Phenol, paraalkylation products with C12-rich branched olefins derived from propene oligomerisation, reaction products with sulphur monochloride and decene, reaction products with Benzoic acid, 2-hydroxy-,C14-18 alkyl dervis., polybutenyl benzenesulphonic acid, carbon dioxide and calcium hydroxide

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation / corrosion

Remarks:

other: In Vitro study conducted in accordance with OECD 439 guidance

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 January 2012 to 01 June 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in accordance with current guidelines and GLP compliant.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD (2010), In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method, OECD Guidelines for the Testing of Chemicals No. 439, OECD, Paris.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Phenol, paraalkylation products with C12-rich branched olefins derived from propene oligomerisation, reaction products with sulphur monochloride and decene, reaction products with Benzoic acid, 2-hydroxy-,C14-18 alkyl dervis., polybutenyl benzenesulphonic acid, carbon dioxide and calcium hydroxide

EC Number:

903-162-9

IUPAC Name:

Phenol, paraalkylation products with C12-rich branched olefins derived from propene oligomerisation, reaction products with sulphur monochloride and decene, reaction products with Benzoic acid, 2-hydroxy-,C14-18 alkyl dervis., polybutenyl benzenesulphonic acid, carbon dioxide and calcium hydroxide

Details on test material:

- Name of test material (as cited in study report): EC-903-162-9
- Physical state: liquid
- Expiration date of the lot/batch: 19 December 2013
- Storage condition of test material: Ambient
- Lot/batch No.: LN07006926

*EC 903-162-9 is exclusively synthesised and handled in solvent oil. Therefore testing was conducted on a sample that contained 40.8% Base oil and 59.2% EC 903-162-9.

Results and discussion

In vivo

Results

Remarks on result:

other: EpiSkin viability of 103.09% ±13.43% (mean of triplicate values). EC 903-162-9 was demonstrated to be non-irritant when tested in the EpiSkin in vitro irritation assay.

Irritant / corrosive response data:

EpiSkin viability of 103.09% ±13.43% (mean of triplicate values)

Any other information on results incl. tables

MTT Direct Reduction Test (Preliminary Assay)

 

The test was scored by visual assessment of the formation of purple-coloured formazan. The negative control (PBS) did not reduce MTT to formazan. The positive control (eugenol) and EC 903-162-9 were reduced MTT to formazan.

 

EpiSkin^®Irritation Test

 

Percentage Viability of EpiSkin^® Tissues After ca 41h Recovery Time.

Treatment	Mean viability per tissue (%)	Mean viability per test item (%)	SD (%)
EC 903-162-9 (corrected)	91.31	103.09	13.43
	100.26
	117.71
Aqueous SDS Solution (5%, w/v) (Positive Control)	12.82	13.22	0.35
	13.39
	13.44
PBS Solution (Negative Control)	105.31	100.00	7.75
	91.10
	103.59

Negative Controls

 

The negative control results were similar for the three viable EpiSkin^®units dosed with

Dulbecco’s PBS. Exposure to Dulbecco’s PBS resulted in a mean EpiSkin^®viability of

100.00% ±7.75%.

 

Positive Controls

 

The positive control results were similar for the three viable EpiSkin^® units dosed with

aqueous SDS solution (5%, w/v). Exposure to aqueous SDS solution (5%, w/v) resulted in amean EpiSkin^® viability of 13.22% ±0.35%.

 

EC 903-162-9

 

Non-Viable Controls

 

The mean absorbance value of the three replicate non-viable tissues dosed with EC 903-162-9 was 0.148 ± 0.034. The mean absorbance value of the three replicate un-dosed non-viable tissues was 0.098±0.016. Therefore, the absorbance value for the effect of the un-removed test item was 0.050. This value was subtracted from the absorbance value of the viable tissues dosed with EC-903-162-9.

 

Viable Tissues

 

The results were similar for the three viable EpiSkin^®units dosed with EC 903-162-9.

Exposure to EC 903-162-9 resulted in a mean EpiSkin^®viability (corrected for effect of un-removed test item) of 103.09% ± 13.43% of the negative control value.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

In conclusion, EC 903-162-9 was demonstrated to be non-irritant when tested in the EpiSkin in vitro irritation assay.

Executive summary:

Evaluation of skin irritation is part of the Human Health Hazard Assessment required for registration of a chemical. In this study, the irritation potential of EC 903-162-9 was evaluated using the SkinEthic EpiSkin in vitro irritation assay. Prior to the conduct of the irritation assay, a preliminary test was conducted to assess the intrinsic ability of the test item to reduce methylthiazoldiphenyl-tetrazolium bromide (MTT) to formazan. The test item was capable of MTT reduction. Therefore, non-viable control tissues were dosed in parallel with the irritation assay to quantify this effect and the results were corrected accordingly. The dermal irritation potential was assessed by applying an aliquot ca 10 µL of EC 903-162-9 to the exposed surface of three EpiSkin reconstructed human epidermis (RhE) units for 15 min. The surface area of the EpiSkin was 0.38 cm², therefore the application rate was 26.3 µL/cm². After the 15 min exposure period, the test item was washed from the surface of the EpiSkin using Dulbecco’s phosphate-buffered saline (PBS) and tissue swabs. The EpiSkin was then incubated for a recovery period of ca 41 h in a humidified incubator set to maintain temperature and CO₂ levels of 37°C and 5%, respectively. Following incubation, the EpiSkin units were transferred to assay medium containing MTT (0.3 mg/mL) and returned to the incubator for 3 h. Biopsies of the EpiSkin membranes were then removed, added to acidified isopropanol, and refrigerated for ca 69 h in order to extract the formazan. The formazan production (cell viability) was assessed by measuring the optical density of the extracts at a wavelength of 550 nm. Three replicates of the positive control, aqueous sodium dodecyl sulphate (SDS) solution (5%, w/v) (10 µL), and the negative control, PBS (10 µL) were tested in parallel to demonstrate the efficacy of the assay. The viability of each individual EpiSkin tissue was calculated as a percentage of the mean negative control viability (defined as 100%). Exposure to EC-903-162-9 resulted in a mean EpiSkin viability of 103.09% ± 13.43% of the negative control value. Exposure to the positive control, aqueous SDS solution (5%, w/v), resulted in a mean EpiSkin viability of 13.22% ± 0.35% of the negative control value. In conclusion, EC 903-162-9 was demonstrated to be non-irritant when tested in the EpiSkin in vitro irritation assay.