Quaternary ammonium compounds, bis(hydrogenated tallow alkyl)dimethyl, acetates

Endpoint summary

Description of key information

Skin Sensitisation: OECD 406; guinea pig; not sensitising. Reliability = 2 (read-across substance)

Respiratory Sensitisation: no data

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Study comparable to OECD guideline 406 with acceptable restrictions. Not GLP but a statement that the report and the study were audited by the Quality Assurance Unit of the testing laboratory was included in the report. No positive control included in the assay. It has to be noted that 3 readings instead of 2 were performed.

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:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

GLP compliance:

no

Remarks:

but a statement that the report and the study were audited by the Quality Assurance unit is included.

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Data performed before Reach guidance.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: D. Hall, Newchurch, Staffordshire
- Age at study initiation: 4-6 weeks
- Weight at study initiation: 400± 50 g
- Housing: in suspended cages with wire mesh floors
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21°C
- Humidity (%): 30-70%
- Air changes (per hr): 15 air changes per hour
- Photoperiod: 12 hrs dark / 12 hrs light

Route:

intradermal and epicutaneous

Vehicle:

water

Concentration / amount:

Main study:
-Induction:
intradermal injection: 0.1% v/v in water for injection.
topical application: 10% v/v in distilled water.
-Challenge:
2.5% and 1% v/v in distilled water

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

Main study:
-Induction:
intradermal injection: 0.1% v/v in water for injection.
topical application: 10% v/v in distilled water.
-Challenge:
2.5% and 1% v/v in distilled water

No. of animals per dose:

Test group of 15 animals and a control group of 15 in the main study.

Details on study design:

RANGE FINDING TESTS:
A preliminary study was conducted to determine the intradermal and topical irritancy of a range of aqueous solutions of Querton 442 (results are not included in the report).
MAIN STUDY

A. INDUCTION EXPOSURE
- No. of exposures: 1 intradermal, 1 epicutaneous
- Exposure period: epicutaneous: 48 hours

-> TEST GROUPS:
Intradermal exposure
Three injections of 0.1 mL were injected into each side of the animal as follows:
. Freund's complete adjuvant (FCA) diluted to 50% with water for injection
. TS at 0.1% w/v in vehicle
. TS at 0.1% w/v in a 50/50 (v/v) mixture of FCA and water for injection

Epicutaneous exposure
Application of 2 x 4cm patch saturated with the TS at 10% w/v in distilled water applied to the scapular region and held in place for 48 hours using an occlusive dressing.

-> CONTROL GROUP:
Intradermal exposure
Three injections of 0.1 mL were injected into each side of the animal as follows:
. Freund's complete adjuvant (FCA) diluted to 50% with water for injection
. vehicle
. A mixture 50/50 (v/v) FCA diluted to 50% with water for injection, and vehicle

Epicutaneous exposure
Application of 2 x 4cm patch saturated with the vehicule applied to the scapular region and held in place for 48 hours using an occlusive dressing.

- Site:
Intradermal exposure
6 injections in the clipped area (4 x 6 cm) in the scapular region

Epicutaneous exposure
4 x 6 cm area over the scapulae

- Frequency of applications:
One intradermal injection and one epicutaneous application 8 days after on the same site.

- Duration:
Epicutaneous exposure: 48 hours

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 22
- Exposure period: 24 hours

-> TEST GROUPS:
0.2 mL of the TS at 2.5% w/v in distilled water on the anterior left flank and 0.2 mL of the TS at 1% w/v in vehicule to a second site on the posterior left flank (occlusive epicutaneous application)

-> CONTROL GROUPS:
Same treatment as test group
- Site: anterior and posterior left flank

- Evaluation (hr after challenge): 24 , 48 hours and 72 hours after removal of the dressing according to the method of Draize.

Challenge controls:

During the induction period the control animals were treated similarly to the test animals with the exception that the test substance was omitted from the intradermal injections and topical application.

Positive control substance(s):

not specified

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

challenged with test substance 2.5% v/v in distilled water

No. with + reactions:

7

Total no. in group:

15

Clinical observations:

4 and 3 animals with erythema grade 1 and 2 respectively.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

challenged with test substance 2.5% v/v in distilled water

No. with + reactions:

4

Total no. in group:

15

Clinical observations:

1 and 3 animals with erythema grade 1 and 2 respectively. One animal with erythema grade 2 showed also oedema grade 1.

Reading:

other: 3rd reading

Hours after challenge:

72

Group:

negative control

Dose level:

challenged with test substance 2.5% v/v in distilled water

No. with + reactions:

3

Total no. in group:

15

Clinical observations:

1 and 2 animals with erythema grade 1 and 2 respectively. 1animal with erythema grade 2 showed also oedema grade 2.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

challenged with test substance 2.5% v/v in distilled water

No. with + reactions:

10

Total no. in group:

15

Clinical observations:

6 and 4 animals with erythema grade 1 and 2 respectively. 1animal with erythema grade 2 showed also oedema grade 2.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

challenged with test substance 2.5% v/v in distilled water

No. with + reactions:

7

Total no. in group:

15

Clinical observations:

4 and 3 animals with erythema grade 1 and 2 respectively. 2 animals with erythema grade 2 showed also oedema (grade 2 for one and grade 1 for the other).

Reading:

other: 3rd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

challenged with test substance 2.5% v/v in distilled water

No. with + reactions:

5

Total no. in group:

15

Clinical observations:

3 and 2 animals with erythema grade 1 and 2 respectively. 2 animals with erythema grade 2 showed also oedema (grade 2 for one and grade 1 for the other).

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

challenged with test substance 1% v/v in distilled water

No. with + reactions:

1

Total no. in group:

15

Clinical observations:

1 animal with erythema grade 1.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

challenged with test substance 1% v/v in distilled water

No. with + reactions:

0

Total no. in group:

15

Reading:

other: 3rd reading

Hours after challenge:

72

Group:

negative control

Dose level:

challenged with test substance 1% v/v in distilled water

No. with + reactions:

0

Total no. in group:

15

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

challenged with test substance 1% v/v in distilled water

No. with + reactions:

3

Total no. in group:

15

Clinical observations:

2 and 1 animals with erythema grade 1 and 2 respectively.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

challenged with test substance 1% v/v in distilled water

No. with + reactions:

2

Total no. in group:

15

Clinical observations:

2 animals with erythema grade 1.

Reading:

other: 3rd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

2 and 1 animals with erythema grade 1 and 2 respectively.

No. with + reactions:

1

Total no. in group:

15

Clinical observations:

1 animal with erythema grade 2.

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Remarks on result:

not measured/tested

The dermal reactions observed in the test animals were considered to be similar to or less marked than the maximum reaction observed in animals of the control group. Some localised dermal reactions were observed, and dryness and sloughing of the epidermis.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions of this study, the substance does not induce delayed contact hypersensitivity in guinea pigs.

Executive summary:

The potential of the substance to induce delayed contact hypersensitivity was assessed in guinea pigs using the technical grade dehydrogenated tallow dimethylammonium chloride (78% active in isopropanol/water). The study was performed according to a method equivalent to the OECD guideline 406 and a statement that the report and the study were audited by the Quality Assurance Unit of the testing laboratory was provided. The induction phase was realized both by intradermal route on day 1 (Test substance 0.1 % v/v in water for injection) and by cutaneous route on day 8 (Test substance 10% v/v in distilled water) in 2 groups of guinea pigs: 15 males for control group and 15 males for treated group. The challenge phase was realized on day 22 by cutaneous application of the test substance at 2.5% and 1% v/v in distilled water on two sites on the left flank. the cutaneous reactions were scored 24 ,48 and 72 hours after the challenge phase.

 

After the challenge application with 2.5 % v/v of the test substance in distilled water, erythema grade 1 or 2 were observed in 7, 4 and 3 controls animals at the 24, 48 and 72-hours readings respectively. 1 control animal showed also oedema. In the treated group, the incidence and severity of the cutaneous reactions were quite similar: 10, 7 and 5 treated animals showed erythema grade 1 or 2 at the 24, 48 and 72-hours readings. 2 treated animals showed oedema.

 

After the challenge application with 1% v/v of the test substance, an erythema (grade 1 or 2) was observed in 3, 2 and 1 out the 15 animals of the treated group at the 24, 48 and 72-hour readings respectively. In the control group, 1animal showed an erythema grade 1at the 24-hour reading. The persistent cutaneous reactions observed in 3/15 animals of the treated group after the challenge application may be attributable to delayed contact hypersensitivity but as the cutaneous reactions were more confined to the 24-hour examination, they were more consistent with irritation than delayed contact hypersensitivity. Therefore, the cutaneous reactions observed in 3/15 animals (20%) of the treated groups were considered as irritative response and the substance was not considered as a skin sensitiser.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

No skin sensitisation 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 skin sensitisation. 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.

 

The potential of the substance to induce delayed contact hypersensitivity was assessed in guinea pigs using the technical grade dihydrogenatedtallowdimethylammonium chloride (78% active in isopropanol/water). The study was performed according to a method equivalent to the OECD guideline 406. The induction phase was realized both by intradermal route on day 1 (Test substance 0.1 % v/v in water for injection) and by cutaneous route on day 8 (Test substance 10% v/v in distilled water) in 2 groups of guinea pigs: 15 males for control group and 15 males for treated group. The challenge phase was realized on day 22 by cutaneous application of the test substance at 2.5% and 1% v/v in distilled water on two sites on the left flank. the cutaneous reactions were scored 24, 48 and 72 hours after the challenge phase. After the challenge application with 2.5 % v/v of the test substance in distilled water, erythema grade 1 or 2 were observed in 7, 4 and 3 controls animals at the 24, 48 and 72-hours readings respectively. 1 control animal showed also oedema. In the treated group, the incidence and severity of the cutaneous reactions were quite similar: 10, 7 and 5 treated animals showed erythema grade 1 or 2 at the 24, 48 and 72-hours readings. 2 treated animals showed oedema. After the challenge application with 1% v/v of the test substance, an erythema (grade 1 or 2) was observed in 3, 2 and 1 out the 15 animals of the treated group at the 24, 48 and 72-hour readings respectively. In the control group, 1 animal showed an erythema grade 1 at the 24-hour reading. The persistent cutaneous reactions observed in 3/15 animals of the treated group after the challenge application may be attributable to delayed contact hypersensitivity but as the cutaneous reactions were more confined to the 24-hour examination, they were more consistent with irritation than delayed contact hypersensitivity. Therefore, the cutaneous reactions observed in 3/15 animals (20%) of the treated groups were considered as an irritative response and the substance was not considered as a skin sensitizer.

 

A human repeat insult patch test performed with a 75% aqueous dilution of the test substance according to the method based on that of Shelanski and Shelanski. A total of 84 volunteers took part in the test. The panel included 77 women and 7 men ageing from 18 to 61 years and more. 73 panelists completed the full test. A patch loaded with 0.5 mL of 2% v/v test material in distilled water was applied down the dorsal surface of the upper arm of each subject. Subjects were instructed to keep the patches dry and to remove and discard them after 24 hours. Patches were applied on Monday, Wednesday and Friday of the first 3 weeks, called the "induction" or "insult" period. Patches were re-applied to the same site unless reaction to a substance or adhesive necessitated relocation, in which case an adjacent location was chosen. The test sites were scored before application of each subsequent patch and on the fourth Monday after the final insult patch.14 days after the final insult patch, challenge patches were applied to both arms of each subject. Scoring were done 48 and 96 hours after patch removal. The test material caused some degree of irritation in most volunteers and therefore it was concluded that test conditions were conducive to the induction of a sensitised state should such be likely to occur in the panelists. The concentration of the material was maintained throughout the study. The test material produced no reaction at challenge which was indicative of skin sensitisation. Therefore, it was concluded that the substance was not sensitising in humans.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

The read-across substance was negative for skin sensitisation in an in vivo guinea pig study and did not produce a sensitisation response in humans. Therefore, no classification is required for skin sensitisation according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.