Disodium 6-(4,6-dichloro-1,3,5-triazin-2-ylamino)-1-hydroxy-2-(4-(2-(sulfonatooxy)ethylsulfonyl)phenylazo)naphthalene-3-sulfonate

Endpoint summary

Description of key information

The test substance showed no evidence for sensitizing properties in guinea pigs.

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:

From Sep. 30, 1997 to Dec. 09, 1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Study was already available

Species:

guinea pig

Strain:

Pirbright-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: HARLAN WINKELMANN Gartenstr. 27, D-33178 Borchen, SPF breeding colony
- Weight at study initiation: 369 g
- Housing: Macrolon cages (type 4) on soft wood granulate in groups of 5 animals
- Diet: ssniff Ms-H (V2233), ad libitum
- Water: Tap water in plastic bottles, ad libitum
- Acclimation period: At least 7 d

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

IN-LIFE DATES: From: Sep. 30, 1997 To: Oct. 31, 1997

Route:

intradermal

Vehicle:

water

Concentration / amount:

5% / 0.1 mL per injection

Day(s)/duration:

Day 1

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

25 % / 0.5 mL

Day(s)/duration:

Day 8 / 48 h

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

25 % / 0.5 mL

Day(s)/duration:

Day 22 / 24 h

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

- Determination of the primary non-irritant concentration: 3
- Determination of the tolerance of the intradermal injections: 2
- Control group: 5
- Treatment group: 10

Details on study design:

RANGE FINDING TESTS:
Determination of the primary non-irritant concentration:
The following test concentrations were administered to the flanks of two guinea pigs:
25.0 % in deionised water
5.0 % in deionised water
1.0 % in deionised water
0.5 mL of the test substance preparation was administered to a 2 x 2 cm cellulose patch, which was fixed to the flank and covered occlusively for 24 h with a bandage and film. 24 h after removal of the patches, the treated skin areas were examined for erythema and edema.

Determination of the tolerance of the intradermal injections:
The following preparations were administered twice by intradermal injection to 2 guinea pigs:
Site 1. 2X0.1 mL 5.0 % in deionised water
Site 2. 2X0.1 mL 1.0 % in deionised water
Site 3. 2X0.1 mL 0.2 % in deionised water

24, 48, 72 and 96 h after administration the injection sites were examined for local tolerance.

MAIN STUDY
A. INDUCTION EXPOSURE (Intradermal)
- No. of exposures: 2
- Exposure period: 1-7 d
- Test groups: Yes
- Control group: Yes, (50 % Freund's Adjuvant)
- Site: Dorsal area
- Frequency of applications: Single on Day 1 (2 intradermal injections/animal)
- Duration: 7 d
- Concentrations: 5.0 % test substance in deionised water

INDUCTION EXPOSURE (Dermal)
- No. of exposures: 0.5 mL of the test substance preparation as cellulose patch
- Test groups: Yes, 25.0 % test substance in deionised water
- Control group: Yes, deionised water
- Site: Dorsal area
- Frequency of applications: Single on Day 8
- Exposure period: 48 h under an occlusive bandage covered with an impermeable film and an elastic bandage.
- Concentrations: 25.0 % test substance in deionised water

B. CHALLENGE EXPOSURE (Dermal)
- No. of exposures: One
- Day(s) of challenge: Day 22
- Exposure period: 24 h under an occlusive bandage covered with an impermeable film and an elastic bandage.
- Test groups: 25.0 % test substance in deionised water
- Control group: 25.0 % test substance in deionised water
- Site: Left flank
- Concentrations: 25 % test substance in deionised water
- Evaluation (hr after challenge): 24 and 48 h after removal of the patches

OTHER:

The substance is considered to be sensitizing if 30 % or more of the animals in the treatment group definitely showed a positive skin reaction and at the same time no irritant effects have

Positive control substance(s):

yes

Remarks:

The validity of the test system Is confirmed by the periodically conducted positive control test using benzocain for the maximization test (Hoechst Marion Roussel, Preclinical Development Germany, Drug Safely) DyStar, TO-ECO, Germany, Report number 97.032

Positive control results:

The validity of the test system Is confirmed by the periodically conducted positive control test using benzocain for the maximization test (Hoechst Marion Roussel, Preclinical Development Germany, Drug Safely) DyStar, TO-ECO, Germany, Report number 97.0326, 1997-06-02

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

25 %

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

25 %

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

no indication of skin sensitisation

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

25 %

No. with + reactions:

0

Total no. in group:

5

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

25 %

No. with + reactions:

0

Total no. in group:

5

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Remarks on result:

positive indication of skin sensitisation

Determination of the primary non-irritant concentration

No signs of irritation occurred after administration of the different test concentrations.

Based on these results, a concentration of 25.0 % test substance in deionised water was chosen for the challenge at Day 22.

Determination of the tolerance of the intradermal injections

The intradermal injections with the 5.0 % preparation caused severe erythema and oedema as well as indurations and encrustations.

Based on this preliminary test, a 5.0 % preparation was selected for the intradermal injections in the main test.

 

Main test for the sensitizing properties

Body weight gains and clinical signs: 

The body weight gains of the animals were not impaired.

The treated animals showed no clinical signs of intoxication throughout the study

Intradermal induction treatment

The intradermal injections with Freund's Adjuvant (with and without test substance) caused severe erythema and edema as well as indurations and encrustations. The administration sites treated with the test substance in deionized water showed well defined erythema and edema. Intradermal injections of the vehicle alone exhibited no signs of irritation.

Due to these strong irritation reactions of the skin, 10% sodium dodecylsulfate was not administered at Day 7.

 

Dermal induction treatment

After the removal of the patches at Day 10, severe erythema and edema, indurated, scabbed and encrusted skin as well as necrosis were observed at the sites previously treated with Freund's Adjuvant. The administration sites treated with the test substance or vehicle alone exhibited no signs of irritation.

Dermal challenge treatment

No skin reactions were observed in the control and the treatment group 24 and 48 h after removal of the occlusive bandage.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the test substance showed no evidence for sensitizing properties in guinea pigs.

Executive summary:

A study was conducted to assess the sensitizing potential of the test substance in female Pirbright-White guinea pigs (Magnusson & Kligman method) according EU Method B.6. and OECD guideline 406 in compliance with GLP.

Intradermal induction was performed using 5 % test substance in deionised water. Dermal induction and challenge treatment were carried out with 25 % test substance in deionised water.

None of ten animals of the treatment group showed a positive skin response after the challenge exposure.

The validity of the test system Is confirmed by the periodically conducted positive control test using benzocain for the maximization test.

Under the test conditions, the test substance showed no evidence for sensitizing properties in guinea pigs.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

A study was conducted to assess the sensitizing potential of the test substance in female Pirbright-White guinea pigs (Magnusson & Kligman method) according OECD guideline 406 and EU Method B.6.

Intradermal induction was performed using 5 % test substance in deionised water. Dermal induction and challenge treatment were carried out with 25 % test substance in deionised water. The validity of the test system is confirmed by the periodically conducted positive control test using benzocain for the maximization test.

None of ten animals of the treatment group showed a positive skin response after the challenge exposure.

The test substance showed no evidence for sensitizing properties in guinea pigs (Dr. Bury D, 1997).

 

The test substance was negative in in vivo skin sensitisation test and therefore no classification is required for sensitisation according to EC criteria (67/548/EEC) and CLP criteria (EC 1272/2008).

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

The test substance was negative in in vivo skin sensitisation test and therefore no classification is required for sensitisation according to EC criteria (67/548/EEC) and CLP criteria (EC 1272/2008).