Benzyltriethylammonium chloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01-04-1986 - 08-04-1986

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Experimental test result performed using standard test guidelines

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1986

Report date:

1986

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Principles of method if other than guideline:

The purpose of the study was to evaluate the ability of the test chemical to produce ocular irritation/corrosion in rabbits following a single application.

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Benzyltriethylammonium chloride

EC Number:

200-270-1

EC Name:

Benzyltriethylammonium chloride

Cas Number:

56-37-1

Molecular formula:

C13H22N.Cl

IUPAC Name:

N-benzyl-N,N-diethylethanaminium chloride

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material :Benzyltriethylammonium chloride
- Molecular formula :C13H22N.Cl
- Molecular weight :227.777 g/mol
- Smiles notation :[N+](CC)(CC)(Cc1ccccc1)CC.[ClH-]
- InChl :1S/C13H22N.ClH/c1-4-14(5-2,6-3)12-13-10-8-7-9-11-13;/h7-11H,4-6,12H2,1-3H3;1H/q+1;/p-1
- Substance type:Organic
- Physical state:Solid (Powder)

Specific details on test material used for the study:

- Name of test material (as cited in study report): Benzyltriethylammonium chloride
- Molecular formula: C13H22NCl
- Molecular weight: 227.777 g/mol
- Smiles notation: [N+](CC)(CC)(Cc1ccccc1)CC.[ClH-]
- InChl: 1S/C13H22N.ClH/c1-4-14(5-2,6-3)12-13-10-8-7-9-11-13;/h7-11H,4-6,12H2,1-3H3;1H/q+1;/p-1
- Substance type: Organic
- Physical state: Solid
- Chemical name: Benzyltriethyl ammonium chloride
- Trade name/code: BETEC-DRY
- Batch No. HH-85-102A
- CAS No.: 56-37-1
- Purity: 100.1-100.2% based on Cl-
- Solubility: Water, ethanol
- Melting point:185°C dec.
- Appearance: Off-white crystalline powder
- Storage: At ambient temperature in the dark
- Comments: Material is hygroscopic, avoid undue exposure to moist air

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: The Broekman Institute, Someren, The Netherlands
- Age at study initiation: 8 to 10 weeks old
- Weight at study initiation: Three young adult female rabbits weighed as 2851, 2907 and 2873 grams
- Housing: individually housed in metal cages with perforated floors (RUCO, Valkenswaard)
- Diet (e.g. ad libitum): fed standard laboratory animal diet (100 g per day), obtained from Hope Farms, Woerden (LK-01, pellet diameter 4 mm)
- Water (e.g. ad libitum): free access to tap-water
- Acclimation period: 16 weeks of acclimation

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 22°C
- Humidity (%): 55 - 75 per cent
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): 12 hours light, 12 hours dark

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 69 mg (equivalent to 0.1 ml)
- Concentration (if solution): not applicable

VEHICLE
- Amount(s) applied (volume or weight with unit): not applicable
- Concentration (if solution):not applicable
- Lot/batch no. (if required): not applicable
- Purity: not applicable

Duration of treatment / exposure:

The test substance is applied in a single dose to one of the eyes of the experimental animals

Observation period (in vivo):

the eyes were examined approximately 1, 24, 4 and 72 hours, and 7 days after instillation of the test substance.

Duration of post- treatment incubation (in vitro):

Not applicable

Number of animals or in vitro replicates:

Not applicable

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Not specifed
- Time after start of exposure: Not specifed

TOOL USED TO ASSESS SCORE: fluorescein

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 1/24/48/72 hrs and 7 days

Score:

0

Max. score:

4

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

mean

Time point:

24/48/72 h

Score:

1.3

Max. score:

3

Reversibility:

fully reversible within: Seven days after dosing the conjunctival effects had disappeared.

Remarks on result:

other: in one of the eyes of three female rabbits, slight to moderate redness (persisting only for 72 hours) was observed

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.6

Max. score:

4

Reversibility:

fully reversible within: Seven days after dosing the conjunctival effects had disappeared.

Remarks on result:

other: In one of the eyes of three female rabbits, acute severe swelling of the conjunctivae was observed which diminished after 24 hrs;

Irritant / corrosive response data:

Table 1 shows the scores for irritation of the cornea, iris and conjunctivae approximately 1, 24, 48 and 72 hours, and 7 days after instillation of the test substance. Immediately after dosing the (animals were observed to squeeze their eyelids together, and lacrimation was excessive. One hour later slight conjunctival redness was observed in all animals, as well as moderate to severe swelling of the eyelids. At this time lacrimation was still increased, but this effect disappeared during the next twenty-four hours. The swelling of the conjunctivae diminished during this period, whereas the erythema became more pronounced. Forty-eight hours after dosing the blood vessels in the nictating membrane were injected, and in one animal (no. 251) this membrane was still slightly swollen. These effects were still present 72 hours after dosing, but disappeared completely during the next four days. Adverse effects on the cornea and iris were not observed in any of the rabbits during the entire observation period. Treatment of the eyes with fluorescein 24 hours after instillation of the test substance did not reveal any epithelial damage. Signs of systemic intoxication were not observed.

Any other information on results incl. tables

Table1:

EYE IRRITATION SCORES FOR BETEC-DRY IN THE RABBIT

Rabbit no. and sex	Body weight (grams)	Observation time after     instillation (hours)	Cornea opacity	Iris lesion	Conjunctivae
					redness	swelling (chemosis)
249; Female	2851	1	0	0	1	3
		24	0	0	2	1
		48	0	0	1	0
		72	0	0	1	0
		Day 7	0	0	0	0
Subtotal^a)/ Mean value^c)			0/0	0/0	4/1.3	1/0.3

 

Rabbit no. and sex	Body weight (grams)	Observation time after     instillation (hours)	Cornea opacity	Iris lesion	Conjunctivae
					redness	swelling (chemosis)
250; Female	2907	1	0	0	1	2
		24	0	0	1	1
		48	0	0	1	0
		72	0	0	1	0
		Day 7	0	0	0	0
Subtotal^a)/ Mean value^c)			0/0	0/0	3/1.0	1/0.3

 

Rabbit no. and sex	Body weight (grams)	Observation time after     instillation (hours)	Cornea opacity	Iris lesion	Conjunctivae
					redness	swelling (chemosis)
251; Female	2873	1	0	0	1	3
		24	0	0	2	1
		48	0	0	2	1
		72	0	0	1	1
		Day 7	0	0	0	0
Subtotal^a)/ Mean value^c)			0/0	0/0	5/1.7	3/1.0
Total^b)/ Mean value^c)			0/0	0/0	12/1.3	5/0.6

a) Subtotal = sum of 24-, 48- and 72-hours scores for each animal individually

b) Total = sum of 24-, 48- and 72-hours scores calculated over all animals.

c) Mean value = mean score of 24-, 48- and 72-hours reading time.

Applicant's summary and conclusion

Interpretation of results:

other: Not irritating

Conclusions:

Immediately after dosing the (animals were observed to squeeze their eyelids together, and lacrimation was excessive. One hour later slight conjunctival redness was observed in all animals, as well as moderate to severe swelling of the eyelids. These effects were still present 72 hours after dosing, but disappeared completely during the next four days. Adverse effects on the cornea and iris were not observed in any of the rabbits during the entire observation period. Treatment of the eyes with fluorescein 24 hours after instillation of the test substance did not reveal any epithelial damage. Signs of systemic intoxication were not observed.
Based on these observations and applying the EEC criteria for classification and labelling of dangerous substances (Annex VI of the EEC Council directive 67/548/EEC as amended by Directive 83/467/EEC), the test chemical need not be labelled as an Eye Irritant.

Executive summary:

The purpose of the study was to evaluate the ability of the test chemical to produce ocular irritation/corrosion in rabbits following a single application. The study was performed according to OECD 405 Guidelines and evaluated according to EEC criteria for classification and labelling of dangerous substances (Annex VI of the EEC Council directive 67/548/EEC as amended by Directive 83/467/EEC).3 young adult female New Zealand White rabbits were used for the study.

Before dosing the test substance was ground to a fine powder using a mortar and pestle. The bulk density of the test substance was determined by NOTOX and amounted to 0.698 g/ml. On the day of dose administration, three portions of 69±1.5 mg of the powder were dispensed in glass containers with screw caps. One portion of the dispensed amount (equivalent to 0.1 ml) was instilled into the conjunctival sac of the right eye of the rabbit using a spatula. The lids were then gently held together for two seconds and then released immediately after instillation of the test substance, the animals were observed and abnormalities were recorded. The eyes were examined approximately 1, 24, 48, 72 hours and 7 days after instillation of the test chemical. Immediately after treatment, the animals were transferred to metal cages. The left eye remained untreated and served as control. Immediately after instillation of the test substance, the animals were observed and abnormalities were recorded. The eyes were examined approximately 1, 24, 48, 72 hours and 7 days after instillation of the test chemical. Approximately 24 hours after instillation of the test substance (immediately after scoring the corneal opacity and the alterations of the iris and conjunctivae), a solution of 2% sodium fluorescien in water (pH adjusted to 7.0) was applied to both eyes of the test animals to examine quantitatively the potential for corneal injury. The brightly green staining area indicating epithelial damage was estimated as a percentage of total corneal area. Any observed local effects other than those indicated above were recorded. The ocular lesions were scored according to OECD 405 Guidelines and the results were evaluated according to the EEC criteria for classification and labeling of dangerous substances (Annex VI of the EEC Council directive 67/548/EEC as amended by Directive 83/467/EEC).

 

Immediately after dosing the (animals were observed to squeeze their eyelids together, and lacrimation was excessive. One hour later slight conjunctival redness was observed in all animals, as well as moderate to severe swelling of the eyelids. At this time lacrimation was still increased, but this effect disappeared during the next twenty-four hours. Forty-eight hours after dosing the blood vessels in the nictating membrane were injected, and in one animal (no. 251) this membrane was still slightly swollen. These effects were still present 72 hours after dosing, but disappeared completely during the next four days. Adverse effects on the cornea and iris were not observed in any of the rabbits during the entire observation period. Treatment of the eyes with fluorescein 24 hours after instillation of the test substance did not reveal any epithelial damage. Signs of systemic intoxication were not observed.

Based on these observations and applying the EEC criteria for classification and labelling of dangerous substances (Annex VI of the EEC Council directive 67/548/EEC as amended by Directive 83/467/EEC), the test chemical need not be labelled as an Eye Irritant.