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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2013

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes (incl. QA statement)
Type of study:
Buehler test
Justification for non-LLNA method:
Data from the in vivo non-LLNA study was already available prior start of present registration for regulatory purposes outside EU.

Test material

Constituent 1
Chemical structure
Reference substance name:
2-{2-[(2-ethylhexyl)oxy]ethoxy}ethyl prop-2-enoate
Cas Number:
117646-83-0
Molecular formula:
C15H28O4
IUPAC Name:
2-{2-[(2-ethylhexyl)oxy]ethoxy}ethyl prop-2-enoate
Test material form:
liquid
Details on test material:
Test substance name: MI RAMER M1086
Appearance : Clear liquid
Manufacturing date: 2012-03-29
Supplier:Miwon Specialty Chemical Co., Ltd.
Receipt day: 2012-05-23
Deli very amount: 399.223 g(Gross)
Cas No.:117646-83-0
Lot No.:120329JY1
Purit y: 95.55 %
Expiration date: 2013-03-28 (Manufactured after 1 year)
Storage condition: Room temperature [(15 ~ 25)°C

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: CrlOri :HA
Sex:
female
Details on test animals and environmental conditions:
Animal strain, species and sex : CrlOri :HA, Guinea pig, Female, SPF
Supplier: Orient Bio Inc. (699-13, Mokdong-Ri, Buk-Myeon, Gapyeong-Gun, Gyeonggi-Do, Korea)
Number of animals: receipt - 50 animals, treatment - 40 animals (Main study)
Age at start of treatment : approximately 6 weeks old
Body weight at the time of treatment: 308.79 ~ 349.35 g (Main study)

The animal facility was maintained at (20 ± 3)°, relative humidity of (50 ± 20) %, air ventilation of (10~15) changes/hr, 12 hours light /dark cycle (light during 8:00~20:00) at (150 ~ 300) Lux.

The temperature and relative humidity were monitored automatically every half-hour by automatic instrument and other environmental condition (illuminance,
ammonia concentration, noise of animal room, etc) were measured periodically (1 t imes/quarter) by SOP of Korea Testing & Research Institute.
There were no influenceable variations for study in environmental measurements.

Five animals per cage were housed in stainless steel cages [700(W) × 475(D) × 200(H), Daejong Instrumental Industry Co. Ltd, Korea] for the study periods
including quarantine and acclimation.

The animals were fed pellet diet for experimental guinea pig including 0.01 % ascorbic acid (Cargi l Agri Purina, I nc. 56-4, Soryong-dong, Gunsan-si, Jeoll abuk-do, Korea) and given the filtered and UV irradiated water ad libitum.

The diet was considered not to contain any contaminant based on the periodical analysis results report of manufacturer and water did not contain any contaminant based on periodical analysis in accordance with the SOP of Korea Testi ng & Research I nsti tute.

Study design: in vivo (non-LLNA)

Induction
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
12.5 %(v/v)
Day(s)/duration:
6 hours
Challenge
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
6.25 %(v/v
Day(s)/duration:
6 hours
No. of animals per dose:
In the preliminary test using 2 animals per concentration, skin reactions (moderate response) were observed at 25 %(v/v) and observed at 12.5 %(v/v, milde response).
Based on this result, concentration of 12.5 %(v/v, induction phase) and 6.25 %(v/v, challenge phase) was chosen for exposure of the main test.
Details on study design:
PRE-SCREEN TESTS:
Dose levels
The concentrations of test substance were 4 types, 50 %(v/v), 25 %(v/v), 12.5 %(v/v) including the highest concentration [100 %, test substance itsel f] with vehicles by using a common ratio 2.
The one flank for 2 animals per concentration was closely clipped [(4 × 6)cm] with hair clipper before the treatment day. The patch [(2.5 × 2.5)cm , Aceband-S,Youngchemical Co. Ltd., Korea] was fully-loaded with test substance (0.5 mL) and applied to the clipped region and maintained using elastic bandage (Coban, 3M, USA)for 6 hours.

Determination of the dose levels
In the preliminary test using 2 animals per concentration, skin reactions (moderate response) were observed at 25 %(v/v) and observed at 12.5 %(v/v, milde response).
Based on this result, concentration of 12.5 %(v/v, induction phase) and 6.25 %(v/v, challenge phase) was chosen for exposure of the main test.

MAIN STUDY

Induction exposure: Day 0, 7, 14
The fur on one flank [(4 × 6)cm²] was closely clipped with a hair clipper before the treatment day. On the treatment day, 0.5 mL each of the test substance preparation, absorbed in a patch [(2.5 × 2.5)cm², Aceband-S, Youngchemical Co., Ltd., Korea] was attached to cover the test area and maintained using elastic bandage (Coban,3M, USA) for 6 hours. In the control group, 80 % ethanol or 1 %(w/v) DNCB was applied in a similar manner to the test area. The same application as on day 0 was carried out on the same test area of the same flank on day 7 and again on day 14.

Challenge exposure: Day 28
The untreated flank [(4 × 6)cm2] of treated and control animals were clipped on the day prior to the challenge patch application. To the treated and negative control animals, patches [(2.5 × 2.5)cm2, Aceband-S, Youngchemical Co. Ltd., Korea] which absorbed 0.5 mL of the test substance preparation were attached to the test area and maintained using elastic bandage (Coban, 3M, USA) for 6 hours. In the positive control group, 1 %(w/v) DNCB was applied in a similar man
ner to the test area.
Positive control substance(s):
yes
Remarks:
Name : 1,chloro-2,4-dinitrobenzene(DNCB) Manufacturer : Sigma-Aldrich Corporation Lot No. : S46383V Cas No. : 97-00-7

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
6.25%(v/v)
No. with + reactions:
0
Total no. in group:
20
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
6.25%(v/v)
No. with + reactions:
0
Total no. in group:
20
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
6.25%(v/v)
No. with + reactions:
0
Total no. in group:
10
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
6.25%(v/v)
No. with + reactions:
0
Total no. in group:
10
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
1% (w/v) DNCB
No. with + reactions:
10
Total no. in group:
10
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
1%(w/v) DNCB
No. with + reactions:
6
Total no. in group:
10

Applicant's summary and conclusion

Conclusions:
This study was conducted to evaluate the skin sensitisation of MIRAMER M1086 in guinea pigs by Buehler test. Parameters measured during the study period were mortality, clinical signs, body weight changes and skin reactions. The following results were obtained.

- The test substance-related clinical signs and moribund (or dead) animals in negative (or positive) group were not observed during the study period. In
treatment group, the coloring of skin and crust formation was observed in 13 animals. These animals were recovered after observation was terminated. These clinical signs were observed in test area and is considered to be result by application of test substance.
- No treatment-related clinical signs were observed in any treated animals.
- All living animals showed a normal increase of body weight.
- At the 24 and 48 hours after challenge with test substance of 6.25 %(v/v), the sensitisation index (mean score of skin reacti on) and frequency index
(sensitisation rate) were ' 0.0', ' 0.0 %' for the test substance, equal to the negative control . In posi tive control, the sensitisation index (mean score of skin reaction) and frequency index (sensitisationrate) were ' 1.2', '0.7' and '100.0 %', ' 60.0 %' at 24 and 48 hours after challenge, respectively.
These results indicate that this substance i s considered to have no skin sensitizing potency of MIRAMER M1086 in guinea pigs by Buehler test under the conditi ons of t his study (Class I ).
Executive summary:

This study was conducted to evaluate the skin sensitisation of MIRAMER M1086 in guinea pigs by Buehler test. Parameters measured during study period were mortality, clinical signs, body weight changes and skin reactions.

No treatment-related mortality or clinical signs in negative (or positive) group were observed in any treated animals.

In treatment group, the clinical signs (coloring of skin, crust formation) was observed in 13 animals and is considered to be result by application of test substance.

All the living animals showed a normal increase of body weight during study period. After challenge completion with 6.25 %(v/v) concentration of test substance, no skin reactions were observed at 24 and 48 hours. The sensitisation index (mean score of skin reaction) and frequency index (sensitisation rate) were‘0.0’,‘0.0 %’, respectively.

In the positive control with 1 %(w/v) DNCB dissolved in corn oil, there were observed skin reaction of erythema. The sensitisation index (mean score of skin reaction) was '1.2 (24 hours)’,‘0.7 (48 hours)’and frequency index (sensitisation rate) was‘100.0 % (24 hours)’,‘60.0 % (48 hours)’, respectively.

These results indicate that this substance is considered to have no skin sensitizing potency (Class I) of MIRAMER M1086 in guinea pigs by Buehler test under the conditions of this study.