Registration Dossier

Toxicological information

Eye irritation

Currently viewing:

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
16 to 18 October 2012
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP study, no specific guideline followed (no guideline available for this test)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
The test is based on the hypothesis that irritant chemicals are able to penetrate the corneal epithelial tissue and are sufficiently cytotoxic to cause cell death. Cytotoxicity is measured with the MTT reduction assay.
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with 2-propanamine
EC Number:
284-664-9
EC Name:
Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with 2-propanamine
Cas Number:
84961-74-0
IUPAC Name:
Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs.-, compd. with 2-propanamine (1:1)
Test material form:
liquid: viscous
Details on test material:
- Name of test material (as cited in study report): Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with 2-propanamine (1:1)
- Substance type: technical product (BIO-SOFT 411-E)
- Physical state: viscous liquid
- Analytical purity: 97%
- Lot/batch No.: 50148
- Expiration date of the lot/batch: 30 June 2013
- Storage condition of test material: room temperature in the dark

Test animals / tissue source

Species:
other: not applicable
Details on test animals or tissues and environmental conditions:
SkinEthic reconstructed Human Corneal Epithelium model (HCE, SkinEthic Laboratories, Lyon, France)

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: not applicable
Amount / concentration applied:
see below details on study design
details on negative control (Solution A); compostition for 1 L: Na2HPO4 0.142 g/l, glucose 1.802 g/l, HEPES 7.149 g/l, KCl 0.224 g/l and NaCl 7.597 g/l
positive control: sodium dodecyl sulphate (SDS) 2% w/v
Details on study design:
Pre-test: Test MTT reduction by the test material: 30 ul test material + 1 ml 0.5 mg/ml MTT solution incubated for 3 h. The blue colour indicates reduced MTT.
MAIN TEST
7-day old tissues were transferred into wells and treated with 30 ul test item
Exposure period: 10 min
Negative control: 30 ul solution A
Positive control: 30 ul of 2% w/v SDS
Washing: DPBS without Ca++ and Mg++
Placement of each tissue into a 24-well plate with 300 ul maintenance medium
MTT loading plates: 300 ul of a 0.5 mg/ml MTT solution
Incubation: 3 hours
Washing: phosphate buffered saline
Isopropanol was applied for the formation of formazan crystals
Optical density (OD) was measured at 540 nm with Anthos 2001 microplate reader

Tissue histopathology: one tissue/treatment group was kept for histopathological examination

Results and discussion

In vitro

Results
Irritation parameter:
other: mean viability %
Value:
ca. 16.6
Remarks on result:
positive indication of irritation

Any other information on results incl. tables

Table 1: Assessment of Eye Irritation Potential- Viability of HCE tissues

Item

OD540of Individual Tissue

Mean OD540

Relative Mean Viability (%)

Negative Control

0.965

0.926

100

0.887

Positive Control

0.022

0.022

2.4

0.021

Test item

0.141

0.154

16.6

0.167

 

Tissue histopathological examination was not performed cause it was not considered necessary.

Applicant's summary and conclusion

Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: other: criteria of the test
Conclusions:
Based on the results of the study, BIO-SOFT 411-E and hence, benzenesulfonic acid, 4-C10-13-sec-alkyl derivs.-, compd. with 2-propanamine (1:1), are both eye irritants.
Executive summary:

The purpose of this study was to determine the eye irritation potential of BIO-SOFT 411 -E using the SkinEthic reconstructed Human Corneal Epithelium model (HCE, SkinEthic Laboratories, Lyon, France) after a treatment period of 10 minutes. The test is based on the hypothesis that irritant chemicals are able to penetrate the corneal epithelial tissue and are sufficiently cytotoxic to cause cell death. Cytotoxicity is measured with the MTT reduction assay.

Triplicate tissues were exposed to the test material and thereafter, to MTT. The optical density was measured at 540 nm.Data were presented in the form of percentage viability (MTT conversion relative to negative controls).The relative mean viability of the test item treated tissues after a 10 min exposure period was 16.6 %, i.e. the test the test item is an eye irritant.