Sorbitan, tridocosanoate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 - 28 Sep 2017

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)

Version / remarks:

28 July 2015

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Sorbitan, tridocosanoate

EC Number:

301-037-8

EC Name:

Sorbitan, tridocosanoate

Cas Number:

93980-59-7

Molecular formula:

C72H138O8

IUPAC Name:

Sorbitan tridocosanoate

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Source strain:

other: Keratinocyte Strain: 4F0219

Justification for test system used:

reccomended in TG OECD 439

Vehicle:

unchanged (no vehicle)

Remarks:

Tissue was wetted with 25 µL of PBS prior to exposure

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Epiderm TM Reconstructed Human Epidermis, EPI-200, EPI-212
- Tissue batch number(s): 27124
- Production date: 27 Sep 2017 (Date of Certificate of Analysis)
- Date of initiation of testing: 25 Sep 2017

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37°C, humid, 5% CO2 (incubator), then room temperature
- Temperature of post-treatment incubation: not provided

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: The tissue was rinsed 15 times with PBS(-), then they were 3 times completely submerged in 150 mL PBS(-) and once rised from inside and outside with PBS(-); Remaining PBS(-) was removed from the inside and outside of the tissue insert with a sterile cotton swab before transferring the tissues into the wells with fresh medium.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 180 ± 5 min
- Spectrophotometer: FLUOstar OPTIMA, BMG LABTECH
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: 1.802 ± 0.057
- Barrier function: test was passed
- Morphology: well-differentiated epidermis consisting of a viable basal layer, intermediate spinous and granular layers and a functional stratum corneum
- Contamination: no

NUMBER OF REPLICATE TISSUES: 3

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA:
- The test substance is considered to be corrosive or irritant to skin if the mean relative viability after 60 minutes exposure is less or equal to 50% of the negative control.
- The test substance is considered to be non-corrosive or non irritant to skin if the viability after 60 minutes exposure is greater than 50%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied: 25 mg

NEGATIVE CONTROL
- Amount(s) applied: 30 µL
- Concentration: undiluted

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

Duration of treatment / exposure:

60 min (35 min at 37°C and 25 min at room temperature)

Duration of post-treatment incubation (if applicable):

24 ± 2 h, after transfer to lower wells containing fresh medium further 18 ± 2 h

Number of replicates:

3

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean value of positive control (5% SDS)

Value:

1.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean value of test substance

Value:

98.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: no
- Direct-MTT reduction: no

DEMONSTRATION OF TECHNICAL PROFICIENCY: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes (mean OD = 2.348)
- Acceptance criteria met for positive control: yes (mean viability = 1.9%)
- Acceptance criteria met for variability between replicate measurements: yes (0.2 - 3.2%)

Any other information on results incl. tables

Table 1: Results MTT assay after 3 h ± 5 min exposure

Substance	Tissue No.	OD570	OD570 (mean per tissue)	OD570 (mean of 3 tissues)	Cell viability [%]	Cell viability (mean) [%]	Standard Deviation of cell viability [%]
Negative Control	1	2.409	2.385	2.348	101.6	100.0	1.4
		2.36
	2	2.376	2.338		99.6
		2.299
	3	2.364	2.322		98.9
		2.280
Positive Control	1	0.046	0.046	0.044	2.0	1.9	0.2
		0.046
	2	0.041	0.041		1.7
		0.04
	3	0.047	0.045		1.9
		0.042
Test substance	1	2.381	2.348	2.313	100.0	98.5	3.2
		2.314
	2	2.389	2.364		100.7
		2.338
	3	2.261	2.227		94.8
		2.192

 

Applicant's summary and conclusion

Interpretation of results:

other: CLP/ EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008