval provided the highest mean score (8.0) for 100% DDC (diallyl diglycol carbonate) and was therefore used as the primary irritation index and classified the test compound as minimally irritating using the method of Draize. Instillation of water into the treated eyes of three rabbits did not change the classification but decreased the mean score to 3.0. The test material is considered to be slightly irritating to eyes.

Justification for classification or non-classification

Eye irritation

According to Regulation (EC) No 1272/2008 (CLP Regulation) Diallyl Diglycol Carbonate does not meet the criteria for classification as eye irritant.

Skin irritation

The classification as skin irritant is warranted according to the criteria of Regulation (EC) No 1272/2008.

GHS: Skin irritation Cat 2., H315