Diphosphoric acid, copper salt

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2013-01-14 to 2013-02-04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

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:

Diphosphoric acid, copper salt

EC Number:

233-279-4

EC Name:

Diphosphoric acid, copper salt

Cas Number:

10102-90-6

Molecular formula:

Cu.xH4O7P2

IUPAC Name:

diphosphoric acid, copper salt

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Sponsor's identification: IP 37: Copper (II)-pyrophosphate
Description: pale blue powder
Batch number: MV500
Purity: >80% (based on Cu content >34%)
Impurities: <18% water, <2% free orthophosphate, pyrophosphate
Date received: 24 September 2012
Expiry date: 12 February 2015
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 UK Ltd., Leicestershire, UK.
- Age at study initiation: 12-20 weeks
- Weight at study initiation: 2.48 or 2.59 kg
- Housing: animals were individually housed in suspended cages.
- Diet (e.g. ad libitum): ad libitum (2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK)
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23°C
- Humidity (%): 30 to 70%
- Air changes (per hr): at least 15 per hour
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml of the test item (weighing approximately 83 mg as measured by gently compacting the required volume into an adapted syringe).

Duration of treatment / exposure:

Animals were observed for 7 days post-dosing.

Observation period (in vivo):

7 Days

Number of animals or in vitro replicates:

2

Details on study design:

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

The test material was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released.

After consideration of the ocular responses produced in the first treated animal, a second animal was treated. In order to minimise pain on application of the test item, one drop of local anaesthetic (Tetracaine hydrochloride 0.5%, Bausch & Lomb, Surrey, UK) was instilled into both eyes of the second animal 1 to 2 minutes before treatment.

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.

ADDITIONAL OBSERVATIONS:
Any clinical signs of toxicity, if present, were also recorded.
An additional observation was made on Day 7 to assess the reversibility of the ocular effects.
Individual bodyweights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable as no effects noted

Remarks on result:

other: IPR = 3

Results 2

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable as no effects noted

Remarks on result:

other: IPR = 0 (anaesthetic administered)

Results 3

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.66

Max. score:

2

Reversibility:

fully reversible within: 72 hours

Results 4

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Results 5

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 6

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 7

Irritation parameter:

chemosis score

Remarks:

(conjunctivae)

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 8

Irritation parameter:

chemosis score

Remarks:

(conjunctivae)

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Individual scores for ocular irritation are given in Table 1. The individual and mean scores as required for EU and GHS classification and labelling are presented in Table 2.
No corneal effects were noted during the study.
Iridial inflammation was noted in both treated eyes one hour after treatment and at the 24 and 48-Hour observations and in one treated eye at the 72-Hour observation.
Moderate conjunctival irritation was noted in both treated eyes one hour after treatment and at the 24 and 48-Hour observations. Moderate conjunctival irritation was noted in one treated eye and minimal conjunctival irritation was noted in the other treated eye 72-Hour observation.
Both treated eyes appeared normal at the 7-Day observation.

Other effects:

Individual bodyweights and bodyweight change 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 Scores for Ocular Irritation

Rabbit number and Sex	72877 Male					72918 Male
	IPR = 3					IPR = 0+
Time after treatment	1 Hrs	24 Hrs	48 Hrs	72 Hrs	7 days	1 Hrs	24 Hrs	48 Hrs	72 Hrs	7 days
Cornea Degree of opacity	0	0	0	0	0	0	0	0	0	0
Area of cornea involved	0	0	0	0	0	0	0	0	0	0
Iris	1	1	1	0	0	1	1	1	1	0
Conjuctivae Redness	2	2	2	2	0	2	2	2	2	0
Chemosis	3	2	2	1	0	2	2	2	2	0
Discharge	1	2	2	0	0	3	1	1	1	0

 

IPR = Initial pain reaction

+ = One drop of local anaesthetic instilled into both eyes 1 to 2 minutes before treatment

Table 2. Individual and Mean Scores for Cornea, Iris and Conjunctivae

Rabbit number and sex	Time after treatment	Corneal opacity	Iridial inflammation	Conjuntival redness	Conjunctival chemosis
72877 Male	24 hours 48 hours 72 hours	0 0 0	1 1 0	2 2 2	2 2 1
Mean:		0.0	0.7	2.0	1.7
72918 Male	24 hours 48 hours 72 hours	0 0 0	1 1 1	2 2 2	2 2 2
Mean:		0.0	1.0	2.0	2.0

 

 

Table 3. Individual Bodyweights and Bodyweight Change

Rabbit number and sex	Individual bodyweight (kg)		Bodyweight change (kg)
	Day 0	Day 7
72877 Male	2.48	2.60	0.12
72918 Male	2.59	2.75	0.16

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

The test item was classified as irritating to eyes (Category 2) according to Regulation (EC) No 1272/2008.

Executive summary:

The test item was classified as irritating to eyes (Category 2) according to Regulation (EC) No 1272/2008. It is reasonable to assume that the signal word “Warning” and the Hazard Statement “H319: Causes serious eye irritation” are therefore required. This study is considered to be adequate and reliable for classification and labelling in accordance with Regulation (EC) No. 1272/2008 (EU CLP) and as a key study in accordance with Regulation (EC) No. 1907/2006 (REACH).