Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
13 June 2012 to 10 August 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed in accordance with OECD, EU, US EPA and JMAFF guidelines in compliance with GLP and reported with a valid GLP certificate.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Qualifier:
according to guideline
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No. 8147, April 2011; including the most recent partial revisions.
GLP compliance:
yes (incl. QA statement)
Type of study:
Buehler test

Test material

Constituent 1
Reference substance name:
71786-47-5
EC Number:
615-420-5
Cas Number:
71786-47-5
IUPAC Name:
71786-47-5
Constituent 2
Reference substance name:
Benzenesulfonic Acid mono and dialkylderivs magnesium salt, neutral, indiluents oil TBN = 20
IUPAC Name:
Benzenesulfonic Acid mono and dialkylderivs magnesium salt, neutral, indiluents oil TBN = 20
Test material form:
liquid: viscous
Details on test material:
Test substance information
Identification: 71786-47-5 Benzenesulfonic Acid mono and dialkylderivs magnesium salt, neutral, indiluents oil TBN = 20
Molecular formula: UVCB
Molecular weight: UVCB
CAS Number: 71786-47-5
Description: Clear brown viscous liquid (determined at WIL Research Europe)
Batch: Not indicated
Purity/Composition: UVCB, 100%
Test substance storage: At room temperature in the dark
Stability under storage conditions: Stable
Expiry date: 31 May 2013

Study specific test substance information
Purity/composition correction factor required: No
Hygroscopic: No
Volatile: No
Stability at higher temperatures: Not indicated
Stability in vehicle:
Kaydol White Mineral Oil: Stable
Solubility in vehicle:
Kaydol White Mineral oil: Miscible

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
Species: Albino guinea pig, Dunkin Hartley strain, (SPF-quality). Recognized by international guidelines as the recommended test system (e.g. OECD, EC).
Source: Charles River Deutschland, Kisslegg, Germany.
Number of animals
Experimental group: 20 females.
Control group: 10 females.
(females were nulliparous and non-pregnant).
Age: Young adult animals (approx. 5 weeks old) were selected.
Identification: Ear tattoo.
Health inspection: A health inspection was performed prior to treatment, to ensure that the animals are in a good state of health. Special attention was paid to the skin to be treated, which was intact and free from any abnormality.

Animal husbandry
Conditions: Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, approximately 15 room air changes/hour, and a 12-hour light/12-hour dark cycle. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.
Accommodation: Group housing of maximally 5 animals per labeled Noryl cage (Tecniplast; 74 cm x 54 cm x 25 cm height) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and shelters (CS3B02A Play tunnels (90 mm x 5 mm x 125 mm), Datesand, Manchester, UK) as cage enrichment. The acclimatization period was at least 5 days before the start of treatment under laboratory conditions.
Diet: Complete breeding diet for guinea pigs (SSNIFF® MS-Z, V2273; SSNIFF® Spezialdiäten GmbH, Soest, Germany). Hay (TecniLab-BMI BV, Someren, The Netherlands) was provided at least twice a week.
Water: Free access to tap water.

Results of analysis for diet, sawdust, shelters and water were assessed and did not reveal any findings that were considered to have affected the study integrity. All certificates and results of analysis are retained in the WIL Research Europe archives.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Kaydol White Mineral Oil
Concentration / amount:
INDUCTION: 0.5 mL of the undiluted test substance concentration
CHALLENGE: 50% (w/w) test substance concentration and the vehicle (0.1 mL of each)
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Kaydol White Mineral Oil
Concentration / amount:
INDUCTION: 0.5 mL of the undiluted test substance concentration
CHALLENGE: 50% (w/w) test substance concentration and the vehicle (0.1 mL of each)
No. of animals per dose:
10 control, 20 experimental group.
Details on study design:
Preliminary irritation study: A preliminary irritation study was conducted in order to select test substance concentrations to be used in the Main Study. The selection of concentrations was based on the following criteria:
-The concentrations are well-tolerated by the animals.
-For the induction exposures: the highest possible concentration that produced mild irritation (grades 2).
-For challenge exposure: the maximum non-irritant concentration.
A series of test substance concentrations was tested. Practical feasibility of administration determined the highest starting-concentration. The test system, procedures and techniques were identical to those used during the main study, unless otherwise specified. The animals selected were between 4 and 9 weeks old. No bodyweights were determined.

Epidermal application: A series of four test substance concentrations was used, the highest concentration being the maximum concentration that could technically be applied. Two different concentrations were applied (0.5 mL each) per animal to the clipped flank using Metalline patches (2x3 cm) mounted on Medical tape, which will be held in place with Micropore tape# and subsequently Coban elastic bandage.
After 6 hours, the dressings were removed and the skin cleaned of residual test substance with Kaydol White Mineral Oil. The resulting dermal reactions were assessed for irritation 24 and 48 hours after removal of the last dressing.

Based on the results, two additional animals were treated each with two test substance concentrations of 50 and 75% to verify the correct challenge concentration.

Main study
INDUCTION - Experimental animals
Days 1, 8 and 15: The left side of the scapular region was clipped and subsequently epidermally treated with 0.5 mL of the undiluted test substance concentration, using Metalline patches (2x3 cm) mounted on Medical tape, which was held in place with Micropore tape and subsequently Coban elastic bandage.
After 6 hours, the dressings were removed and the skin cleaned of residual test substance using Kaydol White Mineral Oil. Immediately after removal of the last induction application on Day 15, the treated skin area was assessed for irritation.

INDUCTION - Control animals
The control animals were treated as described for the experimental animals, except that, instead of the test substance, vehicle alone was administered.

FIRST CHALLENGE - All animals
Day 29: The right flank of all animals was clipped and subsequently treated epidermally with a 50% (w/w) test substance concentration and the vehicle (0.1 mL of each), using Patch Test Plasters (Curatest®, Lohmann, Almere, The Netherlands). The patches were held in place with Micropore tape and subsequently Coban elastic bandage.
After 6 hours, the dressings were removed and the skin cleaned of residual test substance and vehicle using Kaydol White Mineral Oil. The treated sites were assessed for challenge reactions 24 and 48 hours after removal of the dressings.

SECOND CHALLENGE - All animals
Day 36: Based on the results and at request of the sponsor, a second challenge was performed in the same way the first challenge was performed.

After termination, animals were sacrificed using isoflurane.

Observations
Mortality/Viability: Twice daily.
Toxicity: At least once daily.
Bodyweights: Prior to start and at termination of the study.
Necropsy: No necropsy was performed according to protocol.
Irritation: Skin reactions were graded according to the following numerical scoring systems. Furthermore, a description of all other (local) effects was recorded.
Challenge controls:
The control animals were treated as described for the experimental animals, except that, instead of the test substance, vehicle alone was administered.
Positive control substance(s):
no

Results and discussion

Positive control results:
Positive controls not used within this experiment.

In vivo (non-LLNA)

Resultsopen allclose all
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:
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:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. 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:
test chemical
Dose level:
100%
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: 100%. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
100%
No. with + reactions:
3
Total no. in group:
20
Clinical observations:
Scaliness in 2/20 animals
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 100%. No with. + reactions: 3.0. Total no. in groups: 20.0. Clinical observations: Scaliness in 2/20 animals.
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
100%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
100%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 20.0.

Any other information on results incl. tables

Table 1: PRELIMINARY IRRITATION STUDY

SKIN RECATIONS AFTER EPIDERMAL EXPOSURE

Animal number

Conc. %

24 hours after exposure

48 hours after exposure

Erythema (grade)

Oedema (grade)

Erythema (grade)

Oedema (grade)

1

100

0

1

1

0

50

0

0

0

0

2

100

0

1

0

0

50

0

0

0

0

3

20

0

0

0

0

10

0

0

0

0

4

20

0

0

0

0

10

0

0

0

0

1

75

0

0

1

0

50

0

0

0

0

2

75

0

0

0

0

50

0

0

0

0

Note: It was noted that the identification of the animals used in the preliminary study was not unique. The raw data of the study specifies that the animals were of separate deliveries from the supplier and treated on different occasions.

 

Table 2: INDUCTION AND FIRST CHALLENGE READINGS

Animal No.

Induction

First Challenge

Day 15

Day 30

Day 31

 

24 Hour Reading

48 Hour Reading

Readings

Readings

Readings

100% #

50%

Vehicle

50%

Vehicle

Er

Oe

 

 

 

 

Control

91

0

0

0

0

0

0

92

0

0

0

0

0

0

93

0

0

0

0

0

0

94

0

0

0

0

0

0

95

0

0

0

0

0

0

96

0

0

0

0

0

0

97

0

0

0

0

0

0

98

0

0

0

0

0

0

99

0p

0

0

0

0

0

100

0

0

0

0

0

0

Experimental

101

1

0

0

0

0

0

102

1p

0

0

0

0

0

103

1

0

0

0

0

0

104

1p

0

0

0

0p

0

105

2

0

0

0

0

0

106

1

0

0

0

0

0

107

2p

0

0

0

1

0

108

0

0

0

0

0

0

109

1

0

0

0

0

0

110

1

0

0

0

0

0

111

1p

0

0

0

0

0

112

1

0

0

0

0

0

113

2

0

0

0

0

0

114

1p

0

0

0

0

0

115

1

0

0

0

0

0

116

1

0

0

0

0

0

117

1

0

0

0

0

0

118

1

0

0

0

0

0

119

2p

0

0

0

0

0

120

1

0

0

0

0p

0p

#. Test substance concentration (experimental animals) or vehicle (control animals).

p. Scaliness

Er: Erythema.

Oe: Oedema.

Vehicle: Kaydol White Mineral Oil.

 

Table 3: SECOND CHALLENGE READINGS

Animal No.

Challenge

Day 37

Day 38

Readings

Readings

50%

Vehicle

50%

Vehicle

Control

91

0

0

0

0

92

0

0

0

0

93

0

0

0

0

94

0

0

0

0

95

0

0

0

0

96

0

0

0

0

97

0

0

0

0

98

0

0

0

0

99

0

0

0

0

100

0

0

0

0

Experimental

101

0

0

0

0

102

0

0

0

0

103

0

0

0

0

104

0

0

0

0

105

0

0

0

0

106

0

0

0

0

107

0

0

0

0

108

0

0

0

0

109

0

0

0

0

110

0

0

0

0

111

0

0

0

0

112

0

0

0

0

113

0

0

0

0

114

0

0

0

0

115

0

0

0

0

116

0

0

0

0

117

0

0

0

0

118

0

0

0

0

119

0

0

0

0

120

0

0

0

0

#. Test substance concentration (experimental animals) or vehicle (control animals).

Er: Erythema.

Oe: Oedema.

Vehicle: Kaydol White Mineral Oil.

 

Table 4: BODYWEIGHTS (GRAM)

SEX/DOSE LEVEL

ANIMAL

DAY 1

DAY 38

FEMALES CONTROL

 

91

309

449

 

92

302

448

 

93

310

525

 

94

342

557

 

95

323

475

 

96

345

526

 

97

336

509

 

98

350

524

 

99

319

467

 

100

346

509

 

MEAN

328

499

 

ST.DEV.

18

37

 

N

10

10

FEMALES EXPERIMENTAL

 

101

337

493

 

102

352

561

 

103

334

510

 

104

359

534

 

105

306

493

 

106

340

552

 

107

321

478

 

108

333

550

 

109

323

456

 

110

348

558

 

111

280

519

 

112

353

507

 

113

333

510

 

114

306

459

 

115

310

497

 

116

339

572

 

117

334

525

 

118

332

485

 

119

326

543

 

120

332

544

 

MEAN

330

517

 

ST.DEV.

19

34

 

N

20

20

 

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Skin reactions were observed in response to a 50% concentration of 71786-47-5 benzenesulfonic acid mono and dialkylderivs magnesium salt, neutral, indiluents oil TBN = 20 in three of the twenty experimental animals (one grade 1 skin reaction and two observations of scaliness) in the first challenge phase. These skin reactions were not confirmed at second challenge with the same concentration. No skin reactions were observed in the control animals in response to the 50% first and second challenge exposure.
Executive summary:

Assessment of Contact Hypersensitivity to 71786-47-5 Benzenesulfonic Acid mono and dialkylderivs magnesium salt, neutral, indiluents oil TBN = 20 in the Albino Guinea Pig (Buehler Test).

 

The study was carried out based on the guidelines and test method described in:

OECD No. 406 (1992), "Skin Sensitization"

EC No 440/2008; B6: "Skin Sensitization: Buehler Test”.

EPA OPPTS 870.2600 (2003) “Skin Sensitization”

JMAFF: Japanese Test Guidelines (2000) including the most recent partial revisions.

 

The Buehler type of sensitization test is selected at the request of the sponsor since this substance is a surfactant. Studies have shown that the Local Lymph Node Assay, which is typically used, over predicts sensitization potential of these substances.

 

Test substance concentrations selected for the Main study were based on the results of a preliminary study.

In the Main study, twenty experimental animals were epidermally treated on three occasions (Days 1, 8 and 15) with the undiluted test substance and ten control animals were similarly treated, but with vehicle alone (Kaydol White Mineral Oil).

Two weeks after the last induction exposure, all animals were challenged with a 50% test substance concentration and the vehicle. A second challenge was performed one week later with the same test substance concentrations and the vehicle.

 

RESULTS

First Challenge: Skin reactions of grade 1 were observed in one experimental animal and scaliness was observed in two experimental animals in response to the 50% test substance concentration. No skin reactions were evident in the control animals.

 

Second challenge: To confirm the results of the first challenge, a second challenge was performed one week later. No skin reactions were evident after the second challenge exposure in the experimental and control animals.

 

CONCLUSION: Skin reactions were observed in response to a 50% concentration of 71786-47-5 benzenesulfonic acid mono and dialkylderivs magnesium salt, neutral, indiluents oil TBN = 20 in three of the twenty experimental animals (one grade 1 skin reaction and two observations of scaliness) in the first challenge phase.  These skin reactions were not confirmed at second challenge with the same concentration. No skin reactions were observed in the control animals in response to the 50% concentration of the test material in the first and second challenge exposures.  

 

The six-month reliability check with Alpha-hexylcinnamicaldehyde indicates that the Buehler Test as performed at WIL Research Europe is an appropriate model for testing for contact hypersensitivity.

Categories Display