Registration Dossier

Administrative data

Description of key information

In both GPMT study conducted in compliance with OECD guideline 406 with TMPTMA, no animals showed any sign of skin reaction after challenge. Based on these data, TMPTMA is considered to be not skin sensitizing.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From April 29 to June 15, 1996
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
no certificate of analysis, no data on animal gender and purity of test substance
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The guinea-pigs study was performed before the REACH regulation.
Species:
guinea pig
Strain:
Hartley
Sex:
not specified
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: David Hall Limited, Burton-on-Trent, Staffordshire, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 312-490 g
- Housing: Housed singly or in pairs in solid-floor polypropylene cages
- Diet (e.g. ad libitum): Guinea Pig FD1 Diet (Special Diets Services Limited, Witham, UK), ad libitum
- Water (e.g. ad libitum): Mains tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-23 °C
- Humidity (%): 45-68%
- Air changes (per hour): 15/hour
- Photoperiod (hours dark / hours light): 12 hours dark / 12 hours light
Route:
intradermal and epicutaneous
Vehicle:
arachis oil
Concentration / amount:
Range finding test:
- Intradermal induction exposure: 1, 5, 10 or 25% w/v in arachis oil
- Topical induction exposure: 25, 50, 75 or 100% w/v in arachis oil
- Topical challenge exposure: 25, 50, 75 or 100% w/v in arachis oil

Main test:
- Intradermal induction exposure: 25% w/v in arachis oil
- Topical induction exposure: Undiluted
- Topical challenge exposure: 75 or 100% w/v in arachis oil
Route:
epicutaneous, occlusive
Vehicle:
arachis oil
Concentration / amount:
Range finding test:
- Intradermal induction exposure: 1, 5, 10 or 25% w/v in arachis oil
- Topical induction exposure: 25, 50, 75 or 100% w/v in arachis oil
- Topical challenge exposure: 25, 50, 75 or 100% w/v in arachis oil

Main test:
- Intradermal induction exposure: 25% w/v in arachis oil
- Topical induction exposure: Undiluted
- Topical challenge exposure: 75 or 100% w/v in arachis oil
No. of animals per dose:
Range finding test:
- Intradermal induction exposure: 1 animal/dose
- Topical induction and challenge exposure: 2 animals/dose

Main test: 10 and 20 animals for control and test, respectively
Details on study design:
RANGE FINDING TESTS:
- Intradermal induction exposure: Guinea pigs (1/dose) received injections (0.1 mL) of test material at concentrations of 1, 5, 10 or 25% w/v in arachis oil and observed for systemic toxicity and erythema scores (Draize scoring system) at 24, 48 and 72 hours and 7 days.
- Topical induction exposure: Two guinea pigs [intradermally injected with Freund's Complete Adjuvant (FCA) 16 days earlier] applied with test material at concentrations of 25, 50, 75 or 100% v/v in arachis oil for 48 hours and observed for erythema and oedema at 1, 24 and 48 hours.
- Topical challenge exposure: Two guinea pigs applied with test material at concentrations of 25, 50, 75 or 100% v/v in arachis oil for 24 hours and observed for erythema and oedema at 1, 24 and 48 hours.


MAIN STUDY
A. INDUCTION EXPOSURE: INTRADERMAL
- No. of exposures: One
- Test groups: Intradermally injected with 0.1 mL of FCA, 25% w/v test material in arachis oil and 25% w/v test material emulsion in FCA on Day 0
- Control group: Intradermally injected with FCA, arachis oil or 50% w/v arachis oil in FCA on Day 0
- Site: Shoulder region on each side of mid-line
- Duration: Days 0-6

B. INDUCTION EXPOSURE: TOPICAL
- No. of exposures: One
- Exposure period: 48 hours
- Test groups: Filter paper patch saturated with undiluted test material topically applied on Day 7 via occlusive patch
- Control group: Only filter paper patch topically applied on Day 7 via occlusive patch
- Site: Shoulder region on each side of mid-line
- Frequency of applications: Single application
- Duration: Days 7-21

C. CHALLENGE EXPOSURE: TOPICAL
- No. of exposures: One
- Day of challenge: Day 21
- Exposure period: 24 hours
- Test groups: Filter paper patch saturated with 75 or 100% v/v test material in arachis oil topically applied on Day 21 via occlusive patch
- Site: 75 or 100% v/v in arachis oil was applied to left or right shorn flank, respectively
- Evaluation (hour after removal of challenge patch): 24 or 48 hours
Challenge controls:
No data
Positive control substance(s):
yes
Remarks:
Historical data: Ethyl 4-aminobenzoate 98%; 2,4-Dinitrochlorobenzene; Neomycin sulphate; 2-Mercaptobenzothiazole
Positive control results:
Historical data (August 1994 to February 1996):
- Incidence of sensitisation for 98% ethyl 4-aminobenzoate, 2,4-dinitrochlorobenzene, neomycin sulphate, 2-mercaptobenzothiazole were 39% (7/18), 100% (9/9), 100% (10/10), 60% (12/20) or 70% (7/10), respectively
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
75 or 100% v/v in arachis oil
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
None
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
75or 100% v/v on arachis oil
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
None
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Group:
positive control
No. with + reactions:
7
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation
Remarks:
(2-mercaptobenzothiazole)

Table 1: Intradermal sighting test - summary of results

 

Animal identification

Time of observation

Concentration of test material (% w/v)

Grade of erythema at Injection Sites

Evidence of systemic toxicity

A

24 hours

1

1

None

48 hours

2

None

72 hours

3

None

7 days

0

None

B

24 hours

5

1

None

48 hours

2

None

72 hours

3

None

7 days

0

None

E

24 hours

10

1

None

48 hours

1

None

72 hours

1

None

7 days

1

None

F

24 hours

25

2

None

48 hours

2

None

72 hours

2

None

7 days

1

None

Conclusion: The concentration of the test material selected for the intradermal induction stage of the main study was 25% w/v in arachis oil BP

  

Table 2: Topical sighting test for induction application (48-hour exposure) - individual skin reactions

 

Animal identification

Concentration of test material (% v/v)

Skin Reactions (Hours After Removal of Patches)

1

24

48

Er

Oe

Other

Er

Oe

Other

Er

Oe

Other

G

100

2

1

-

1

0

-

0

0

-

75

2

1

-

1

0

-

0

0

-

50

2

0

-

0

0

-

0

0

-

25

1

0

-

0

0

-

0

0

-

H

100

2

0

-

2

0

-

1

0

-

75

2

0

-

1

0

-

0

0

-

50

1

0

-

0

0

-

0

0

-

25

0

0

-

0

0

-

0

0

-

Conclusion: The undiluted test material was selected for the main study topical induction

 

Table 3: Topical sighting test for challenge application (24-hour exposure) - individual skin reactions

Animal identification

Concentration of test material (% v/v)

Skin Reactions (Hours After Removal of Patches)

1

24

48

Er

Oe

Other

Er

Oe

Other

Er

Oe

Other

I

100

1

0

-

0

0

-

0

0

-

75

1

0

-

0

0

-

0

0

-

50

0

0

-

0

0

-

0

0

-

25

0

0

-

0

0

-

0

0

-

J

100

1

0

-

0

0

-

0

0

-

75

1

0

-

0

0

-

0

0

-

50

0

0

-

0

0

-

0

0

-

25

0

0

-

0

0

-

0

0

-

Er = erythema; Oe = oedema; - = no other reactions noted

 

Conclusion: The concentrations of the test material selected for the main study topical challenge were 100 and 75% v/v in arachis oil BP

Interpretation of results:
GHS criteria not met
Remarks:
not skin sensitizing
Conclusions:
Under these test conditions, TMPTMA is not a skin sensitiser and is not classified according to the CLP Regulation (EC) N° (1272-2008).
Executive summary:

In a Magnusson & Kligman maximisation study (GPMT) performed according to OECD guideline 406 and in compliance with GLP, groups of 20 albino Dunkin Hartley guinea pigs were intradermally induced with three injections of 0.1 mL of FCA, 25% w/v TMPTMA in arachis oil and 25% w/v TMPTMA emulsion in FCA on Day 0 on shoulder region on each side of mid-line. After one week the same area were topically induced with 0.2 mL of undiluted test material via occluded filter paper patch for 48 hours. After 2 weeks of rest period, a challenge patch of 75 or 100% v/v test material in arachis oil was applied to left or right shorn flank, respectively. Control groups of 10 animals was included and treated with FCA, arachis oil or 50% w/v arachis oil in FCA. The test concentrations for the main study were determined from a sighting study using two animals.

 

Intradermal and topical induction indicated evidence of dermal irritation. No skin reactions were noted at the challenge sites of the test or control group animals at the 24 or 48-hour observations. TMPTMA produced a 0% (0/20) sensitisation rate and was considered to be a nonsensitiser to guinea pig skin. Historical data on positive controls (ethyl 4-aminobenzoate 98%; 2,4-dinitrochlorobenzene; neomycin sulphate; 2-mercaptobenzothiazole) exhibited evidence of sensitisation, thus confirming the susceptibility of this group of animals to dermal sensitisation.

 

Under these test conditions, TMPTMA is not classified according to the Annex VI to the Directive 67/548/EEC and the CLP Regulation (EC) N° (1272-2008).

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
April/May 1989
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The guinea-pigs study was performed before the REACH regulation.
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Shamrock Bio Service (France)
- Age at study initiation: no data
- Weight at study initiation: males = 336+/-14 g, females = 328+/-8 g
- Housing: individually housed in sterilizable polycarbonate cages
- Diet (e.g. ad libitum): Certified pellet diet, ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-3°C
- Humidity (%): 50+/-20
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light):12/12
Route:
intradermal and epicutaneous
Vehicle:
paraffin oil
Concentration / amount:
A preliminary test allowed to define the dose of the test substance to be administered in the main study. The sensitization of the animals by intradermal route was performed with the test substance at the concentration of 10% in the vehicle which is a non-irritant dose.
The sensitization of the animals by cutaneous route and challenge was made with the test substance in its original form, which is the aximum non-irritant dose, and was covered by an occlusive dressing for 24 hours.
Route:
epicutaneous, occlusive
Vehicle:
paraffin oil
Concentration / amount:
A preliminary test allowed to define the dose of the test substance to be administered in the main study. The sensitization of the animals by intradermal route was performed with the test substance at the concentration of 10% in the vehicle which is a non-irritant dose.
The sensitization of the animals by cutaneous route and challenge was made with the test substance in its original form, which is the aximum non-irritant dose, and was covered by an occlusive dressing for 24 hours.
No. of animals per dose:
Control group : 5 males and 5 females
Treated group: 10 males and 10 females
Details on study design:
During a 10-day sensitisation period (D1 to D11), the test substance was administered by intradermal injection (D1) and by cutaneous application (D9). The intradermal administrations of the test substance and vehicle were performed in the presence of an adjuvant in order tp maximize any potential cutaneous sensitization reactions. After a 15-day rest period, another cutaneous application of the test substance was performed (Day 26) in order to induce any potential cutaneous sensitization reactions.
In all animals, the application site was clipped just before each treatment, on day 1 and day 8 on the scapular area (4x2 cm) and clipped again and shaved on day 25 on each flank (2x2 cm).

The animals were observed daily in order to record any eventual clinical signs, which appeared during the study and to check for mortality.
On day 29, the animals were weighed and then sacrified after CO2 inhalation in excess after the 48-hour scoring period. Cutaneous samples were taken from the challenge application site of each animal and were preserved in 10% buffered formalin.
No histological examinations were performed.
Challenge controls:
no
Positive control substance(s):
no
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
No. with + reactions:
0
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
No. with + reactions:
0
Total no. in group:
10

No special symptoms were observed throughout the study. No mortality occurred.

The bodyweight gain of the treated animals was normal when compared to that of the control animals.

On day 11, after removal of the dressing, a cutaneous irritation in the control animals and a slight necrosis in the treated animals were noted at the intradermal injection sites.

No cutaneous reactions were observed on the left flanks (vehicle) and right flanks (test substance) in all control and treated animals.

Interpretation of results:
GHS criteria not met
Conclusions:
Under these experimental conditions and according to the maximalisation method of Magnusson and Kligman, no cutaneous reactions likely to have been caused by the sensitization potential of the test substance TMPTMA were observed in the Guinea-pigs.
Executive summary:

The sensitization potential of the test substance TMPTMA was evaluated in the Guinea-pig by intradermal injection and cutaneous application. The evaluation was performed after a 10 -day induction period, during the animals treated with the vehicle (control group, 5 males and 5 females) or the test substance (treated group, 10 males and 10 females). On day 1, in the presence of Freund's adjuvant, 0.1 ml of the test substance was administered by intradermal route at a concentration of 10% in paraffin oil. On Day 9, 0.5 ml of the test susbtance in its original form was applied by cutaneous route. After a period of 15 days without treatment, a challenge cutaneous application of 0.5 ml of the vehicle (left flank) was then administered to all animals. The substances were held in place for 24 hours by means of an occlusive dressing. The cutaneous reactions were then evaluated at the challenge application site, 24 and 48 hours after removal of the dressing. After the final scoring period, the animals were sacrified and cutaneous samples were taken from the challenge appplication sites in all animals. Due to the absence of any "doubtful" macroscopic cutaneous reactions, no histological examination was performed on the cutaneous samples.

No symptoms were observed throughout the study. After the challenge application of the vehicle or the test substance, no cutaneous reactions were observed in the animals from the control and the treated groups.

Under these experimental conditions and according to the maximalisation method of Magnusson and Kligman, no cutaneous reactions likely to have been caused by the sensitization potential of the test substance TMPTMA were observed in the Guinea-pigs.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

Key study (Allen 1996)

In a Magnusson & Kligman maximisation study (GPMT) performed according to OECD guideline 406 and in compliance with GLP, groups of 20 albino Dunkin Hartley guinea pigs were intradermally induced with three injections of 0.1 mL of FCA, 25% w/v TMPTMA in arachis oil and 25% w/v TMPTMA emulsion in FCA on Day 0 on shoulder region on each side of mid-line. After one week the same area were topically induced with 0.2 mL of undiluted test material via occluded filter paper patch for 48 hours. After 2 weeks of rest period, a challenge patch of 75 or 100% v/v test material in arachis oil was applied to left or right shorn flank, respectively. Control groups of 10 animals was included and treated with FCA, arachis oil or 50% w/v arachis oil in FCA. The test concentrations for the main study were determined from a sighting study using two animals.

Intradermal and topical induction indicated evidence of dermal irritation. No skin reactions were noted at the challenge sites of the test or control group animals at the 24 or 48-hour observations. TMPTMA produced a 0% (0/20) sensitisation rate and was considered to be a nonsensitiser to guinea pig skin. Historical data on positive controls (ethyl 4-aminobenzoate 98%; 2,4-dinitrochlorobenzene; neomycin sulphate; 2-mercaptobenzothiazole) exhibited evidence of sensitisation, thus confirming the susceptibility of this group of animals to dermal sensitisation.

Supporting study (Clouzeau 1989):

The sensitization potential of the test substance TMPTMA was evaluated in the Guinea-pig by intradermal injection and cutaneous application. The evaluation was performed after a 10 -day induction period, during the animals treated with the vehicle (control group, 5 males and 5 females) or the test substance (treated group, 10 males and 10 females). On day 1, in the presence of Freund's adjuvant, 0.1 ml of the test substance was administered by intradermal route at a concentration of 10% in paraffin oil. On Day 9, 0.5 ml of the test susbtance in its original form was applied by cutaneous route. After a period of 15 days without treatment, a challenge cutaneous application of 0.5 ml of the vehicle (left flank) was then administered to all animals. The substances were held in place for 24 hours by means of an occlusive dressing. The cutaneous reactions were then evaluated at the challenge application site, 24 and 48 hours after removal of the dressing. After the final scoring period, the animals were sacrified and cutaneous samples were taken from the challenge appplication sites in all animals. Due to the absence of any "doubtful" macroscopic cutaneous reactions, no histological examination was performed on the cutaneous samples.

No symptoms were observed throughout the study. After the challenge application of the vehicle or the test substance, no cutaneous reactions were observed in the animals from the control and the treated groups.

Under these experimental conditions and according to the maximalisation method of Magnusson and Kligman, no cutaneous reactions likely to have been caused by the sensitization potential of the test substance TMPTMA were observed in the Guinea-pigs.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on the available data, no classification for skin sensitisation is required for TMPTMA according to the CLP Regulation (EC) N° 1272-2008.