Potassium hexafluorophosphate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-08-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Potassium hexafluorophosphate

EC Number:

241-143-0

EC Name:

Potassium hexafluorophosphate

Cas Number:

17084-13-8

Molecular formula:

F6P.K

IUPAC Name:

potassium hexafluoro-λ⁵-phosphanuide

Test material form:

solid

Details on test material:

Aggregate State at RT: solid
Colour: white
Storage conditions: room temperature protected from light
Physical State: solid (crystalline powder)
Purity: > 99%

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: abattoir A. Moksel AG, Buchloe, Germany
- Number of animals: not specified
- Characteristics of donor animals (e.g. age, sex, weight): not specified
- Storage, temperature and transport conditions of ocular tissue: on the test day, fresh eyes were collected from the slaughterhouse and were transported in HBSS containing Pen/Strep on ice to the laboratories.
- Time interval prior to initiating testing: Immediately after arrival of the eyes, cornea preparation was initiated.
- indication of any existing defects or lesions in ocular tissue samples: The eyes were carefully examined for defects and any defective eyes were discarded.
- Indication of any antibiotics used: penicillin/streptomycin in HBSS during transport

Test system

Vehicle:

unchanged (no vehicle)

Remarks:

The test item was administered directly and moistened with physiological saline 0.9% NaCl (B. Braun Melsungen, lot no. 17185401, expiry date: 04/2020).

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 750 mg

Duration of treatment / exposure:

4 hours ± 5 minutes at 32 ± 1 °C.

Observation period (in vivo):

n.a.

Duration of post- treatment incubation (in vitro):

90 minutes at 32 ± 1 °C.

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS: eyes were carefully examined for defects and any defective eyes were discarded.
QUALITY CHECK OF THE ISOLATED CORNEAS: corneas have been visually examined for defects and any defective cornea had been discarded.
NUMBER OF REPLICATES: three
NEGATIVE CONTROL USED: yes, 750 µL physiological saline
POSITIVE CONTROL USED: yes, 750 µL imidazole 20% in physiological saline
APPLICATION DOSE AND EXPOSURE TIME: Dose: 750 mg. Exposure time: 4 hours ± 5 minutes at 32 ± 1 °C.
TREATMENT METHOD: control substance: closed chamber. Test item: open chamber
REMOVAL OF TEST SUBSTANCE: epithelium was 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).
POST-EXPOSURE INCUBATION: 90 minutes at 32 ± 1 °C.
METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: illuminance measurement.
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of UV/VIS spectrophotometry (OD490); Jenway 6405 UV/VIS.
- Others: each cornea was observed visually and pertinent observations were recorded.
SCORING SYSTEM: In Vitro Irritancy Score (IVIS)
DECISION CRITERIA: according to guideline.
For further details, see section “any other information on materials and methods incl. tables”.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

ca. 9.49

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

not determinable

Remarks:

no prediction can be made

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: not reported
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Range of historical values: for detailed information, see tables 4 and 5 in section “any other information on results incl. tables”.

Any other information on results incl. tables

The eye irritancy potential of the test substance was investigated in the bovine corneal opacity and permeability assay.

The test item was administered directly and moistened with physiological saline 0.9 % NaCl.

All 3 corneas treated with the test substance showed slight opacity of the tissue.

The following mean in vitro irritation score was calculated: 9.49

No prediction can be made regarding the classification of the test substance according to the evaluation criteria.

The in vitro irritation 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.

The negative control responses resulted in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.

For detailed data see tables 1-3 below.

INDIVIDUAL DATA

Table 1: Opacity. MV= mean value.

Cornea Nr. 1	Test item	Initial opacity	Final opacity	Change of opacity value	Corrected opacity value
1	Negative control	3.11	3.34	0.23
2		2.65	4.39	1.74
3		3.38	4.18	0.80
MV		3.05	3.97	0.92
4	Positive control	5.01	127.80	122.78	121.86
5		5.96	123.14	117.17	116.25
6		5.10	107.75	102.65	101.73
MV		5.36	119.56	114.20	113.28
7	Test item	1.94	12.13	10.19	9.27
8		2.05	8.24	6.18	5.26
9		1.94	10.91	8.97	8.04
MV		1.98	10.43	8.45	7.52

Table 2: permeability. MV= mean value.

Cornea Nr.	Test item	OD490	Corrected OD490 value
1	Negative control	0.008
2		0.008
3		0.098
MV		0.038
4	Positive control	0.662	0.624
5		1.220	1.182
6		2.260	2.222
MV		1.381	1.343
7	Test item	0.160	0.122
8		0.190	0.152
9		0.157	0.119
MV		0.169	0.131

 Table 3: in vitro irritation score. MV= mean value.

Cornea Nr.	Test item	Corrected opacity	Corrected OD490 value	IVIS
1	Negative control	0.23	0.008	1.49
2		1.74	0.008
3		0.80	0.098
MV		0.92	0.038
4	Positive control	121.86	0.624	133.42
5		116.25	1.182
6		101.73	2.222
MV		113.28	1.343
7	Test item	9.27	0.122	9.49
8		5.26	0.152
9		8.04	0.119
MV		7.52	0.131

 Table 4: Historical mean in vitro irritation score of the positive control

	IVIS Positive Control
Mean value (MV)	125.20
Standard deviation (SD)	17.75
MV – 2xSD	89.71
MV + 2xSD	160.70
Number of replicates providing historical mean: 27

 Table 5: Historical mean in vitro irritation score of the negative control

	IVIS Positive Control
Mean value (MV)	1.23
Standard deviation (SD)	0.79
MV – 2xSD	-0.35
MV + 2xSD	2.80
Number of replicates providing historical mean: 27

 

Applicant's summary and conclusion

Interpretation of results:

other: Expert judgement: not Category 1

Conclusions:

The eye irritancy potential of the test substance was investigated in the bovine corneal opacity and permeability assay. The following mean in vitro irritation score was calculated: 9.49. No prediction can be made regarding the classification of the test substance to the evaluation criteria. For safety reasons, the test substance is classified as Category 2.

Executive summary:

In a primary ex vivo eye irritation guideline study (bovine corneal opacity and permeability assay) under GLP conditions,750 mg of neat potassium hexafluorophosphate was applied on corneas for 4 hours. Irritation was scored by the method recommended in guideline OECD 437.

In this study, the mean in vitro irritation score was 9.49. According to GHS, no prediction can be made using this value. For precautionary measures, potassium hexafluorophosphate is classified voluntarily as mildly irritating to eyes (GHS Category 2).