Dicalcium pyrophosphate

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 02 January 2013 and 10 January 2013.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no minor deviations from standard test guidelines and no minor methodological deficiencies. This study is conducted according to an appropriate validated in vitro guideline and under the conditions of GLP and therefore the study is considered to be acceptable and to adequately satisfy both the guideline requirement and the regulatory requirement as a key study for this endpoint. In addition, the data is considered to be adequate and reliable for classification and labelling in accordance with Regulation (EC) No. 1272/2008 (EU CLP).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

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

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of GLP inspection: 10 July 2012; Date of Signature on GLP certificate: 30 November 2012

Test material

Test material information

Constituent 1

Reference substance name:

Dicalcium pyrophosphate

EC Number:

232-221-5

EC Name:

Dicalcium pyrophosphate

Cas Number:

7790-76-3

Molecular formula:

Ca2O7P2

IUPAC Name:

dicalcium (phosphonatooxy)phosphonate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Sponsor's identification : IP 26: Dicalcium pyrophosphate
CAS number: 7790-76-3
Description: white powder
Batch number: MV3603
Date received: 24 September 2012
Expiry date: 03 May 2014
Storage conditions: room temperature in the dark

Test animals / tissue source

Species:

rabbit

Strain:

other: New Zealand White (Hsdlf:NZW)

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan UK Ltd., Leicestershire, UK.

- Age at study initiation: Twelve to twenty weeks old

- Weight at study initiation: 2.61 or 2.93 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 (2030C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd, 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


IN-LIFE DATES:
From: day 1 To:day 3

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): 100 mg 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.

VEHICLE

- Amount(s) applied (volume or weight with unit):Not applicable

- Concentration (if solution):Not applicable

- Lot/batch no. (if required):Not applicable

- Purity: Not applicable

Duration of treatment / exposure:

Up to 21 days (test item was not removed from the eyes).

Observation period (in vivo):

Assessment of ocular damage/irritation was made 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:

2 animals were tested in total. (After consideration of the ocular responses produced in the first treated animal, one additional animals was treated).

Details on study design:

REMOVAL OF TEST SUBSTANCE

- Washing (if done): Not applicable

- Time after start of exposure: Not applicable

INTERPRETATION OF RESULTS
The numerical values corresponding to each animal, tissue and observation time were recorded. The data relating to the conjunctivae were designated by the letters A (redness), B (chemosis) and C (discharge), those relating to the iris designated by the letter D and those relating to the cornea by the letters E (degree of opacity) and F (area of cornea involved). For each tissue the score was calculated as follows:

Score for conjunctivae = (A + B + C) * 2
Score for iris = D * 5
Score for cornea = (E * F) * 5

Using the numerical data obtained a modified version of the system described by Kay JH and Calandra J C (1962), J. Soc. Cosmet. Chem. 13, 281-289 (see below) was used to classify the ocular irritancy potential of the test item. This was achieved by adding together the scores for the cornea, iris and conjunctivae for each time point for each rabbit. The group means of the total scores for each observation were calculated. The highest of these group means (the maximum group mean score) together with the persistence of the reactions enabled classification of the eye irritancy potential of the test item.

If evidence of irreversible ocular damage is noted, the test item will be classified as corrosive to the eye.

The results were also interpreted according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

72824 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: No effects observed

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

72839 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: No effects observed

Results 3

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

72824 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0.25

Max. score:

2

Reversibility:

fully reversible within: 24 hours

Results 4

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

72839 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0.25

Max. score:

2

Reversibility:

fully reversible within: 24 hours

Results 5

Irritation parameter:

other: redness

Basis:

animal #1

Remarks:

72824 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0.5

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Results 6

Irritation parameter:

other: redness

Basis:

animal #2

Remarks:

72839 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0.5

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Results 7

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

72824 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0.5

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

72839 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0.75

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritant / corrosive response data:

Individual and group mean scores for ocular irritation are given in Table 1 and Table 2.

No corneal effects were noted during the study.

Iridial inflammation was noted in both treated eyes one hour after treatment.

Moderate conjunctival irritation was noted in one treated eye and minimal conjunctival irritation was noted in the other treated eye one hour after treatment. Minimal conjunctival irritation noted in both treated eyes at the 24-hour observation.

Both treated eyes appeared normal at the 48-hour observation.

Other effects:

Individual bodyweights and bodyweight changes are given in Table 3.
Both animals showed expected gain in bodyweight during the study.

Any other information on results incl. tables

Table 1. Individual Score and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex	72824 Male				72839 Male
	IPR = 1				IPR = 2
Time After Treatment	1 Hour	24 Hours	48 Hours	72 Hours	1 Hour	24 Hours	48 Hours	72 Hours
CORNEA
Degree of Opacity	0	0	0	0	0	0	0	0
Area of Cornea Involved	0	0	0	0	0	0	0	0
IRIS	1	0	0	0	1	0	0	0
CONJUNCTIVAE
Redness	1	1	0	0	1	1	0	0
Chemosis	1	1	0	0	2	1	0	0
Discharge	1	0	0	0	2	1	0	0

 

IPR =  Initial pain reaction

Table 2. Individual bodyweights and bodyweight changes

Rabbit number and sex	Individual bodyweight (kg)		Bodyweight change (kg)
72824 Male	Day 0	Day 3	0.07
	2.93	3.00
72839 Male	Day 0	Day 21	0.07
	2.61	2.68

 

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures.

This study is conducted according to the appropriate guidelines (OECD 405) and under the conditions of GLP and therefore the study is considered to be acceptable and to adequately satisfy both the guideline requirement and the regulatory requirement for REACH (Regulation (EC) No.1907/2006) as a key study for this endpoint. Study is sufficient for classification and labelling purposes, in accordance with Regulation (EC) No. 1272/2008 (EU CLP).