(hexadecylamidopropyl)trimethylammonium chloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

February 24, 1987

Deviations:

yes

Remarks:

observation time only 7 d, thus, reversibility cannot finally be judged / test substance was used as 30% solution

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

(hexadecylamidopropyl)trimethylammonium chloride

EC Number:

257-104-6

EC Name:

(hexadecylamidopropyl)trimethylammonium chloride

Cas Number:

51277-96-4

Molecular formula:

C22H47N2O.Cl

IUPAC Name:

(3-hexadecanamidopropyl)trimethylazanium chloride

Constituent 2

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

water

Test material form:

solid - liquid: aqueous solution

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS- Age at study initiation: young adult- Weight at study initiation: Males: 2.175-2.500 kg; Females: 2.250-2.650 kg- Housing: individually in suspended wire bottom stainless steel cages- Diet (e.g. ad libitum): PM1 Rabbit Diet #5321; presented in measured amounts- Water (e.g. ad libitum): municipal water supply, ad libitum- Acclimation period: 5 dENVIRONMENTAL CONDITIONS- Temperature (°C): 72 ± 5°F (= 22°C)- Humidity (%): 30 - 80- Air changes (per hr): 10 - 12- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The untreated left eyes served as controls.

Amount / concentration applied:

TEST MATERIAL- Amount(s) applied (volume or weight with unit): 0.1 mL

Duration of treatment / exposure:

3 animals: treated eyes were washed after 30 s, 6 animals: treated eyes were washed after 24 h

Observation period (in vivo):

7 d (the study was terminated on Day 7)

Number of animals or in vitro replicates:

9 (3 washed after 30 s, 6 washed after 24 h)

Details on study design:

REMOVAL OF TEST SUBSTANCE- Washing (if done): Three of the treated eyes (“washed eyes”) were each washed with room temperature deionized water for one minute beginning 30 seconds after treatment. All treated eyes were washed with room temperature deionized water for one minute immediately after recording the 24-hour observation.SCORING SYSTEM: Kay & Calandra ScaleTOOL USED TO ASSESS SCORE: white room lighting (daylight-type fluorescent tubes) + additional source of white light / fluorescein

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 d

Remarks on result:

positive indication of irritation

Remarks:

eyes washed after 24 h; 24 h unable to score due to severity of chemosis

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #4

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 d

Remarks on result:

positive indication of irritation

Remarks:

eyes washed after 24 h;24 h unable to score due to severity of chemosis

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #5

Time point:

24/48/72 h

Score:

1.5

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 d

Remarks on result:

positive indication of irritation

Remarks:

eyes washed after 24 h; 24 h unable to score due to severity of chemosis

Results 4

Irritation parameter:

cornea opacity score

Basis:

animal #6

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 d

Remarks on result:

positive indication of irritation

Remarks:

eyes washed after 24 h; 24 h unable to score due to severity of chemosis

Results 5

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: eyes washed after 24 h; 24 h unable to score due to severity of chemosis

Results 6

Irritation parameter:

iris score

Basis:

animal: #4

Time point:

24/48/72 h

Score:

0.5

Max. score:

2

Reversibility:

fully reversible within: 4 d

Remarks on result:

other: eyes washed after 24 h; 24 h unable to score due to severity of chemosis; constricted pipils still present at day 7

Results 7

Irritation parameter:

iris score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0.5

Max. score:

2

Reversibility:

not fully reversible within: 7 d

Remarks on result:

other: eyes washed after 24 h; 24 h unable to score due to severity of chemosis

Results 8

Irritation parameter:

iris score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0.5

Max. score:

2

Reversibility:

fully reversible within: 4 d

Remarks on result:

other: eyes washed after 24 h; 24 h unable to score due to severity of chemosis

Results 9

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not reversible

Remarks:

within 7 d

Remarks on result:

positive indication of irritation

Remarks:

eyes washed after 24 h

Results 10

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2.7

Max. score:

3

Reversibility:

not reversible

Remarks:

within 7 d

Remarks on result:

positive indication of irritation

Remarks:

eyes washed after 24 h

Results 11

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

not reversible

Remarks:

within 7 d

Remarks on result:

positive indication of irritation

Remarks:

eyes washed after 24 h

Results 12

Irritation parameter:

conjunctivae score

Basis:

animal: #4, #5, #6

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not reversible

Remarks:

within 7 d

Remarks on result:

positive indication of irritation

Remarks:

eyes washed after 24 h

Results 13

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #4, #5, #6

Time point:

24/48/72 h

Score:

2.3

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 d

Remarks on result:

positive indication of irritation

Remarks:

eyes washed after 24 h

Results 14

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2.7

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 d

Remarks on result:

positive indication of irritation

Remarks:

eyes washed after 24 h

Irritant / corrosive response data:

Washing after 30 s: Fluorescein staining was observed in all three eyes at 24 hours after treatment and persisted in two eyes through Day 7 after treatment. Corneal opacity was not possible to score after 1 h in 2/3 animals due to severity of chemosis. A slight dulling of normal luster (graded as 0.5) was present in all 3 animals from 24 h after treatment to day 4 or 7, respectively, after treatment. In 1/3 animals corneal opacity, grade1 was observed on day 7. Iris scores were 1 in all three animals 1 h and 24 h after treatment, and zero thereafter. Constricted pupils were observed in 2/3 animals up to day 4. Conjunctial redness grade 2 was present in all three animals after 1 h, advancing to grade 3 in 2/3 animals after 24 h. Grade 1-2 conjunctival redness was observed after 48 h, 72 h and after 4 d, and was still present in 2/3 animals at the end of the observation period of 7 d.Grade 3 chemosis was observed in 3/3 animals at the 1 h observation, grade 2 chemosis in 3/3 animals at the 24 h observation, grade 1-2 chemosis was observed after 48 h, 72 h and after 4 d, and was still present in 2/3 animals at the end of the observation period of 7 d.Grade 3 discharge was present in 3/3 animals after 1 h, receding to grade 2 after 2 h and grade 1 thereafter. Slight discharge (grade 1) was still present on day 7 in 1/3 animals.Washing after 24 h: Fluorescein staining was observed in all six eyes at 48 hours after treatment and persisted in all six eyes through Day 7 after treatment.Corneal opacity and iris were not possible to score after 1 and 24 h in all 6 animals due to severity of chemosis. Corneal opacity, grade 1 (4/6 animals) and 2 (2/6 animals) was observed after 48 h. From day 3 to 7, corneal opacity grade 2 was present in 3/6 animals and grade 1 in 3/6 animals. Iris scores were 0 in all six animals 48 h after treatment, 1 at the 72 h observation in 3/6 animals and was still present (grade 1) in 1/6 animals after 7 d. Constricted pupils were observed in 5/6 animals at the 48 h observation, in 4/6 animals at the 72 h observation, in 3/6 animals on day 4 and was still present in 2/6 animals on day 7.Conjunctial redness grade 2 was present in all three animals after 1 h. Grade 2-3 conjunctival redness was present at the 24, 48, 72 and 4 d observations. Grade 2 conjunctival redness was still observed after 7 d in all six animals. Grade 3 chemosis was present in all six animals at the 1 h and 24 h observations, and in 1/6 animals at the 48 h observation. Grade 2 chemosis was present in 5/6 animals at the 48 h observation, and in all six animals at the 72 h and 4 d observation. Grade 1 (1/6 animals) – 2 (5/6 animals) chemosis was still present in all six animals at the end of the observation period of 7 d.Grade 1 – 3 discharge was observed throughout the study period and was still present at the end of the observation time in 5/6 animals.

Any other information on results incl. tables

Irritant/corrosive response data for each animal at each observation time up to removal of each animal from the test

Washed after 30 s

Score at time point / Reversibility	Cornea	Iris	Conjunctivae	Chemosis	Discharge
	Max. score: 4	Max. score: 2	Max. score: 3	Max. score: 4	Max. score: 3
60 min	+/f/f	1/1/1	2/2/2	3/3/3	3/3/3
24 h	+/+/+	1/1/1	2/3/3	2/2/2	2/2/2
48 h	+/+/+	0c/0c0c/	1/1/2	1/2/1	1/1/1
72 h	+/+/+	0c/0c/0	1/2/2	1/2/1	1/1/1
4 d	+/+/+	0c/0c/0	1/2/2	1/2/1	0/1/1
7 d	+/1e/0	0/0/0	1/2/0	1/2/0	0/1/0
Average 24h, 48h, 72h	0.5/0.5/0.5	0.3/0.3/0.3	1.3/2/2.3	1.3/2/1.3	1.3/1.3/1.3

 

Washed after 24 h

Score at time point / Reversibility	Cornea	Iris	Conjunctivae	Chemosis	Discharge
	Max. score: 4	Max.score: 2	Max. score: 3	Max. score: 4	Max. score: 3
60 min	f/f/f/f/f/f	f/f/f/f/f/f	2/2/2/2/2/2	3/3/3/3/3/3	3/3/2/3/3/3
24 h	f/f/f/f/f/f	f/f/f/f/f/f	3/3/2/3/3/3	3/3/3/3/3/3	2/3/2/3/3/3
48 h	1/1/1/2/1/2	0/0c/0c/0c/0c/0c	3/3/2/3/3/3	2/2/3/2/2/2	2/2/2/2/2/2
72 h	1/1/1/2/2/2	0/0/0c/1c/1c/1c	3/2/2/3/3/3	2/2/2/2/2/2	2/2/2/2/2/2
4 d	1/1/1/2/2/2	0/0/0c/0c/0c/0	3/2/2/3/2/3	2/2/2/2/2/2	2/1/1/2/2/2
7 d	1/1/1/2/2/2	0/0/0/0c/1c/0	2/2/2/2/2/2	2/2/1/2/2/2	2/1/0/2/2/2
Average 24h, 48h, 72h	1/1/1/2/1.5/2	0/0/0/0.5/0.5/0.5	3/2.7/2/3/3/3	2.3/2.3/2.7/2.3/2.3/2.3	2/2.3/2/2.3/2.3/2.3

 

+Slight dulling of normal luster, scored as 0.5

f) unable to score due to severity of chemosis

c) constricted pupils

e) apparent invasion of cornea by blood vessels

 

Fluorescein staining: 

	Eyes washed after 30 s	Eyes washed after 24 h
24 h	D/D/C	f/f/f/f/f/f
48 h	D/C/C	D/D/D/D/D/D
72 h	D/C/B	D/D/C/D/D/D
4 d	C/B/B	C/C/B/C/C/C
7 d	B/B/0	C/B/A/C/C/B

 

 

Area of cornea involved

No fluorescein staining: 0

One quarter (or less), but not zero: A

Greater than one quarter, but less than half: B

Greater than half, but less than three quarters: C

Greater than three quarters, up to whole area: D

 

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

In this study, C16 Alkylamidopropyltrimethylammonium Chloride has irreversible effects on the eyes (Category 1) based on the effects being not reversible at the end of the observation period of 7 d.

Executive summary:

In a primary eye irritation study according to OECD guideline 405 (February 24, 1987) 0.1 mL of C16 Alkylamidopropyltrimethylammonium Chloride (30% a.i.) was instilled into the conjunctival sac of the right eye of 9 young adult New Zealand White rabbits. The untreated left eyes served as controls. In 3 animals, the treated eyes were rinsedwith room temperature deionized water for one minute beginning 30 seconds after treatment. In the remaining 6 animals, the treated eyes were rinsedwith room temperature deionized water for one minute immediately after recording the 24-hour observation.

Animals were then observed for 7 days.  Irritation was scored by the method of Kay and Calandra.

 

Washing after 30 s:

Fluorescein staining was observed in all three eyes at 24 hours after treatment and persisted in two eyes through Day 7 after treatment.

Corneal opacity was not possible to score after 1 h in 2/3 animals due toseverity of chemosis.A slight dulling of normal luster (graded as 0.5) was present in all 3 animals from 24 h after treatment to day 4 or 7, respectively, after treatment. In 1/3 animals corneal opacity, grade1was observed on day 7.

Iris scores were 1 in all three animals 1 h and 24 h after treatment, and zero thereafter. Constricted pupils were observed in 2/3 animals up to day 4.

Conjunctial redness grade 2 was present in all three animals after 1 h, advancing to grade 3 in 2/3 animals after 24 h. Grade 1-2 conjunctival redness was observed after 48 h, 72 h and after 4 d, and was still present in 2/3 animals at the end of the observation period of 7 d.

Grade 3 chemosis was observed in 3/3 animals at the 1 h observation, grade 2 chemosis in 3/3 animals at the 24 h observation, grade 1-2 chemosis was observed after 48 h, 72 h and after 4 d, and was still present in 2/3 animals at the end of the observation period of 7 d.

Grade 3 discharge was present in 3/3 animals after 1 h, receding to grade 2 after 2 h and grade 1 thereafter. Slight discharge (grade 1) was still present on day 7 in 1/3 animals.

Score at time point / Reversibility	Cornea	Iris	Conjunctivae	Chemosis	Discharge
	Max. score: 4	Max. score: 2	Max. score: 3	Max. score: 4	Max. score: 3
60 min	+/f/f	1/1/1	2/2/2	3/3/3	3/3/3
24 h	+/+/+	1/1/1	2/3/3	2/2/2	2/2/2
48 h	+/+/+	0c/0c0c/	1/1/2	1/2/1	1/1/1
72 h	+/+/+	0c/0c/0	1/2/2	1/2/1	1/1/1
4 d	+/+/+	0c/0c/0	1/2/2	1/2/1	0/1/1
7 d	+/1e/0	0/0/0	1/2/0	1/2/0	0/1/0
Average 24h, 48h, 72h	0.5/0.5/0.5	0.3/0.3/0.3	1.3/2/2.3	1.3/2/1.3	1.3/1.3/1.3

 

Washing after 24 h:

Fluorescein staining was observed in all six eyes at 48 hours after treatment and persisted in all six eyes through Day 7 after treatment.

Corneal opacity and iris were not possible to score after 1 and 24 h in all 6 animals due toseverity of chemosis.

Corneal opacity, grade 1 (4/6 animals) and 2 (2/6 animals) was observed after 48 h. From day 3 to 7, corneal opacity grade 2 was present in 3/6 animals and grade 1 in 3/6 animals.

Iris scores were 0 in all six animals 48 h after treatment, 1 at the 72 h observation in 3/6 animals and was still present (grade 1) in 1/6 animals after 7 d. Constricted pupils were observed in 5/6 animals at the 48 h observation, in 4/6 animals at the 72 h observation, in 3/6 animals on day 4 and was still present in 2/6 animals on day 7.

Conjunctial redness grade 2 was present in all three animals after 1 h. Grade 2-3 conjunctival redness was present at the 24, 48, 72 and 4 d observations. Grade 2 conjunctival redness was still observed after 7 d in all six animals.

Grade 3 chemosis was present in all six animals at the 1 h and 24 h observations, and in 1/6 animals at the 48 h observation.

Grade 2 chemosis was present in 5/6 animals at the 48 h observation, and in all six animals at the 72 h and 4 d observation. Grade 1 (1/6 animals) – 2 (5/6 animals) chemosis was still present in all six animals at the end of the observation period of 7 d.

Grade 1 – 3 discharge was observed throughout the study period and was still present at the end of the observation time in 5/6 animals.

 

Score at time point / Reversibility	Cornea	Iris	Conjunctivae	Chemosis	Discharge
	Max. score: 4	Max.score: 2	Max. score: 3	Max. score: 4	Max. score: 3
60 min	f/f/f/f/f/f	f/f/f/f/f/f	2/2/2/2/2/2	3/3/3/3/3/3	3/3/2/3/3/3
24 h	f/f/f/f/f/f	f/f/f/f/f/f	3/3/2/3/3/3	3/3/3/3/3/3	2/3/2/3/3/3
48 h	1/1/1/2/1/2	0/0c/0c/0c/0c/0c	3/3/2/3/3/3	2/2/3/2/2/2	2/2/2/2/2/2
72 h	1/1/1/2/2/2	0/0/0c/1c/1c/1c	3/2/2/3/3/3	2/2/2/2/2/2	2/2/2/2/2/2
4 d	1/1/1/2/2/2	0/0/0c/0c/0c/0	3/2/2/3/2/3	2/2/2/2/2/2	2/1/1/2/2/2
7 d	1/1/1/2/2/2	0/0/0/0c/1c/0	2/2/2/2/2/2	2/2/1/2/2/2	2/1/0/2/2/2
Average 24h, 48h, 72h	1/1/1/2/1.5/2	0/0/0/0.5/0.5/0.5	3/2.7/2/3/3/3	2.3/2.3/2.7/2.3/2.3/2.3	2/2.3/2/2.3/2.3/2.3

 

Reversibility cannot finally be judged as thestudy was terminated on day 7.

Thus, C16 Alkylamidopropyltrimethylammonium Chloride (30% a.i.), produced irreversible effects to the eye in this study and thus has to be classified as Category 1 (irreversible effects on the eye).