Tetradonium bromide

Endpoint summary

Description of key information

Based on positive response in dermal testing with tetradonium bromide classification with Skin Irrit 2 can be applied. The potential for dermal irritation is further supported by read-across to dermal testing on cetrimounium chloride in 24-30% aqueous solutions in two OECD 404 studies.
Only slight eye irritation was noted in experimental tests with tetradonium bromide up to a concentration of 1.1%. However, severe eye irritation response corresponding to Eye Dam 1 was observed in two OECD 405 studies using cetrimonium chloride in water at concentrations of 24-30% . Using read-across to tetradonium bromide this justifies classification with Eye Dam 1.
The substance should be considered as a respiratory irritant and should be classified as STOT SE3, H335. (see study section 7.2.2)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

September 1980

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with no GLP compliance.

Qualifier:

according to guideline

Guideline:

other: Appraisal of the Safety of Chemicals in Foods, Drugs, and Cosmetics

Deviations:

yes

Remarks:

Test performed with three rabbits and three preparations

Principles of method if other than guideline:

The method used for testing primary skin irritation was described by J.H. Draize in "Appraisal of the Safety of Chemicals in Foods, Drugs, and Cosmetics", The Association of Food and Drug Officials of the United States, 1975 (pp. 46-48).

GLP compliance:

no

Specific details on test material used for the study:

White powder

Species:

rabbit

Strain:

other: SPF-Russian strain

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Versuchstierzuchtanstalt WIGA, D-8741 Sulzfeld
- Weight at study initiation: 1600-1750 g
- Housing: Single cages, 45x90 cm with a floor of wire netting
- Diet: free access to feed pellets (green rabbit KFK 7112)
- Water: free access to fresh water
- Acclimation period: one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19±2
- Humidity (%): 50±10

Type of coverage:

other: 2 layers of gauze, rabbits hereafter wrapped in a plastic-coated cellulose material fastened by tape

Preparation of test site:

other: clipped, half of test sites were scratched by a 18G needle

Vehicle:

not specified

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g to each test field
- Concentration (if solution): 0.1, 0.5 and 2.5 % (w/w)

Duration of treatment / exposure:

24 hours, treated skin was washed with water and left uncovered 48 hours

Observation period:

24 hours, 72 hours

Number of animals:

9

Details on study design:

TEST SITE
- Area of exposure: 10x15 cm, divided into six fields,
- Type of wrap if used: 2 layers of gauze, rabbits hereafter wrapped in a plastic-coated cellulose material fastened by tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, after 24 hours of exposure
- Time after start of exposure: 24 hours

SCORING SYSTEM: Erythema and eschar; oedema

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3

Time point:

other: 24 hours

Score:

0

Max. score:

4

Remarks on result:

other: Intact and scratched, 0.1%

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

other: 24 hours

Score:

2

Max. score:

4

Remarks on result:

other: Scratched, 0.5%

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

other: 24 hours

Score:

1

Max. score:

4

Remarks on result:

other: Intact and scratched, 0.5%

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

other: 24 hours

Score:

0

Max. score:

4

Remarks on result:

other: Intact and scratched, 0.5%

Irritation parameter:

erythema score

Basis:

animal: #7, #8, #9

Time point:

other: 24 hours

Score:

0

Max. score:

4

Remarks on result:

other: Intact and scratched, 2.5%

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 72 hours

Score:

1

Max. score:

4

Remarks on result:

other: Scratched, 0.1 %

Irritation parameter:

erythema score

Basis:

animal: #2, #3

Time point:

other: 72 hours

Score:

0

Max. score:

4

Remarks on result:

other: Intact and scratched, 0.1%

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

other: 72 hours

Score:

2

Max. score:

4

Remarks on result:

other: Scratched, 0.5%

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

other: 72 hours

Score:

2

Max. score:

4

Remarks on result:

other: Intact and scratched, 0.5%

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

other: 72 hours

Score:

1

Max. score:

4

Remarks on result:

other: Intact and scratched, 0.5%

Irritation parameter:

erythema score

Basis:

animal: #7, #8

Time point:

other: 72 hours

Score:

3

Max. score:

4

Remarks on result:

other: Intact and scratched, 2.5%

Irritation parameter:

erythema score

Basis:

animal: #9

Time point:

other: 72 hours

Score:

2

Max. score:

4

Remarks on result:

other: Intact and scratched, 2.5%

Irritation parameter:

edema score

Basis:

animal: #1, #2, #3

Time point:

other: 24 and 72 hours

Score:

0

Max. score:

4

Remarks on result:

other: Intact and scratched, 0.1%

Irritation parameter:

edema score

Basis:

animal: #4, #5, #6

Time point:

other: 24 and 72 hours

Score:

0

Max. score:

4

Remarks on result:

other: Intact and scratched, 0.5%

Irritation parameter:

edema score

Basis:

animal: #7

Time point:

other: 24 hours

Score:

0

Max. score:

4

Remarks on result:

other: Intact and scratched, 2.5%

Irritation parameter:

edema score

Basis:

animal: #8

Time point:

other: 24 hours

Score:

1

Max. score:

4

Remarks on result:

other: Intact and scratched, 2.5%

Irritation parameter:

edema score

Basis:

animal: #9

Time point:

other: 24 hours

Score:

1

Max. score:

4

Remarks on result:

other: Scratched, 2.5%

Irritation parameter:

edema score

Basis:

animal: #7, #8

Time point:

other: 72 hours

Score:

2

Max. score:

4

Remarks on result:

other: Intact and scratched, 2.5%

Irritation parameter:

edema score

Basis:

animal: 9

Time point:

other: 72 hours

Score:

1

Max. score:

4

Remarks on result:

other: Intact and scratched, 2.5%

Table 1: Scores for primary irritation of cetrimide BP Powder, batch no. 8820

Concentration 0.1% (w/w)
			Erythema-Eschar observation		Oedema observation		Index
			24 hours	72 hours	24 hours	72 hours
Rabbit no.	Weight, g	Skin
361	1650	Intact Scratched	0 0	0 1	0 0	0 0	0.25
362	1700	Intact Scratched	0 0	0 0	0 0	0 0	0.00
363	1600	Intact Scratched	0 0	0 0	0 0	0 0	0.00
Concentration 0.5% (w/w)
364	1650	Intact Scratched	0 2	1 2	0 0	0 0	1.25
365	1650	Intact Scratched	1 1	2 2	0 0	0 0	1.5
366	1650	Intact Scratched	0 0	1 1	0 0	0 0	0.5
Concentration 2.5% (w/w)
367	1750	Intact Scratched	3 3	3 3	0 0	2 2	4.00
368	1600	Intact Scratched	3 3	3 3	1 1	2 2	4.50
369	1600	Intact Scratched	3 3	2 2	0 1	1 1	3.25

 

Interpretation of results:

Category 2 (irritant)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The skin irritant effect of the test substance is investigated at different concentrations on nine albino male rabbits. Accordingly, at 0.1% (w/w) the substance was not a skin irritant, at 0.5% (w/w) the substance was non to mildly irritant, and at 2.5% (w/w) the susbtance was a moderate irritant giving a response complying with the CLP criteria for classification as Skin Irrit. 2.

Executive summary:

The skin irritant effect of the test substance is investigated at different concentrations on nine albino male rabbits. Accordingly, at 0.1% (w/w) the substance was not a skin irritant, at 0.5% (w/w) the substance was non to mildly irritant, and at 2.5% (w/w) the substance was a moderate irritant complying with the CLP criteria for classification as Skin Irrit. 2. A specific concentration limit for classification of 2.5% would be appropriate.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Jun-Jul 1984; Feb-Mar 1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Read-across to tetradonium bromide from data on cetrimonium chloride (with data reliability value of 1).

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes

Remarks:

Except study with 28-30% cetrimonium chloride, which was performed before EU GLP-guidelines

Species:

rabbit

Strain:

New Zealand White

Vehicle:

water

Remarks:

28-30% cetrimonium chloride in water; 24-26% cetrimonium chloride in wate

Controls:

other: The untreated eye served as control.

Amount / concentration applied:

0.1 ml of test substance

Observation period (in vivo):

Ocular reactions were evaluated 1, 24, 48, and 72 hours after instillation of the test article. When ocular reactions were noted at 72 hours, additional examinations were performed at 7, 14 and 21 days after the instillation.

Number of animals or in vitro replicates:

3

Details on study design:

A volume of 0.1 ml of the test substance was placed into the conjunctival sac of one eye of each animal. For 28-30% cetrimonium chloride treated eyes were rinsed thoroughly with physiological saline warmed to 37 °C after 24 hours. No informaiton on rinse of treated eyes was not given for the study with 24-26% cetrimonium chloride.

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

mean

Time point:

other: 24, 48, 72 hours

Score:

1.9

Remarks on result:

other: 28-30% cetrimonium chloride

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24, 48, 72 hours

Score:

1

Remarks on result:

other: 28-30% cetrimonium chloride

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 24, 48, 72 hours

Score:

2.3

Remarks on result:

other: 28-30% cetrimonium chloride

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24, 48, 72 hours

Score:

3.7

Remarks on result:

other: 28-30% cetrimonium chloride

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

mean

Time point:

other: 24, 48, 72 hours

Score:

2.8

Remarks on result:

other: 24-26% cetrimonium chloride

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 24, 48, 72 hours

Score:

2.4

Remarks on result:

other: 24-26% cetrimonium chloride

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24, 48, 72 hours

Score:

4

Remarks on result:

other: 24-26% cetrimonium chloride

Irritant / corrosive response data:

For 28-30% cetrimonium chloride: Grade 1-2 corneal opacity was found at time points between 1 and 72 hours. At 7, 14 and 21 days, grade 3 corneal opacity was noted. Grade 1 iritis was found at all time points between 1 hour and 7 days. No iritis was reported at 14 days and grade 1 iritis in 1/3
animals at 21 days. Conjunctival irritation was evident as grade 1-3 redness and grade 3-4 swelling at time points 24, 48 and 72 hours and still persisted as grade 1-2 redness and grade 2 swelling at 21 days.
For 24-26% cetrimonium chloride: The behaviour and physical condition of the rabbits were normal throughout the study. Grade 1 corneal opacity was observed at 1 hour and grade 3-4 opacity was found at all later time points including day 21. The iris could not be evaluated due to the corneal opacity. Conjunctival irritation was evident as grade 2-3 redness and grade 3-4 swelling at all time points including day 21.

Observation time	Observations
	28-30% cetrimonium chloride	24-26% cetrimonium chloride
1 hour	Grade 1-2 corneal opacity Grade 1 iritis	Grade 1 corneal opacity Grade 2-3 redness Grade 3-4 swelling
24 hours	Grade 1-2 corneal opacity Grade 1 iritis Grade 1-3 redness Grade 3-4 swelling	Grade 3-4 corneal opacity Grade 2-3 redness Grade 3-4 swelling
48 hours	Grade 1-2 corneal opacity Grade 1 iritis Grade 1-3 redness Grade 3-4 swelling	Grade 3-4 corneal opacity Grade 2-3 redness Grade 3-4 swelling
72 hours	Grade 1-2 corneal opacity Grade 1 iritis Grade 1-3 redness Grade 3-4 swelling	Grade 3-4 corneal opacity Grade 2-3 redness Grade 3-4 swelling
7 days	Grade 3 corneal opacity Grade 1 iritis	Grade 3-4 corneal opacity Grade 2-3 redness Grade 3-4 swelling
14 days	Grade 3 corneal opacity	Grade 3-4 corneal opacity Grade 2-3 redness Grade 3-4 swelling
21 days	Grade 3 corneal opacity Grade 1 iritis 1/3 Grade 1-2 redness Grade 2 swelling	Grade 3-4 corneal opacity Grade 2-3 redness Grade 3-4 swelling

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on the observations in the two studies it is concluded that cetrimonium chloride in aquaous solutions in the cocentration range from 24-30% cause irreversible ocular damage, corneal opacity and conjunctival irritation which persisted throughout the test period until day 21.
This imply classification with Eye Dam 1.

Executive summary:

The SCCS opinion document refers to two studies with cetrimonium chloride: 28-30% cetrimonium chloride in water and 24-26% cetrimonium chloride in water. The studies were carried out according to OECD TG 405 with New Zealand white albino rabbits. A volume of 0.1 ml of the test substance was placed into the conjunctival sac of one eye of each animal. The untreatred eye served as control. Ocular reactions were evaluated 1, 24, 48, and 72 hours after instillation of the test article. When ocular reactions were noted at 72 hours, additional examinations were performed at 7, 14 and 21 days after the instillation.

For 28 -30% cetrimonium chloride, the mean score values of the 24, 48 and 72 hour readings were 1.9 for opacity, 1.0 for iritis, 2.3 for conjunctival redness and 3.7 for conjunctival chemosis. Corneal opacity persisted throughout the test period until day 21. Conjunctival irritation and swelling persisted as grade 1 -2 redness and grade 2 swelling at 21 days.

For 24 -26% cetrimonium chloride, the mean score values of the 24, 48 and 72 hour readings were 2.8 for opacity, 2.4 for conjunctival redness and 4.0 for conjunctival chemosis. Corneal opacity persisted throughout the test period until day 21. Conjunctival irritation was evident as redness and sweling at all time points including day 21.

Based on the observations in the two studies it is concluded that cetrimonium chloride causes irreversible ocular damage, corneal opacity and conjunctival irritation which persisted throughout the test period until day 21.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Additional information

Justification for selection of skin irritation / corrosion endpoint:
Test performed with tetradonium bromide applicable for classification as Skin Irrit. 2

Justification for selection of eye irritation endpoint:
Eye damage observed in eye irritation tests with the read-across subtance cetrimonium chloride 24-30%

Effects on skin irritation/corrosion: moderately irritating

Effects on eye irritation: moderately irritating

Effects on respiratory irritation: irritating

Justification for classification or non-classification

Based on positive response in dermal testing with tetradonium bromide classification with Skin Irrit 2 with a SCL of 2.5% can be applied. The potential for dermal irritation is further supported by dermal testing on cetrimonium chloride in 24-29% aqueous solutions in two OECD 404 studies.

Only slight eye irritation was noted in experimental tests with tetradonium bromide up to a concentration of 1.1%. However, severe eye irritation response corresponding to Eye Dam 1 was observed in two OECD 405 studies using cetrimonium chloride in water at concentrations of 24-30%. Using read-across to tetradonium bromide this justifies classification with Eye Dam 1.

The substance should be considered as a respiratory irritant and should be classified as STOT SE3, H335. (see study section 7.2.2)