Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
June 06, 2018 to November 03 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2018

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: TG 406
Version / remarks:
"Skin Sensitisation Study" (406) published by the Ministry of Environmental Protection of People's Republic of China 2013.
Deviations:
no
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
According to the Guidelines for the testing of chemicals "Skin Sensitisation Study" (406) published by the Ministry of Environmental Proection of People's Republic of China in the year of 2013, guinea pig is the preferred strain for the heredity characters, stability and available background data. Based on the Skin Sensitization result of positive control test item DNCB, this strain is reliable.

Test material

Constituent 1
Chemical structure
Reference substance name:
(2-fluoro-3-methoxyphenyl)boronic acid
EC Number:
609-099-0
Cas Number:
352303-67-4
Molecular formula:
FC6H3(OCH3)B(OH)2
IUPAC Name:
(2-fluoro-3-methoxyphenyl)boronic acid
Test material form:
solid: particulate/powder

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
female
Details on test animals and environmental conditions:
Species: Albino Guinea pigs
Strain: Hartley
Grade: SPF
Supplier: Beijing Vital River Laboratory Animal Technology Co., Ltd.
License No.: SCXK. (Jing) 2016-0011
Certification No.: 11400700312017
Animal Amount: 32
Sex: Female, nulliparous and non-pregnant.
Age and Body Weight of Animals on Arrival: the ages were between 4 and 6 weeks (28~4I) days and the body weights were between 210 and 275 g.
Age and Body Weight of Animals of the Main Skin Sensitization Test before Administration: the ages were between 7 and 10 weeks (55~68 days) and the body weights were between 285 and 400 g.
Physical Examination and Acclimatization: All animals were acclimatized in room Al 18. The acclimatization period was 27 days for the health young adult test animals. All animals· were observed daily for common status during the acclimatization periods. A health examination was conducted, and all animals were weighted and marked by special animal markers beginning from No.l and number written on the cage cards within 24 hours after arrival.

Justification of Test System
According to the Guidelines for the testing of chemicals "Skin Sensitisation. Study" (406) published by the Ministry of Environmental Protection of People's Republic of China in the year of 2013, guinea pig is the preferred strain for the heredity characters, stability and available background data. Based on the Skin Sensitization result of positive control test item DNCB (2,4-Dinitrochlorobenzene), this strain is reliable.

Test conditions
Husbandry condition
Husbandry: Animals were raised in plastic cage (L80cmxW50cmxH20cm) and each held no more than 12 animals with com cob bedding during the study period. Animals used for preliminary irritation test was housed in a separate cage after grouping. There was a cage card of each cage bearing cage number, group number, animal ID etc.

Environmental Controls
Temperature and humidity were controlled automatically and recorded daily. The temperature values in the animal room were 17-23°C. The relative humidity values were 40%-70%. The lighting sequence was 12 h light and 12 h dark.

Food and Water
Animals were provided with pellet diet with complete nutrition supplied by Beijing Keaoxieli Feed Co., Ltd.. Analysis reports of diet were supplied by the supplier. Water was purified by HT-ROl000 purity system and was analyzed annually. Food and water was available to the animals ad libitum during test. The diet and water were considered have no ingredients that affect the test.

Animal Welfare
Animal usage . complied with national animal welfare laws and regulations (instructive notions with respect to caring for laboratory animals) (2006, Ministry of Science, P.R.C.). The animal care and use activities required for conduct of this study were reviewed and approved by the testing facility Animal Care and Use Committee (IACUC).

Study design: in vivo (non-LLNA)

Induction
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
0.301 g raw test item/animal
Day(s)/duration:
On day 0, 7, and 14. 6 hours of exposure
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
0.302 g raw test item/animal
Day(s)/duration:
On day 28 with exposure period of 6 hours
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
2 for preliminary study, 10 for control group, 20 for treated group
Details on study design:
Preliminary irritation test Method
Two animals were randomly selected from all the animals. The test item was moistened with the minimum ultra-purified water and then was applied to the animals in the preliminary group to test if it has a skin irritating effect. In case of such effect was not found, the over saturate highest concentration (mass/volume) of the test item was selected for both induction and challenging.

Test Item Preparation:
Fixed amount test item was weighed and placed in a small container; a little vehicle was added, and the test ite and vehicle were mixed to form an even mixture. The amount of the vehicle was calculated. The average dose of three induction phases for each animal was 0.301 g and the volume of vehicle for each animal was 0.5 mL. The average dose of challenging phase for each animal was 0.302 g and the volume of vehicle for each animal was 0.5 mL. The test item was administered via a vehicle, and the homogeneity, concentration and stability of the test item in the vehicle were not determined because they were not considered to have impacts on the quality and integrity ofthe study.

Animal preparation:
For treated group animals, in the morning of the day before each induction day, the left flanks of guinea pigs will be clipped free of hair. For both treated group and control group animals, in the morning of the day before each challenging day, the right flanks of guinea pigs were clipped free of hair. The clipped area was about 4cmx4 cm.

Administration: The test item was homogeneously spreaded on one layer filter paper. The paper (about 2cmx2cm)was on two layers gauze (about 4cmx4cm), and the gauze was stick to one layer non-allergenic medical adhesive tape which contained one layer polyethylene film. The patch was placed on each side of clipped area of the animal. The animal was wrapped with the non-allergenic medical adhesive tape for 6 hours.

Animal Management after Exposure: After 6 hours of exposure, the patches were removed and the test sites were gently cleansed of any residual test item with water.

Induction Phase:
Treated Group: On day O (first induction date), day 7 and day 14, dosing the same depilation areas of the left flanks of the animals.
Control Group: No treatment.
Clinical Observation: At approximately 24 h after the patch was removed, each dosing site was observed for erythema and edema.

Challenging Phase:
Control group and Treated Group: On day 28, dosing the right flanks of treated and control group animals. For the control group, the challenge site was right ribbed abdomen of each animal. For the treated group, the challenge site was also right flank but different from the previous one of each animal.

Clinical Observation and Scoring: At approximately 24 hand 48 h after exposure, the application sites were observed and scored for erythema and edema.

Moribund /Mortality Observation: All guinea pigs were observed once each day for mortality and moribund status during the test period.

Body Weights: Animals were weighed on arrival day (within 24 hours), on the grouping day and at the end of the study.
Challenge controls:
none
Positive control substance(s):
yes
Remarks:
2,4-Dinitrochlorobenzene

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
0.302g
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
0.302g
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:
0.302g
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:
0.302g
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
No. with + reactions:
16
Total no. in group:
20
Remarks on result:
positive indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
No. with + reactions:
16
Total no. in group:
20
Remarks on result:
positive indication of skin sensitisation

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Based on above results, FMPBA is not a skin sensitizer to guinea pigs.