D-Glucopyranose, oligomeric, butyl glycoside

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation / corrosion

Remarks:

other: in-vitro

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP - Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD Guidelines for Testing of Chemicals, Guideline No. 439: In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method (adopted 22 July 2010)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: European Community (EC). Commission regulation (EC) No. 440/2008, Part B: Methods for the Determination of Toxicity and other health effects, Guideline B.46 “In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test "

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: EpiSkin™ SOP, Version 1.8 (February 2009), ECVAM Skin Irritation Validation Study: Validation of the EpiSkin™ test method 15 min - 42 hours for the prediction of acute skin irritation of chemicals

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Food and Consumer Product Safety Authority (VWA), GG Utrecht, The Netherlands

Test material

Test material information

Constituent 1

Reference substance name:

D-Glucopyranose, oligomeric, butyl glycoside

EC Number:

939-389-5

Molecular formula:

not applicable, UVCB substance

IUPAC Name:

D-Glucopyranose, oligomeric, butyl glycoside

Details on test material:

- Name of test material (as cited in study report): LIN11001-butylglucoside
- Physical state: beige chunks
- Analytical purity: > 98%
- Expiration date of the lot/batch: 2013-02-06
- Storage condition of test material: at room temperature in the dark
- Other: stability under storage conditions: stable

Test animals

Species:

human

Strain:

other: EPISKIN Small ModelTM (EPISKIN-SMTM), reconstructed three-dimensional human epidermis model

Details on test animals or test system and environmental conditions:

TEST SKIN MODEL
- Source: SkinEthic Laboratories, Lyon, France

TEST METHOD
This model is a three-dimensional human epidermis model, which consists of adult human-derived epidermal keratinocytes which have been seeded on a dermal substitute consisting of a collagen type I matrix coated with type IV collagen. The keratinocytes were cultured for 13 days, which results in a highly differentiated and stratified epidermis model comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum. Irritant materials are identified by their ability to penetrate the stratum corneum and to damage the underlaying cell layers which is determined through a decrease in cell viability as determined by MTT reduction assay.

ADAPTATION TO CELL CULTURE CONDITIONS
On the day of receipt, the tissues were transferred to 12-well plates and preincubated with prewarmed Maintenance Medium (SkinEthic Laboratories, Lyon, France) for 24 h at 37 °C.

INCUBATION CONDITIONS (INCUBATOR)
- Temperature (°C): 36.2-37.6
- CO2 gas concentration (%): 5.0 ± 0.5
- Humidity (%): 80-97

Test system

Type of coverage:

other: open (in vitro system)

Preparation of test site:

other: intact reconstructed human epidermis

Vehicle:

unchanged (no vehicle)

Controls:

other: concurrent control tissues treated with PBS served as negative controls, positive controls were exposed to 5% SDS

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 11.9-13.6 mg on the skin tissue moistened with 5 μL of Milli-Q water (Millipore Corp., Bedford, Mass., USA)

POSITIVE CONTROL SUBSTANCE:
- Positive control substance: SDS (Sigma Aldrich, Zwijndrecht, The Netherlands), 5% (v/v) in PBS

Duration of treatment / exposure:

15 min

Observation period:

Not applicable. Post-treatment incubation period: 44 h

Number of animals:

Not applicable. The test was performed in triplicates for each treatment and control group.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: the tissues were washed with PBS to remove residual test substance.
- Time after start of exposure: 15 min
- Post-treatment incubation period: 44 h

CELL VIABILITY MEASUREMENTS
For determining alterations in cell viability, MTT reduction assays were performed 44 h after the incubation period. Therefore, tissues were incubated in 2 mL prewarmed MTT solution (0.3 mg/mL) for approximately 3 h at 37 °C and 5% CO2. After aspiration of the MTT solution, tissues were placed on blotting paper for drying. Extraction of the formazan product was carried out in 0.5 mL isopropanol (Merck, Darmstadt, Germany). The optical density was measured at 570 nm wave length in a 96 wells plate using a plate spectrophotometer (TECAN Infinite® M200 Pro Plate Reader).

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

other: cell viability (% of negative control)

Basis:

other: mean value of negative controls (sterile Milli-Q water)

Time point:

other: 15 min

Score:

100

Reversibility:

other: reversibility: not applicable

Results 2

Irritation parameter:

other: cell viability (% of negative control)

Basis:

other: mean value of positive controls (5% SDS)

Time point:

other: 15 min

Score:

5

Reversibility:

other: reversibility: not applicable

Results 3

Irritation parameter:

other: cell viability (% of negative control)

Basis:

other: mean value of the test item (100%)

Time point:

other: 15 min

Score:

99

Reversibility:

other: reversibility: not applicable

Other effects:

No further effects were observed.

Any other information on results incl. tables

Table 3. Results of MTT assay after 15 min exposure

	Negative control			Positive control			Test item
Tissue sample	1	2	3	1	2	3	1	2	3
OD570	0.908 0.947	0.941 0.968	0.970 1.001	0.045 0.051	0.039 0.065	0.042 0.057	0.957 0.982	0.951 0.845	0.988 0.979
OD570 (mean)	0.928	0.955	0.986	0.048	0.052	0.050	0.970	0.898	0.984
SD	0.028	0.019	0.022	0.004	0.018	0.011	0.018	0.075	0.006
OD550 (mean values of replicates ± SD)	0.956 ± 0.029			0.050 ± 0.002			0.950 ± 0.046
Viability (%)	100			5			99

OD = optical density; SD = standard deviation

In this table, the values are corrected for background absorption (0.042). Isopropanol was used to measure the background absorption.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

CLP: not classified
DSD: not classified