Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2016-03-07 until 2016-03-07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report date:
2016

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Version / remarks:
To further establish the classification, histopathological observations were made
on two sections of each of the 3 corneas treated with test item (6 sections).
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
3,9-dimethyl-2,4,8,10-tetraoxa-3,9-diphosphaspiro[5.5]undecane 3,9-dioxide
EC Number:
221-088-9
EC Name:
3,9-dimethyl-2,4,8,10-tetraoxa-3,9-diphosphaspiro[5.5]undecane 3,9-dioxide
Cas Number:
3001-98-7
Molecular formula:
C7H14O6P2
IUPAC Name:
3,9-dimethyl-2,4,8,10-tetraoxa-3lambda5,9lambda5-diphosphaspiro[5.5]undecane-3,9-dione
Test material form:
solid: particulate/powder
Details on test material:
AFLAMMIT® PCO 962

Test animals / tissue source

Species:
chicken
Strain:
other: Strain of chicken: ROSS 308; Source: TARAVIS KFT. 9600 Sárvár, Rábasömjéni út. 129.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes
yes, concurrent vehicle
yes, concurrent positive control
Amount / concentration applied:
Three eyes were treated with 30 mg of powdered test item.
Duration of treatment / exposure:
After 10 seconds, the surface of the eyes was
rinsed with physiological saline solution.
Duration of post- treatment incubation (in vitro):
The control and test eyes were evaluated pre-treatment and at approximately 30, 75,
120, 180 and 240 minutes after the post-treatment rinse.
Details on study design:
At the end of the evaluation procedures, the corneas were carefully removed from the eyes and
placed individually into labelled containers of preservative fluid (10% neutral buffered
formalin, Manufacturer: Reanal, Batch number: KTM14051, Expiry date: October
2017 for potential histopathology [9,10] and stored at room temperature.

Results and discussion

In vitro

Results
Irritation parameter:
other: TEST ITEM
Run / experiment:
Overall ICE Class:1xI 2xII
Vehicle controls validity:
not valid
Negative controls validity:
valid
Remarks:
Overall ICE Class: 3xI
Positive controls validity:
valid
Remarks:
Overall ICE Class: 1xIII, 2xIV
Remarks on result:
other: Based on this in vitro eye irritation assay in the isolated chicken eyes test with AFLAMMIT PCO 962, the test item is not classified as a severe irritant and not classified as non-irritant.
Remarks:
It is concluded that further information is required for classification.I
Other effects / acceptance of results:
HISTOPATHOLOGY
Semi-quantitative microscopic evaluation was performed on the cornea in the ICET.
The classification of histopathology findings was performed based on two publications,
M.K. Prinsen et all. (Toxicology in Vitro 25 (2011), 1475-1479) and Elodie Cazelle et
all/ Toxicology in Vitro 28 (2014), 657-666.
The negative control 0.9 % Sodium chloride (Salsol solution) cornea showed no
abnormalities. Positive control, Imidazole was associated with moderate epithelial
erosion. No compromised Bowman’s or basement membrane as well as no endothelial
changes were recorded.
AFLAMMIT PCO 962 produced very slight vacuolation (15A, 15B) or very slight
erosion (16A) of the corneal epithelium. The 16B, 17A and 17B samples did not show
significant changes No stromal and endothelial changes were observed as well as no
effects on integrity of basement, Bowman’s and Descemet’s membranes.

Applicant's summary and conclusion

Interpretation of results:
other: CLP criteria are not met.
Conclusions:
Based on this in vitro eye irritation assay in the isolated chicken eyes test with AFLAMMIT PCO 962, the test item is not classified as a severe irritant. It is concluded that further information is required for classification.
To further establish the classification, histopathological observations were made on two sections of each of the 3 corneas treated with test item (6 sections). Microscopic evaluation showed very slight vacuolation (15A and 15B) or very slight erosion (16A) of the corneal epithelium. The 16B, 17A, 17B samples did not show significant changes. No stromal or endothelial changes were observed as well as no effects on integrity of basement, Bowman’s and Descemet’s membranes.
Based on the data the CLP criteria for classification are not met.