Registration Dossier

Administrative data

Description of key information

Skin sensitization

Everzol SR98 was non-sensitizingto skin (OECD TG406).

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 December 06, 2017 to April 03, 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
The guinea pig Buehler test is already available for the registration outside of EU. Therefore, LLNA is not conducted.
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
- Source: Laboratorio Nacional Agropecuana Lanagro S.P.
- Age at study initiation: 12-15 weeks old
- Housing: ten animals per sex were grouped inside polypropylene cages
- Acclimation period: at least 5 days
- Temperature (°C): 18.0-23.0 °C
- Humidity (%): 48-70 %
- Photoperiod: 12-hrs dark / 12-hrs light
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
100%
Day(s)/duration:
Days 0-14
Adequacy of induction:
highest technically applicable concentration used
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
100%
Day(s)/duration:
Day 14-28
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
Two for pilot study, twenty for treatment group and ten for control group
Positive control substance(s):
yes
Remarks:
α-Hexylcinnamaldehyde (HCA)
Positive control results:
The result of α-Hexylcinnamaldehyde (HCA) was 50% of the animals showed positive response of sensitivity.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
100%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
100%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
100%
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:
positive control
Dose level:
100%
No. with + reactions:
10
Total no. in group:
20
Remarks on result:
positive indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
100%
No. with + reactions:
10
Total no. in group:
20
Remarks on result:
positive indication of skin sensitisation

Table 1. Skin Reactions Observed in the Pilot Study.

Concentration Agreed (%)

Weight (g)

Animals #

Sex

Skin Reaction

Initial

Final

After 24 hours

After 48 hours

100

503

512

1

0

0

75

497

509

2

0

0

Table 2. Skin Reactions Observed in Test Group at Inductions Period.

Animal #

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

Sex

Initial weight (g)

480

560

548

566

646

502

584

558

494

536

427

550

474

591

603

463

563

569

558

527

1st Induction

24hrs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2nd Induction

24hrs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3rd Induction

24hrs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Table 3. Skin Reactions Observed in Control Group at Inductions Period.

Animal #

23

24

25

26

27

28

29

30

31

32

Sex

Initial weight (g)

550

479

573

507

517

510

585

550

508

517

1st

Induction

24hrs

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

2nd

Induction

24hrs

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

3rd

Induction

24hrs

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

Table 4. Skin Reactions Observed in Test Group at Challenge Period.

Animal #

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

Final weight (g)

690

595

697

642

671

533

628

582

713

586

626

644

677

605

600

595

692

612

690

631

24hrs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Table 5. Skin Reactions Observed in Control Group at Challenge Period.

Animal #

23

24

25

26

27

28

29

30

31

32

Final weight (g)

707

696

670

561

681

505

637

636

550

612

24hrs

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

Table 6. Clinical and Behavioral Alterations Observed in the Animals of Test Group.

Group

Test

Animal #

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

Pilot

24hrs

0

0

 -

-

-

-

48hrs

0

0

 -

1st Induction

24hrs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

48hrs

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2nd Induction

24hrs

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3rd Induction

24hrs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

48hrs

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Challenge

24hrs

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Table 7. Clinical and Behavioral Alterations Observed in the Animals of Control Group.

Group

Control

Animal #

23

24

25

26

27

28

29

30

31

32

1st

Induction

24hrs

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

2nd

Induction

24hrs

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

3rd

Induction

24hrs

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

Challenge

24hrs

0

0

0

0

0

0

0

0

0

0

48hrs

0

0

0

0

0

0

0

0

0

0

0: None visual alterations observed.

Interpretation of results:
GHS criteria not met
Conclusions:
According to OECD 406 test method, Everzol SR98 showed that non-sensitizing to guinea pigs. Therefore, Everzol SR98 was not met a category based on GHS criteria.
Executive summary:

This test using the procedures outlined in the BIOAGRI Study Plan for 16264.467.103.17.The results of this OECD 406 test for Everzol SR98 show that test reliability criteria was met. The concentration agreed as 100% was applied in the inductions and in the challenge, according to the results obtained in the pilot study and inductions periods. The quantity of 0.5 mL of the test item homogenized in purified water was held in contact with the skin of the flank region for a 6-hours exposure period. Thirty-two (females nulliparous and non-pregnant) guinea pigs were tested, two for pilot study, twenty for treatment group and ten for control group, maintained under controlled environmental conditions during the acclimation and observation periods. Each one of the twenty treatment animals was exposed to the test item in both inductions and challenge, although all ten control animals were exposed to the test item only during the challenge period. Skin reactions after inductions and after challenge period were appraised according to the Magnusson & Kligman’s grading scale. Reactions were evaluated 24 and 48 hours after occlusive bandages removes, approximately. When applied on the guinea pigs’ skin, the test item did not cause any alteration in challenge period. Therefore, Everzol SR98 was classified as non-sensitizing to guinea pigs.

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

The results of this OECD 406 test for Everzol SR98 show that test reliability criteria was met. The concentration agreed as 100% was applied in the inductions and in the challenge, according to the results obtained in the pilot study and inductions periods. The quantity of 0.5 mL of the test item homogenized in purified water was held in contact with the skin of the flank region for a 6-hours exposure period. Thirty-two (females nulliparous and non-pregnant) guinea pigs were tested, two for pilot study, twenty for treatment group and ten for control group, maintained under controlled environmental conditions during the acclimation and observation periods. Each one of the twenty treatment animals was exposed to the test item in both inductions and challenge, although all ten control animals were exposed to the test item only during the challenge period. Skin reactions after inductions and after challenge period were appraised according to the Magnusson & Kligman’s grading scale. Reactions were evaluated 24 and 48 hours after occlusive bandages removes, approximately. When applied on the guinea pigs’ skin, the test item did not cause any alteration in challenge period. Therefore, Everzol SR98 was classified as non-sensitizing to guinea pigs.

Justification for classification or non-classification