Hydrogen (glycinato-N,O)[sulphato(2-)-O,O']ferrate(1-)

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

- 2020-10-31

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Version / remarks:

adopted 18 June 2019

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Hydrogen (glycinato-N,O)[sulphato(2-)-O,O']ferrate(1-)

EC Number:

241-221-4

EC Name:

Hydrogen (glycinato-N,O)[sulphato(2-)-O,O']ferrate(1-)

Cas Number:

17169-60-7

Molecular formula:

C2H5FeNO6S

IUPAC Name:

2-aminoacetate;hydron;iron(2+);sulfate

Test material form:

solid

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Cell source was not reported

Justification for test system used:

This test method is able to detect chemicals that cause skin irritation, i.e. produce reversible damage to the skin and allows for hazard identification in accordance with UN GHS “Category 2”. Depending on the regulatory framework it can also be used to identify non-classified chemicals.

Vehicle:

other: sterile DBPS

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: reconstituted three-dimensional human skin model EpiDerm™ (MatTek)
- Tissue batch number(s):
- Date of initiation of testing:

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 25 ± min at room temperature and 35 ± 1min at 37°C
- Temperature of post-treatment incubation (if applicable): 37°C for 42 h

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: 15 times with DBPS and subsequently submerged three-times with 150 µL DBPS and shaken to remove rests of the test item
- Observable damage in the tissue due to washing:
- Modifications to validated SOP:

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3h ± 5 min
- Spectrophotometer: plate spectrophotometer
- Wavelength: 570 nm
- Filter bandwidth: with a filter band pass of maximum ± 30 nm without reference wavelength


FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability:
- Barrier function:
- Morphology:
- Contamination:
- Reproducibility:

NUMBER OF REPLICATE TISSUES: triplicate

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Fresh tissues / killed tissues
- Procedure used to prepare the killed tissues (if applicable):
- N. of replicates :
- Method of calculation used:

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION:

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive /irritant to skin if the viability after 1 hour exposure is less than 50%.
- The test substance is considered to be non-corrosive/irritant to skin if the viability after 1 hour exposure is greater than or equal to 50%.

Control samples:

yes, concurrent no treatment

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg and 25 µL to moistened the tissue

VEHICLE
- Amount(s) applied (volume or weight with unit): 25 µL DBPS
- Lot/batch no. (if required): Cat. No.: 14040-091

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL DBPS

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL SDS solution
- Concentration (if solution): 5%

Duration of treatment / exposure:

1h

Duration of post-treatment incubation (if applicable):

42 h

Number of replicates:

triplicate

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean of three treatments with the test material

Value:

76

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: Not reported
- Direct-MTT reduction:
- Colour interference with MTT:

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Please refer to the 'any other information on results incl. tables' section
- Acceptance criteria met for positive control: Please refer to the 'any other information on results incl. tables' section
- Acceptance criteria met for variability between replicate measurements: Please refer to the 'any other information on results incl. tables' section
- Range of historical values if different from the ones specified in the test guideline:

Any other information on results incl. tables

Results are draft report results which are reported in more detail on the next update

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In the present in vitro test conducted according to OECD guideline 439, 25 mg ferrous monoglycinate sulfate were applied to human keratinocytes for 1h. Afterwards the cell viability was measured with MTT at 570 nm. The mean measured viability as compared to the negative controls for the test item was 76%, thus the test item is not classified according to Regulation (EU) No. 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS)

Executive summary:

In a dermal irritation study performed in accordance with OECD Guideline 439 (In Vitro Skin Irritation) (adopted 18 June, 2019), ferrous monoglycinate sulfate (100% a.i.) was applied to the three-dimensional human epidermis model tissue for an exposure period of 1 h in triplicates. 25 μL of DBPS were topically applied to the epidermal surface in order to improve further contact between the solid and the epidermis. Each approximately 25 mg of the test item were applied to the wetted tissues. The test item was spread to match the surface of the tissue.

After 25 minutes exposure at room temperature and 35 minutes at 37°C, the tissues were washed with DBPS to remove any residual test material. Subsequently the tissue constructs were incubated for 42 h at 37°C. Cytotoxicity (irritancy) was expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment.

The positive (5% SDS) and negative (DBPS) control gave responses that were within the acceptance criteria and as such demonstrated the validity of the study.

The relative mean tissue viability obtained after 1h treatment with ferrous monoglycinate sulfate compared to the negative control tissues was 76%. Since the mean relative tissue viability for the test substance was above 50%, ferrous monoglycinate is identified to be not irritating.