Ethyltris(2-hydroxyethyl)ammonium ethyl sulphate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

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:

equivalent or similar 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:

Ethyltris(2-hydroxyethyl)ammonium ethyl sulphate

EC Number:

250-796-0

EC Name:

Ethyltris(2-hydroxyethyl)ammonium ethyl sulphate

Cas Number:

31774-90-0

Molecular formula:

C6H15NO3 (TEA) C16H37NO7S (Triethoxy-TEA ethylsulfate)

IUPAC Name:

ethyltris(2-hydroxyethyl)azanium ethyl sulfate

impurity 1

Reference substance name:

2,2',2''-nitrilotriethanol

EC Number:

203-049-8

EC Name:

2,2',2''-nitrilotriethanol

Cas Number:

102-71-6

Molecular formula:

C6H15NO3

IUPAC Name:

2,2',2''-nitrilotriethanol

Test material form:

solid: bulk

Details on test material:

Name: Ethyltris(2-hydroxyethyl)ammonium ethyl sulphate
Product Description: Triethanolamine DES Quat
CAS No.: 31774-90-0
Physical state: white solid at 20 °C
Batch No.: PFS-755-175
Re-certification date of batch: 21 April 2018
Purity: 100 % (mono constituent substance, water content 0.33 % (w/w))
pH, 5% in water 7.85
Acid Value , mg KOH/g 38.45
Moisture, % 0.33
Total Amine, mg/g 33.10
Viscosity,cps, #4@60,25C 1580
Stability: stable under test conditions
Storage condition of test material: Room temperature, protected from light

Specific details on test material used for the study:

Name: Ethyltris(2-hydroxyethyl)ammonium ethyl sulphate
Product Description: Triethanolamine DES Quat
CAS No.: 31774-90-0
Physical state: colourless to yellow viscous liquid at 20 °C
Batch No.: PFS-755-175
Re-certification date of batch: 21 April 2018
Purity: 100 % (UVCB, water content 0.33 % (w/w))
pH, 5% in water 7.85
Acid Value , mg KOH/g 38.45
Moisture, % 0.33
Total Amine, mg/g 33.10
Viscosity,cps, #4@60,25C 1580
Stability: stable under test conditions
Storage condition of test material: Room temperature, protected from light

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

other: normal human epidermal keratinocytes (NHEK)

Cell source:

other: certificate of analysis of supplier available

Source strain:

not specified

Details on animal used as source of test system:

not applicable

Vehicle:

unchanged (no vehicle)

Details on test system:

The test item was applied undiluted. 30 µL (47 µL/cm2) of the test item was dispensed directly atop the EpiDerm tissue. The test item was gently spread to match size of the tissue using a bulb-headed Pasteur pipette.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

The test item was applied undiluted. 30 µL (47 µL/cm2) of the test item was dispensed directly atop the EpiDerm tissue. The test item was gently spread to match size of the tissue using a bulb-headed Pasteur pipette.

Duration of treatment / exposure:

In the present study the test item was applied topically to the EpiDerm tissue for 60 min followed by a 42 h post-incubation period and immediate determination of cytotoxic effects via MTT reduction assay.

Duration of post-treatment incubation (if applicable):

In the present study the test item was applied topically to the EpiDerm tissue for 60 min followed by a 42 h post-incubation period and immediate determination of cytotoxic effects via MTT reduction assay.

Number of replicates:

3 replicate tissues per group

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean relative tissue viability [%]

Value:

85

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Pre-Experiments

The mixture of 30 µL test item per 1 mL MTT medium showed no reduction of MTT compared to the solvent. The mixture did not turn blue/purple. Therefore, NSMTT equalled 0%. The mixture of 30 µL of the test item per 300 µL aqua dest. and per 300 µL isopropanol showed no colouring detectable by unaided eye-assessment. Therefore, NSC equalled 0%.

Results

Table 1: Result of the Test Item

Name	NC			PC			Test item
Tissue	1	2	3	1	2	3	1	2	3
absolute OD570	1.923 1.901	2.198 2.146	2.195 2.236	0.100 0.099	0.125 0.128	0.132 0.131	1.691 1.634	2.015 1.933	1.751 1.726
OD570 (blank-corrected)	1.882 1.860	2.156 2.195	2.154 2.195	0.059 0.058	0.084 0.087	0.091 0.090	1.650 1.593	1.974 1.892	1.710 1.685
mean OD570 of the duplicates (blank-corrected)	1.871	2.131	2.174	0.059	0.085	0.090	1.621	1.933	1.697
total mean OD570 of 3 replicate tissues (blank-corrected)	2.059*			0.078			1.751
relative tissue viability [%]	90.9	103.5	105.6	2.8	4.1	4.4	78.8	93.9	82.5
SD OD570	0.164			0.017			0.163
mean relative tissue viability [%]	100.0			3.8**			85.0
SD tissue viability [%]***	8.0			0.8			7.9
CV [% viabilities]	8.0			21.8			9.3

* Blank-corrected mean OD570 nm of the negative control corresponds to 100% absolute tissue viability.

** Mean relative tissue viability of the three positive control tissues is <= 20%.

*** Standard deviation (SD) obtained from the three concurrently tested tissues is ≤ 18%

Discussion

The potential of the test item to induce skin irritation was analysed by using the three-dimensional human epidermis model EpiDerm comprising a reconstructed epidermis with a functional stratum corneum. In the present study the test item was applied topically to the EpiDerm tissue for 60 min followed by a 42 h post-incubation period and immediate determination of cytotoxic effects via MTT reduction assay. Irritant potential of the test item was predicted from the relative mean tissue viabilities obtained compared to the corresponding negative control tissues concurrently treated with DPBS. The test item showed no non-specific reduction of MTT compared to the solvent. Therefore, NSMTT equalled 0%. The mixture of 30 µL of the test item per 300 µL aqua dest. and per 300 µL isopropanol showed no colouring detectable by unaided eye-assessment. Therefore, NSCliving equalled 0%.The test item showed no irritant effects. The mean relative tissue viability (% negative control) was > 50% (85.0%) after 60 min treatment and 42 h post-incubation. The controls confirmed the validity of the study. The mean absolute OD570 of the three negative control tissues was >= 0.8 and ≤ 2.8. The mean relative tissue viability (% negative control) of the positive control was <= 20% (3.8%). Standard deviation of viability of replicate tissues of all dose groups was ≤ 18% (0.8% - 8.0%).

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In this in vitro skin irritation 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”.

Executive summary:

In this in vitro skin irritation study (OECD 439) 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”.