Registration Dossier

Administrative data

Endpoint:
eye 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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: ICCVAm Test Method Evaluation Report: Appendix G -> ICCVAM recommended HET-CAM Method Protocol (Nov. 2006)
Deviations:
no
Principles of method if other than guideline:
Additional information was taken from:
- Background Review Document; "Current Status of In Vitro Test Methods for Identifying Ocular Corrosives and Severe Irritants: Hen's Egg Test - Chorioallantoic Membrane Test Method (March 2006, NIH Publication No. 06-4515)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
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 / tissue source

Species:
other: chicken eggs
Strain:
other: Lohmann selected Leghorn chicken´s eggs
Details on test animals or tissues and environmental conditions:
TEST SYSTEM
- Source: LSL Rhein Main Geflügelvermehrungsbetrieb (Dieburg, Germany)
- Age at study initiation: 9 days
- Weight at study initiation: 50-60 g
- Housing: incubator

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 38.5 °C
- Humidity (%): 58-60 %
- Air changes (per hr): no air change
- Photoperiod (hrs dark / hrs light): not necessary

Test system

Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.3 mL
- Concentration (if solution): pure
Duration of treatment / exposure:
not applicable
Observation period (in vivo):
5 minutes
Number of animals or in vitro replicates:
not applicable

Results and discussion

In vivo

Results
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 5 min
Score:
10.83
Reversibility:
other: absence of the possibility to assess reversibility (HET CAM test)
Other effects:
no

Any other information on results incl. tables

1) Observations

The observations which were made are given in the following table:

Table: Experimental Observations

Test Group Amount applied Time until Haemorrhage Time until Lysis Time until Coagulation
  [µL] [s] seconds seconds
Negative Control 300 301 301 301
300 301 301 301
300 301 301 301
Positive Control 0.1-N NaOH 300 9 21 15
300 7 20 15
300 5 20 13
Positive Control 1% SDS 300 10 23 301
300 12 25 301
300 11 21 301
Reaction mass of biphenyl-2-yl diphenyl phosphate and triphenyl phosphate and bis(biphenyl-2-yl) phenyl phosphate and tris 2-biphenylyl phosphate 300 28 30 301
300 27 31 301
300 27 32 301
300 28 32 301
300 29 32 301
300 30 33 301

2) Calculation of Irritation Score (IS)

The irritation score was calculated from the following equation:

IS = [(301 - time until haemorrhage)/300]*5 + [(301 - time until lysis)/300]*7 + [(301 - time until coagulation)/300]*9

The calculated IS for each replicate and the corresponding means are presented in the following table:

Table: Experimental Observations

Test Group Irritation Score Mean Irritation Score
Negative Control 0.00 0.00
0.00
0.00
Positive Control 0.1-N NaOH 19.98 20.05
20.04
20.13
Positive Control

Na-dodecylsulphate, 1 %

11.34 11.32
11.26
11.37
Reaction mass of biphenyl-2-yl diphenyl phosphate and triphenyl phosphate and bis(biphenyl-2-yl) phenyl phosphate and tris 2-biphenylyl phosphate 10.87 10.83
10.87
10.84
10.83
10.81
10.77

3) Classification

Classification of the irritation potential is made as stated in the following table:

Table: Classification

Irritation Score (IS) Classification
0-0.9 not irritant
1 -4.9 slightly irritant
5-8.9 moderately irritant
9-21 severely irritant

In the negative control, no signs of irritation were observed. Both positive controls showed severe irritation.

The test item 'Reaction mass of biphenyl-2-yl diphenyl phosphate and triphenyl phosphate and bis(biphenyl-2-yl) phenyl phosphate and tris 2-biphenylyl phosphate' showed severe irritation.

Applicant's summary and conclusion

Interpretation of results:
corrosive
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test item 'Reaction mass of biphenyl-2-yl diphenyl phosphate and triphenyl phosphate and bis(biphenyl-2-yl) phenyl phosphate and tris 2-biphenylyl phosphate' showed severe effects on the blood vessels of the CAM. The calculated mean irritation score is 10.83.
In conclusion, it can be stated that the test item possesses severe irritation potential.