GOLDEN YELLOW SCJ 1006

Endpoint summary

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:

1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline study under GLP without deviations

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Version / remarks:

92/69/EEC (Maximisation test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

Species Dunkin Hartley Crl: (HA)BR albino guinea pig (SPF-quality), recognised by international guidelines as the recommended test system (e.g. OECD, EEC), Source : Charles River, Germany
Number of animals: Experimental group: 20 females, Control group: 10 females
Age at start of study: Approx. 5 weeks
Body weight prior to start: 356 - 433 grams
Identification: Ear tattoo
Test system check: A positive control experiment is performed once every six months as a sensitivity check of the test system. The most recent test is summarised in the original report.
ANIMAL HUSBANDRY
Conditions: Air-conditioned room with approximately 15 air changes per hour and the environment controlled with optimal conditions considered as being a temperature of 21°C and a relative humidity of 50%. Fluctuations from these optimal conditions were noted, but were considered not to have affected study integrity. Lighting was 12 hours artificial fluorescent light and 12 hours dark per day.
Accommodation: Housing of 1 or 2 animals per labelled metal cage with wire-mesh floors and equipped with an automatic drinking system (ITL, Bergen, The Netherlands). The acclimatisation period was at least 5 days before start of treatment under laboratory conditions.
Diet: Free access to standard guinea pig diet, including ascorbic acid (1600 mg/kg); LC 23-B, pellet diameter 4mm (Hope farms, Woerden, The Netherlands). Certificates of analysis were examined and retained in the NOTOX archives. In addition, hay (B.M.I., Helmond, The Netherlands) was provided once a week.
Water: Free access to tap water, diluted with decalcified water. Certificates of analysis were examined and retained in the NOTOX archives.

Route:

intradermal

Vehicle:

CMC (carboxymethyl cellulose)

Concentration / amount:

Concentration of test material and vehicle used at induction: Intradermal with 5% in 1% aqueous carboxymethylcellulose Topical: 25% in 1% aqueous carboxymethylcellulose Concentration of test material and vehicle used for each challenge: 25% topically

Route:

epicutaneous, occlusive

Vehicle:

CMC (carboxymethyl cellulose)

Concentration / amount:

Concentration of test material and vehicle used at induction: Intradermal with 5% in 1% aqueous carboxymethylcellulose Topical: 25% in 1% aqueous carboxymethylcellulose Concentration of test material and vehicle used for each challenge: 25% topically

No. of animals per dose:

Number of animals in test group: 20 females
Number of animals in negative control group: 10 females

Details on study design:

PRELIMINARY IRRITATION STUDY
Prior to the start of the Main study, the intradermal and epidermal irritancy ofFAT 40543/A was investigated to select test substance concentrations suitable for the induction and challenge phase of the Main Study. The test system, procedures and techniques were identical to those used during the main study, unless otherwise mentioned. The animals were selected from stock and were between 5 and 9 weeks old. Body weights were determined prior to treatment.
Intradermal injections: A 5% and a 2% concentration were injected in one animal into two injections sites (0.1 mL each) in the left and right clipped shoulder regions, respectively. A 1% concentration was injected in a similar manner in a second animal. The resulting dermal reactions were assessed 24 and 48 hours after treatment.
Epidermal application: A 25% concentration (0.5 ml) was applied epidermally on a clipped flank of one animal, using a Scotchpak-non-woven patch (2.5 x 2.2 cm) on Micropore tape and held in place by Coban elastic bandage. A 10% concentration was applied in a similar manner on a second animal. Four concentrations of the test substance (25%, 10%, 5% and 2%, 0.05 ml each) were applied occlusively on a clipped flank of each of two other animals using Square chambers (v.d. Bend, Brielle, The Netherlands), mounted on Micropore tape and held in place by Coban elastic bandage. After 24 hours, the dressings were removed and the skin cleaned of residual test substance. Red staining of the treated skin-areas had occurred by epidermal exposure to the test substance. To remove the staining caused by the test substance and to facilitate scoring, the epidermal application areas were cleaned by depilation prior to skin reading. Approximately 21 hours after patch removal, an approved depilatory cream (VEET cream, Reckitt & Colman bv, Naarden, The Netherlands) was placed on the application sites and surrounding areas for 3 - 5 minutes. Thereafter, the cream was gently removed using a plastic spatula. The skin was further cleaned by washing with lukewarm, running tap-water. Subsequently, the animals were dried with a disposable towel and returned to their cages. The treated skin-sites were assessed 24 and 48 hours after exposure.
MAIN STUDY
INDUCTION - EXPERIMENTAL ANIMALS
Day 1 Three pairs of intradermal injections (0.1 mL/site) were made in the clipped scapular region as follows:
A) Freunds' Complete Adjuvant (Difco, Detroit, U.S.A.), 50:50 with water for injection (Ferensius AG, Bad Homburg, Germany).
B) The test substance at a 5% concentration.
C) The test substance, at twice the concentration used in (B), emulsified in a 50:50 mixture of Freunds' Complete Adjuvant.
Note: One of each pair was on each side of the midline and from cranial A) to caudal C).
Day 3 The dermal reactions caused by the intradermal injections were assessed for erythema (see scale above) or, in case of necrosis, the diameter of necrosis.
Day 7: The clipped scapular area was rubbed with 10% sodium-dodecyl-sulfate (SDS, Boom, Meppel, The Netherlands) in vaseline using a spatula. This concentration of SDS provokes a mild inflammatory reaction.
Day 8 The clipped area between the injection sites was treated with 0.5 mL of a 25% test substance concentration using a Scotchpak-non-woven patch ( 2 x 4 cm) mounted on Micropore tape and secured with Coban elastic bandage.
Day 10 After 48 hours, the dressings were removed and the skin cleaned of residual test substance. Subsequently, all dermal reactions caused by the epidermal exposure were assessed according the scale above.
INDUCTION - CONTROL ANIMALS
The control animals received similar treatment as described for the experimental animals, except that the vehicle only was administered.
CHALLENGE
Day 22 All animals were treated epidermally with a 25% test substance concentration and the vehicle (0.5 ml each) on a clipped flank, using the same type of occlusive dressings used in the induction phase.
Day 23 After approximately 24 hours, the dressings were removed and the skin cleaned of residual test substance and vehicle. The treated sites were assessed for erythema and swelling 24 and 48 hours after removal of the dressings, using the numerical grading system below. To remove the staining caused by the test substance and to facilitate scoring, an approved depilatory cream (VEET cream, Reckitt & Colman bv, Naarden, The Netherlands) was placed on the patch sites and surrounding areas for 5 minutes. The cream was then gently removed using a plastic spatula. The skin was further cleaned by washing with lukewarm, running tap-water. Subsequently, the animals were dried with a disposable towel and returned to their cages.
No visible change: 0
Discrete or patchy erythema: 1
Moderate and confluent erythema: 2
Moderate erythema and swelling: 3
Intense erythema and swelling: 4
At the end of the study period all animals were killed by oxygen/carbon dioxide asphyxiation.

Challenge controls:

The control animals received similar treatment as described for the experimental animals, except that the vehicle only was administered.

Positive control substance(s):

yes

Remarks:

Alpha-hexylcinnamicaldehyde

Positive control results:

A positive control experiment is carried out at regular intervals to check the reliability of the test system as used by NOTOX. In this study, performed in July 1995, the Dunkin Hartley guinea pig was checked for the sensitivity to ALPHA-HEXYLCINNAMICALDEHYDE, TECH. 85%. The study was carried out in accordance with the OECD Guideline No. 406, the EEC Directive 92/69/EEC, Part B.6 and in accordance with the method described by Magnusson and Kligman, "Allergic Contact Dermatitis in the Guinea Pig - Identification of Contact Allergens".
ALPHA-HEXYLCINNAMICALDEHYDE, TECH. 85%, was fabricated under lot no. 80281 and the purity was 99.4% (glc) (Aldrich Chemicals Co., Germany).
Concentrations selected for this study were: Intradermal induction: 5% solution in physiological saline (w/w). Epidermal induction: undiluted. Challenge: a 25, 10 and 5% solution in distilled water (w/w).
CONCLUSION
The skin reactions observed in response to all three test substance concentrations in all ten experimental animals in the challenge phase were considered indicative of sensitisation, based on the absence of any response in the control animals. These results lead to a sensitisation rate of 100 per cent. From these results, it was concluded that the Albino Dunkin Hartley strain of guinea pig is an appropriate animal model for the performance of studies designed to evaluate the sensitising potential of a substance. The raw data and report from this study are kept in the NOTOX archives. The test described above was performed in accordance with NOTOX Standard Operating Procedures and the report was audited by the QA-unit.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

0 %

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

25 %

No. with + reactions:

1

Total no. in group:

20

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25 %. No with. + reactions: 1.0. Total no. in groups: 20.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

0 %

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

25 %

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25 %. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0 %

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

25 %

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 25 %. No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

0 %

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

25 %

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 25 %. No with. + reactions: 0.0. Total no. in groups: 10.0.

Maximum concentration not causing irritating effects in preliminary test: 25 % 

Signs of irritation during induction: Irritation could not be determined due to substance-related staining of the skin. Evidence of sensitisation of each challenge concentration: 25%: One single skin reaction of grade 1 in one animal.

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

FAT 40543/A was tested for skin sensitisation in a guinea pig maximisation test and found to be non-sensitising to skin.

Executive summary:

The study was carried out in accordance with the OECD Guideline No. 406, "Skin Sensitisation", the EEC Directive 92/69/EEC, Part B.6, "Skin. Sensitisation" and based on the method described by Magnusson and Kligman, "Allergic Contact Dermatitis in the Guinea Pig - Identification of Contact Allergens".

Test substance concentrations (in 1% aqueous carboxymethyl cellulose) selected for the main study were based on the results of a preliminary study. In the main study, twenty experimental animals were intradermally injected with a 5% concentration and epidermally exposed to 25% concentration. Ten control animals were similarly treated, but with omission of the test substance. Approximately 24 hours before the epidermal induction exposure all animals were treated with 10% SDS. Two weeks after the epidermal application all animals were challenged with a 25% test substance concentration and the vehicle. To remove the staining caused by the test substance and to facilitate scoring, the skin was cleaned by depilation prior to skin reading. A single skin reaction of grade 1 was observed in one experimental animal in response to the 25% test substance concentration. No skin reactions were evident in the control animals. No signs of systemic toxicity were observed in any of the animals during the study period. The skin reaction observed in the challenge phase was considered indicative of sensitisation, based on the absence of any response in the control animals. These results lead to a sensitisation rate of 5 per cent. Applying the rating of allergenicity described by Kligman A.M. (1966) on the results obtained in this test, FAT 40543/A is considered to have weak sensitising properties. However, classification for skin-sensitisation is not required according to CLP or DSD.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

A study in accordance with the OECD Guideline No. 406, "Skin Sensitisation", the EEC Directive 92/69/EEC, Part B.6, "Skin Sensitisation" and based on the method described by Magnusson and Kligman, "Allergic Contact Dermatitis in the Guinea Pig - Identification of Contact Allergens" is available.

Test substance concentrations (in 1% aqueous carboxymethyl cellulose) selected for the main study were based on the results of a preliminary study. In the main study, twenty experimental animals were intradermally injected with a 5% concentration and epidermally exposed to 25% concentration. Ten control animals were similarly treated, but with omission of the test substance. Approximately 24 hours before the epidermal induction exposure all animals were treated with 10% SDS. Two weeks after the epidermal application all animals were challenged with a 25% test substance concentration and the vehicle. To remove the staining caused by the test substance and to facilitate scoring, the skin was cleaned by depilation prior to skin reading. A single skin reaction of grade 1 was observed in one experimental animal in response to the 25% test substance concentration. No skin reactions were evident in the control animals. No signs of systemic toxicity were observed in any of the animals during the study period. The skin reaction observed in the challenge phase was considered indicative of sensitisation, based on the absence of any response in the control animals. These results lead to a sensitisation rate of 5 per cent. Applying the rating of allergenicity described by Kligman A.M. (1966) on the results obtained in this test, FAT 40543/A is considered to have weak sensitising properties. However, classification for skin-sensitisation is not required according to CLP or DSD.

Migrated from Short description of key information:
OECD406 test (maximisation test) with FAT 40543/A for skin sensitisation was negative

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the negative outcome of a skin sensitisation study the substance is not subject to classification for skin sensitisation according to CLP (Regulation EC No 1272/2008) or DSD (Directive 67/548/EEC). Data on respiratory sensitisation are not available.