Benzene, di-C10-14-alkyl derivs., sulfonated, sodium salts

Endpoint summary

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation - in vitro

A study was conducted to determine the in vitro skin irritation potential of the test substance according to OECD Guideline 439 and EU Method B.46 (Reconstructed Human Epidermis Test Method), in compliance with GLP. Human three dimensional epidermal models (triplicates) were exposed to 25 µL undiluted test substance for 15 min. After a 42 h post-incubation period, a determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity was expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment. Skin irritation was expressed as the remaining cell viability after exposure to the test substance. The positive control (25 µL 0.5% SDS) had a mean cell viability of 16% after 15 ± 0.5 min exposure. The absolute mean OD570 (optical density at 570 nm) of the negative control (25 µL PBS) tissues was within the laboratory historical control data range. The relative mean tissue viability obtained after 15 ± 0.5 min treatment with the test substance compared to the negative control tissues was 87%. The standard deviation value of the percentage viability of three tissues treated identically was less than 16%, indicating that the test system functioned properly. The experiment was considered valid. Since the mean relative tissue viability for the test substance was above 50% after 15 ± 0.5 min treatment, the test substance was considered to be non-irritant to human skin under the study conditions (Eurlings, 2016).

Eye irritation - in vitro

A study was conducted to determine the in vitro eye irritation potential of the test substance according to OECD Guideline 437 (bovine corneal opacity and permeability (BCOP) test), in compliance with GLP. Bovine corneas were exposed to the test substance for a period of 10 min (followed by a post-incubation of 2 h with fresh medium). Opacity and permeability were measured and an in vitro irritancy score was then established. Physiological saline and ethanol were used as negative and positive controls, respectively. The corneas treated with the test substance showed opacity values ranging from -0.7 to 3.7 and permeability values ranging from 0.241 to 0.604. The corneas were clear (two out of three) or slightly translucent with a test substance spot (one out of three) after the 10 min of treatment with the test substance. No pH effect of the test substance was observed on the rinsing medium. The in vitro irritancy scores ranged from 3.4 to 12.8 after 10 min of treatment. The individual in vitro irritancy scores ranged from -1.3 to -0.4 for the negative control and from 53 to 73 for the positive control. The corneas treated with the positive control were turbid after the 10 min of treatment. The negative and positive control values were within the expected range; hence the experiment was considered valid. Under the study conditions, the test substance induced ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of 7.5 after 10 min of treatment. Since the test substance induced an IVIS > 3 ≤ 55, no prediction on the classification could be made (Eurlings, 2016).

A study was conducted to determine the in vitro eye irritation potential of the test substance according to OECD Guideline 492 (EpiOcular™ Cornea Epithelial Model), in compliance with GLP. The EpiOcular tissue construct is a nonkeratinized epithelium prepared from normal human keratinocytes. It models the cornea epithelium with progressively stratified, but not cornified cells. The test consisted of an application of the test substance (50 µL) to the surface of the cornea epithelial construct for 30 min. After exposure, the cornea epithelial construct was thoroughly rinsed to remove the test substance and transferred to fresh medium for an immersion incubation. After transfer to fresh medium for a 2 h incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) at the end of the treatment (mean absorption measurement - optical density reading at 570 nm).  Eye hazard potential is expressed as the remaining cell viability after exposure to the test substance. The test substance showed colour interference in aqueous conditions. In addition to the normal procedure, two tissue were treated with test substance. Instead of MTT solution these tissues were incubated with assay medium. The non-specific colour of the test substance was 0.43% of the negative control tissues. True tissue viability was calculated as the percent tissue viability obtained with living tissues exposed to the interfering test chemical and incubated with MTT solution minus the percent non-specific colour obtained with living tissues exposed to the interfering test chemical and incubated with medium without MTT. The relative mean tissue viability obtained after 30 ± 2 min treatment with the test substance compared to the negative control tissues was 54%. Since the relative tissue viability for the test substance was spread over two categories (65 and 43%, respectively). The experiment was repeated. In the repeat experiment, the non-specific colour of the test substance was 1.00% of the negative control tissues. The OD of the treated tissues without MTT assay was subtracted from the ODs of the test substance treated viable tissues with MTT assay. The relative mean tissue viability obtained after 30 ± 2 min treatment with the test substance compared to the negative control tissues was 66%. Again, the relative tissue viability for the test substance was spread over two categories (55 and 77%, respectively). In both tests equivocal results were obtained after 30 min treatment. The positive control had a mean cell viability of 15 and 4% after 30 ± 2 min exposure, in the initial and repeat experiments, respectively. The absolute mean OD570 values (optical density at 570 nm) of the negative control tissues were within the acceptability range from > 0.8 to < 2.5. The tests functioned properly. Under the study conditions, no conclusion could be made about the eye hazard potential of the test substance (Verbaan, 2017).

Eye irritation - in vivo

A study was conducted to determine the in vivo eye irritation potential of the test substance according to OECD Guideline 405, EU Method B.5 and EPA OPPTS 870.2400, in compliance with GLP. A single sample of 0.1 mL of the test substance were instilled into one eye of one male New-Zealand rabbit. Observations (corneal opacity, iris irritation and conjunctivae redness and chemosis) were made 1, 24, 48 and 72 h and 7, 14 and 21 d after instillation. Mortality, systemic toxicity and body weight were also recorded. Instillation of approximately 0.1 mL of the unchanged test substance resulted in effects on the cornea, iris and conjunctivae. The corneal injury consisted of opacity and epithelial damage. As a result of the corneal injury, pannus (neovascularization of the cornea) was apparent from 7 d after instillation onwards. The corneal injury did not resolve within 21 d. Iridial irritation was observed and resolved within 48 h. The irritation of the conjunctivae consisted of redness, chemosis and discharge and did not completely resolve within 21 d. There was no evidence of ocular corrosion. No staining of (peri) ocular tissues by the test substance was observed and no test substance remnants were seen. No signs of systemic toxicity were observed in the animal during the test period and no mortality occurred. Based on the severity and persistence of the effects until termination in the first animal, there was no need to dose two additional animals. Under the study conditions, the test substance induced severe irritation to the rabbit eye (van Sas, 2017).

Justification for classification or non-classification

Skin irritation

Based on the results of an in vitro study in Reconstructed Human Epidermis, the substance does not require classification for skin irritation according to CLP (Regulation 1272/2008) criteria.

Eye irritation

Based on the results of an in vivo eye irritation study in rabbit, the substance requires classification as Eye Damage 1 - H318 (Causes serious eye damage) for eye irritation according to CLP (Regulation 1272/2008) criteria.​