Registration Dossier

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-11-2 to 2017-12-12
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
GLP compliance:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
α-d-Glucopyranoside, β-d-fructofuranosyl, benzoate
EC Number:
235-795-5
EC Name:
α-d-Glucopyranoside, β-d-fructofuranosyl, benzoate
Cas Number:
12738-64-6
Molecular formula:
C12-H22-O11.x-C7-H6-O2; x=5-8
IUPAC Name:
α-d-Glucopyranoside, β-d-fructofuranosyl, benzoate
Specific details on test material used for the study:
Name of substance : MIRAMER SB
α-d-Glucopyranoside, β-d-fructofuranosyl,
benzoate
Supplier : Miwon Specialty Chemical Co.,Ltd
KTR code : TS-00832
Cas No. : 12738-64-6
Lot No. : 161228BO5
Purity : 100 %
Physical description : Flake (Solid)
Storage condition : Room temperature[(1 – 30 C°)]

In vitro test system

Test system:
other: Reconstructed human epidermal (RHE) model
Source species:
human
Justification for test system used:
SkinEthic RHE tissue model is commonly used for in vitro skin irritation test,
Details on test system:
Quality assurance results for viability, barrier function and morphology were provided by manufacturer of skin (EPISKIN)
Culture condition:
Temperature 37 ± 1 °C
CO2 gas 5 ± 1 %
Humidity Under moist atmosphere
Incubator CO2 incubator (Thermo, Forma 3121)
The temperature and humidity during the test period were measured by COBEX recorder of a CO2 incubator.

Culture medium:
The culture medium used during the test period was provided by the manufacturer (EPISKIN)
Name Maintenance medium
Lot No. 17 SMM 022

Name Growth medium
Lot No. 17 SGM 033

SkinEthic RHE tissue model observation:
Before the beginning of incubation, each insert containing the RHA tissue was checked whether tissue surface were even and excess moisture were present and the seleted a tissue that was free of defects.

Tissue preparation:
Each insert containing the RHE tissue was removed from the agarose gel and transferred to the 6-well plate with 1 ml of growth medium. the plate was incubated at 37 °Cm 5% CO2 incubator for 2 hours.

Application of test substance:
18 ul or 10 ul water with 16 mg test substance was topically applied to the upper epithelial surface og the tissue with intervals of 1 minutes

Rinsing of test substances
According to the test substance application interval, treated RHE tissues was washed with PBS solution. The RHE tissue was washed 25 times (1 ml/time) to remove all residual test subtsnace and nylon mesh.

Post incubation:
Washed RHE tissue was transferred to the 6-well plate with 2 ml of groeth medium. The plate was incubated at 37 °C, 5% CO2 incubator for 42 hours.

Cell viabilty tests were performed on cells
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
16 mg 100% sucrose benzoate applied with an interval of 1 minute
Duration of treatment / exposure:
42 minutes
Number of replicates:
3

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
RHE model
Value:
72.5
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation

Any other information on results incl. tables

The absorbance value of the negative control substance was 1.079. The result was within the acceptable range (0.8 ≤ OD ≥ 3.0) under the skin iiritaion test using SkinEhic RHE model.

The cell viability of the positive control group was 1.6 ± 0.2%

The cell viability of the test subtance group was 72.5 ± 4.5 %

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Sucrose benzoate is not a skin irritant
Executive summary:

This study was conducted to assess the toxic potential of skin irritation of MIRAMER SB in SkinEhic RHETM (Reconstructed Human Epidermis) Model. The test results were evaluated as follows.

The test was performed in three groups; negative control, positive control, and test substance group.

Direct dyeing and non-specific reduction of MTT were not observed on the Test substance - The absorbance value of the negative control group and the cell viability value of the positive control group were satisfied the acceptance criterion.

- The cell viability of positive control group (Group 2) was calculated as 1.6 ± 0.2 %, and for the test substance group (Group 3), test substance was calculated as 72.5 ± 4.5 %.

As a result of in vitro skin irritation test in MIRAMER SB in SkinEhic RHETM (Reconstructed Human Epidermis) Model, cell viability has exceeded the reference value of 50 %. Therefore It was considered to be a non-irritant substance corresponding to 'No category' based on the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).