Tetrasodium enneatitanium icosaoxide hydrate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

5.5.2015-9.6.2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

The study was conducted in accordance with recommended Guidelines (OECD Guideline for the Testing of Chemicals No. 405 “Acute Eye Irritation/Corrosion” (adopted 02 October 2012) and Method B5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No. 440/2008) and GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

Adopted 02 October 2012

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

(EC) No. 440/2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Certificate as an attachment to the study report

Test material

Test material information

Constituent 1

Reference substance name:

nonatitanium(4+) tetrasodium hydrate icosaoxidandiide

EC Number:

601-472-6

Cas Number:

117314-20-2

Molecular formula:

Sodium form: Na4Ti9O20 × n H2O Sodium/hydrogen form: 50 % Na4Ti9O20 × n H2O, 50 % Na2H2Ti9O20 × n H2O

IUPAC Name:

nonatitanium(4+) tetrasodium hydrate icosaoxidandiide

Test material form:

solid: particulate/powder

Remarks:

powder

Details on test material:

- Substance type: commercial product (pure active substance)
- Physical state: Solid powder
- Storage condition of test material: Ambient temperature, humidity and pressure. Stored in sealed containers in darkness.
- Stability under test conditions: Stable
- Purity: ca 100 %
- Particle size distribution: 2.92% <100μm
- Crystal structure: TiO6-octaedra
- Density: 2.83 x 10^3 kg/m3
- pH value: the pH value of the substance in an aqueous solution is appr. 11

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 to 20 weeks old
- Weight at study initiation: 2.74 or 3.29 kg
- Housing: The animals were individually housed in suspended cages.
- Diet (e.g. ad libitum): 2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK
- Water (e.g. ad libitum): tap water
- Acclimation period: at least 5 days

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

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL (weight approximately 95 mg, as measured by gently compacting the required volume into an adapted syringe)

Duration of treatment / exposure:

1 treatment. The test item was left in the eyefor the 14-day assessment period.

Observation period (in vivo):

14 days. 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 and 14 to assess the reversibility of the ocular effects. Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

Number of animals or in vitro replicates:

2 (males)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No washing

SCORING SYSTEM:
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 described by Kay J.H. and Calandra J.C. (1962) 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.

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:

overall irritation score

Remarks:

Considers: cornea, iris and conjunctivae

Basis:

mean

Time point:

14 d

Score:

12.5

Max. score:

110

Reversibility:

fully reversible within: 14 days

Remarks on result:

positive indication of irritation

Remarks:

Iridial inflammation was noted in one treated eye and moderate conjunctival irritation was noted in both treated eyes. No corneal effects were noted during the study. The effects were fully reversible.

Results 2

Irritation parameter:

cornea opacity score

Remarks:

Considers: degree of opacity, area of cornea involved

Basis:

animal #1

Remarks:

Male

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: no indication of irritation

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

cornea opacity score

Remarks:

Considers: degree of opacity, area of cornea involved

Basis:

animal #2

Remarks:

Male

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: no indication of irritation

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

Male

Time point:

24/48 h

Score:

5

Max. score:

5

Reversibility:

fully reversible within: 72 hours

Remarks on result:

positive indication of irritation

Remarks:

Iridial inflammation was noted in one treated eye.

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

Male

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: no indication of irritation

Remarks on result:

no indication of irritation

Results 6

Irritation parameter:

conjunctivae score

Remarks:

Considers redness, chemosis and discharge

Basis:

animal #1

Remarks:

Male

Time point:

24/48/72 h

Score:

10.6

Max. score:

12

Reversibility:

fully reversible within: 14 days

Remarks on result:

positive indication of irritation

Remarks:

Moderate conjunctival irritation was noted in the treated eye.

Results 7

Irritation parameter:

conjunctivae score

Remarks:

Considers redness, chemosis and discharge

Basis:

animal #2

Remarks:

Male

Time point:

other: 1 h

Score:

12

Max. score:

12

Reversibility:

fully reversible within: 14 days

Remarks on result:

positive indication of irritation

Remarks:

Moderate conjunctival irritation was noted in the treated eye.

Results 8

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

Male

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 14 days

Results 9

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

Male

Time point:

other: 1 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 14 days

Irritant / corrosive response data:

No corneal effects were noted during the study.

Iridial inflammation was noted in one treated eye at the 24 and 48-Hour observations.

Moderate conjunctival irritation was noted in both treated eyes 1 and 24 hours after treatment. Moderate conjunctival irritation was noted in one treated eye and minimal conjunctival irritation was noted in the other treated eye at the 48 and 72-Hour observations with minimal conjunctival irritation noted in both treated eyes at the 7-Day observation.

Both treated eyes appeared normal at the 14-Day observation. See Tables 1 and 2, Any other information on results incl. tables.

Other effects:

No other effects. Both animals showed expected gain in body weight during the study (see Table 3, Any other information on results incl. tables).

Any other information on results incl. tables

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

Number and Sex	75059 Male						75072 Male
	Initial pain reaction = 0						Initial pain reaction = 0
Time After Treatment	1 hour	24 hours	48 hours	72 hours	7 days	14 days	1 hour	24 hours	48 hours	72 hours	7 days	14 days
CORNEA
E = Degree of Opacity	0	0	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	0	0
Score (E x F) x 5	0	0	0	0	0	0	0	0	0	0	0	0
IRIS
D	0	1	1	0	0	0	0	0	0	0	0	0
Score (D x 5)	0	5	5	0	0	0	0	0	0	0	0	0
CONJUNCTIVAE
A = Redness	2	2	2	2	1	0	2	2	2	2	1	0
B = Chemosis	2	2	2	2	1	0	2	1	1	1	1	0
C = Discharge	2	2	1	1	0	0	2	1	0	0	0	0
Score (A + B + C) x 2	12	12	10	10	4	0	12	8	6	6	4	0
Total Score	12	17	15	10	4	0	12	8	6	6	4	0

Initial pain reaction 0 = No response: No initial pain

Cornea: 

(E) Degree of Opacity (most dense area used)

0 = No opacity

Iris:

(D) Values

0 = Normal

1 = Folds above normal, congestion, swelling, circumcorneal injection (any or all of these or combination of any thereof) iris still reacting to light (sluggish reaction is positive)

Conjunctivae:

(A) Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)

0 =Vessels normal

1 = Vessels definitely injected above normal

2 = More diffuse, deeper crimson red, individual vessels not easily discernible

(B) Chemosis

0 = No swelling

1 = Any swelling above normal (includes nictitating membrane)

2 = Obvious swelling with partial eversion of lids

(C) Discharge

0 = No discharge

1 = Any amount different from normal (does not include small amounts observed in inner canthus of normal animals)

2 = Discharge with moistening of the lids and hairs just adjacent to lids

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	14 days
75059 Male	12	17	15	10	4	0
75072 Male	12	8	6	6	4	0
Group Total	24	25	21	16	8	0
Group Mean Score	12.0	12.5	10.5	8.0	4.0	0.0

Table 3. Individual Body Weights and Body Weight Change

Rabbit Number and Sex	Individual Body Weight (kg)		Body Weight Change (kg)
	Day 0	Day 3
75058 Male	2.74	3.17	0.43
75063 Male	3.29	3.54	0.25

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Remarks:

Eye irrit. 2 H319. Criteria used for interpretation of results: EU

Conclusions:

The test results are sufficient to make conclusions related to the eye irritation effects of the test item.

The test item produced a maximum group mean score of 12.5 and was classified as a mild irritant (Class 4 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system. According to EC No 1272/2008 the results indicate that the test item is an eye irritant (category 2, H319).

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.

Results

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 14-Day observation.

Conclusion

The test item produced a maximum group mean score of 12.5 and was classified as a mild irritant (Class 4 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.