Phosphonic acid, P-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)-, sodium salt (1:1)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Arrival of the Test Item: 16 June 2011 Date of Final Report: 06 September 2011

Reliability:

1 (reliable without restriction)

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD 437

Principles of method if other than guideline:

This study followed the procedures indicated by internal BSL BIOSERVICE SOPs and the following internationally accepted guidelines and recommendations:

OECD Guideline for the Testing of Chemicals, number 437 “Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants” (adopted: 7 September 2009) [3]

EPA Health Effects Test Guidelines, OPPTS 870.1000 “Acute toxicity testing background”, EPA 712-C-02-l 89, December 2002 [4]

Ministry of Health and Welfare (MHW, Japan): Japanese Guidelines for Nonclinical Studies of Drugs Manual 1995, Yakuji-Nippo Co. Ltd., 1995 (unofficial translation) [5]

Japanese Ministry of Agriculture, Forestry and Fisheries (JMAF’F), Guidelines for Preparation of Study Results, Eye Irritation Studies, Guideline 2-1-5. Notification 12 NohSan No. 8147, as partly revised in 16-Shouan-9260, on March 2005. English translation by IAI:ACIS on 17 October 2005 [6]

[3] OECD Guideline for the Testing of Chemicals, number 437 “Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants” (adopted: 07 September 2009

[4] EPA Acute Toxicity Testing - Background. Health Effects Test Guidelines, OPPTS 870.1000. United States, EPA 7l2-C-02-189, December 2002, United States, Environmental Protection Agency, Prevention, Pesticides and Toxic Substances (7101)

[5] Ministry of Health and Welfare (MHW, Japan): Japanese Guidelines for Nonclinical Studies of Drugs Manual 1995, Yakuji-Nippo Co. Ltd., 1995 (unofficial translation)

[6] Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Guidelines for Preparation of Study Results, Eye Irritation Studies, Guideline 2-1-5. Notification 12 NohSan No. 8147, as partly revised in 16-Shouan-9260, on March 2005. English translation by lAI:ACIS on 17 October 2005

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

sodium hydrogen (3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)phosphonate

EC Number:

700-812-1

Cas Number:

1189052-95-6

Molecular formula:

C8H5F13O3PNa

IUPAC Name:

sodium hydrogen (3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)phosphonate

Details on test material:

- Name of test material (as cited in study report): FHP-OHS
- Lot/batch No.: Z1601

Component 1:
- Chemical Name: 3,3,4,4,5,5,6,6,7,7,8,8,8-Tridecafluorooctyl-
phosphonic acid sodium salt (1:1)
- CAS Number: 1189052-95-6
- Formula: C8H5F13O3P*Na
- Analytical purity: 93%

Component 2:
- Chemical Name: 3,3,4,4,5,5,6,6,7,7,8,8,8-Tridecafluorooctyl-
phosphonic acid
CAS No.: 252237-40-4
- Formula: C8H6F1303P
- Analytical Purity: 7%

Date of Manufacture: 9 June 2011
Expiration Date: 3 years
Colour: white to yellow
Storage Conditions: at room temperature
Date of Analysis: 10 June 2011 (Certificate of Analysis see
Appendix 2 of attached report)
Safety Precautions: The routine hygienic procedures were sufficient to assure personnel health and safety.

Test animals / tissue source

Species:

other: Bovine corneal opacity and permeability assay

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Preparation of the Corneas:

The assay uses isolated corneas obtained as a by-product from an abattoir from freshly slaughtered animals.

The eyes were carefully examined for defects and any defective eyes were discarded.

The tissue surrounding the eyeball was carefully pulled away and the cornea was excised leaving a 2 to 3 mm rim of sclera. The isolated corneas were stored in a petri dish containing HBSS. Corneas were then mounted in corneal holders (MC2, Clermont, France) with the endothelial side against the 0-ring of the posterior chamber. The anterior chamber was then positioned on top of the cornea and tightened with screws. The chambers of the corneal holder were then filled with RPMI (without phenol red) containing 1% FBS and 2 mM L-glutamine (complete RPMI). The posterior chamber was always filled first. The corneas were incubated for one hour at 32 ± 1°C in a water bath.

Treatment of the Corneas:

After the incubation period, the medium was removed from both chambers and replaced with fresh Complete RPMI. An initial opacity measurement was performed on each of the corneas using an opacitometer (MC2, Clermont, France). Three corneas with opacity readings approximately equivalent to the median opacity of all corneas were selected as negative-control corneas. The opacity of each cornea was read against an air-filled chamber and recorded. Corneas that have an initial opacity reading above 7 units were not dosed. The medium was removed from the anterior chamber and replaced with the test item or control.

750 µL of the test item preparation or the control substance was introduced into the anterior chamber. After 4 hours ± 5 minutes incubation at 32 ± 1°C either the test substance or the control substance was removed and the epithelium washed at least three times with MEM (containing phenol red). Once the medium was free of test substance, the cornea was finally rinsed with complete RPMI (without phenol red). The anterior chamber was refilled with complete RPMI and an opacity measurement was performed.
After the opacity measurement the medium was removed from both chambers of the holder. The posterior chamber was refilled with fresh complete RPMI. 1 mL of a 5 mg/mL sodium fluorescein solution was added to the anterior chamber and the corneas were incubated for 90 minutes at 32 ± 1°C. Then the medium from the posterior chamber was removed and its optical density at 490 nm (OD490) was determined, using a spectrophotometer.

Test Groups:

- 3 corneas for the test item
- 3 corneas as negative controls treated with physiological saline 0.9% NaCI
- 3 corneas as positive control treated with imidazole 20% in physiological saline 0.9% NaCI
- The BCOP assay is considered to be valid if the in vitro score obtained with the positive control falls within the two standard deviations of the current historical mean.

Test system

Vehicle:

physiological saline

Controls:

other: 3 corneas for positive control and negative controls

Amount / concentration applied:

20% solution in saline

Duration of treatment / exposure:

750 µL of the test item preparation or the control substance was introduced into the anterior chamber. After 4 hours ± 5 minutes incubation at 32 ± 1°C either the test substance or the control substance was removed and the epithelium washed at least three times with MEM (containing phenol red). Once the medium was free of test substance, the cornea was finally rinsed with complete RPMI (without phenol red). The anterior chamber was refilled with complete RPMI and an opacity measurement was performed.

After the opacity measurement the medium was removed from both chambers of the holder. The posterior chamber was refilled with fresh complete RPMI. 1 mL of a 5 mg/mL sodium fluorescein solution was added to the anterior chamber and the corneas were incubated for 90 minutes at 32 ± 1°C. Then the medium from the posterior chamber was removed and its optical density at 490 nm (OD490) was determined, using a spectrophotometer.

Observation period (in vivo):

N/A

Number of animals or in vitro replicates:

3 corneas for Test Item

Details on study design:

See attached report

Results and discussion

In vivo

Irritant / corrosive response data:

Severe irritant

Other effects:

N/A

Any other information on results incl. tables

The eye irritancy potential of FHP-OHS was investigated in the bovine corneal opacity and permeability assay.   The test item was dissolved in physiological saline 0.9% NaCI to gain a 20% concentration.   The following mean in vitro score was calculated: 59.54   Therefore the test item was classified as severe irritant.   Thein vitroscore obtained with the positive control fell within the two standard deviations of the current historical mean and therefore this assay is considered to be valid.   For detailed data see Tables 2 - 4 in Appendix 1 of attached report.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Conclusion: According to the evaluation criteria the test item FHP-OHS is classified as severe eye irritant.

According to GHS (Globally Harmonized Classification System) [3] [9] the test item FHP-OHS is classified into Category 1.

[3] OECD Guideline for the Testing of Chemicals, number 437 “Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants” (adopted: 07 September 2009

[9] Globally Harmonized System of Classification and Labelling of Chemicals (GHS), New York & Geneva: United Nations Publications, UN (2007)

Executive summary:

Summary Results: The eye irritancy potential of FHP-OHS was investigated in the bovine corneal opacity and permeability assay. - Preparation of the test item: dissolved in physiological saline 0.9% NaCI - Test item concentration: 20% - Mean in vitro score: 59.54 - Classification: severe irritant - The in vitro score obtained with the positive control fell within the two standard deviations of the current historical mean and therefore this assay is considered to be valid.   - Conclusion: According to the evaluation criteria the test item FHP-OHS is classified as severe eye irritant.   According to GHS (Globally Harmonized Classification System) [3] [9] the test item FHP-OHS is classified into Category 1.   [3] OECD Guideline for the Testing of Chemicals, number 437 “Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants” (adopted: 07 September 2009   [9] Globally Harmonized System of Classification and Labelling of Chemicals (GHS),&: United Nations Publications, UN (2007)