Cyclohexanaminium, N,N,N-trimethyl-, sulfate (2:1)

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation / corrosion

Remarks:

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:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Trimethylcyclohexylammonium sulfate

IUPAC Name:

Trimethylcyclohexylammonium sulfate

Constituent 2

Reference substance name:

bis(N,N,N-trimethylcyclohexanaminium) sulfate

EC Number:

807-085-0

Cas Number:

1004297-30-6

Molecular formula:

2 C9 H20 N .O 4S

IUPAC Name:

bis(N,N,N-trimethylcyclohexanaminium) sulfate

Details on test material:

- Name of test material (as cited in study report): Trimethylcyclohexylammonium sulfate
- Physical state: Solid / yellowish
- Analytical purity: 95.9 wt.%
- Lot/batch No.: VUONGH-00104

Test animals

Species:

other: in vitro

Strain:

other: in vitro

Test system

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

25 μL de-ionized water was applied first. Thereafter, a bulk volume of 25 μL (about 17 mg) of the solid test substance

Duration of treatment / exposure:

Corrosion test: 3 min and 1 h

Details on study design:

The EpiDermTM model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multi layered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDermTM tissues (surface 0.6 cm²) are cultured on specially prepared cell culture inserts (MILLICELLs, 10 mm ∅) and commercially available as kits (EpiDerm™ 200), containing 24 tissues on shipping agarose.
Corrosion test
From the day of arrival in the laboratory, tissues were kept in the refrigerator. At least 1 hour but not more than 1.5 hours before test-substance application, tissues were transferred to 6- well plates with 0.9 mL assay medium and preconditioned in the incubator at 37°C. The preincubation
medium was replaced with fresh medium immediately before application. Two tissues per exposure time (3 minutes at room temperature or 1 hour in the incubator, as a rule) and test group (test material, negative control and positive control; 12 tissues per test) were used.
25 μL de-ionized water was applied first. Thereafter, a bulk volume of 25 μL of the solid test material was applied with a sharp spoon and homogeneously distributed with the water.
Control tissues were concurrently treated with 50 μL of de-ionized water (NC) or with 50 μL of 8 N potassium hydroxide (PC) The tissues were washed with PBS to remove residual test material 3 minutes or 1 hour after start of the application treatment.
Rinsed tissues were kept in 24-well plates (holding plates) at room temperature on assay medium until all tissues per application time were dosed and rinsed. The assay medium was then replaced by MTT solution and tissues were incubated for 3 hours.
After incubation, the tissues were washed with PBS to stop the MTT-incubation. The formazan that was metabolically produced by the tissues was extracted by incubation of the tissues in isopropanol. The optical density at a wavelength of 570 nm (OD570) of the extracts was determined spectrophotometrically. Blank values were established of 4 microtiter wells filled with isopropanol for each microtiter plate.

Results and discussion

Any other information on results incl. tables

Viability of the test-substance treated tissues determined after an exposure period of 3 minutes was 105.4%. Viability of the test-substance treated tissues determined after an exposure period of 1 hour was 90.5%. Based on the observed results and applying the evaluation criteria cited in chapter 3.8 it was concluded, that Trimethylcyclohexylammonium sulfate does not show a corrosion potential in the EpiDerm™ skin corrosion test under the test conditions chosen.