2-nitrophenyl acetate

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)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

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)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: MatTek Protocol for: INVITRO EpiDermTM SKIN IRRITATION TEST For use with MatTek Corporation’s Reconstructed Human Epidermal Model EpiDerm, Model EPI- 200-SIT, Rev. 26/3/2012,1-37

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Acetic acid, 2-nitrophenyl ester

IUPAC Name:

Acetic acid, 2-nitrophenyl ester

Constituent 2

Reference substance name:

2-nitrophenyl acetate

EC Number:

210-233-1

EC Name:

2-nitrophenyl acetate

Cas Number:

610-69-5

Molecular formula:

C8H7NO4

IUPAC Name:

2-nitrophenyl acetate

Test material form:

solid: crystalline

Details on test material:

- Name of test material (as cited in study report): 2-Nitrophenyl Acetate

- Physical state: solid light yellow
- Analytical purity: 95 % w/w

- Impurities (identity and concentrations):
Phenol <5.0 % (w/w)
2,4-dinitrophenol <5.0 % (w/w)
2,6-dinitrophenol <5.0 % (w/w)
Acetic acid <1.0 % (w/w)
Water <5.0 % (w/w)
2-nitrophenol <5.0 % (w/w)

- Lot/batch No.: 201408
- Expiration date of the lot/batch: unlisted

- Storage condition of test material: The test substance will be stored in dry room in dark in
closed container at the room temperature

Test animals

Species:

other: reconstructed human epidermal model EpiDerm™ (EPI-200 ver. 2.0, MatTek, Ashland, USA); Lot No. 21646, kit A

Test system

Type of coverage:

occlusive

Preparation of test site:

not specified

Vehicle:

not specified

Duration of treatment / exposure:

60 minutes

Results and discussion

Any other information on results incl. tables

OD570 measuring was performed after 2-hour extraction with shaking. Results are given in the Table 1. Extracts as well as tissues after extraction are given in Figure 2.

Table 1: OD570 values obtained at the MTT test, their means, standard deviations and relative viabilities (%)

	Treatment	OD570	Mean	SD	Relative viability
	1	2	3	(% NC)
NC	PBS	1.874	1.976	2.015	1.955	0.059	100.0
	viability (% NC)	95.86	101.07	103.07	100.00	3.041
C2	66/15	2.309	2.010	2.020	2.113	0.139	108.1
	viability (% NC)	118.11	102.81	103.32	108.08	7.092
PC	5% SDS	0.049	0.045	0.045	0.046	0.002	2.4
	viability (% NC)	2.51	2.30	2.30	2.37	0.096

NC = negative control

PC = positive control

C2 = test substance

mean = arithmetic mean

SD = standard deviation calculated from individual % tissue viabilities

viability (%) = viability of single tissues compared with negative control, mean NC viability is defined as 100

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The effect of the test substance was negative in EpiDermTM model.

Executive summary:

Test substance, 2-Nitrophenyl Acetate, was assayed for the in vitro skin irritation in human epidermal model EpiDermTM. The test was performed according to the Method B.46. In vitro skin irritation: Reconstructed human epidermis model test and Protocol for: In Vitro EpiDermTM Skin Irritation Test For use with MatTek Corporation’s Reconstructed Human Epidermal Model EPI-200-SIT (see par. 1.4, (1), (3)).

After pre-incubation of tissues, 25 mg of the test was placed directly atop to the previously moistened tissue so it covered all tissue surfaces. Length of exposition was 60 minutes. Three tissues were used for every concentration and controls.

After removal of the test substance from tissues, tissues were post incubated for 42 hours due to leave of damage reparation. Three hours incubation with MTT and two hours extraction period with shaking followed then. Optical density (OD570) of isopropyl alcohol extracts was measured on a spectrophotometer. Relative cell viability was calculated for each tissue as % of the mean viability of the negative control tissues.

Under the above-described experimental design, average viability of tissues treated by the test substance was 108.1 %, i.e.viability was > 50 %.

The effect of the test substance was negative in EpiDermTM model (the tissue was not damaged).

According to the classification criteria given in chapter 3.9., the test substance is considered to have no category in regard to skin irritation.