Registration Dossier

Skin: OECD 404, rabbit. Not irritating. Reliability = 2 (read-across substance)

Eye: OECD 405; rabbit. Severely irritating. Reliability = 2 (read-across substance)

Reference

Endpoint:

skin irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From 1986-02-11 to 1986-02-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Although this study is guideline and GLP compliant and would normally be assigned a reliability of 1 (reliable without restrictions), this study is used for read-across and therefore has been assigned a reliability of 2 (reliable with restrictions).

Justification for type of information:

The primary component of all substances provide complete coverage of 68334-33-8. 68334-33-8 shares high structural similarity with 61789-80-8, 68783-78-8, 107-64-2, and 112-02-7. As 68334-33-8 is a UVCB its components encapsulate the other substances except for the counter ion (Cl-). In solution, the counter ions will dissociate from the parent structures. Therefore, we are comparing substances of equivalent nature. CAS 107-64-2 represents the C18 boundary of the 61789-80-8. Ignoring the salt component CAS 61789-80-8 is equivalent to CAS 68334-33-8. 68783-78-8 is a worst case of both 68334-33-8 and 61789-80-8 since it is unsaturated and the SP2 carbon-carbon bonds are of higher reactivity and a more likely site of metabolic activation. The primary component of CAS 112-02-7 is a substructure of all the other substances. Additional documentation, provided within the IUCLID Assessment Reports section, supports the read-across approach.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hoechst AG, Kastengrund
- Age at study initiation: 3-5 months
- Weight at study initiation: 2.2-3.4 kg
- Housing: individually
- Diet: ad libitum
- Water : ad libitum
- Acclimation period: 1 week prior to the start of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3°C
- Humidity (%): 50±20%
- Photoperiod : 12 hrs dark / 12 hrs light

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

physiological saline

Controls:

other: each animal served as its own control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied : 0.5 g
- Concentration (if solution):

VEHICLE
- Amount(s) applied : 0.9ml isotonic saline.
0.5 g test substance pasted with 0.9 ml isotonic saline.

Duration of treatment / exposure:

4 hours

Observation period:

30 min, 60 min, 24, 48, 72 hrs and 7d, 14d after removal the patch

Number of animals:

3 rabbits

Details on study design:

TEST SITE
- Area of exposure: skin of the back
- % coverage: 2.5 cm x 2.5 cm
- Type of wrap if used: semi-occlusive

REMOVAL OF TEST SUBSTANCE
- Washing (if done): residual test substance was removed from skin with warm tap water
- Time after start of exposure: 4 hours

SCORING SYSTEM: according to OECD guideline 404 (draize scoring system)

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 2 days

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

30-60 minutes and 24 hours after patch removal the animals showed very slight to well defined erythema. 48 hours to 7 days after patch removal two rabbits showed a very slight or well defined erythema. 14 days after application erythema and oedema formation were reversed. Additionally, the treated skin areas were sporadically dry-rough, discoloured light brown and fine or coarse scaly during the observation period.

Individual skin irritation scores according to the OECD 404 Guideline
Time after patch removal	30-60 minutes			24 h			48 h			72 h			7 d			14 d
Animal no.	1	2	3	1	2	3	1	2	3	1	2	3	1	2	3	1	2	3
Erythema	2	1	1	2	1	1	2	1	0	2	1	0	2	1	0	0	0	0
Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

Skin surface
- dry rough						X	X		X	X	X	X	X	X	X	X	X
- fine scaly						X								X			X
- coarse scaly													X			X
- large disol. light brown										X

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions of this study and according to the criteria laid down in CLP (Reg 1272/2008/EC) and directive 67/548/EEC, the test item dimethyldioctadecylammonium chloride (purity 97%) is not classified.

Executive summary:

The potential of Dimethyldioctadecylammonium chloride (purity 97%) to induce skin irritation was assessed in 3 rabbits according to the OECD guideline 404 and to the EU Method B.4. The study was conducted in compliance with the principles of Good Laboratory Practice regulations.

A single dose of 0.5 g pasted with 0.9 ml isotonic saline was applied to the skin of the rabbits under a semi-occlusive dressing for a 4-hour exposure period. Skin reactions were observed approximately 0.5, 1, 24, 48 and 72 hours after removal of the dressing and then on day 7, 14 in order to observe their reversibility. The mean values of the scores for erythema and oedema were calculated for each animal.

The mean scores over 24, 48 and 72 hours for individual animals were 2.0, 1.0, and 0.3 for erythema and 0.0, 0.0, and 0.0 for oedema and all reactions reversed within 14 days. Oedema were not observed. In addition, the treated skin sites were sporadically dry-rough, discoloured light brown and fine or coarse scales occurred during the observation period of 14 days. Based on the study results, the test material is not subject to labelling requirements with regard to skin irritation.

Endpoint conclusion:

no adverse effect observed (not irritating)

Reference

Endpoint:

eye irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

from 1990-12-13 to 1990-12-14

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Although this study is guideline and GLP compliant and would normally be assigned a reliability of 1 (reliable without restrictions), this study is used for read-across and therefore has been assigned a reliability of 2 (reliable with restrictions).

Justification for type of information:

The primary component of all substances provide complete coverage of 68334-33-8. 68334-33-8 shares high structural similarity with 61789-80-8, 68783-78-8, 107-64-2, and 112-02-7. As 68334-33-8 is a UVCB its components encapsulate the other substances except for the counter ion (Cl-). In solution, the counter ions will dissociate from the parent structures. Therefore, we are comparing substances of equivalent nature. CAS 107-64-2 represents the C18 boundary of the 61789-80-8. Ignoring the salt component CAS 61789-80-8 is equivalent to CAS 68334-33-8. 68783-78-8 is a worst case of both 68334-33-8 and 61789-80-8 since it is unsaturated and the SP2 carbon-carbon bonds are of higher reactivity and a more likely site of metabolic activation. The primary component of CAS 112-02-7 is a substructure of all the other substances. Additional documentation, provided within the IUCLID Assessment Reports section, supports the read-across approach.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage cunicole de Val de Selle, Prouzel, France.
- Weight at study initiation: 2,5 kg.
- Housing: individually in polystyren cage.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3 °C
- Humidity (%): 50 ± 20 %
- Photoperiod: 12 hrs dark /12 hrs light

Controls:

other: each animal served as its own control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied : 0.1ml previously melted at 20 - 25°C.

Duration of treatment / exposure:

Single exposure

Observation period (in vivo):

1 hour and 24 hours after instillation

Number of animals or in vitro replicates:

1 male

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing : eye not rinsed

SCORING SYSTEM: according to OECD guideline 405 (draize scoring system)

Irritation parameter:

overall irritation score

Basis:

animal #1

Time point:

other: 1-hour and 24-hour readings

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks on result:

other: the ocular lesions noted after 24 hours in the rabbit were so severe that the animal was sacrificed for ethical grounds

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24 h

Remarks on result:

probability of severe irritation

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24 h

Remarks on result:

probability of severe irritation

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24 h

Remarks on result:

not determinable

Remarks:

the animal was sacrificed for ethical grounds before the nature of the reactions concerning the iris and the cornea could have been checked

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24 h

Remarks on result:

not determinable

Remarks:

the animal was sacrificed for ethical grounds before the nature of the reactions concerning the iris and the cornea could have been checked

Irritant / corrosive response data:

1 hour then 24 hours after instillation of the test substance, the ocular reactions were severe and the conjunctival chemosis (score of 4) obscured the evaluation of the reactions at the iris and cornea. The flowing of a whitish purulent substance was also noted.
Some severe ocular lesions were expected at the next reading times but taking into consideration the severity of the lesions noted after 24 hours, the animal was sacrificed for ethical grounds before the nature of the reactions concerning the iris and the cornea could have been checked.

Interpretation of results:

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

Conclusions:

Under the experimental conditions, the test material was considered as severely irritating when administered by ocular route to rabbits.

Executive summary:

The potential of the technical grade dehydrogenated tallow dimethylammonium, chlorides (78% active in isopropanol/water) to induce eye irritation was assessed in 1 rabbit according to the OECD guideline 405 in compliance with the principles of Good Laboratory Practice regulations. Taking into account the highly irritating effects observed in the acute dermal irritation/ corrosion test previously performed in rabbits, it was decided for ethical considerations, to perform the assay with one animal. A single dose of 0.1 mL of the test material was instilled into one eye, the other eye was not treated and served as control. The eyes were not rinsed after administration of the test item. Ocular reactions were observed 1 hour and 24 hours later.

1 hour then 24 hours after instillation of the test substance, the ocular reactions were severe and the conjunctival chemosis (score of 4) obscured the evaluation of the reactions at the iris and cornea. The flowing of a whitish purulent substance was also noted. Some severe ocular lesions were expected at the next reading times but taking into consideration the severity of the lesions noted after 24 hours, the animal was sacrificed for ethical grounds before the nature of the reactions concerning the iris and the cornea could have been checked. Under these experimental conditions, the test material was considered as severely irritating when administered by ocular route to rabbits.

Endpoint conclusion:

adverse effect observed (irreversible damage)

Endpoint conclusion:

no study available

No skin or eye irritation studies with the test substance are available. Therefore, a study with DHTDMAC (CAS 61789-80-8) was used as read across to fulfil the data gap for eye irritation and DODMAC (CAS 107-64-2) was used as read across to fulfil the data gap for skin irritation. The primary component of all substances provide complete coverage of 68334-33-8. 68334-33-8 shares high structural similarity with 61789-80-8, 68783-78-8, 107-64-2, and 112-02-7. As 68334-33-8 is a UVCB its components encapsulate the other substances except for the counter ion (Cl-). In solution, the counter ions will dissociate from the parent structures. Therefore, we are comparing substances of equivalent nature. CAS 107-64-2 represents the C18 boundary of the 61789-80-8. Ignoring the salt component CAS 61789-80-8 is equivalent to CAS 68334-33-8. 68783-78-8 is a worst case of both 68334-33-8 and 61789-80-8 since it is unsaturated and the SP2 carbon-carbon bonds are of higher reactivity and a more likely site of metabolic activation. The primary component of CAS 112-02-7 is a substructure of all the other substances. Additional documentation, provided within the IUCLID Assessment Reports section, supports the read-across approach.

Pure DODMAC (97% in water) applied to the skin of 3 rabbits under a semi-occlusive dressing, was only slightly irritating to skin following a 4-hour exposure period according to OECD Guideline 404. In this study, skin reactions were observed approximately 0.5, 1, 24, 48 and 72 hours after removal of the dressing and then on day 7, 14 in order to observe their reversibility. The mean scores over 24, 48 and 72 hours for individual animals were 2.0, 1.0, and 0.3 for erythema and 0.0, 0.0, and 0.0 for oedema. Whereas no oedema was observed following treatment, mild to moderate erythema occurred which reversed fully within the observation period of 14 days.

The potential of the technical grade DHTDMAC (78% active in isopropanol/water) to induce eye irritation was assessed in 1 rabbit according to the OECD Guideline 405. As severe ocular effects were anticipated, it was decided for ethical considerations to perform the assay with one animal. A single dose of 0.1 mL of the test material was instilled into one eye, the other eye was not treated and served as control. The eye was not rinsed after administration of the test item. Ocular reactions were observed 1 hour and 24 hours later. 1 hour then 24 hours after instillation of the test material the ocular reactions were severe and the conjunctival chemosis (score of 4) obscured the evaluation of the reactions at the iris and cornea. The flowing of a whitish purulent substance was also noted. Some severe ocular lesions were expected at the next reading times but taking into consideration the severity of the lesions noted after 24 hours, the animal was sacrificed for ethical grounds before the nature of the reactions concerning the iris and the cornea could have been checked. Under these experimental conditions, the test material was considered as severely irritating when administered by ocular route to rabbits.

Based on results from in vivo tests in rabbits with read-across substance, the test substance is unlikely to be irritating to the skin, but severely irritating to the eye. Therefore, the substance does not need to be classified for skin irritation, but is classified as Cat 1 (H318: causes serious eye damage) according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.