Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with 2-propanamine

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2012-10-16 to 2013-01-15

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

no guideline followed

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

Test material information

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

Molecular formula:

N/A

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

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:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 30 µL

Duration of treatment / exposure:

10 min

Number of animals or in vitro replicates:

2

Details on study design:

Pre-test: Test MTT reduction by the test material: 30 µL 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 µL test item
Exposure period: 10 min
Negative control: 30 µL solution A
Positive control: 30 µL of 2% w/v SDS
Washing: DPBS without Ca++ and Mg++
Placement of each tissue into a 24-well plate with 300 µl maintenance medium
MTT loading plates: 300 µL 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

Results 1

Irritation parameter:

other: OD at 540 nm

Run / experiment:

1

Value:

ca. 0.141

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 2

Irritation parameter:

other: OD at 540 nm

Run / experiment:

2

Value:

ca. 0.167

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 3

Irritation parameter:

other: mean percent tissue viability  (migrated information)

Run / experiment:

mean of 2 tissues

Value:

ca. 16.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

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 because it was not considered necessary.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

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:

In anin vitro eye irritation study, 30 µL of BIO-SOFT 411-E and hence, benzenesulfonic acid, 4-C10-13-sec-alkyl derivs.-, compd. with 2-propanamine (1:1) (97% purity) was applied in the SkinEthic reconstructed Human Corneal Epithelium model (HCE, SkinEthic Laboratories, Lyon, France) for 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 was 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 controls confirmed the validity of the study.

The test item showed irritant effects. The mean relative tissue viability of the three replicates was < 60% (16.6% ). Therefore, the test item is considered to be irritating to the eyes in accordance with UN GHS “Category 2”.