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

Endpoint summary

Description of key information

Two in vitro tests were performed in order to assess the skin corrosive/irritating potential of LAS MIPA. Both studies gave a negative result, and hence the substance is not irritating to the skin.  The in vitro eye irritating test suggests that the substance is irritating to the eye. However, since the test is not so far officially accepted for regulatory purposes, it is proposed that the test material is classified as damaging to the eye.

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October 2012

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP study. No specific guideline followed.

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)

Species:

other: not applicable

Details on test animals or tissues and environmental conditions:

SkinEthic reconstructed Human Corneal Epithelium model (HCE, SkinEthic Laboratories, Lyon, France)

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 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

Remarks on result:

not measured/tested

Other effects / acceptance of results:

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

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

Item	OD540of Individual Tissue	Mean OD540	Relative Mean Viability (%)
Negative Control	0.727	0.698	100
	0.669
Positive Control	0.019	0.026	3.7
	0.033
Test item	0.058	0.058	8.3
	0.057

 

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

Interpretation of results:

irritating

Remarks:

Migrated information irritating Criteria used for interpretation of results: other: criteria of the test

Conclusions:

Based on the results of the study, Marlon AMI 80 and hence, benzenesulfonic acid, 4-C10-13-sec-alkyl derivs.-, compd. with isopropanolamine (1:1), are both eye irritants.

Executive summary:

The purpose of this study was to determine the eye irritation potential of Marlon AMI 80 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 8.3%, i.e. the test the test item is an eye irritant.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

The potential skin effects of MARLON AMI 80 (78% benzenesulfonic acid, 4-C10-13-sec-alkyl derivs.-, compd. with isopropanolamine (1:1) in  1,2-propylene glycol) were tested in two in vitro studies with the use of reconstructed human epidermis models (EPISKIN^TM). The principle of the assays is based on the measurement of cytotoxicity in the human epidermal cultures following topical exposure to the test material, with the MTT reduction assay. Both test gave negative results, suggesting that MARLON AMI 80, and hence, LAS MIPA, has no skin corrosive or irritating properties.

 

Eye irritation

An in vitro eye irritation test was performed with the use of the SkinEthic Human Corneal Epithelium model. Currently, the aforementioned test is not accepted for regulatory purposes. Therefore, the classification of the test item cannot be based solely on this test. Since no additional information is available on the eye irritation potential of benzenesulfonic acid, 4-C10-13-sec-alkyl derivs.-, compd. with isopropanolamine (1:1), the substance shall be classified and labelled as corrosive to the eye.

Justification for selection of skin irritation / corrosion endpoint:

Weight of evidence approach applied

Justification for selection of eye irritation endpoint:

Only one study available

Effects on eye irritation: corrosive

Justification for classification or non-classification

According to the classification criteria laid down on EC Regulation 1272/2008, LAS MIPA shall not be classified for skin irritation. The in vitro eye irritation test suggests that the substance is irritating to the eye. However, hitherto the test is not officially accepted for regulatory purposes, and hence, it is proposed that LAS MIPA is classified as damaging to the eye, Eye Dam.1, H318.