Magnesium bis(dihydrogenorthophosphate)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Remarks:

In vivo. Study was performed prior to the update of the REACH Regulation and prior to the validation of in vitro test methods.

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was performed between 02 January 2013 and 21 January 2013.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

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:

Magnesium bis(dihydrogenorthophosphate)

EC Number:

236-004-6

EC Name:

Magnesium bis(dihydrogenorthophosphate)

Cas Number:

13092-66-5

Molecular formula:

H3O4P.1/2Mg

IUPAC Name:

magnesium bis(dihydrogenorthophosphate)

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Sponsor's identification : IP 31: Magnesium bis(dihydrogenorthophosphate)
CAS number : 13092-66-5
Description : white powder
Batch number : Laboratory preparation 11.10.11
Purity : >98%
Date received : 02 October 2012
Expiry date : not supplied
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: Twelve to twenty weeks old

- Weight at study initiation: 2.70 or 2.83 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 (2930 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 7

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

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

VEHICLE

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

Duration of treatment / exposure:

72 hours

Observation period (in vivo):

Approximately 1 hour and 24, 48 and 72 hours following treatment. An additional observation was made on Day 7 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 animal was 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: 72826 Male

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: No effects observed

Remarks on result:

other: Initial pain reaction = 2

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal: 72873 Male

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: No effects observed

Remarks on result:

other: Inital pain reaction = 2

Results 3

Irritation parameter:

iris score

Basis:

animal: 72826 Male

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: No effect observed

Results 4

Irritation parameter:

iris score

Basis:

animal: 72873 Male

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: No effect observed

Results 5

Irritation parameter:

conjunctivae score

Basis:

animal: 72826 Male

Time point:

24/48/72 h

Score:

1.66

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 6

Irritation parameter:

conjunctivae score

Basis:

animal: 72873 Male

Time point:

24/48/72 h

Score:

1.66

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 7

Irritation parameter:

chemosis score

Basis:

animal: 72826 Male

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 8

Irritation parameter:

chemosis score

Basis:

animal: 72873 Male

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

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 one treated eye one hour after treatment.
Moderate conjunctival irritation was noted in both treated eyes one hour after treatment
and at the 24 and 48-Hour observations with minimal conjunctival irritation noted at the
72-Hour observation.
Both treated eyes appeared normal at the 7-Day observation.

Other effects:

Bodyweight
Both animals showed expected gain in bodyweight during the study.

Any other information on results incl. tables

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) x 2
Score for iris                            =         D x 5
Score for cornea                      =         (E x F) x 5

Using the numerical data obtained a modified version of the system ( Modified Kay and Calandra Interpretation of Eye Irritation Test was used to classify the ocular irritancy potential of the test material. This was achieved by adding together the scores for the cornea, iris and conjunctivae for each time point for each rabbit. 

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

 

Table1               Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex	72826 Male					72873 Male
	IPR= 2					IPR = 2
Time After Treatment	1 Hour	24 Hours	48 Hours	72 Hours	7 Days	1 Hour	24 Hours	48 Hours	72 Hours	7 days
CORNEA
E = Degree of Opacity	0	0	0	0	0	0	0	0	0	0
F = Area of Cornea Involved	0	0	0	0	0	0	0	0	0	0
Score (E x F) x 5	0	0	0	0	0	0	0	0	0	0
IRIS
D	1	0	0	0	0	0	0	0	0	0
Score (D x 5)	5	0	0	0	0	0	0	0	0	0
CONJUNCTIVAE
A = Redness	2	2	2	1	0	2	2	1	1	0
B = Chemosis	2	2	1	1	0	2	2	1	1	0
C = Discharge	3	2	1	0	0	2	1	1	0	0
Score (A + B + C) x 2	14	12	8	4	0	12	10	8	4	0
Total Score	19	12	8	4	0	12	10	8	4	0

 

IPR=  Initial pain reaction

 

Table 2               Individual Total Scores and Group Mean Scores for Ocular Irritation

Rabbit Number and Sex	Individual Total Scores At:
	1 Hour	24 Hours	48 Hours	72 Hours	7 Days
72826 Male	19	12	8	4	0
72873 Male	12	10	8	4	0
Group Total	31	22	16	8	0
Group Mean Score	15.5	11.0	8.0	4.0	0.0

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test material did not meet the criteria for classification as irritant according to Regulation (EC) No. 1272/2008 (EU CLP). 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 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).

Executive summary:

Introduction

The study was performed to assess the irritancy potential of the test item to the eye of the New Zealand White rabbit. The method was designed to be compatible with 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

Result

A single application of the test item to the non-irrigated eye of two rabbits produced iridial inflammation and moderate conjunctival irritation. Both treated eyes appeared normal at the 7-Day observation.

Conclusion

The test item produced a maximum group mean score of 15.5 and was classified as a moderate irritant (Class 5 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.

The test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.