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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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
Study period:
1 (reliable without restriction)

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
Diethyl ethylphosphonate
EC Number:
EC Name:
Diethyl ethylphosphonate
Cas Number:
Molecular formula:
diethyl phosphonate
Test material form:

Test animals / tissue source

other: slaughterhouse Müller Fleisch GmbH, Enzstr. 2-4, 75217 Birkenfeld, Germany
Details on test animals or tissues and environmental conditions:
The cattle were between 12 and 60 months old. The eyes were transported to the test facility in Hanks’ Balanced Salt Solution with 1% Penicillin-Streptomycin solution (Penicillin 100 U/mL, Streptomycin 100 µg/mL) in a suitable cooled container.

Test system

not specified
Amount / concentration applied:
750 µL
Duration of treatment / exposure:
10 minutes at 32 ± 1 °C
Duration of post- treatment incubation (in vitro):
2 hours at 32 ± 1 °C

Results and discussion

In vitro

Irritation parameter:
in vitro irritation score
ca. 68.18
Negative controls validity:
Positive controls validity:
Other effects / acceptance of results:
In the negative control, no signs of eye irritation were observed.
The positive control induced serious eye damage, which would be classified as GHS category I.
The test item Diethyl ethylphosphonate induced serious eye damage on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) is 68.18.

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
This in vitro study was performed to assess corneal damage potential of Diethyl ethylphosphonate by quantitative measurements of changes in opacity and permeability in a bovine cornea.
The test item Diethyl ethylphosphonate was brought onto the cornea of a bovine eye which previously had been incubated with cMEM without phenol red at 32 ± 1 °C for 1 hour and whose opacity had been determined. The test item was incubated on the cornea for 10 minutes at 32 ± 1 °C. After removal of the test item and 2 hours post-incubation, opacity and permeability values were measured.

The test item was tested pure.
Under the conditions of this test, the test item Diethyl ethylphosphonate induced serious eye damage on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) is 68.18.
According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS > 55 induces serious eye damage, that should be classified as UN GHS Category I.

The negative control (HBSS) and the positive control (undiluted dimethylformamide) have met the validity criteria.

No observations were made which might cause doubts concerning the validity of the study outcome. The test is considered valid.