Reaction mass of biphenyl-2-yl diphenyl phosphate and triphenyl phosphate and bis(biphenyl-2-yl) phenyl phosphate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: EU-Method B.46

Deviations:

no

Principles of method if other than guideline:

- Commission Regulation (EC) No. 761/2009, Method B.46: "In Vitro Skin Irritation: Reconstructed human epidermis model test" adopted 23. July 2009
- OECD Guideline for the Testing of Chemicals, Draft Proposal for a New Guideline, In Vitro Skin Irritation: Reconstructed Human Epidermis (RhE) Test Method, Version 7.6, 9 September 2009
- ECVAM international validation study on in vitro tests for acute skin irritation: Report on the validity of the EPISKIN and EpiDerm assays and on the Skin Integrity Function Test (Altern Lab Anim. 2007 Dec; 35 (6): 559-601).
- Protocol for IN VITRO EpiDerm™ SKIN IRRITATION TEST (EPI-200-SIT), Rev. 3/23/2009, MatTek Corporation, Ashland, MA 01721, USA

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of biphenyl-2-yl diphenyl phosphate and triphenyl phosphate and bis(biphenyl-2-yl) phenyl phosphate

EC Number:

700-393-5

IUPAC Name:

Reaction mass of biphenyl-2-yl diphenyl phosphate and triphenyl phosphate and bis(biphenyl-2-yl) phenyl phosphate

Details on test material:

- Lot/batch No.: F11101
- Expiration date of the lot/batch: Feb 2011
- Stability under test conditions: yes
- Storage condition of test material: Room Temperature: (20 ± 5°C); keep away from light and humidity.

Test animals

Species:

human

Strain:

other: not applicable

Test system

Type of coverage:

open

Preparation of test site:

other: EpiDerm tissues are cultured on specially prepared cell cultures inserts

Vehicle:

other: serum-free DMEM medium

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 µL

VEHICLE
- Lot/batch no. (if required): 09051OTTD

Duration of treatment / exposure:

1 h

Observation period:

35 - 60 min

Number of animals:

-

Details on study design:

TEST SITE
- % coverage: 100 %

REMOVAL OF TEST SUBSTANCE
- Washing (if done): DPBS-Buffer
- Time after start of exposure: 35 min.. 60 min

SCORING SYSTEM: formazan production is calculated as % photometric absorption compared with the negative control

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

other: other: % Formazan production

Value:

118.6

Remarks on result:

other:

Remarks:

Basis: mean. Reversibility: no data. Remarks: threshold for irritation (>50% of negative control = non-irritant). (migrated information)

Any other information on results incl. tables

Comparison of Formazan Production

For the test item and the positive control, the following percentage values of formazan production were calculated in comparison to the negative control:

Table: Formazan production (%)

Designation	Reaction mass of biphenyl-2-yl diphenyl phosphateand triphenyl phosphate and bis(biphenyl-2-yl) phenylphosphate and tris 2-biphenylyl phosphate	Positive Control
% Formazan production (Tissue 1)	115.1%	9.9%
% Formazan production (Tissue 2)	132.4%	10.7%
% Formazan production (Tissue 3)	108.2%	12.2%
% Formazan production Mean	118.6%	10.9%
Assessment: <50% of negative control = irritant; >50% of negative control = non-irritant

Irritation Potential of the Test Item

The relative absorbance values were increased to 118.6% after the treatment. This value is above the threshold for irritation (50%). Therefore, the test item is considered as not irritant. Note: optical density values of more than 100% of the negative control are not unusual.

Validity and Acceptability

Validity criteria and results are stated in the following table:

Criterion	Demanded	Found
OD of negative control	between 1.0 and 2.5	1.347
% Formazan production of positive control	≤20 % of negative control	10.9%
Variation within replicates (RSD)	< 18%	12.0 % (negative control) 11.1 % (positive control) 10.5% (test item)

The value for the negative control was not within the range of historical data of the test facility (see annex 2, page 20). This is considered as uncritical for the following reason: The deviation was only 4.8 % below the respective range of the historical data. Variation of biological systems within this order of magnitude is not unusual; furthermore, for the preparation of different batches of the cell cultures, different cell donors may have been used. Therefore the experiment is considered valid.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The multi-constituent substance 'Reaction mass of biphenyl-2-yl diphenyl phosphate and triphenyl phosphate and bis(biphenyl-2-yl) phenyl phosphate and tris 2-biphenylyl phosphate' is considered as not irritant in the Human Skin Model Test.