Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
05 May 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guideline
Qualifier:
according to guideline
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)
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
N,N'-[dithiobis[2-(2-hydroxyethyl)-1-methylvinylene]]bis[N-[(4-amino-2-methyl-5-pyrimidinyl)methyl]formamide
EC Number:
200-644-4
EC Name:
N,N'-[dithiobis[2-(2-hydroxyethyl)-1-methylvinylene]]bis[N-[(4-amino-2-methyl-5-pyrimidinyl)methyl]formamide
Cas Number:
67-16-3
Molecular formula:
C24H34N8O4S2
IUPAC Name:
N-[(4-amino-2-methylpyrimidin-5-yl)methyl]-N-[(2E)-3-{[(2E)-2-{N-[(4-amino-2-methylpyrimidin-5-yl)methyl]formamido}-5-hydroxypent-2-en-3-yl]disulfanyl}-5-hydroxypent-2-en-2-yl]formamide
Test material form:
solid: particulate/powder
Remarks:
Pale yellow powder
Details on test material:
Batch : 18117240
Dose calculation will not be adjusted to purity
Specific details on test material used for the study:
Batch N°18117240
CAS No.: 67-16-3
Purity: 98.2 %, dose calculation was not adjusted to purity
Appearance: Pale yellow powder

Test animals / tissue source

Species:
cattle
Details on test animals or tissues and environmental conditions:
Freshly isolated bovine cornea (at least 9 month old donor cattle)

Test system

Vehicle:
physiological saline
Remarks:
The test item was tested as a 20% suspension (w/v) in saline.
Amount / concentration applied:
The test item was tested as a 20% suspension (w/v) in saline.
Duration of treatment / exposure:
The incubation time lasted 240 minutes.
Number of animals or in vitro replicates:
Negative Control : 3
Positive Control : 3
Test Item : 3

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Remarks:
Mean IVIS
Value:
0
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation

In vivo

Irritant / corrosive response data:
Not categorized

Any other information on results incl. tables

Results after 240 Minutes Incubation Time

Test Group

Opacity value =

Difference (t240-t0) of Opacity

Permeability at 490 nm (OD490)

IVIS

Mean IVIS

Proposedin vitroIrritancy Score

 

 

Mean

 

Mean

 

 

 

Negative Control

0

0.00

0.079

0.078

1.19

1.17

Not categorized

0

0.079

1.19

0

0.076

1.14

Positive Control

93.00*

0.177*

95.66

109.92

Category 1

116.00*

0.181*

118.72

111.00*

0.292*

115.38

5007 TDS

HYDRATE SEC

-1.00*

0.011*

-0.84

0.00

Not categorized

0.00*

0.006*

0.09

0.00*

0.043*

0.65

*corrected values 

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
In conclusion, according to the current study and under the experimental conditions reported, 5007 TDS HYDRATE SEC is not categorized (GHS).
Executive summary:

This in vitro study was performed to assess the corneal damage potential of 5007 TDS HYDRATE SEC by means of the BCOP assay using fresh bovine corneae.

After a first opacity measurement of the fresh bovine corneae (t0), the 20% (w/v) suspension in saline (0.9% (w/v) NaCl in deionised water) of the test item 5007 TDS HYDRATE SEC, the positive and the negative controls were applied to corneae fixed in an incubation chamber in horizontal position and incubated for 240 minutes at 32 ± 1 °C. The posterior chamber contained incubation medium. After the incubation phase the test item, the positive, and the negative controls were each rinsed from the corneae and opacity was measured again (t240).

After the opacity measurements permeability of the corneae was determined by measuring spectrophotometrically the transfer of sodium fluorescein after incubation in a horizontal position for 90 minutes at 32 ± 1 °C.

With the negative control (physiological saline) neither an increase of opacity nor permeability of the corneae could be observed.

The positive control (10% (w/v) Benzalkonium chloride in saline) showed clear opacity and distinctive permeability of the corneae corresponding to a classification as serious eye damage (CLP/EPA/GHS (Cat 1)).

Relative to the negative control, the test item 5007 TDS HYDRATE SEC did not cause any increase of the corneal opacity or permeability. The calculated mean in vitro irritancy score was 0.00. According to OECD 437 (see table in chapter 3.8.3) the test item is not categorized (GHS).