Bismuth oxide salicylate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 April 2012 to 20 April 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

The relative humidity occasionally was out of the target range, bw recorded at start not on treatment day

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Bismuth oxide salicylate

EC Number:

238-953-1

EC Name:

Bismuth oxide salicylate

Cas Number:

14882-18-9

Molecular formula:

C7H5BiO4

IUPAC Name:

2-hydroxy-4H-1,3,2-benzodioxabismin-4-one

Details on test material:

Name: Bismuth subsalicylate

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Species and strain: New Zealand White rabbits
Source: S&K-LAP Kft.
2173 Kartal, Császár út 135, Hungary
Justification of strain: The New Zealand White rabbit is one of the standard strains used for acute irritation toxicity studies.
Animal health: Only animals in acceptable health condition were used for the test. Both eyes of each animal provisionally selected for testing were examined prior to starting the study. Animals showing eye irritation, ocular defects or pre-existing corneal injury were not used.
Number of animals: 3 animals
Age of animals at treatment: ~11 weeks old (adult)
Sex: Male
Body weight range at the beginning of the experiment: 2766 – 2875 g
before euthanasia: 3046 – 3100 g
Date of receipt: 04 April 2012
Acclimatization time: at least 8 days
Animal identification: The individual identification was by engraved ear tag. The cages were marked with individual identity cards with information about study code, sex, dose, cage number and individual animal number.

HUSBANDRY
Animal health: Only healthy animals were used for the test. The veterinarian certified health status.
Number of animal room: 607
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature : 20 ± 3°C
Relative humidity: 24 – 62 %
Housing/Enrichment: Rabbits were individually housed in AAALAC approved metal wire rabbit cages. Cages were of an open wire structure and cages were placed together to allow some social interaction with rabbit(s) in adjoining cages
Ventilation: 15-20 air exchanges/hour
The environmental parameters were recorded twice daily during the study.

FOOD AND FEEDING
Animals received diet for rabbits produced by AGRIBRANDS Europe Hungary PLC, H-5300 Karcag, Madarasi road, Hungary, ad libitum (Batch number: 0030 03 12 Expiry date: 31 May 2012, Batch number: 0060 04 12 Expiry date: 05 July 2012). The details of the diets used will be archived with the raw data and are not reported.

WATER SUPPLY AND QUALITY CONTROL OF WATER
The animals received municipal tap water, as for human consumption, ad libitum, from an automatic system. The quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives of CiToxLAB Hungary Ltd.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The contra lateral eye served as the control.

Amount / concentration applied:

A single dose of 0.1 g of the solid test item Bismuth subsalicylate was administered to each animal.

Duration of treatment / exposure:

1 hour

Observation period (in vivo):

1 week

Number of animals or in vitro replicates:

3 male animals

Details on study design:

Application of the Test Item
Three male animals in acceptable health condition were selected for the test. Care was taken to select only those animals that had a normal eye condition and any with ocular lesions were rejected.
An initial test was performed using one animal. The test item was instilled into the conjunctival sac of the left eye. The eyelids were held closed for a few seconds to prevent the loss of the test item. The contra lateral eye served as the control. Immediately after the administration of the test item, an assessment of the initial pain reaction was made according to the six point scale.
After consideration of the ocular responses produced in the first animal, approximately 24 hour after the treatment of the first animal, two additional animals were treated.

Duration of Exposure
The eyes of the test animals were washed out at 1 hour after application of test item.

OBSERVATIONS AND SCORING

Clinical Observations
The eyes were examined at 1, 24, 48, 72 hours and 1 week after treatment. The duration of the observation period was sufficient to identify reversibility or irreversibility of changes. Any clinical signs of toxicity or signs of ill-health during the study were recorded. At the end of the observation period, each animal was sacrificed by intramuscular injections of CP-Ketamin 10% and CP-Xylazin 2% followed by iv. Euthasol® 40% anaesthesia. Death was verified by checking pupil and corneal reflex and the absence of respiration.

Scoring and Assessment of Local Reaction
The eye irritation scores were evaluated according to the scoring system by Draize (1977) and OECD 405 (24 April 2002).

Classification of the Test Items
Individual reactions of the animals were recorded at each observation time. The nature, severity and duration of all lesions observed were described.
Results were presented and interpreted according to Regulation (EC) No 1272/2008, as follows:

Irreversible effects on the eye/serious damage to eyes (Category 1)
Substances that have the potential to seriously damage the eyes are classified in Category 1 (irreversible effects on the eye). These observations include animals with grade 4 cornea lesions and other severe reactions (e.g., destruction of cornea) observed at any time during the test, as well as persistent corneal opacity, discoloration of the cornea by a dye substance, adhesion, pannus, and interference with the function of the iris or other effects that impair sight.

Category for irreversible eye effects
If, when applied to the eye of an animal, a substance produces:
— at least in one animal effects on the cornea, iris or conjunctiva that are not expected to reverse or have not fully reversed within an observation period of normally 21 days;
and/or
— at least in 2 of 3 tested animals, a positive response of:
o corneal opacity ≥ 3 and/or
o iritis > 1.5
calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material.

Reversible effects on the eye/irritating to eyes (Category 2)
Substances that have the potential to induce reversible eye irritation are classified in Category 2 (irritating to eyes).
Category for reversible eye effects
If, when applied to the eye of an animal, a substance produces:
— at least in 2 of 3 tested animals, a positive response of:
o corneal opacity ≥ 1 and/or
o iritis ≥ 1, and/or
o conjunctival redness ≥ 2 and/or
o conjunctival oedema (chemosis) ≥ 2

— calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material, and which fully reverses within an observation period of 21 days.

Measurement of Body Weight
Individual body weight was recorded at the beginning of the experiment and before euthanasia.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #1

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Results 8

Irritation parameter:

conjunctivae score

Basis:

animal #2

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 1 week

Results 9

Irritation parameter:

conjunctivae score

Basis:

animal #3

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 1 week

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Results 13

Irritation parameter:

other: discharge

Basis:

animal #1

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Results 14

Irritation parameter:

other: discharge

Basis:

animal #2

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Results 15

Irritation parameter:

other: discharge

Basis:

animal #3

Remarks:

(mean)

Time point:

other: Overall at 24, 48 and 72 hours after treatment

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Irritant / corrosive response data:

The eyes were examined at 1, 24, 48, 72 hours and 1 week after the application.
Initial Pain Reaction (IPR) (score 1 or 2) was observed in all animals.
One hour after the application: Conjunctival redness (score 1) and discharge (score 1) were found in all animals. In addition conjunctival chemosis (score 1) was observed in two animals.
At 24 hours after the application: Conjunctival redness (score 1) was found in all animals.
At 48 hours after the application: Conjunctival redness (score 1) was found in two animals.
At 72 hours after the application: Conjunctival redness (score 1) was found in two animals.
At 1 week after treatment no signs of eye irritation or other clinical signs were observed.
As all signs of eye irritation had fully reversed the study was terminated after a period of 1 week observation.

Other effects:

There was no mortality observed during the study.
The body weight and body weight change were considered to be normal with no indication of a treatment related effect.
There were no clinical signs observed that could be related to treatment.

Any other information on results incl. tables

Abbreviations: R = Redness                                         OD = Opacity degree of density

CH = Chemosis                                                             OE = Extent of opaque area

D = Discharge                                                               IPR = Initial pain reaction

Time	Animal No.	Score of irritation								IPR
		Conjunctivae			Opacity of cornea		Iris	Control eye	Other sign
		R	CH	D	OD	OE	R
1 hour	00519	2	0	2	0	0	0	0	-	2
	00520	2	1	2	0	0	0	0	-	2
	00577	2	1	2	0	0	0	0	-	2

 

 

Time	Animal No.	Score of irritation
		Conjunctivae			Opacity of cornea		Iris	Control eye	Other sign
		R	CH	D	OD	OE	R
24 hours	00519	1	0	0	0	0	0	0	-
	00520	1	0	0	0	0	0	0	-
	00577	1	0	0	0	0	0	0	-

 

 

Time	Animal No.	Score of irritation
		Conjunctivae			Opacity of cornea		Iris	Control eye	Other sign
		R	CH	D	OD	OE	R
48 hours	00519	1	0	0	0	0	0	0	-
	00520	1	0	0	0	0	0	0	-
	00577	0	0	0	0	0	0	0	-

 

 

Time	Animal No.	Score of irritation
		Conjunctivae			Opacity of cornea		Iris	Control eye	Other sign
		R	CH	D	OD	OE	R
72 hours	00519	1	0	0	0	0	0	0	-
	00520	0	0	0	0	0	0	0	-
	00577	0	0	0	0	0	0	0	-

 

 

Time	Animal No.	Score of irritation
		Conjunctivae			Opacity of cornea		Iris	Control eye	Other sign
		R	CH	D	OD	OE	R
1 week	00519	0	0	0	0	0	0	0	-
	00520	0	0	0	0	0	0	0	-
	00577	0	0	0	0	0	0	0	-

 

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item Bismuth Subsalicylate, applied to rabbit eye mucosa, caused conjunctival irritant effects at one hour which were reduced at 72 hours after application. The effects were fully reversible within 1 week.
According to Regulation (EC) No 1272/2008, Bismuth Subsalicylate does not require classification as an eye irritant.

Executive summary:

An acute eye irritation study of the test item Bismuth Subsalicylate was performed in New Zealand White rabbits. The irritation effects of the test item were evaluated according to the Draize method (OECD No.: 405, 2002).

The test item was placed into the conjunctival sac of the left eye of each animal. The untreated right eye served as control. An amount of 0.1 g of the test item was administered as a single dose.

Individual body weight was recorded at the beginning of the experiment and before euthanasia. Morbidity and clinical signs of toxicity were checked daily. The eyes were examined at 1, 24, 48, 72 hours and 1 week after the application.

Initial Pain Reaction (IPR) (score 1 or 2) was observed in all animals.

One hour after the application: Conjunctival redness (score 1) and discharge (score 1) were found in all animals. In addition conjunctival chemosis (score 1) was observed in two animals.

At 24 hours after the application: Conjunctival redness (score 1) was found in all animals.

At 48 hours after the application: Conjunctival redness (score 1) was found in two animals.

At 72 hours after the application: Conjunctival redness (score 1) was found in two animals.

At 1 week after treatment no signs of eye irritation or other clinical signs were observed.

As all signs of eye irritation had fully reversed the study was terminated after a period of 1 week observation.

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.

The body weights of all rabbits were considered to be within the normal range of variability.

The animal’s individual mean scores (considering readings at 24, 48 and 72 hours after the treatment) were as follows:

chemosis : 0.00, 0.00, 0.00

discharge : 0.00, 0.00, 0.00

redness : 0.33, 1.00, 1.00

cornea opacity : 0.00, 0.00, 0.00

iris : 0.00, 0.00, 0.00

The test item Bismuth Subsalicylate, applied to rabbit eye mucosa, caused conjunctival irritant effects at one hour which were reduced at 72 hours after application. The effects were fully reversible within 1 week.

According to Regulation (EC) No 1272/2008, Bismuth Subsalicylate does not require classification as an eye irritant.