Registration Dossier

Administrative data

Description of key information

No LLNA was performed due to the surfactant property of the registration substance.

Instead the skin sensitization property of the registration substance was investigated according to the Guideline OECD 406 in guinea pigs. The test substance corresponded to ca 45% of registration substance in water. For the intradermal and the topical induction the animals were treated at concentration of 25% and 100% of test material respectively, and for the challenge the animals were treated at concentration of 100 % of test material. None of the treated animals responded. The registration substance is not skin sensitizing.

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:
19/12/2016 - 24/03/2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according 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 registration substance is a surfactant.
Specific details on test material used for the study:
The test material corresponded to the approximately 45% of the registration substance.
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Geniron Biolabs Pvt. Ltd., No.93, Solur, Thally Road, Anekal, Bengaluru - 562106, India

- Age at study initiation: 15 to 17 weeks at intra-dermal induction
- Weight at study initiation: Males : 396.74 to 489.13 g, Females : 410.05 to 474.02 g

- Housing: Animals were housed individually in standard polysulfone cages (Size: approximately L 425 x B 266 x H 185 mm)
- Diet (e.g. ad libitum): Guinea pig feed manufactured by Pranav agro Industries Ltd, Pune, Maharashtra
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in ‘Aquaguard’ on-line water filter-cum-purifier manufactured by Eureka Forbes Ltd., Mumbai 400 001, India
- Acclimation period: Start : 28 January 2017 End : 04 February 2017


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 23°C
- Humidity (%): 48 to 67%
- Air changes (per hr): 12 to 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark

IN-LIFE DATES: From: 28 January 2017 To: 01 March 2017
Route:
intradermal
Vehicle:
propylene glycol
Remarks:
The test material corresponds ca 45% of the registration substance in water
Concentration / amount:
Site 1: 0.1 mL of 50% Freund's Adjuvant in vehicle
Site 2: 0.1 mL of the test item in vehicle (25% w/v)
Site 3: 0.1 mL of 1:1 mixture of test item in vehicle (50% w/v) and Freund's Adjuvant in vehicle (50%)
Day(s)/duration:
On day 1
Adequacy of induction:
non-irritant substance, but skin pre-treated with 10% SDS
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
0.5 mL of test item
Day(s)/duration:
On day 8
Adequacy of induction:
non-irritant substance, but skin pre-treated with 10% SDS
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Day(s)/duration:
On day 22
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
10 (5 male and 5 female) for control group
20 (10 male and 10 female) for treatment group
Positive control substance(s):
yes
Remarks:
2-Mercaptobenzothiazole
Positive control results:
Skin sensitization rate of 60% at 24h reading and 50% at 48h reading
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no effect
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no effect
Remarks on result:
no indication of skin sensitisation

TABLE 1.             Summaryof Skin Reactions

INDUCTION

 

Group

Observation

Concentration (%w/v)

Application site

INDUCTION PERIOD

Time (hours)

Injection

site

Skin reaction scores and response*

Erythema

Edema

G1

(Vehicle Control)

Intradermal

Induction

‘0’

Shoulder region

24

1

2

3

10**/10

0/10

10**/10

10Ψ/10

0/10

10Ψ/10

48

1

2

3

10**/10

0/10

10**/10

10Ψ/10

0/10

10Ψ/10

Topical application

‘0’

Shoulder region

1@

 

NA

0/10

0/10

24@

 

NA

0/10

0/10

 

 

 

 

G2 (Treatment)

Intradermal

Induction

25

Shoulder region

24

1

2

3

20**/20

20**/20

20**/20

20Ψ/20

19Ψ/20

20Ψ/20

48

1

2

3

20**/20

16**/20

20**/20

20Ψ/20

0Ψ/20

20Ψ/20

Topical application

0.5 mL of undiluted test item

Shoulder region

1@

NA

0/20

0/20

24@

NA

0/20

0/20

@ : Observation post removal of test patch;                 NA: Not Applicable

*  : Represents the number of animals showing skin reactions out of the total number

      of animals in each group

** : Skin reaction score of 1-very slight erythema (barely perceptible)

Ψ : Skin reaction score of 1-very slight edema (barely perceptible)

0  : No erythema / No edema – correspondingly


 TABLE 1 contd. Summary of Skin Reactions

 

CHALLENGE

Group

Concentration (%w/v)

Application site

CHALLENGE PERIOD

Time (hours)

Skin reaction

Response

Sensitization rate (%)

G1

(Vehicle Control)

 

0.5 mL of undiluted test item

Left flank

24

 

0*/10

0

48

 

0*/10

0

 

G2 (Treatment)

 

0.5 mL of undiluted test item

Left flank

24

 

0*/20

0

48

 

0*/20

0

 

*

:

Represents the number of animals showing sensitisation response out of the total number of animals challenged in each group

 


Interpretation of results:
GHS criteria not met
Conclusions:
The skin sensitization property of the registration substance was treated according to the Guideline OECD 406. Based on the result obtained no classification is warranted.
Executive summary:

The skin sensitization property of the registration substance was investigated according to the Guideline OECD 406 in guinea pigs. The test substance corresponded to ca 45% of registration substance in water. For the intradermal and the topical induction the animals were treated at concentration of 25% and 100% of test material respectively, and for the challenge the animals were treated at concentration of 100 % of test material. None of the treated animals responded. The registration substance is not skin sensitizing.

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

Comments on the reliability of the obtained results in three in-vitro assays:

For the animal welfare reasons, three in-vitro assays were performed prior to performance of the in-vivo study. Two positive responses (KeratinoSens and h-CLAT) and one negative response (DPRA) were obtained. The registration substance is synthesized using unsaturated fatty acids that are not human-sensitizer but activated dendritic cells and/or keratinocytes (Frohwein TA et al. 2016. Arch Toxicol apr; 90(4):927 -36). Although two responses out of three were positive, the obtained results in in-vitro assays were considered not sufficiently reliable to claim the registration substance as a skin-sensitizer.

Justification for classification or non-classification

The skin sensitization property of the registration substance was investigated according to the Guideline OECD 406 in guinea pigs. The test substance corresponded to ca 45% of registration substance in water. For the intradermal and the topical induction the animals were treated at concentration of 25% and 100% of test material respectively, and for the challenge the animals were treated at concentration of 100 % of test material. None of the treated animals responded. The registration substance is not skin sensitizing. No classification is warranted.