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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP, comparable to guidelines, in vitro method

Data source

Reference Type:
study report
Report date:

Materials and methods

Principles of method if other than guideline:
HET-CAM in vitro corrosion test (GLP)
There are no official guidelines for the HET-CAM Test at date; however, the study was conducted according to the following publications:
- Lüpke N.P. (1985): Hen’s Egg Chorio allantoic Membrane Test for Irritation Potential. Fd. Chem. Toxic. 23, pp. 287 – 291.
- Spielmann, H. (1995): HET-CAM Test. In: Methods in Molecular Biology, 43 (eds.: O’Hare, S. and Atterwill, C. K.) pp. 199 – 204.
- Spielmann, H. et al. (1996): Results of a Validation Study in Germany on Two In Vitro Alternatives to the Draize Eye Irritation Test the HET-CAM Test and the 3T3 NRU Cytotoxicity Test. ATLA 24, pp. 741 – 858.
In addition the study follows the testing strategy for determination of eye irritation/corrosion as given in the following guideline:
- OECD Guideline for Testing of Chemicals No. 405, April 24, 2002 (“Acute Eye Irritation/Corrosion”).
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
hexafluoro-λ⁵-phosphanuide; tris({4-[(4-acetylphenyl)sulfanyl]phenyl})sulfanium
EC Number:
Cas Number:
Molecular formula:
C42 H33 O3 S4 . F6 P
hexafluoro-λ⁵-phosphanuide; tris({4-[(4-acetylphenyl)sulfanyl]phenyl})sulfanium

Test animals / tissue source

other: in vitro [HET-CAM]

Test system

unchanged (no vehicle)
Amount / concentration applied:
- Amount(s) applied (volume or weight with unit): A bulk volume of 25 μL (about 10 mg) per egg of the solid test substance was applied on approximately the half of the membrane area
- Concentration: undiluted
Duration of treatment / exposure:
100 seconds
Number of animals or in vitro replicates:
3 eggs
Details on study design:
- Washing (if done): The test substance was removed by washing with 0.9 % aqueous NaCl-solution. A final assessment of the CAM was performed immediately after washing.
- Time after start of exposure: 100 seconds

- Preparation and opening of the eggs: The eggs were candled on the day of application to ensure viability and in order to mark the location of the air chamber with a felt pen. The eggshell was cut along the marking line with an electric drill and removed exposing the egg membrane, which forms the inner barrier between egg content and air chamber. This membrane was moistened with warm physiological saline and the eggs were then placed in the incubator again until they were used for testing (maximum of 30 minutes between opening the eggs and application).
- Selection of the eggs: After removal of the egg membrane the CAM was investigated for signs of pre-existing damage, which would exclude the egg from the assay. Only eggs with an adequate vascular system and even CAM surface were used for the study.

After application of the test substance the chorioallantoic membrane was observed by means of a stereomicroscope until unambiguous irritation reactions were detected or up to a maximum time period of 100 seconds, respectively.
The evaluation of the reactions was performed according to the following grading:
0 No visible change
1 Slight reaction
2 Moderate reaction
3 Severe reaction

EVALUATION OF RESULTS: The mean time until appearance of reaction over the eggs of a treatment group was calculated (mean time to coagulation = mtc in seconds).

Positive controls: aqueous solution of 0.1 M NaOH and 10 % SDS (Sodium dodecyl sulfate), which are known to cause moderate to severe irritation were tested, in order to demonstrate the sensitivity of the method.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
other: Haemorrhagia
Time point:
other: 100 seconds
Max. score:
Remarks on result:
other: No effects observed
Irritation parameter:
other: Coagulation
Time point:
other: 100 seconds
Max. score:
Remarks on result:
other: No effects observed
Irritant / corrosive response data:
The test substance does not produce changes indicative for serious eye damage in the HET-CAM Test under the test conditions chosen.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Migrated information Criteria used for interpretation of results: other: EU-GHS