1,4:3,6-dianhydro-2,5-bis-O-(diphenoxyphosphoryl)-D-glucitol

Endpoint summary

Description of key information

- Skin irritatio/ corrosion: OECD TG 431 & 439 (BASF SE, 2011; in vitro), OECD TG 402 (BASF SE, 2011; rats); not corrosive, not irritating to skin.
- Eye Irritation: OECD TG 437 (BASF SE, 2011), EpiOcularTM test (BASF SE, 2011); no serious eye damage, not irritating to the eye.

Additional information

Skin irritation / corrosion

- In vitro tests

The potential of the test substance to cause dermal corrosion/irritation was assessed by a single topical application of 25 μL bulk volume (about 19 mg) of the test substance to a reconstructed three dimensional human epidermis model (EpiDerm™), according to the OECD TG 431 & 439 (BASF SE, 2011).

For the corrosion test 2 EpiDerm™ tissue samples were incubated with the test substance for 3 minutes and 1 hour, respectively. The irritation test was performed with 3 EpiDerm™ tissue samples, which were incubated with the test substance for 1 hour followed by a 42-hours post-incubation period. Tissue destruction was determined by measuring the metabolic activity of the tissue after exposure/post-incubation in a colorimetric test, using the reduction of mitochondrial dehydrogenase activity (measured by reduced formazan production after incubation with a tetrazolium salt [MTT] in comparison to the negative control tissues) as endpoint. The quotient of the test substance and negative control values indicates the relative tissue viability.

Following results were obtained: (1) the test substance is not able to reduce MTT directly; (2) in the corrosion test; the mean viability of the test-substance treated tissues was 103% after 3 min exposure, and 106% after 1 hour exposure; (3) in the irritation test; the mean viability of the test-substance treated tissues was 100% after 1 hour exposure and ca. 42 hours post-incubation.

This Test may be used for the hazard identification of corrosive (OECD TG 431) or irritant (OECD TG 439) chemicals in accordance with UN GHS and EU CLP Category 2. Based on the observed results and applying the evaluation criteria, the test substance does not show a skin irritation potential in the EpiDerm™ skin corrosion/irritation test under the test conditions chosen. Therefore, the test substance is not irritating according to EU CLP. Additionally taken into account that no irritation reaction was observed in the acute demal toxicity study conducted according to the OECD TG 402 in rats up to a dose level of 2000 mg/kg bw, the likelihood of a mildly irritating potential also can be excluded (the test substance is therefore not irritating according to GHS).

- In vivo tests

No data available 

 

Eye irritation

- In vitro tests

In a first in vitro study conducted according to the OECD TG 437 (BASF SE, 2011), the potential of the test substance to cause serious damage to the eyes was assessed by a single topical application of 750 μL of a 20% test substance preparation in highly de-ionized water to the epithelial surface of isolated bovine corneas, for an exposure period of 4 hours (Val 1).

Corneal opacity was then measured quantitatively as the amount of light transmission through the cornea, as well as permeability as the amount of sodium fluorescein dye that passes across the full thickness of the cornea. Both measurements were further used to calculate an In Vitro Irritancy Score (IVIS = mean opacity value + 15 * mean permeability OD value) of the test substance relative to the control corneas. A risk of serious damage to the eyes is predicted if the IVIS per treatment group was greater than 55, and no risk of serious damage to the eyes was predicted otherwise (if the IVIS per treatment group is ≤55. The calculated mean IVIS was 7.5, 2.5 and 120.5, respectively in the test substance group, the negative control group and the positive control group. Based on the observed results and applying the evaluation criteria, the test substance does not cause serious eye damage in the Bovine Corneal Opacity and Permeability Test (BCOP Test) under the test conditions chosen.

 

The potential of the test substance to cause eye irritation was further evaluated in a second study, also conducted following the testing strategy for determination of eye irritation/corrosion as given in the OECD TG 405 and using the three dimensional human cornea model, based on the observation that irritant chemicals produce cytotoxicity in human reconstructed cornea after a short term topical exposure (BASF SE, 2011). In the well conducted GLP study (Val 1), 50 μL bulk volume (about 25 mg) of the test substance was applied to a reconstructed three dimensional human cornea model (EpiOcular^TM; 2 tissue sample per treatment) and the treated tissue was incubated for 90 minutes, washed out, and post-incubated under normal medium and culture conditions for 18 hours. Tissue destruction was then determined by measuring the metabolic activity of the tissue after exposure/post-incubation using a colorimetric test. The reduction of mitochondrial dehydrogenase activity, measured by reduced formazan production after incubation with a tetrazolium salt (MTT) was chosen as endpoint. The formazan production of the test substance treated epidermal tissues is compared to that of negative control tissues. The quotient of the values indicates the relative tissue viability. The test substance was considered "irritant", if the mean relative tissue viability with a test material is less than or equal to 50%, “non-irritant” or inconclusive otherwise.

The test substance was not able to reduce MTT directly, and the mean viability of the test-substance treated tissues was 123%. Therefore, as all quality criteria of the test were met, it was considered to be valid despite the high %-viability determined for the test-substance treated tissues. Based on the observed results and applying the evaluation criteria, it was concluded, that the test substance does not show an eye irritation potential in the EpiOcular™ eye irritation test under the test conditions chosen.

 

- In vivo tests

No data

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available experimental test data are considered reliable and suitable for the purpose of classification (skin irritation). Based on the criteria for classification of Directive 67/548/EEC, as amended for the 28th time in Directive 2001/59/EC, no classification is warranted for skin and eye irritation.

 

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for the purpose of classification (skin irritation). No classification is warranted for skin and eye irritation.