Benzyltris(dimethylaminato)phosphorus(1+) tetrafluoroborate(1-)

Eye irritation

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:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Benzyltris(dimethylaminato)phosphorus(1+) tetrafluoroborate(1-)

EC Number:

302-080-5

EC Name:

Benzyltris(dimethylaminato)phosphorus(1+) tetrafluoroborate(1-)

Cas Number:

94088-77-4

Molecular formula:

C13H25BF4N3P

IUPAC Name:

benzyltris(dimethylamino)phosphanium; tetrafluoroboranuide

Test material form:

solid: crystalline

Test animals / tissue source

Species:

human

Details on test animals or tissues and environmental conditions:

Commercially available EpiOcularTM kit.
The EpiOcularTM tissue consists of normal, human-derived keratinocytes which have been cultured to form a stratified squamous epithelium similar to that found in the human cornea. It consists of highly organized basal cells. These cells are not transformed or transfected with genes to induce an extended life span. The EpiOcularTM tissues are cul-tured in specially prepared cell culture inserts with a porous membrane through which nutrients can pass to the cells. The tissue surface is 0.6 cm2.

EpiOcularTM tissues were procured from MatTek In Vitro Life Science Laboratories, Mylnské Nivy 73, 82105 Bratislava, Slovakia.
Designation of the kit: OCL-212-EIT
Day of delivery: 06. Sep. 2016
Batch no.: 23732

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

Tissue 1 50.5 mg
Tissue 2 49.7 mg

Duration of treatment / exposure:

5 h 53 min.

Duration of post- treatment incubation (in vitro):

17 h 58 min.

Results and discussion

In vitro

Results

Irritation parameter:

other: Optical density (Absorbance Values)

Run / experiment:

1-2

Value:

0.475 - 0.588

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

 Measured Values

As blank, the optical density of isopropanol was measured in eight wells of the 96-well-plate. The measured values and their mean are given in the following table:

Table9.1‑a      Absorbance Values Blank Isopropanol (OD at 570 nm)

Replicate	1	2	3	4	5	6	7	8	Mean
Absorbance	0.036	0.036	0.037	0.038	0.042	0.037	0.036	0.040	0.038

 

The absorbance values of negative control, test item and positive control are given in the following table:

Table9.1‑b      Absorbance Values Negative Control, Positive Control and Test Item (OD at 570 nm)

Designation	Measurement	Negative Control	Positive Control	Tecnoflon BA 104
Tissue 1	1	1.339	0.364	0.517
	2	1.410	0.362	0.509
Tissue 2	1	1.355	0.250	0.629
	2	1.133	0.248	0.622

 

From the measured absorbances, the mean of each tissue was calculated, subtracting the mean absorbance of isopropanol as given in table 9.1-a (= corrected values).

Table9.1‑c      Mean Absorbance Negative Control, Positive Control and Test Item

Designation	Negative Control	Positive Control	Tecnoflon BA 104
Mean – blank (Tissue 1)	1.337	0.325	0.475
Mean – blank (Tissue 2)	1.206	0.211	0.588

1.2     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:

Table9.2‑a      % Viability Positive Control and Test Item

Designation	Positive Control	Tecnoflon BA 104
% Viability (Tissue 1)	25.6%	37.4%
% Viability (Tissue 2)	16.6%	46.2%
% Viability Mean	21.1%	41.8%

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

One valid experiment was performed.
The test item Tecnoflon BA 104 was applied to a three-dimensional human cornea tissue model in duplicate for an exposure time of 5 hours and 53 minutes.
The solid test item was applied to each tissue.
After treatment, the respective substance was rinsed from the tissue; then, cell viability of the tissues was evaluated by addition of MTT, which can be reduced to formazan. The formazan production was evaluated by measuring the optical density (OD) of the resulting solution.
Demineralised water was used as negative control; Methyl acetate was used as positive control.
The controls showed the following results: After treatment with the negative control, the absorbance values were within the required acceptability criterion of 0.8 ≤ mean OD ≤ 2.5, OD was 1.3. The positive control showed clear eye irritating effects, the relative absorb-ance value was reduced to 21.1 % (< 50%).
Variation within tissue replicates was acceptable (< 20%).

After treatment with the test item, the relative absorbance values were reduced to 41.8 %.
This value is below the threshold for eye irritation potential (≤ 60%).

Under the conditions of the test system, Tecnoflon BA 104 is considered as
eye irritant in the EpiOcularTM Eye Irritation Test.