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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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
other: in vitro
Type of information:
experimental study
Adequacy of study:
key study
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well-documented study report which meets basic scientific principles. Study was carried out with reference to selected rules of GLP

Data source

Reference Type:
study report

Materials and methods

Test guideline
according to guideline
other: HET-CAM Test
Principles of method if other than guideline:
With the HET-CAM the irritation property of a test item after contact with mucous membranes or the eye can be determined based on the occurrence of alterations at the CAM and the adjacent capillary system of fertilized and incubated chicken eggs.
The chorioallantoic membrane (CAM) is a vital vascular membrane with a closed blood vessel system. After application of a test item on the membrane the underlying blood vessels could become damaged. In particular three events are observed:
- hemorrhage (H) (bleeding out from the vessels of the vascularized CAM)
- vessel lysis (L) (disappearance of small vessels)
- protein coagulation (C) (intravasal or extravasal)
Two different methods can be used depending on the transparency of the test item, the reaction-time method or the end-point assessment. Since 2-phenylethyl cyanoacetate is an intransparent solid substance, the end-point assessemnt was applied.

The study procedures were based on the following document:
- The Ocular Toxicity Working Group (OTWG) of the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) and the National Interagency Centre for the Evaluation of Alternative Toxicological Methods (NICEATM), Background Review Document (BRD): current status of in vitro test methods for identifying ocular corrosives and severe irritants: The Hen’s Egg Test - Chorioallantoic Membrane (HET-CAM) test method, March 2006.
GLP compliance:

Test material

Constituent 1
Chemical structure
Reference substance name:
2-phenylethyl 2-cyanoprop-2-enoate
EC Number:
Cas Number:
Molecular formula:
C12 H11 N O2
2-phenylethyl 2-cyanoprop-2-enoate
Constituent 2
Reference substance name:
[TN]PhECA[/TN][SPEC][/SPEC][AM] > 98.8 % [/AM]
[TN]PhECA[/TN][SPEC][/SPEC][AM] > 98.8 % [/AM]
Test material form:
solid: crystalline
Details on test material:
2-Phenylethyl cyanoacrylate, Batch 3635-59, > 98.8 %

Test animals / tissue source

other: HET-CAM
other: not applicable

Test system

unchanged (no vehicle)
other: not applicable
Amount / concentration applied:
300 mg test item
0.3 ml positive control (1% sodium dodecylphosphate prepared in physiological saline) and negative control (physiological
Duration of treatment / exposure:
300 s
Observation period (in vivo):
5 m
Number of animals or in vitro replicates:
Details on study design:
Fertilized eggs were incubated until day 9, then the air cell of the eggs were marked and cut off with a rotating saw blade. The uncovered inner membrane was moistened with physiological saline and subsequently the egg was kept at room temperature. Subsequently the physiological saline was removed and the inner egg membrane carefully eliminated using forceps to expose the CAM.
Three eggs were treated for 300 seconds with 300 mg 2-phenylethyl cyanoacrylate or for 30 seconds with 0.3 ml Texapon ASV (5% active substance) as a positive controle. After the treatment period the eggs were carefully rinsed with approximately 5 ml of physiological saline. The appearance of haemorrhage, vessel lysis and coagulation on the CAM was monitored and recorded over the following 280-second period.
Results for the three test method endpoints were evaluated separately for each egg and the S-score was calculated.

Classification scheme of the HET-CAM End-Point Assessement
S [sum of single value] Evaluation
0 – 5 slightly irritating
6 – 12 moderately irritating
13 – 15 irritating
16 – 18 severely irritating (R 41)

Results and discussion

In vivo

Irritation parameter:
other: HET-CAM S-score
Time point:
other: 600 s
Max. score:
not specified
Remarks on result:
other: individual scores: see below
Other effects:
no other effects

Any other information on results incl. tables

Test results:

Test substance

Irritation value




Assessment [S]


PhECA, Charge 3635-59



slightly irritating

Reference item

Texapon ASV, 5% AS


moderately irritating


It was observed that the test substance partially polymerized on the CAM, leaving a sticky cover that could not be rinsed but removed with forceps. Removing the substance sheet like this did not result in a problematic damage of the CAM.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Migrated information Criteria used for interpretation of results: other: ICCVAM
The mean in vitro irritancy score obtained with the positive control (Texapon ASV) was within the historical control data range. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.
The mean in vitro irritancy score obtained after a treatment period of 300 seconds and an observation period of 280 seconds with 2-phenylethyl cyanoacrylate was 0.
Finally, it is concluded that this test is valid and that 2-phenylethyl cyanoacrylate is not irritant in the Hen’s Egg Test – Chorioallantoic Membrane Test under the described experimental conditions.
Executive summary:

The possible ocular irritancy of 2 -phenylethyl cyanoacrylate was tested with an HET-CAM through topical application for 300 seconds using the endpoint assessment method. The test substance was applied undiluted (300 mg) directly on top of the chorioallantoic membrane.

The mean S-score obtained with the positive control (5 % Texapon ASV) was 11 and within the historical control data range. It was therefore concluded that the test conditions were adequate and that the test system functioned properly. The mean S-score obtained with 2 -phenylethyl cyanoacrylate was 0.

Thus, it is concluded that this test is valid and that 2 -phenylethyl cyanoacrylate is not irritant in the Hen’s Egg Test – Chorioallantoic Membrane Test under the experimental conditions described in this report.