N,N'-1,4-phenylenebis[3-oxobutyramide]

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well performed GLP study according to recent OECD TG.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Version / remarks:

24 April 2002

Deviations:

no

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-1,4-phenylenebis[3-oxobutyramide]

EC Number:

246-438-8

EC Name:

N,N'-1,4-phenylenebis[3-oxobutyramide]

Cas Number:

24731-73-5

Molecular formula:

C14H16N2O4

IUPAC Name:

3-oxo-N-[4-(3-oxobutanamido)phenyl]butanamide

Details on test material:

- Name of test material (as cited in study report): Diacetessig-p-phenylendiamid (DAEP)

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

Test system Mice, CBA/CaOlaHsd
Rationale: Recognised as the recommended test system
Source: Harlan Netherlands, B.V. Postbus 6174, NL - 5960 AD Horst / The Netherlands
Number of animals for the pre-test (non-GLP): 2 females
Number of animals for the main study: 16 females
Number of animals per group: 4 females (nulliparous and non-pregnant)
Number of test groups: 3
Number of control (vehicle) groups: 1
Age: 6 - 7 weeks (beginning of acclimatization)
Identification: Single caging. The animals will be distributed into the test groups at random and identified by cage number.
Acclimatisation: Under test conditions after health examination. Only animals without any visible signs of illness will be used for the study.

The animals were kept conventionally. The experiment was conducted under standard laboratory conditions.
Housing: single
Cage Type: Makrolon Type I, with wire mesh top (EHRET GmbH, D-79302 Emmendingen)
Bedding: granulated soft wood bedding (Harlan Winkelmann GmbH, D-33178 Borchen)
Feed: pelleted standard diet, ad libitum (Harlan Winkelmann GmbH, D-33178 Borchen)
Water: tap water, ad libitum (Gemeindewerke, D-64380 Rossdorf)
Environment: temperature 22 + 3°C
Relative humidity 30-70%
Artificial light 6.00 a.m. - 6.00 p.m.

Study design: in vivo (LLNA)

Vehicle:

dimethyl sulphoxide

Concentration:

Group 1 (vehicle control): 0%
Group 2: 5 % (w/v)
Group 3: 10 % (w/v)
Group 4: 20 % (w/v)

No. of animals per dose:

4

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:

EC3 = 18.2 % (w/v)

EC3 = Estimated concentration for a S.I. of 3.

Any other information on results incl. tables

Calculation and Results of Individual Data

Vehicle: DMSO

Test item concentration % (w/v)	Group	Measurement DPM	Calculation			Result
			DPM-BG^a)	number of lymph nodes	DPM per lymph node^b)	S.I.
---	BG I	18.80	---	---	---	---
---	BG II	101.60	---	---	---	---
---	CG 1	3924.46	3864.30	8	483.00
5	TG 2	4819.49	4759.30	8	594.90	1.23
10	TG 3	5621.87	5561.70	8	695.20	1.44
20	TG 4	5692.13	5631.90	8	704.00	1.46

 

BG = Background (1 ml 5% trichloroacetic acid) in duplicateCG = Control Group

TG = Test Group

S.I. = Stimulation Index

a)     = The mean value was taken from the figures BG I and BG II

b)     = Since the lymph nodes of the animals of a dose group were pooled, DPM/node was determined by dividing the measured value by the number of lymph nodes pooled

The EC3 Value could not be calculated, since all stimulation indices are below 3.

Viability / Mortality

No deaths occurred during the study period.

Clinical Signs

The animals did not show any clinical signs of toxicity after the first and second application. After the third application the highest dose (20%) induced redness of the ear skin of all 4 animals of the group. At the mid dose of 10% the ear skin of two animals was only slighly reddend.

Body Weights

The body weight of the animals, recorded prior to the first application and prior to treatment with ³HTdR, was within the range commonly recorded for animals of this strain and age.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

The test item Diacetessig-p-phenylendiamid (DAEP) was not a skin sensitiser under the described conditions.

Executive summary:

In the study the test item Diacetessig-p-phenylendiamid (DAEP) dissolved in DMSO was assessed for its possible contact allergenic potential.

For this purpose a local lymph node assay was performed using test item concentrations of 5, 10 and 20%.

The animals did not show any clinical signs of toxicity after the first and second application. After the third application the mid dose (10 %) induced slight redness of the ear skin of two animals of the group. At the high dose of 20 % the ear skin of all 4 animals was reddend after the third application. No cases of mortality were observed.

In this study Stimulation Indices (S.I.) of 1.23, 1.44 and 1.46 were determined with the test item at concentrations of 5, 10 and 20% (w/v) in DMSO, respectively.

The test item Diacetessig-p-phenylendiamid (DAEP) was not a skin sensitiser under the described conditions.