Reaction mass of 2,6-dimethylpyridinium chloride and 3-methylpyridinium chloride and 4-methylpyridinium chloride

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

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:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 2,6-dimethylpyridinium chloride and 3-methylpyridinium chloride and 4-methylpyridinium chloride

EC Number:

941-122-2

Molecular formula:

not yet assigned

IUPAC Name:

Reaction mass of 2,6-dimethylpyridinium chloride and 3-methylpyridinium chloride and 4-methylpyridinium chloride

Test material form:

semi-solid (amorphous): gel

Remarks:

migrated information: paste

Test animals

Species:

human

Strain:

other: human skin model EPISKIN-SM™

Details on test animals or test system and environmental conditions:

The EPISKIN-SMTM tissues are provided as kits (SkinEthic), consisting of the following components relevant for this study:
1x EPISKIN-SM™ plate containing 12 reconstructed epidermis units (area: 0.38 cm2);
each reconstructed epidermis is attached to the base of a tissue culture insert with an O-ring set and maintained on nutritive agar for transport (Lot: 14-EKIN-033; 14-EKIN-038)
1x 12-well assay plate
1x flask of sterile maintenance medium (basic medium for incubations, Lot: 14-MAIN3-037; 14-MAIN3-043)
1x flask of sterile assay medium (basic medium for use in MTT assays, Lot: 14-ESSC-035; 14-ESSC-041)

Test system

Vehicle:

water

Controls:

other: positive and negative control with EPISKIN-SMTM tissues (SkinEthic)

Amount / concentration applied:

Test Item: 10 mg + 5 μL aqua dest.
Negative control: 10 μL Phosphate Buffered Saline
Positive control: 10 μL 5% Sodium Dodecyl Sulfate in aqua dest.

Duration of treatment / exposure:

15 minutes

Observation period:

42 h post-incubation

Number of animals:

3 tissues per dose group

Details on study design:

TEST SITE
- Area of exposure: 0.38 cm2
- Type of wrap if used: no wraps, open in incubators

REMOVAL OF TEST SUBSTANCE
- Washing: washed with Phosphate Buffered Saline to remove any residual test item. Excess Phosphate Buffered Saline was removed by blotting bottom with blotting paper.
- Time after start of exposure: 15 ± 0.5 min.

The test meets ACCEPTANCE CRITERIA if:
- OD 550 nm of the blank is < 0.1
- mean OD 550 nm of the three negative control tissues is ≥ 0.6 and ≤ 1.5
- mean relative tissue viability of the three positive control tissues is ≤ 40%
- the standard deviation (SD) of relative tissue viability obtained from each three concurrently tested tissues is < 18%.

Results and discussion

Any other information on results incl. tables

Experimental Results; 1st experiment

Name	Negative Control			Positive Control			Test Item
Tissue	1	2	3	1	2	3	1	2	3
absolute OD550	0.754 0.756	0.875 0.845	0.798 0.782	0.096 0.093	0.148 0.152	0.098 0.094	0.614 0.596	0.320 0.320	0.568 0.561
OD550 (blankcorrected)	0.710 0.712	0.831 0.801	0.754 0.738	0.052 0.049	0.104 0.108	0.054 0.050	0.570 0.552	0.276 0.276	0.524 0.517
mean OD550 of the duplicates (blankcorrected)	0.711	0.816	0.746	0.05	0.106	0.052	0.561	0.276	0.521
total mean OD550 of 3 replicate tissues (blankcorrected)	0.758*			0.069			0.453
SD OD550	0.053			0.032			0.154
relative tissue viabilities [%]	93.9	107.7	98.4	6.6	14.0	6.9	74.1	36.4	68.7
mean relative tissue viability [%]	100.0			9.2**			59.8
SD tissue viability [%]***	7.0			4.2			20.4
CV [% viability]	7.0			45.9			34.1

* Corrected mean OD550 of the negative control corresponds to 100% absolute tissue viability.

** Mean relative tissue viability of the three positive control tissues is ≤ 40%.

*** The standard deviation (SD) obtained from the three concurrently tested control tissues is < 18%. This criterion failed for the test substance.

Experimental Results; 2nd experiment

Name	Negative Control			Positive Control			Test Item
Tissue	1	2	3	1	2	3	1	2	3
absolute OD550	1.118 1.156	1.038 1.144	0.959 1.046	0.135 0.138	0.104 0.117	0.100 0.109	1.020 1.123	0.873 0.974	0.960 1.021
OD550 (blankcorrected)	1.075 1.113	0.995 1.101	0.915 1.003	0.092 0.095	0.060 0.073	0.056 0.065	0.976 1.079	0.829 0.930	0.916 0.978
mean OD550 of the duplicates (blankcorrected)	1.094	1.048	0.959	0.093	0.067	0.061	1.028	0.880	0.947
total mean OD550 of 3 replicate tissues (blankcorrected)	1.033*			0.074			0.951
SD OD550	0.068			0.017			0.074
relative tissue viabilities [%]	105.8	101.4	92.8	9.0	6.4	5.9	99.5	85.1	91.6
mean relative tissue viability [%]	100.0			7.1**			92.1
SD tissue viability [%]***	6.6			1.7			7.2
CV [% viability]	6.6			23.4			7.8

* Corrected mean OD550 of the negative control corresponds to 100% absolute tissue viability.

** Mean relative tissue viability of the three positive control tissues is ≤ 40%.

*** The standard deviation (SD) obtained from the three concurrently tested tissues is < 18%.

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Remarks:

Migrated information

Conclusions:

In this study under the given conditions the test item showed no irritant effects. The test item is therefore classified as “non-irritant” in accordance
with UN GHS “No Category”.