2-[tert-butyl(phenylmethyl)amino]-1-[4-hydroxy-3-(hydroxymethyl)phenyl] hydrochloride

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 January - 25 February 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Done by OECD and GLP standards

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

GLP compliance:

yes

Remarks:

OECD Principles on Good Laboratory Practice (revised 1997, ENV/MC/CHEM(98)17); and are in accordance with, and implement, the requirements of Directives 87/18/EEC (as amended by Directive 1999/11/EC) and 88/320/EEC (as amended by Directive 1999/12/EC).

Type of study:

mouse local lymph node assay (LLNA)

Species:

mouse

Strain:

other: CBA/Ca (CBA/CaBkl)

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: B & K Universal Ltd, Hull, UK
- Age at study initiation: 8 - 12 weeks
- Weight at study initiation: 15 - 23 g
- Housing: individually housed in suspended solid-floor polypropylene cages furnished with softwood woodflakes
- Diet: Certified Rat and Mouse Diet (Code 5LF2) supplied by International Product Supplies Limited, Wellingborough, Northants, UK. ad libitum.
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25°C
- Humidity: 30 to 70%
- Air changes (per hr): 15 approx
- Photoperiod (hrs dark / hrs light): twelve hours continuous light (06.00 to 18.00) and twelve hours darkness

Route:

epicutaneous, open

Vehicle:

other: Dimethyl formamide

Route:

intradermal

Vehicle:

other: Dimethyl formamide

No. of animals per dose:

4

Vehicle:

dimethylformamide

Concentration:

0.5%, 5% and 50% w/v in dimethyl formamide

No. of animals per dose:

Groups of four mice were treated with the test material at concentrations of 0.5%, 5% or 50% w/v in dimethyl formamide

Details on study design:

Preliminary Screening Test:
As no toxicological information was available regarding the systemic toxicity/irritancy potential of the test material a preliminary screening test was performed using one mouse. The mouse was treated by daily application of 25 µl of the test material, at a concentration of 50% w/v in dimethyl formamide, to the dorsal surface of each ear for three consecutive days (Days 0, 1, 2). The mouse was observed daily for 5 days. Any signs of toxicity or signs of ill health during this period were recorded. The bodyweight was recorded on Day 0 (prior to dosing) and on Day 5.

Test Material Administration:
Groups of four mice were treated with the test material at concentrations of 0.5%, 5% or 50% w/v in dimethyl formamide. The preliminary screening test suggested that the test material would not produce systemic toxicity or excessive local irritation at the highest suitable concentration. The mice were treated by daily application of 25 µl of the appropriate concentration of the test material to the dorsal surface of each ear for three consecutive days (Days 0, 1, 2). The test material formulation was administered using an automatic micropipette and spread over the dorsal surface of the ear using the tip of the pipette.



The mice were treated by daily application of 25 µl of the appropriate concentration of the test material to the dorsal surface of each ear for three consecutive days (Days 0, 1, 2). The test material formulation was administered using an automatic micropipette and spread over the dorsal surface of the ear using the tip of the pipette.

A further group of four mice received the vehicle alone in the same manner.

Five days following the first topical application of the test material (Day 5) all mice were injected via the tail vein with 250 µl of phosphate buffered saline containing 3H-methyl thymidine (3HTdR: 80 µCi/ml, specific activity 2.0 Ci/mmol, Amersham Pharmacia Biotech UK Ltd) giving a total of 20 µCi to each mouse.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Positive control results:

α-Hexylcinnamaldehyde, was considered to be a sensitiser under the conditions of the test with test control ration obtained for 25% v/v HCA at 8.4

Parameter:

SI

Remarks on result:

other: see Remark

Remarks:

Concentration of test material in the vehicle at 0.5 % resulted in a stimulation index (SI) of 0.96 which is a negative result. Concentration of test material in the vehicle at 5 % resulted in a stimulation index (SI) of 1.54 which is a negative result. Concentration of test material in the vehicle at 50 % resulted in a stimulation index (SI) of 5.44 which is a positive result. .

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: see Remark

Remarks:

Concentration of test material in the vehicle at 0.5 % resulted in a mean disintegration of 10375.52 dpm which is a negative result.Concentration of test material in the vehicle at 5 % resulted in a mean disintegration of 16666.81 dpm which is a negative result. Concentration of test material in the vehicle at 50 % resulted in a mean disintegration of 58856.52 dpm which is a positive result.

A stimulation index of greater than 3 was recorded for the highest concentration of the test material (50% w/v).

 

 A stimulation index of less than 3 was recorded for the two lower concentrations of the test material (5 and 50% w/v).

There were no deaths. No signs of systemic toxicity were noted in the test or control animals during the study.

Interpretation of results:

sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test material was considered to be a sensitiser under the conditions of the test.

Executive summary:

The test material was classified as a sensitiser according to EU labelling regulations Commission Directive 2001/59/EC.

The symbol "Xi",  indication of danger 'irritant' and the risk phrase R43 "May Cause Sensitisation by Skin Contact" are therefore required.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

A single study has been conducted in accordance with GLP and the OECD 429 guideline. The results of the study indicate that the substance is a moderate skin sensitiser in mice with a measured EC3 value of >5% w/w in dimethylformamide. The LLNA EC3 value for induction (21.8%) was calculated according to Kimber et al 2001 Tox Sci 59(2)198-208. This is converted to a dose of 5450µg/cm2 in accordance with ECHA guidance document Chapter R8, in Appendix R.8-10.; this point of departure is considered to be a LOAEL.

Migrated from Short description of key information:

The test material was considered to be a sensitiser under the conditions of the test.

0.5%,  5%  and 50% w/v of test material was used with dimethyl formamide as the vehicle.

Justification for selection of skin sensitisation endpoint:

The test substance was analysed in accordance with OECD 429 and EU method B42. The substance was determined to be a sensitizer (Category 1, sub-category 1B).

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

The substance has been tested in the mouse local lymph node assay in accordance with OECD 429. Application of the classification criteria set out in CLP Regulation Annex I Section 3.4.2.2.3 results in a classification of Skin sensitiser Category 1B for the calculated EC3 value of 21.8% w/v in dimethylformamide.