Calcium bis(dihydrogenorthophosphate)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was performed between 06 May 2010 and 02 June 2010.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Principles of method if other than guideline:

Due to the low pH value of the test material a Rabbit Enucleated Eye Test (REET) was performed prior to the in vivo test. The results are given in Appendix 3 and indicated that the test material was unlikely to cause severe ocular irritancy.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Calcium bis(dihydrogenorthophosphate)

EC Number:

231-837-1

EC Name:

Calcium bis(dihydrogenorthophosphate)

Cas Number:

7758-23-8

Molecular formula:

CaH4O8P2

IUPAC Name:

calcium dihydrogen phosphate

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source:
Harlan UK Limited, Bicester, Oxon, UK

- Age at study initiation:
Twelve to twenty weeks old

- Weight at study initiation:
2.0 to 3.5 kg

- Housing:
The animals were individually housed in suspended cages. The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

- Diet (e.g. ad libitum):
ad libitum (2030 Teklad Global Rabbit diet supplied by Harlan Teklad, Blackthorn, Bicester, Oxon, UK)

- 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:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL

- Amount(s) applied (volume or weight with unit):
A volume of 0.1 mL of the test material was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball.

- Concentration (if solution):
Undiluted and used as supplied

Duration of treatment / exposure:

Test material was administered once on day 1. Observations continued until day 21. The test material was not removed from the eye of the test animal.

Observation period (in vivo):

Approximately 1 hour and 24, 48 and 72 hours following treatment. Additional observations were made on Days 7, 14 and 21 to assess the reversibility of the ocular effects.

Number of animals or in vitro replicates:

1 animal was tested in total. Due to the severity of the ocular responses produced in the first treated animal, and in accordance with UK Home Office guidelines, no additional animals were treated.

Details on study design:

REMOVAL OF TEST SUBSTANCE

- Washing (if done):
Not applicable

- Time after start of exposure:
Not applicable


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).


TOOL USED TO ASSESS SCORE:

Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: 69201 Male

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not reversible

Results 2

Irritation parameter:

iris score

Basis:

animal: 69201 Male

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 Days

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 69201 Male

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

not reversible

Remarks:

Haemorrhage scattered over the majority of the nictitating membrane Day 7 to Day 14, Off white appearance covering one quarter of the upper conjunctival membrane Day 7 to Day 21.

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 69201 Male

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritant / corrosive response data:

Ocular Reactions
Individual scores for ocular irritation are given in Table 1.
Scattered or diffuse corneal opacity was noted in the treated eye at the 24, 48 and 72 hour observations with translucent corneal opacity noted at the 7, 14 and 21 Day observations.
Iridial inflammation was noted in the treated eye one hour after treatment and at the 24, 48, 72 Hour observations.
Moderate conjunctival irritation was noted in the treated eye one hour after treatment and at the 24, 48, 72 Hour, 7 and 14 Day observations with minimal conjunctival irritation at the 21 Day observations.
Haemorrhage scattered over the majority of the nictitating membrane was noted in the treated eye at the 7 and 14 Day observations. Off white appearance covering one quarter of the upper conjunctival membrane was noted in the treated eye at the 7, 14 and 21 Day observations.
The persistence of reactions in the treated eye at the 21 Day observation was considered to be indicative of irreversible ocular damage.

Other effects:

The animal showed expected gain in bodyweight during the study.

Any other information on results incl. tables

Table1              IndividualScores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex	69201Male
	IPR= 2
Time After Treatment	1 Hour	24 Hours	48 Hours	72 Hours	7 Days	14 Days	21 Days
CORNEA
Degree of Opacity	0	1	1	1	2	2	2
Area of Cornea Involved	0	2	2	2	2	1	1
IRIS	1	1	1	1	0	0	0
CONJUNCTIVA
Redness	2	2	2	2	2HP	2HP	1P
Chemosis	2	2	2	2	1	1	0
Discharge	2	2	2	2	1	1	0

IPR=  Initial pain reaction

H =      Haemorrhage scattered over the majority of the nictitating membrane

P =      Off white appearance covering one quarter of the upper conjunctival membrane

Table2              Individual Bodyweights and Bodyweight Change

Rabbit Number and Sex	Individual Bodyweight (kg)		Bodyweight Change (kg)
	Day 0	Day 21
69201Male	2.39	2.79	0.40

Applicant's summary and conclusion

Interpretation of results:

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

Conclusions:

The test material produced irreversible ocular damage and was considered to be CORROSIVE to the rabbit eye (based on one rabbit only). In accordance with the Guidance on the application of Regulation (EC) No. 1272/2008 (EU CLP) the test substance meets the criteria for classification as corrosive to the eyes and no further testing is required.

Executive summary:

Introduction. The study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method was designed to meet the requirements of the following:

OECD Guidelines for the Testing of Chemicals No. 405 "Acute Eye Irritation/Corrosion" (adopted 24 April 2002)

Method B5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No. 440/2008

Initial considerations. Due to the low pH value of the test material a Rabbit Enucleated Eye Test (REET) was performed prior to the in vivo test. The results are given in Appendix 3 and indicated that the test material was unlikely to cause severe ocular irritancy.

Result. A single application of the test material to the non-irrigated eye of one rabbit produced translucent corneal opacity, iridial inflammation and moderate conjunctival irritation. Other ocular effects noted were off white appearance covering one quarter of the upper conjunctival membrane and haemorrhage scattered over the majority of the nictitating membrane. The persistence of reactions in the treated eye at the 21‑Day observation was considered to be indicative of irreversible ocular damage.

Conclusion. The test material produced irreversible ocular damage and was considered to be CORROSIVE to the rabbit eye (based on one rabbit only).