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

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Unavailable - study report dated 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

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

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

Three-dimensional reconstructed human epidermis- EPISKIN model kit purchased from SkinEthic Laboratories (Lyon, France).

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount applied: 10 µL

NEGATIVE CONTROL
- Amount applied: 10 µL DPBS

POSITIVE CONTROL
- Amount applied: 10 µL 5% w/v SDS

Duration of treatment / exposure:

15 min

Duration of post-treatment incubation (if applicable):

42 hours

Number of replicates:

not specified

Test system

Details on study design:

Test MTT reduction by the test material: 10 µL test material + 2 mL 0.3 mg/mL MTT solution incubated for 3 h. The blue colour indicates reduced MTT.

MAIN TEST
DAY 1-Application of test item
Exposure: 15 min
Amount applied: 10 µL
Negative control: 10 µL DPBS
Positive control: 10 µL 5% w/v SDS
Washing: DPBS with Ca++ and Mg++
Post-exposure incubation: 42 h

DAY 3-MTT Loading/Formazan Extraction
Plate shaker: 15 min
MTT solution: 2 mL of 0.3 mg/mL per well
Incubation: 3 h
Biopsy of the epidermis was performed. The tissue was then transfered in micro tubes with 500 µL acidified isopropanol and refrigerated till day 6.

DAY 6-Absorbance/Optical Density
at 540 nm with Anthos 2001 microplate reader

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Remarks:

Relative Mean Viability

Value:

6.8

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other:

Remarks:

See below for detailed results.

Other effects / acceptance of results:

- OTHER EFFECTS:
The MTT solution conatining the test item did not turn blue, suggesting that the test material does not reduce directly MTT.

- ACCEPTANCE OF RESULTS:
Acceptance criteria met for negative and positive controls.

Any other information on results incl. tables

Table 1: Mean OD540 values and percentage viabilities for the negative control Item, positive control Item and test item

Item	OD540 of tissues	Mean OD540 of triplicate tissues	± SD of OD540	Relative individual tissue viability (%)	Relative mean viability (%)	± SD of Relative mean viability (%)
Negative Control Item	0.786	0.753	0.029	104.4	100	3.8
	0.737			97.9
	0.735			97.6
Positive Control Item	0.129	0.110	0.039	17.1	14.6	5.2
	0.065			8.6
	0.136			18.1
Test Item	0.032	0.051	0.017	4.2	6.8	2.3
	0.055			7.3
	0.066			8.8

 

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

Under the conditions of this in vitro test benzenesulfonic acid, 4-C10-13-sec-alkyl derivs.-, compd. with 2-propanamine (1:1) is irritating to the skin.

Executive summary:

In a primary dermal irritation study conducted according to OECD Guideline for the Testing of Chemicals No. 439 “In Vitro Skin Irritation” (adopted 22 July 2010), EPISKIN™ reconstructed human epidermis model was topically exposed to 10 µL of undiluted benzenesulfonic acid, 4-C10-13-sec-alkyl derivs.-, compd. with 2 -propanamine (97% purity) for 15 min and 42 h post incubation period. The principle of the assay was based on the measurement of cytotoxicity in the human epidermal cultures following topical exposure to the test item, with the MTT reduction assay. The negative and positive controls confirmed the validity of the study.

The test item is irritating to the skin. The relative mean tissue viability was ≤ 50% (6.8%) after 15 min treatment and 42 h post-incubation. Based on the results, benzenesulfonic acid, 4-C10-13-sec-alkyl derivs.-, compd. with 2-propanamine is classified to be irritating to the skin in accordance with UN GHS "Category 2".

The study is considered acceptable based on the quality criteria.