Bis(3,5,5-trimethylhexanoyl) peroxide

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Remarks:

in vitro

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2012-03-20 to 2012-03-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Valid without restriction; 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:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Bis(3,5,5-trimethylhexanoyl)peroxide

IUPAC Name:

Bis(3,5,5-trimethylhexanoyl)peroxide

Constituent 2

Reference substance name:

Bis(3,5,5-trimethylhexanoyl) peroxide

EC Number:

223-356-0

EC Name:

Bis(3,5,5-trimethylhexanoyl) peroxide

Cas Number:

3851-87-4

Molecular formula:

C18H34O4

IUPAC Name:

3,5,5-trimethylhexanoyl 3,5,5-trimethylhexaneperoxoate

Details on test material:

- Name of test material (as cited in study report): Peroxan NPO; Bis(3,5,5-trimethylhexanoyl)peroxide
- Substance type: organic peroxide
- Physical state: liquid
- Analytical purity: 75 % (iodometric titration)
- Composition of test material, percentage of components: 75 % Peroxan NPO, 25 % Isododecane
- Lot/batch No.: 10618310
- Expiration date of the lot/batch: 2012-04-20
- Storage condition of test material: -8 to 0°C, protected from light

Test animals

Species:

other: reconstituted three-dimensional human skin model EpiDerm (MatTek)

Strain:

other: not applicable

Details on test animals or test system and environmental conditions:

Test system:
The test was carried out with the reconstituted three-dimensional human skin model EpiDermTM (MatTek). It consists of non-cancerous, human-derived epidermal keratinocytes (NHEK) which have been cultutred to form a multi-layered, highly differentiated model of the human epidermis. The NHEK are cultured are cultured on chemically modified, collagen-coated cell culture inserts (Millicell). The test systewm exhibits in vivo like morphological and growth characteristics and consists of organised basal, spinous, granular and cornified layers analogous to those found in vivo.

Test system

Type of coverage:

other: not applicable

Preparation of test site:

other: not applicable

Vehicle:

unchanged (no vehicle)

Controls:

other: EpiDerm Tissue was treated with distilled water and 8 N KOH

Amount / concentration applied:

The test item was applied undiluted. 50 µl of the test item were dispensed directly onto the EpiDerm tissue. The test item was spread to match size of the tissue.

Duration of treatment / exposure:

3 minutes and 60 minutes exposure

Observation period:

n.a.

Number of animals:

n.a.

Details on study design:

see "any other information on materials and methods incl. tables"

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 min

Value:

90

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 min

Value:

83

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

In vivo

Other effects:

n.a.

Any other information on results incl. tables

Table1: 3 min Exposure

Name	Negative Control		Test item		Positive control
Tissue	1	2	1	2	1	2
Absolute OD550-Werte	2.080	2.087	1.959	1.787	0.309	0.269
	2.082	2.088	1.962	1.784	0.315	0.271
	2.068	2.061	1.945	1.770	0.312	0.269
Mean OD550	2.077	2.079	1.955	1.780	0.312	0.270
SD	0.008	0.015	0.009	0.009	0.003	0.001
Total mean OD550(mean of two replicate tissues)	2.078*		1.868		0.291
Mean relative tissue viability [%]	100		90		14**
Mean inter tissue viability difference [%]***	0.1		9.4		14.5

* mean OD₅₅₀ ≥ 0.8

** mean relative tissue viability of the 3 min positive control ≤ 30 %

*** inter tissue viability difference ≤ 30 %

Table 2: 60 min Exposure

Name	Negative Control		Test item		Positive control
Tissue	1	2	1	2	1	2
Absolute OD550-Werte	1.955	1.763	1.815	1.454	0.114	0.119
	1.983	1.790	1.692	1.376	0.117	0.129
	2.000	1.782	1.706	1.361	0.115	0.121
Mean OD550	1.979	1.778	1.706	1.361	0.115	0.121
SD	0.023	0.014	0.067	0.050	0.002	0.005
Total mean OD550(mean of two replicate tissues)	1.879*		1.567		0.119
Mean relative tissue viability [%]	100		83		6
Mean inter tissue viability difference [%]***	10.7		21.7		6.4

* mean OD₅₅₀ ≥ 0.8

*** inter tissue viability difference ≤ 30 %

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

In this study the test item Peroxan NPO is not considered to be corrosive.

Executive summary:

The potential of the test item to induce skin corrosion was analysed by using the three dimensional human skin model EpiDerm, comprising a reconstructed epidermis with a functional stratum corneum. Peroxan NPO was applied topically to the EpiDerm tissue for 3 min and 60 min, respectively followed by immediate determination of cytotoxic effects via MTT reduction assay.

The test item showed no corrosive potential. The mean relative tissue viability (% negative control) was >= 50% (90%) after 3 min treatment and >= 15% (83%) after 60 min treatment.

The controls confirmed the validity of the study. The mean OD550 of the two negative control tissues was >= 0.8 for each exposure period. The mean relative tissue viability of the positive control was =< (14%) after 3 min treatment. The maximum inter tissue difference of replicate tissues of all dose groups was =< 30 % (0.1%-21.7%).