reaction mass of calcium bis(dihydrogenorthophosphate) and calcium hydrogenorthophosphate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Remarks:

Carried out before the entry into force of the amendments to Annexes VII and VIII

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

In vivo study was carried out before the entry into force of the amendments to Annexes VII and VIII

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

reaction mass of calcium bis(dihydrogenorthophosphate) and calcium hydrogenorthophosphate

EC Number:

914-172-8

Molecular formula:

not available

IUPAC Name:

reaction mass of calcium bis(dihydrogenorthophosphate) and calcium hydrogenorthophosphate

Details on test material:

Sponsor's identification : a reaction mass of monocalcium phosphate and dicalcium phosphate
Description : cream coloured granular solid
Batch number : not supplied
Date received : 05 May 2010
Expiry date : 05 May 2012
Storage conditions : room temperature in the dark

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories U.K. Ltd., Loughborough, UK
- Age at study initiation: twelve to twenty weeks old
- Weight at study initiation: 2.63 kg
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23°C
- Humidity (%): 30 to 70%
- Air changes (per hr): at least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): twelve hours continuous light (06:00 to 18:00) and twelve hours darkness

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The left eye remained untreated and was used for control purposes.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL, weighing approximately 93 mg.

Duration of treatment / exposure:

7 days

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

1

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): None

SCORING SYSTEM:
Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation given in Appendix 2, (from Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49). Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope. Any clinical signs of toxicity, if present, were also recorded. Additional observations were made on Days 7 and 11 to assess the reversibility of the ocular effects. The animal’s bodyweight was recorded on Day 0 (the day of dosing) and at the end of the observation period.

TOOL USED TO ASSESS SCORE: ophthalmoscope

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: a score of 3 for corneal opacity was recorded on day 11

Results 2

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

not fully reversible within: 11 days

Results 3

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

not fully reversible within: 11 days

Results 4

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 11 days

Other effects:

Individual scores for ocular irritation are given in Table 1. Opalescent areas of the cornea were noted in the treated eye at the 11-Day observation.
Iridial inflammation and moderate conjunctival irritation were noted in the treated eye one hour after treatment and at the 24, 48, 72-Hour, 7 and 11-Day observations.
A pale area on the nictitating membrane, approximately 5 mm x 5 mm in size, was noted in the treated eye at 7-Day observation. Additional ocular effects noted in the treated eye at the 11-Day observation were vascularisation, with a generalised ingrowth of vessels for approximately 3 mm, over lower half of cornea, pale areas on the remainder of the nictitating membrane and lower conjunctival membrane, blood stained discharge, a white area on the nictitating membrane, approximately 10 mm x 10 mm in size, and also scattered areas on the lower conjunctival membrane, and a small area of sloughing on lower edge of the nictitating membrane, approximately 1 mm x 4 mm in size, which had almost completely detached. The reactions noted were considered to be indicative of irreversible ocular damage. Due to worsening reactions the animal was killed for humane reasons immediately after the 11-Day observation in accordance with current UK Home Office guidelines.

Any other information on results incl. tables

Table 1: Individual scores for ocular irritation

Rabbit number and sex	69653 male
	IPR = 2
Time after treatment	1 hour	2 hours	48 hours	72 hours	7 days	11 days K
CORNEA
Degree of opacity	0	0	0	0	0	3
Area of cornea involved	0	0	0	0	0	2
IRIS	1	1	1	1	1	1
CONJUNCTIVA
Redness	2	2	2	2	2	2
Chemosis	2	2	2	2	2	2
Discharge	2	3	3	3	1	2
Other reactions	-	-	-	-	P	VBsWP*Sl

 

IPR = Initial pain reaction V = Vascularisation, with a generalised ingrowth of vessels for approximately 3 mm, over lower half of cornea

P = Pale area on nictitating membrane, approximately 5 mm x 5 mm in size

Bs = Blood stained discharge

P* = Pale area on remainder of nictitating membrane and lower conjunctival membrane

W = White area on nictitating membrane, approximately 10 mm x 10 mm in size, and scattered areas on lower conjunctival membrane

Sl = Sloughing, small area on lower edge of nictitating membrane approximately 1 mm x 4 mm in size, almost completely detached

K = Due to the persistence of severe reactions, animal killed for humane reasons in accordance with current UK Home Office guidelines

- = Not applicable

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

In accordance with Regulation (EC) No. 1272/2008 the reaction mass of calcium bis(dihydrogenorthophosphate) and calcium hydrogenorthophosphate is classified as Category 1: irreversible effects on the eye/serious damage to eyes due to the severe reactions noted at day 7 which led to the animal being euthanized for humane reasons.