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Administrative data

Description of key information

When tested for skin irritation/corrosion in vivo, Trihexyltetradecylphosphonium chloride was corrosive and classified in GHS Category 1B.

When tested for eye irritation, Trihexyltetradecylphosphonium chloride induced severe ocular damage and reversibility was not expected. Trihexyltetradecylphosphonium chloride is therefore considered to cause irreversible tissue damage to the eye.

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:
2015-03-27 - 2015-05-29
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Guideline study OECD 404, GLP
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
The dosing regimen was modified due to the severity of the responses observed following the 3-minute and 1-hour exposures on the initial animal.
GLP compliance:
yes
Specific details on test material used for the study:
Trade name: CYPHOS IL 101 Phosphonium Salt
Physical state: liquid
Composition of test material, percentage of components: Trihexyl(tetradecyl)phosphonium chloride (258864-54-9), > 95%
Lot/batch No.: WEC031280
Expiration date of the lot/batch: >1 year when stored at room temperature and protected from direct contact with water (hydrophobic)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Covance Research Products inc. Denver, PA
- Age at study initiation: 18-19 weeks old
- Weight at study initiation: 2.5 kg
- Housing: individually housed in a suspended metal cage
- Diet: Fresh PMI Rabbit Chow (Diet#5321) was provided daily
- Water: ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: temperature controlled
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount applied: A volume of 0.5 mL per site
Duration of treatment / exposure:
Site #1: 3 minutes
Site #2: 1 hour
Residual test article was removed with dry gauze
Observation period:
Site #1: Immediately and at 1 hour following the patch removal for the 3 minute exposure, and at 24, 48 and 72 hours and on Day 7 and 14.
Site #2: At 1 hour following patch removal for the one-hour exposure, and at 24, 48 and 72 hours and on Day 7 and 14.
Number of animals:
1 (female)
Details on study design:
TEST SITE
- Area of exposure: On the day before the test, the dorsal area of the trunk of the animal was clipped free of hair.
- % coverage: Each dose site was approximately 6 square cm (2.5 x 2.5 cm).
- Type of wrap if used: The test material, was introduced under a 2 cm X 3 cm cotton gauze patch. The patch was secured in position with a peice of porous dressing large enough to cover dose site with at least 5 cm square to spare on all sides of the gauze patch. To prevent the animal interfering with patch, the trunk was wrapped with a porous and non-irritating tape, and the animals were returned to their cages for the duration of the exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Both sites were gently wiped with dry gauze to remove residual test article

SCORING SYSTEM: Erythema and edema were scored according to the Draize scoring system.
The skin was also evaluated for ulceration and necrosis or any evidence of tissue destruction.

Erythema formation
No erythema: 0
Very slight erythema (barely perceptible): 1
Well defined erythema: 2
Moderate to severe erythema: 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth): 4
Maximum possible: 4

Edema formation
No edema: 0
Very slight edema (barely perceptible): 1
Slight edema (edges of area well-defined by definite raising): 2
Moderate edema (raised approximately 1.0 mm): 3
Severe edema (raised more than 1.0 mm, extending beyond area of exposure): 4
Maximum possible: 4

The body weight of the animal was recorded pretest and at the study termination. The animal was observed for toxicological and pharmacological effects at each dermal observation period and observed for mortality daily. The animal was humanely sacrificed using CO2 following study termination.
Irritation parameter:
erythema score
Basis:
animal #1
Remarks:
3-minute exposure
Time point:
other: Immediately
Score:
1
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #1
Remarks:
3-minute exposure
Time point:
24/48/72 h
Score:
4
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
edema score
Basis:
animal #1
Remarks:
3-minute exposure
Time point:
24/48/72 h
Score:
4
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
erythema score
Basis:
animal #1
Remarks:
1-hour exposure time
Time point:
24/48/72 h
Score:
4
Max. score:
4
Reversibility:
not reversible
Remarks on result:
other: necrosis
Irritation parameter:
edema score
Basis:
animal #1
Remarks:
1-hour exposure time
Time point:
24/48/72 h
Score:
4
Max. score:
4
Reversibility:
not reversible
Irritant / corrosive response data:
See Table 1. corrosive effects were observed (see Other effects for more details)

- Three-minute exposure: Immediately following the exposure, erythema was very slight and edema was absent. One hour following exposure, erythema was well-defined and edema was slight. 24, 48, and 72 hours post-exposure, erythema and edema were severe. On Day 7 and 14, erythema was severe and edema was moderate; severe eschar formation was observed.

- One-hour exposure: One hour following exposure, erythema was moderate to severe and edema was moderate; shiny areas were observed. 24 hours post-exposure, erythema and edema were severe; pale areas were observed. 48 and 72 hours post-exposure, erythema and edema were severe; moderate eschar formation and pale areas were observed. On Day 7, erythema was severe and edema was moderate, necrotic, cracking, and flaking skin and dark areas were observed. By Day 14, erythema was severe and edema was moderate, necrotic and flaking skin and shiny areas were observed.

- Other adverse systemic effects: No abnormal physical signs were observed. The animal gained weight by study termination

Immediately following patch removal 1 hour 24 hours 48 hours 72 hours 7 days 14 days
Site #1: 3-minute exposure Erythema 1 2 4 4 4 >4s >4s
Edema 0 2 4 4 4 3 3
Site #2: 1-hour exposure Erythema n.o. 3,s 4,p >4m,p >4m,p >4n,c,d,f >4n,f,s
Edema n.o. 3 4 4 4 3 3

n.o. - not observed; >4s = severe eschar; >4m = moderate eschar; >4n = necrosis

c = craking skin; d = dark area; f = flaking skin; p = pale areas; s = shiny areas

Interpretation of results:
Category 1B (corrosive) based on GHS criteria
Conclusions:
Under the conditions of this study (in vivo skin irritation test according to OECD guideline 404), Trihexyltetradecylphosphonium chloride was determined to be corrosive to skin. Based on these results, the test item is considered Category 1B according to GHS critera.
Executive summary:

In a primary dermal irritation study, a young adult New Zealand White rabbit (female) was dermally exposed to 0.5 mL of Trihexyltetradecylphosphonium chloride. Two test sites were covered with a semi-occlusive dressing for an exposure period of three minutes on site#1 and 1 hour on site #2. Skin reactions were assessed immediately and at 1 hour after patch removal. Reactions were scored again at 24, 48 and 72 hours and on Days 7 and 14 following patch removal for both sites.

Three-minute exposure: 24, 48, and 72 hours post-exposure, erythema and edema were severe. On Day 7 and 14, erythema was severe and edema was moderate; severe eschar formation was observed.

One-hour exposure: 24, 48 and 72 hours post-exposure, erythema and edema were severe. On Day 7, erythema was severe and edema was moderate, necrotic, cracking, and flaking skin and dark areas were observed. By Day 14, erythema was severe and edema was moderate, necrotic and flaking skin and shiny areas were observed.

Given these results, Trihexyltetradecylphosphonium chloride is corrosive and classified in GHS Category 1B.

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:
From 27 MAR 2015 to 29 MAY 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP and OECD test guideline-compliant study
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
Trade name: CYPHOS IL 101 Phosphonium Salt
Physical state: liquid
Composition of test material, percentage of components: Trihexyl(tetradecyl)phosphonium chloride (258864-54-9), > 95%
Lot/batch No.: WEC031280
Expiration date of the lot/batch: >1 year when stored at room temperature and protected from direct contact with water (hydrophobic)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Covance Research Products, Inc., Denver, PA
- Age at study initiation: 18 week old
- Weight at study initiation: 2.7 kg
- Housing: individually housed in a suspented cage
- Diet: Fresh PMI Rabbit Chow (Diet#5321) was provided daily
- Water: available ad libitum
- Acclimation period: At 5 five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): temperature controlled
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Test animal was instilled with 0.1 mL 

VEHICLE: none (undiluted)
Duration of treatment / exposure:
Single application, not washed
Observation period (in vivo):
60 min, 24, 48 and 72 hours post dose
Number of animals or in vitro replicates:
1 male
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): not washed

SCORING SYSTEM: Occular reactions were graded according to the numerical Draize technique

TOOL USED TO ASSESS SCORE: Sodium fluorescein dye procedure were used at the 24-, 48-, 72-hour observation intervals
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: 1 Hour
Score:
3
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: 1 Hour
Score:
1
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: 1 Hour
Score:
2
Max. score:
3
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 1 Hour
Score:
3
Max. score:
4
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Reversibility:
fully reversible within: 48h
Remarks on result:
other: Although the score for 48 and 72 hours is 0, pale areas were noted
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
4
Max. score:
4
Remarks on result:
other: No expectation of reversibility after 72-hour post dose
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Remarks on result:
not determinable
Remarks:
Due to chemosis
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Remarks on result:
not determinable
Remarks:
Due to chemosis
Irritant / corrosive response data:
See Table 1
The study was terminated at 72 hours since there was no improvement in chemosis of the treated eye. Reversibility of the occular reactions was not expected.

After 72 hours of exposure, severe ocular lesions were observed, mainly on conjunctiva, with swelling and abundant lachrymal discharge.
Due to chemosis, corneal opacity and iritis could not be determined at the 24-, 48-, 72-hour time points.

The animal lost weight. The animal had diminished food consumption at 72 hours post-dose.

Table 1: Occular findings at each observation time

Tissue Reading 1 Hour 24 Hour 48 Hour 72 Hour
Cornea Opacity Max. Score: 4 3 ? ? ?
Area Max. Score: 4 2 ? ? ?
Iris Max. Score: 2 1 ? ? ?
Conjunctiva Redness Max. Score: 3 2 3 0 p  0 p 
Chemosis Max. Score: 4 3 4 4 4
Discharge Max. Score: 3 3 3 3 3

Details of eye irritation scores (according to Draize)

The control eye appeared normal at all observation periods

? = unable to determine due to chemosis

p = pale areas

Table 2: Systemic Observations

Time periods
Day 1 A/A
Day 2 A/A
Day 3 1/1

A/A = Appeared normal (a.m./p.m. observations)

1/1 = diminished food consumption (a.m./p.m. observations)

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
Trihexyltetradecylphosphonium chloride is considered to cause irreversible tissue damage to the eye. Conjunctival chemosis was still observed at 72-hour post dose without expectation of reversibility
Executive summary:

Trihexyltetradecylphosphonium chloride, has been tested for acute eye irritation on 1 male New Zealand White rabbit, in accordance with OECD test guideline 405, and in compliance with Good Laboratory Practice.

The test article was applied as such in a single conjunctival dose of 0.1 mL in the right eye, the untreated eye being used as a control. Ocular examinations, grading cornea, iris and conjunctiva (redness, chemosis and discharge) reactions, were performed at 1, 24, 48, 72 hours after instillation of the test article.

Within the first hour post dose, the test article induced conjunctival abnormalities including hyperhaemia grade 2 (diffuse, crimson color, individual vessels not easily discernible), edema grade 3 and conjunctivae discharge (grade 3). Corneal opacity and injection of the iris (grade 3 and 1, respectively) were also observed.

At the observations carried out 24, 48 and 72 hours after instillation, hyperhaemia chemosis of grade 4 and discharge of grade 3 were observed. Due to chemosis, corneal opacity and iritis could not be determined at the 24-, 48-, 72-hour time points.

The study was terminated at 72 hours since there was no improvement in chemosis of the treated eye. Reversibility of the ocular reactions was not expected.

 

Based on these results, Trihexyltetradecylphosphonium chloride was considered to cause irreversible tissue damage to the eye.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

Trihexyltetradecylphosphonium chloride was tested in vivo in a Klimisch score 1 study (2015), used as a key study. In this study, the test substance was dermally applied to a young adult New Zealand White rabbit. Two test sites were covered with a semi-occlusive dressing for an exposure period of three minutes on site#1 and 1 hour on site #2. The test sites were examined for evidence of primary irritation and scored according to Draize method.

The three-minute exposure resulted in severe erythema and edema visible 24, 48, and 72 hours post-exposure. On Day 7 and 14, erythema was severe and edema was moderate; severe eschar formation was observed.

One-hour exposure: 24, 48 and 72 hours post-exposure, erythema and edema were severe. On Day 7, erythema was severe and edema was moderate, necrotic, cracking, and flaking skin and dark areas were observed. By Day 14, erythema was severe and edema was moderate, necrotic and flaking skin and shiny areas were observed.

Eye irritation:

Trihexyltetradecylphosphonium chloride was tested in vivo in a Klimisch score 1 study (2015), used as a key study. In this study, the test substance was instilled in the eyes of rabbits observed for up 72 hours. At the observations carried out 24, 48 and 72 hours after instillation, hyperemia chemosis and discharge were observed. The study was terminated at 72 hours since there was no improvement in chemosis of the treated eye. Reversibility of the ocular reactions was not expected.

Justification for selection of skin irritation / corrosion endpoint:
Recent GLP and OECD test guideline-compliant study (Klimisch score 1)

Justification for selection of eye irritation endpoint:
Recent GLP and OECD test guideline-compliant study (Klimisch score 1)

Justification for classification or non-classification

Skin irritation:

In a guideline in vivo skin irritation study, the test substance was corrosive to skin. Hence, the following classification is proposed: Skin Corr. 1B - H314: causes severe skin burns and eye damage according to Regulation (EC) 1272/2008.

 

Eye irritation:

In a guideline in vivo eye irritation/corrosion study, the test substance was considered to cause irreversible tissue damage to the eye. Trihexyltetradecylphosphonium chloride should therefore be classified as Eye Damage 1 - H318: causes serious eye damage according to Regulation (EC) 1272/2008.