Benzethonium chloride

Endpoint summary

Description of key information

Skin corrosion in-vivo / Skin irritation in-vivo

In the key study, skin corrosion was tested in accordance with DOT Skin Corrosion, Department of Transportation CFR Title 49, 173, 1200 on NZW rabbits. Based on the results of this study, the test material is considered to be corrosive to the skin. Due to the corrosive response in this study, no additional in-vivo skin irritation test was conducted. An in-vitro study does not need to be conducted as adequate data from an in-vivo study are available.

In another study on rats, the purpose was to determine if a 1% ( w/v) aqueous solution of benzethonium chloride can be applied dermally once per day for five consecutive days without producing more than slight dermal irritation. In conclusion, a 1% (w/v) aqueous solution of benzethonium chloride can be applied dermally once per day for five consecutive days without producing more than slight dermal irritation.

In further non-guideline studies, Benzethonium Chloride was non-irritating when applied topically (0.1 percent) to the skin of rabbits, although mice had local reactions to the moderate and high doses (35 to 280 mg/kg) of Benzethonium Chloride in a skin-painting study. Benzethonium Chloride was slightly irritating when administered intracutaneously to rabbits and non-irritating in tests of vaginal contraceptives with rabbits, dogs, and monkeys. (Source: CIR-Report (Lit: Journal of the American College of Toxicology, Volume 4, Number 5, 1985, Mary Ann Liebert, Inc., Publishers))

Eye irritation in-vivo

The key study was performed according to methods OECD Guidelines for Testing of Chemicals No. 405 and Method B5 of Commission Directive 92/69/EEC. The test material did not meet the criteria for classification as irritant according to GHS regulations.

In other old, non-guideline eye irritation studies, Benzethonium Chloride was irritating to the rabbit eye at concentrations greater than 0.03 percent. Additionally, Benzethonium Chloride produced ocular irritation in rabbits at a concentration of 0.1 percent and gave a maximum tolerated concentration (no corneal or iridic lesions at 7 days) of 0.5 percent. (Source: CIR-Report (Lit: Journal of the American College of Toxicology, Volume 4, Number 5, 1985, Mary Ann Liebert, Inc., Publishers))

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Jaunary 1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

Principles of method if other than guideline:

DOT Skin Corrosion, Department of Transportation CFR Title 49, 173, 1200

GLP compliance:

no

Specific details on test material used for the study:

- Source and lot/batch No.of test material: Lonza Inc., Fair Lawn, New Jersey
- product name: HYAMINE 1622 Powder (K-11-16)
- description: white powder
- Expiration date of the lot/batch: no data
- Purity test date: no data

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Albino Rabbits from Davidson's Mill Farm, USA
- Age at study initiation: no data
- Weight at study initiation: no data
- Housing: animals were individually housed in stainless steel wire bottomed cages
- Diet (e.g. ad libitum): Purina Rabbit Pellets
- Water (e.g. ad libitum): water ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): room temperature
- Photoperiod (hrs dark / hrs light): 12 hour light/dark cycle

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

no

Amount / concentration applied:

0.78 grams of test material was uniformly mixed with deionized distilled water and applied as a 63.7% suspension (w/w) to give 0.5 grams of material per site

Duration of treatment / exposure:

4 hours

Observation period:

Reading 4 and 48 hours as well as 5 and 8 days after exposure

Number of animals:

6 animals

Details on study design:

A 2.5 cm2 gauze patch was placed over intact skin on each rabbit. Five-tenths of a gram of the testmaterial was placed under each patch. The patches were secured in place with adhesive tape. The entire trunk of each animal was wrapped with a rubberized elastic cloth to retard evaporation and as an aid in maintaining test patch position. The rabbits were then immobilized in head stocks for 4 hours after which time the patches were removed. All test sites were washed to prevent further exposure. Corrosion readings were made at 4 and 48 hours after exposure. Corrosion was considered to have resulted if the test substance caused destruction or irreversible alteration of the tissue. Tissue destruction was considered to have occurred if, at any of the readings, there was ulceration or necrosis. Epidermal sloughing, erythema, edema or fissuring were not considered tissue destruction.

Irritation parameter:

other:

Basis:

mean

Time point:

other: 4 h

Remarks on result:

probability of weak irritation

Irritation parameter:

other:

Basis:

mean

Time point:

48 h

Remarks on result:

positive indication of irritation

Irritation parameter:

other:

Basis:

mean

Time point:

other: 8 d

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

At 4 hours the test site was brownish in color. Thickening of the test site with reduced flexibility was alsp noted. There was no evidence of necrosis, ulceration or sloughing. At 48 hours one rabbit exhibited necrosis. The test sites on the other rabbits remained brown (slightly darker than at 4 hours) and thickened, with reduced flexibility. At 8 days the rabbits exhibited necrosis, however, there were no signs of ulceration or sloughing. In conclusion, the test item needs to be considered as corrosive to rabbit skin.

Interpretation of results:

Category 1B (corrosive) based on GHS criteria

Conclusions:

Based on the 4 and 48 hour scores, the test material is considered to be corrosive to the skin.

Executive summary:

Skin corrosion was tested in accordance with DOT Skin Corrosion, Department of Transportation CFR Title 49, 173, 1200. Six NZW rabbits were treated occlusive with 0.78 g of test item mixed with deionized water for 4 hours. Skin reactions were assessed 4 hours, 48 hours and 8 days after removal of the patch. At 4 hours the test site was brownish in color. Thickening of the test site with reduced flexibility was alsp noted. There was no evidence of necrosis, ulceration or sloughing. At 48 hours one rabbit exhibited necrosis. The test sites on the other rabbits remained brown (slightly darker than at 4 hours) and thickened, with reduced flexibility. At 8 days the rabbits exhibited necrosis, however, there were no signs of ulceration or sloughing. Based on the 4 and 48 hour scores, the test material is considered to be corrosive to the skin.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Jaunary 1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Remarks:

species = rat (instead of rabbit)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Principles of method if other than guideline:

The purpose of this study was to determine if a 1% ( w/v) aqueous solution of benzethonium chloride can be applied dermally once per day for five consecutive days without producing more than slight dermal irritation. Study was performed on rats instead of rabbits; readings on days 1-5

GLP compliance:

no

Specific details on test material used for the study:

- Source: Lonza Inc., Fair Lawn, New Jersey
- Product name: Benzethonium Chloride (BTC)
- Batch: Lonza Batch 80I02, Lot 8K0909
- Expiration date: 3/30/2001
- Description: white powder
- Purity test date: no data

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Sprague Dawley, Inc. (Indianapolis, Indiana)
- Strain: F344
- Age: 11-12 weeks (Date of birth: males Feb 18, 2000; females, Feb 11, 2000)
- Weight: 144 to 183 g
- Number and sex: 6 rats (3 male and 3 female rats)
- Acclimation: 3 days
- Euthanasia: Rats were euthanized by C02 induced hypoxia following the five day observation period.

ENVIRONMENTAL CONDITIONS / ANIMAL HUSBANDRY
- Caging: Rats were individually housed in suspended stainless steel cages
- Feed: Rats were provided rodent feed ad libitum
- Water: Drinking water was provided ad libitum. The water is periodically analyzed for microbial and chemical agents
- Temperature (°C): room temperature

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

no

Amount / concentration applied:

The dosing solution was prepared by dissolving BTC in Sterile Water for Injection to achieve a concentration of 1% BTC (w/v).

Duration of treatment / exposure:

24 hours

Observation period:

Reading on days 1-5

Number of animals:

6 rats (3 male and 3 female rats)

Details on study design:

Three male and three female rats received five consecutive daily topical applications of the test article at an aqueous concentration of 1% (w/v). Approximately 24 hours after each application, the treatment site was observed for signs of erythema and edema and washed with water prior to receiving the next application.
On Day 0 approximately 30 minutes prior to dermal application of the test article, the dorsum of each animal was clipped free of hair using an electric clipper. Beginning on Day 0, three male and three female rats received five consecutive daily topical applications (100 ul per application) of the test article at an aqueous concentration of 1% (w/v). The 100 ul application was applied to the back of rats over a surface area of approximately 10 cm2. Approximately 24 hours following each of the first four applications, the treatment site was washed with water (i.e. after treatment site observation and prior to next application). Approximately 24 hours following each application, the treatment site was evaluated for erythema and edema according to Draize.

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: days 1-5

Score:

0.8

Max. score:

1

Reversibility:

not reversible

Remarks on result:

probability of weak irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

other: days 1-5

Score:

0.07

Max. score:

1

Reversibility:

fully reversible within: 5 d

Remarks on result:

probability of weak irritation

Irritant / corrosive response data:

There was no evidence of irritation in one male rat. One female rat exhibited barely perceptible erythema after the first treatment with no evidence of irritation after the second through the fifth treatment. Two male rats exhibited barely perceptible erythema after each treatment. One female rat developed barely perceptible erythema after the third treatment and nonconfluent well defmed erythema over approximately 10% of the treatment sight with barely perceptible erythema over the remaining treatment site area after the
fourth and fifth treatment. One female exhibited barely perceptible erythema after the first and second treatment, non-confluent well defined erythema and barely perceptible edema after the third and fourth treatment, and non-confluent well defined erythema with no evidence of edema after the fifth treatment.

Interpretation of results:

study cannot be used for classification

Conclusions:

A 1% (w/v) aqueous solution ofbenzethonium chloride can be applied dermally once per day for five consecutive days without producing more than slight dermal irritation.

Executive summary:

The purpose of this study was to determine if a I% ( w/v) aqueous solution of benzethonium chloride can be applied dermally once per day for five consecutive days without producing more than slight dermal irritation. There was no evidence of irritation in one male rat. One female rat exhibited barely perceptible erythema after the first treatment with no evidence of irritation after the second through the fifth treatment. Two male rats exhibited barely perceptible erythema after each treatment. One female rat developed barely perceptible erythema after the third treatment and nonconfluent well defmed erythema over approximately 10% of the treatment sight with barely perceptible erythema over the remaining treatment site area after the fourth and fifth treatment. One female exhibited barely perceptible erythema after the first and second treatment, non-confluent well defined erythema and barely perceptible edema after the third and fourth treatment, and non-confluent well defmed erythema with no evidence of edema after the fifth treatment.

In conclusion, a 1% (w/v) aqueous solution ofbenzethonium chloride can be applied dermally once per day for five consecutive days without producing more than slight dermal irritation.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (corrosive)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

TEST MATERIAL
- Sponsor’s identification: P4123
- Batch number: 3072-S
- Date received: 28 June 1995
- Purity: 99.5%
- Description: white powder
- Stability: Considered to be stable under conditions of shipment, storage, and use in this study.
- Storage Conditions: Room temperature in the dark
- Expiration date: no data

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

- Source: David Percival Ltd., Moston, Sandbach, Cheshire, UK,
- Age at study initiation: 12-16 weeks
- Weight at study initiation: 2.78-2.92 kg
- Housing: individually housed in suspended metal cages
- Diet (e.g. ad libitum): STANRAB SQC Rabbit Diet, Special Diets Services Ltd., Witham, Essex, UK
- Water (e.g. ad libitum): tab water
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-22°C
- Humidity (%): 59-74%
- Air changes (per hr): 15 changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours continuous light and 12 hours darkness

Vehicle:

water

Amount / concentration applied:

For the purpose of this study the test material was diluted to a 0.1% solution in distilled water before use.

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment.

Details on study design:

One rabbit was initially treated. A volume of 0.1 ml of the test material was instilled 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 instillation, to prevent loss of the test material, and then released. The left eye remained untreated and was used for control purposes. Immediately after administration of the test material, an assessment of the initial pain reaction was made. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.1

Max. score:

1

Reversibility:

fully reversible within: 24 h

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No corneal effects were noted during the study. Iridial inflammation was noted in two treated eyes at the 1-hour observation. Minimal conjunctival irritation was noted in all treated eyes at the 1-hour observation and one treated eye at the 24-hour observation. No ocular effects were noted in two treated eyes at the 1-hour observation and the remaining treated eye at the 48-hour observation.

Interpretation of results:

GHS criteria not met

Conclusions:

The test material did not produce positive criteria in any rabbit according to the GHS regulations. No symbol and P phrases are therefore required.

Executive summary:

A study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method used followed that described in the OECD Guidelines for Testing of Chemicals No. 405 and Method B5 of Commission Directive 92/69/EEC. A single instillation of a 0.1% solution of the test material to the non-irrigated eye of three rabbits produced iridial inflammation and minimal conjunctival irritation. No ocular effects were noted in two treated eyes at the 24-hour observation and the remaining treated eye at the 48-hour observation. The test material did not meet the criteria for classification as irritant according to GHS regulations.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

According to the GHS criteria, the test item needs to be classified as "Corrosive to skin Cat. 1B".