Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2004
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Recognised methodology used for testing in circumstances where animal testing is not permitted i.e. cosmetics. Testing carried out by a recognised test facility under GLP conditions by trained laboratory personnel. Test method B.48 / OECD 438 - Isolated Chicken Eye Test (ICE) Method is an in vitro test method that may be used to identify chemicals (substances or mixtures) as either 1) causing “serious eye damage” (CLP category 1), or 2) not requiring classification for eye irritation or serious eye damage according to the CLP. However, this test is only recommended to identify chemicals inducing serious eye damage, i.e. chemicals to be classified as CLP Category 1, without further testing, and also recommended to identify chemicals that do not require classification for eye irritation or serious eye damage • If the results of testing the substance is not classified as Cat 1 under CLP or not classified, more testing is required • May result in false positive predictions for alcohols and false negative predictions for solids and surfactants

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
EU method B.48 (Isolated chicken eye test method for identifying occular corrosives and severe irritants)
Deviations:
not specified
GLP compliance:
yes
Remarks:
Performed under Good Laboratory Practice principles (including govenunent regulations to the extent applicable) and in accordance with standard operating procedures and applicable standard protocols. Quality assessed.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: 5% active aqueous solution
Details on test material:
No batch or lot data.

Test animals / tissue source

Species:
other: hen's egg
Strain:
other: white leghorn
Details on test animals or tissues and environmental conditions:
Not applicable -no live animals used.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
5% active solution
Duration of treatment / exposure:
The test or.reference article, at a dosage of three-tenths of one milliliter (0.3 ml) of a liquid or three-tenths of one gram (0.3 g) of a solid was then administered to each of four (4) CAM's. Twenty seconds later, the test or control article was rinsed from each CAM with five (5) milliliters of physiological saline. All CAM's •were observed immediately prior to test article administration and at 30 seconds. two (2) and five (5) minutes after exposure to the test article. The reactions of the CAM, the blood vessels, including the capillaries, and the albumin were examined and scored for irritant effects.
The numerical, time dependent scores were totaled for each CAM. Each reaction type can be recorded only once for each CAM, therefore the maximum score per CAM is 32. The mean score was determined for all CAM's similarly tested.
Two reference materials were used as a comparison to the ColaMoist 200 (5% active aqueous sample).
These were (1) Johnson's baby shampoo and (2) Prell shampoo (no data on pH of this material). Both reference materials are commercially available shampoos that have a similar % active ingredient level to the ColaMoist 200 (5% active) material
Details on study design:
Fresh, fertile, White Leghorn eggs were obtained from Avian Services in Frenchtown, New Jersey. They were stored at this facility for up to seven (7) days, at 13° C, before being incubated. For incubation the eggs were placed, on their sides, in a Kuhl incubator. The incubator is such that the eggs are automatically rotated once every hour. The temperature was controlled at 99° F (± I 0 with a relative humidity of 60 - 70% for the ten (I0) days of incubation. On day eight (8) the eggs were turned so that the acutely angled end faced down.

On day ten ( I 0) each egg was removed from the incubator and placed in a Plexiglas work enclosure. This enclosure had been preheated and humidified so that its environment approached that of the incubator. A cut was made in the Iaeger end of each egg, where the air sack is located. A Dremel11 Moto-Flex Tool (model 232-5) equipped with a Dremel-,; Cut-Off Wheel (No. 409) was used to make each cut. Forceps were then used to remove the shell down to the shell­
membranc junction. The inner egg membrane was then hydrated with a warm, phyiological saline solution. The saline was removed after a two (2) to five (5) minute exposure. Utilizing pointed forceps, the inner egg membrane was then carefully removed to reveal the CAM.

The test or.reference article, at a dosage of three-tenths of one milliliter (0.3 ml) of a liquid or three-tenths of one gram (0.3 g) of a solid was then administered to each of four (4) CAM's. Twenty seconds later, the test or control article was rinsed from each CAM with five (5) milliliters of physiological saline. All CAM's •were observed immediately prior to test article administration and at 30 seconds. two (2) and five (5) minutes after exposure to the test article. The reactions of the CAM, the blood vessels, including the capillaries, and the albumin were examined and scored for irritant effects.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
overall irritation score
Remarks:
Mean CAM score
Basis:
mean
Score:
2.5
Max. score:
3
Reversibility:
other: not applicable
Remarks on result:
other: ColaMoist 200 (5% active) test sample
Irritation parameter:
overall irritation score
Remarks:
Mean CAM score
Basis:
mean
Score:
11
Max. score:
12
Reversibility:
other: not applicable
Remarks on result:
other: Johnson's baby Shampoo - Reference 1
Irritation parameter:
overall irritation score
Remarks:
Mean CAM score
Basis:
mean
Score:
24
Max. score:
26
Reversibility:
other: not applicable
Remarks on result:
other: Prell shampoo - Reference 2
Irritant / corrosive response data:
Each article was classified using the following scoring system
Irritation Potential - Practically None - Score 0 - 4.9
- Slight - Score 5.0 - 9.9
- Moderate - Score 10.0 - 14.9
- Severe - Score 15 - 32

Any other information on results incl. tables

Previous studies have shown that the CAM of the hen's egg is more sensitive to lquid irritants than is the rabbit eye. Therefore, 50% dilutions of the liquid test and reference articles were used.

Historical in-vivo testing on Johnson's reference product has shown this material to be 'moderatley irritating' eliciting scores approaching 10 at 24 hours when dosed at 100% and tested using the draize ocular irritation method.

The Prell reference product under the same in-vivo Draize testing has given scores approaching 30 at 24 hours when dosed at 100%.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: expert judgment
Conclusions:
The scores from the CAM test measured against known reference materials whose in-vivo eye irritation results are known show that ColaMoist 200 (5% active solution) has little or no irritant effect by the Hen's Egg in-vitro method.