Chromium trichloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study without deviations

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:

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

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Chromium trichloride

EC Number:

233-038-3

EC Name:

Chromium trichloride

Cas Number:

10025-73-7

Molecular formula:

Cl3Cr

IUPAC Name:

chromium trichloride

Test material form:

solid: crystalline

Details on test material:

Name of test item: Chromium trichloride
Test-item No.: 09/0625-1
Batch identification: 03207EJ
CAS No.: 10025-73-7
Purity: 32.4 % Cr
Homogeneity: The test item was homogeneous by visual inspection.
Storage stability: The stability under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.
Storage conditions: Room temperature, dry storage
Physical state / color: Solid / violet
pH (undiluted test item, moistened with water): approx. 4.5

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Test species / strain / quality: Rabbits / New Zealand white A 1077 INRA (SPF)
Reasons for the selection of the test species: This animal species is the worldwide accepted test system forskin irritation / corrosion studies.
Age on day 0: Ca. 5-6 months
Sex: Males and females
Supplier: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
Arrival in the testing facility: Acclimatization for at least 5 days before application
Identification: Ear tattoo (by the rabbit supplier) and cage cards.
Body weight range on day 0: 4.41 kg - 5.38 kg
HOUSING AND DIET
Room temperature / relative humidity: The animals were housed in fully air-conditioned rooms. Central air-conditioning guaranteed a range of 20 °C ± 3 °C for temperature and of 30 - 70% for relative humidity. There were no deviations from these ranges, which influenced the results of the study.
Day / night rhythm: 12 h / 12 h (6.00 a.m. - 6.00 p.m. / 6.00 p.m. - 6.00 a.m.)
Type of cage: Stainless steel wire mesh cages with grating with shallow cage body; floor area: 4225 cm²
Enrichment: Wooden gnawing blocks (Typ KNH E-041); Abedd(R) Lab. And Vet. Service GmbH Vienna, Austria
Number of animals per cage: Single housing
Feeding: STANRAB (P) SQC; SOS Special Diets Services, 67122 Altrip, Germany
Drinking water: Tap water ad libitum
Selection: Before the beginning of application both eyes were investigated for signs of pre-existing irritation. Only animals with intact cornea and conjunctiva were used.
Analysis of feed: The feed used in the study was assayed for chemical and microbial contaminants by the manufacturer in quarterly intervals.
Analysis of drinking water: The drinking water was regularly assayed for contaminants by the municipal authorities of Heidelberg. The German Drinking Water Regulation of Dec. 5, 1990 served as the guideline for maximum tolerable contaminants.
Bedding and enrichment analysis: The bedding and enrichment were regularly assayed for contaminants (chlorinated hydrocarbons and heavy metals).

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated left eye served as control

Amount / concentration applied:

0.1 mL bulk volume (about 41 mg of the comminuted test item)

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

Readings were made approx. 1, 24, 48 and 72 h after application and then in weekly intervals maximally up to day 28.

Number of animals or in vitro replicates:

3

Details on study design:

EXPERIMENTAL PROCEDURE
Conduct of the study
Body weight determination: Just before application of the test item and after the last reading.
Route of application: The test item was applied in a single dose to the conjunctival sac of the right eyelid.
Application volume: 0.1 mL bulk volume (about 41 mg of the comminuted test item)
Rinsing of the eye: About 24 hours after application of the test item the treated eye of the animal(s) was rinsed with 3 to 6 mL of hand warm tap water for 1 to 2 minutes using a syringe with a blunt probe.
Negative control: Untreated left eye
Readings: Approx. 1, 24, 48 and 72 h after application and then in weekly intervals maximally up to day 28. For details about the observation period of each animal see Table: Individual ocular observations and mean values in the Appendix.
Illumination used for reading: Reactions on cornea or iris were observed by using a slit lamp.
Mortality: A check for any dead or moribund animal was made at least once each workday.
Assessment of ocular reactions
The evaluation of eye irritation was performed according to the quoted guidelines. In addition to specific observations recommended by the regulatory authorities, evaluations were made of discharge from the eye and the area of cornea affected by the lesions.
Cornea opacity (op):
Degree of density (the most dense area is taken for reading)
Grading
0 No ulceration or opacity
1 Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible
2 Easily discernible translucent area, details of iris slightly obscured
3 Nacreous area, no details of iris visible, size of pupil barely discernible
4 Opaque cornea, iris not discernible through the opacity
Iris:
Grading
0 Normal
1 Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection, any of these observations or combination of any thereof, iris still reacting to light (sluggish reaction is positive)
2 No reaction to light, haemorrhage, gross destruction (any or all of these)
Conjunctivae redness (red) (Refers to palpebral and bulbar conjunctivae, not to cornea and iris):
Grading
0 Blood vessels normal
1 Some blood vessels definitely hyperemic (injected)
2 Diffuse, crimson color, individual vessels not easily discernible
3 Diffuse beefy red
Chemosis (sw) Lids and/or nictitating membrane:
Grading
0 No swelling
1 Any swelling above normal (includes nictitating membrane)
2 Obvious swelling with partial eversion of lids
3 Swelling with lids about half closed
4 Swelling with lids more than half closed
EVALUATION OF RESULTS
For evaluation, the calculation of the mean values of corneal opacity, iris, conjunctival redness and chemosis for readings 24, 48 and 72 hours were quoted. Additionally, the reversibility of the observed findings was taken into account.
The evaluation is based on the criteria of the OECD Harmonized Integrated Classification System that was in place on the date of report signature. Calculations were performed using Microsoft Excel 2003 and checked with a calculator.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 24 h

Score:

0

Max. score:

4

Results 2

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 48 h

Score:

0

Max. score:

4

Results 3

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 72 h

Score:

0

Max. score:

4

Results 4

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 24 h

Score:

0

Max. score:

2

Results 5

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 48 h

Score:

0

Max. score:

2

Results 6

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 72 h

Score:

0

Max. score:

2

Results 7

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 24 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Results 8

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 48 h

Score:

0

Max. score:

3

Results 9

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 72 h

Score:

0

Max. score:

3

Results 10

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 24 h

Score:

0

Max. score:

4

Results 11

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 48 h

Score:

0

Max. score:

4

Results 12

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Slight conjunctival redness (grade 1) was observed in two animals 1 hour after application and persisted in one animal up 24 hours. Slight discharge of the conjunctiva (grade 1) was noted in all animals 1 hour after application. The ocular reactions were reversible in two animals within 24 hours and in one animal within 48 hours after application.
Mean scores calculated for each animal over 24,48 and 72 hours were 0.0, 0.0 and 0.0 for corneal opacity, 0.0, 0.0 and 0.0 for iris lesions, 0.0, 0.0 and 0.3 for redness of the conjunctiva and 0.0, 0.0 and 0.0 for chemosis.

Other effects:

Additional finding of vascularization of the cornea (scleral vessels injected in a circumscribed area) were noted in two animals 1 hour after application.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Considering the described ocular reactions as well as the average score for irritation, chromium trichloride does not show an eye irritation under the test conditions chosen.

Executive summary:

The potential of chromium trichloride to cause damage to the conjunctiva, iris or cornea was assessed by a single ocular application of 0.1 mL bulk volume (about 41 mg) of the test item to one eye of three White New Zealand rabbits (stepwise procedure starting with one animal and supplementing two additional animals). About 24 hours after application the eye was rinsed with tap water.

The ocular reactions were assessed approximately 1, 24, 48 and 72 hours after application.

Slight conjunctival redness and slight discharge were observed in the animals during the course of the study. Additional findings like vascularization of the cornea in a circumscribed area were noted in the animals during the observation period.

The ocular reactions were reversible in all animals within 48 hours after application.

Mean scores calculated for each animal over 24,48 and 72 hours were 0.0, 0.0 and 0.0 for corneal opacity, 0.0, 0.0 and 0.0 for iris lesions, 0.0, 0.0 and 0.3 for redness of the conjunctiva and 0.0, 0.0 and 0.0 for chemosis.

Considering the described ocular reactions as well as the average score for irritation, Chromium trichloride does not show an eye irritation potential under the test conditions chosen.