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Toxicological information

Skin sensitisation

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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:
31 July 2012 to 02 August 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed in accordance with OECD, EU, US EPA and JMAFF test 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:
OECD Guideline 406 (Skin Sensitisation)
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Qualifier:
according to
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. certificate)
Type of study:
Buehler test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
Test substance information
Identification: 93820-55-4 Benzenesulfonic Acid, di C10-18-alkyl derivs barium salts, neutral, in diluents oil and TBN = <10
Molecular formula: UVCB
Molecular weight: UVCB
CAS Number: 93820-55-4
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
Hygroscopic: No
Volatile: No
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:
Test system
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: undiluted test substance 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: undiluted test substance and the vehicle (0.1 mL of each)
No. of animals per dose:
Control: 10 animals
Experimental test group: 20 animals
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 (grade 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 starting- and subsequent concentrations were taken from the series: 100% (undiluted), 50%, 20%, 10%, and, if needed, further lower concentrations using the same steps. 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 exposure.

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.

CHALLENGE - All animals
Day 29: The right flank of all animals was clipped and subsequently treated epidermally with the undiluted test substance 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.

After termination, animals were sacrificed using isoflurane and an intra-peritoneal injection of Euthasol® 20%.

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. To facilitate scoring, the epidermally treated skin-areas were clipped at least 3 hours before the 48-hour reading of these areas in the preliminary irritation study and challenge phase.
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:
Postive control not utilised in this study.

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:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
100%
No. with + reactions:
7
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: 7.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
100%
No. with + reactions:
3
Total no. in group:
20
Clinical observations:
Scaliness in 2/20
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 .

Any other information on results incl. tables

Table 1: PRELIMINARY IRRITATION STUDY

SKIN REACTIONS AFTER EPIDERMAL EXPOSURE

Animal number

Conc. %

24 hours after exposure

48 hours after exposure

Erythema

Oedema

Erythema

Oedema

(grade)

(grade)

(grade)

(grade)

5

100

0

0

0

0

50

0

0

0

0

6

100

0

0

0

0

50

0

0

0

0

7

20

0

0

0

0

10

0

0

0

0

8

20

0

0

0

0

10

0

0

0

0

 

Table 2: INDUCTION AND CHALLENGE READINGS

Animal No

Induction

Challenge

Day 15

Day 30

Day 31

Readings

24 Hour Readings

48 Hour Readings

100% #

100%

Vehicle

100%

Vehicle

Er

Oe

 

 

 

 

Control

61

0

0

0

0

0

0

62

0

0

0

0

0

0

63

0p

0

0

0

0

0

64

0

0

0

0

0

0

65

0p

0

0

0

0

0

66

0

0

0

0

0

0

67

0

0

0

0

0

0

68

0

0

0

0

0

0

69

0

0

0

0

0

0

70

0

0

0

0

0

0

Experimental

71

1

0

0

0

0

0

72

1p

0

1

0

1

0

73

0

0

0

0

0

0

74

0

0

0

0

0

0

75

1

0

0

0

0

0

76

1

0

0

0

0

0

77

1

0

1

0

0

0

78

0

0

0

0

0

0

79

1

0

1

0

0

0

80

0

0

0

0

0

0

81

1

0

1

0

1p

0

82

1

0

1

0

0

0

83

1

0

0

0

0

0

84

0

0

0

0

0

0

85

1

0

0

0

0

0

86

1p

0

0

0

0

0

87

1p

0

1

0

1

0

88

1

0

1

0

0p

0

89

0p

0

0

0

0

0

90

1

0

0

0

0

0

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

Er: Erythema

Oe: Oedema

p. Scaliness

Vehicle: Kaydol White Mineral Oil.

 

Table 3: BODYWEIGHTS (GRAM)

SEX/DOSE LEVEL

ANIMAL

DAY 1

DAY 31

FEMALES CONTROL

 

61

320

518

 

62

303

439

 

63

322

502

 

64

348

515

 

65

332

513

 

66

326

495

 

67

315

475

 

68

310

486

 

69

301

425

 

70

307

443

 

MEAN

318

481

 

ST.DEV.

14

34

 

N

10

10

FEMALES EXPERIMENTAL

 

71

325

491

 

72

316

471

 

73

341

518

 

74

324

519

 

75

310

416

 

76

324

448

 

77

327

497

 

78

341

495

 

79

295

491

 

80

311

422

 

81

320

447

 

82

311

473

 

83

306

437

 

84

314

446

 

85

340

495

 

86

294

448

 

87

303

471

 

88

345

524

 

89

325

468

 

90

355

539

 

MEAN

321

476

 

ST.DEV.

17

35

 

N

20

20

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The skin reactions observed in response to the undiluted test substance in seven of the twenty, (35%) experimental animals in the challenge phase were considered indicative of sensitization, based on the ≥15% criteria for evidence of skin sensitisation and the absence of any response in the control animals.
Executive summary:

Assessment of Contact Hypersensitivity to 93820-55-4 Benzenesulfonic Acid, di C10-18-alkyl derivs barium salts, neutral, in diluents oil and TBN = <10 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 preferred sensitization test (Local Lymph Node Assay) 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 the undiluted test substance.

 

RESULTS: In the challenge phase, skin reactions of grade 1 were observed in seven experimental animals in response to the undiluted test substance. No skin reactions were evident in the control animals. Scaliness was seen in two of the experimental animals with grade 1 skin reactions.

 

CONCLUSION: The skin reactions observed in response to the undiluted test substance in seven of the twenty (35%) experimental animals in the challenge phase were considered indicative of sensitization, based on the ≥ 15% criteria for evidence of skin sensitisation and the absence of any response in the control animals.