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Toxicological information

Skin irritation / corrosion

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Administrative data

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.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1979
Report date:
1979

Materials and methods

Test guideline
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

Test material

Constituent 1
Chemical structure
Reference substance name:
Diallyl 2,2'-oxydiethyl dicarbonate
EC Number:
205-528-7
EC Name:
Diallyl 2,2'-oxydiethyl dicarbonate
Cas Number:
142-22-3
Molecular formula:
C12H18O7
IUPAC Name:
3-({[2-(2-{[(prop-2-en-1-yloxy)carbonyl]oxy}ethoxy)ethoxy]carbonyl}oxy)prop-1-ene
Details on test material:
- Name of test material (as cited in study report): CR-39 Monomer
- Substance type: pure active substance
- Physical state: clear colourless liquid
- Analytical purity: 100% commercial product
- Impurities (identity and concentrations):

Sample H-6916:
Acrolein before testing 42.5 ppm, after testing 42.0 ppm
Allyl Alcohol before testing 28.2 ppm, after testing 27.4 ppm
Diallyl Carbonate before testing 2399 ppm, after testing 1499 ppm

Sample J-4944:
Acrolein before testing 22 ppm, after testing - no data
Allyl Alcohol before testing 19 ppm, after testing - no data
Diallyl Carbonate before testing 2789 ppm, after testing - no data

- Composition of test material, percentage of components: data not available
- Isomers composition: data not available
- Purity test date: (see above the conversation letter)
- Lot/batch No.: Sample 1: H-6916 (received from PPG Industries on January 2, 1979); Sample 2: J-4944 (received from PPG Industries on February 13, 1979)
- Expiration date of the lot/batch: data not available
- Stability under test conditions: No significant changes in monomer composition during the testing period
- Storage condition of test material: data not available

Test animals

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

Test system

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.

Results and discussion

In vivo

Resultsopen allclose all
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

Any other information on results incl. tables

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.

Applicant's summary and conclusion

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.