Amidinourea phosphate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20-03-2017 to 16-05-2017

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 Ocular Corrosives and Severe Irritants)

Version / remarks:

OECD Guideline for the Testing of Chemicals, number 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” (adopted: 26 July 2013)

Deviations:

yes

Remarks:

minor deviations that did not influence the quality or integrity of the present study.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Amidinourea phosphate

EC Number:

241-659-6

EC Name:

Amidinourea phosphate

Cas Number:

17675-60-4

Molecular formula:

C2H6N4O.xH3O4P

IUPAC Name:

diaminomethylideneurea;phosphoric acid

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Batch No.: 16VL8189
Expiry Date: 03 August 2017
Storage Conditions: room temperature, protected from light

Test animals / tissue source

Species:

other: Bovine eyes were collected from a slaughterhouse.The eyes were examined for defects, 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.

Strain:

not specified

Details on test animals or tissues and environmental conditions:

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. Before the corneas were mounted in corneal holders (Duratec GmbH) with the endothelial side against the O-ring of the posterior chamber, they had been visually examined for defects and any defective cornea had been discarded. 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.

Test system

Vehicle:

physiological saline

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

750 µL of the test item applied (The test item was suspended with physiological saline 0.9% NaCl to give a 20% concentration.

Duration of treatment / exposure:

4 hours ± 5 minutes

Observation period (in vivo):

Not applicable

Duration of post- treatment incubation (in vitro):

90 minutes

Number of animals or in vitro replicates:

3 per group

Details on study design:

See any other information on materials and methods

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

3.58

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

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

Any other information on results incl. tables

In vitro irritation score

Cornea	Test Item	Corrected	Corrected OD490 Value	IVIS
No.		Opacity
1	Negative control	0.43	0.004
2		2.21	0.014
3		1.32	0.016
MV		1.32	0.011	1.49
4	Positive control	133.12	0.861
5		122.92	2.069
6		129.61	1.078
MV		128.55	1.336	148.59
7	Test Item	1.96	0.008
8		3.17	0.011
9		4.45	0.058
MV		3.2	0.025	3.58

MV = mean value

IVIS = in vitro irritation score

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

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

Executive summary:

The eye irritancy potential of GUP was investigated in the bovine corneal opacity and permeability assay. The test item was suspended with physiological saline 0.9% NaCl (see 10.2) to give a

20% concentration. All 3 corneas treated with GUP showed slight opacity of the tissue. The following mean in vitro irritation score was calculated: 3.58.

No prediction can be made regarding the classification of the test substance GUP 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 should result in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.