Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: No guidline study, but conducted according to GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

Materials and methods

Test guideline
Qualifier:
no guideline available
Guideline:
other: HET-CAM Test
Principles of method if other than guideline:
There are no official guidelines for the HET-CAM Test; 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. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
The analyses of the test substance were carried out at the Analytical Department of BASF Aktiengesellschaft, Ludwigshafen, Germany.
Name of test substance: rac-1-(3-Methoxyphenyl)ethylamine
Test-substance No.: 06/0647-1
Batch identification: VSVU0019/VU0962 Fr 6 + Fr 8 LJ 8248/06
Purity: 99.3 area-% (analytical report No.: 06L00246)
Homogeneity: The test substance was homogeneous by visual inspection.
Storage stability: The stability under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.

ADDITIONAL TEST-SUBSTANCE INFORMATION
pH-value: ca. 7 (undiluted test substance); ca. 6 (10% preparation)

Test animals / tissue source

Species:
other: Chicken
Strain:
other: White Leghorn

Test system

Vehicle:
other: Olive oil Ph.Eur./DAB
Controls:
yes
Amount / concentration applied:
undiluted and 10% solution of the test substance in olive oil Ph.Eur./DAB
Duration of treatment / exposure:
A volume of 0.3 mL per egg of the liquid test substance / testsubstance solution was applied with a syringe and the CAM was observed until unambiguous irritation reactions were detected or up to a maximum time period of 3.5 minutes, respectively. It was possible to observe the reactions through the test substance/test-substance solution. The time until appearance of reactions was recorded.
Number of animals or in vitro replicates:
Number of eggs: 3 (undiluted test substance) and 3 (10% test-substance solution)

Results and discussion

Any other information on results incl. tables

The evaluation depends on the solubility of the test substance, the tested concentrations and the time until appearance of unambiguous intravascular coagulation of middle-sized vessels or extravascular coagulation of the treated CAM, respectively. The mean time until appearance of reaction over the eggs of a treatment group was calculated (mean time to coagulation = mtc in seconds):

mtc(100%) < 90 "risk of serious damage to the eyes"

or

mtc(100%) x 0.019 + mtc(10%) x 0.017 < 4.404 "risk of serious damage to the eyes"

mtc(100%) ≥ 90 "no risk of serious damage to the eyes"

Grading

0 No visible change

1 Slight reaction

2 Moderate reaction

3 Severe reaction

Applicant's summary and conclusion

Conclusions:
Based on the results of this study it was concluded, that rac-1-(3-Methoxyphenyl)ethylamine caused serious eye damage in the HET-CAM Test under the test conditions.
Executive summary:

The potential of rac-1-(3-Methoxyphenyl)ethylamine to cause serious damage to the eyes/mucous membranes was assessed by a single topical application of 0.3 mL of the undiluted test substance and 0.3 mL of a 10% test-substance solution in olive oil Ph.Eur./DAB, to the chorionallantoic membrane (CAM) of fertilized and incubated hen eggs. Three eggs per test-substance concentration were observed until unambiguous irritation reactions were detected or up to a maximum time period of 210 seconds. The occurrence of vascular injury or intravascular coagulation in response to the test substance was recorded. The undiluted test substance caused severe intravascular as well as extravascular coagulation and moderate haemorrhagia in all eggs within 9 – 12 seconds after application. The 10% test substance solution in olive oil Ph.Eur./DAB caused moderate intravascular coagulation in all eggs after 49 – 85 seconds and moderate haemorrhagia in all eggs within 11 – 17 seconds. Based on the results of this study it was concluded, that rac-1-(3-Methoxyphenyl)ethylamine causes serious eye damage in the HET-CAM Test under the test conditions.