N,N'-diphenyl-p-phenylenediamine

Specific investigations: other studies

Administrative data

Endpoint:

specific investigations: other studies

Remarks:

embryotoxicity

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference 1

Reference Type:

secondary source

Title:

Embryotoxicity of Sixteen Industrial Amines to the Chicken Embryo

Author:

Korhonen A. et al.

Year:

1983

Bibliographic source:

J. Appl. Toxicol., 3, 112-117, (1983)

Reference 2

Reference Type:

publication

Title:

Toxicity of rubber chemicals towards three-day chicken embryos

Author:

Korhonen A. et al.

Year:

1983

Bibliographic source:

Scand. j. work. environ. health, 9,115-119, (1983)

Materials and methods

Principles of method if other than guideline:

A method utilizing chicken embryos is employed to allow observation of each exposed embryo throughout the whole period of embryonic development. The advantage of continuous observation is that dead embryos can be readily recorded and inspected before maceration.

GLP compliance:

not specified

Type of method:

other: in ovo

Endpoint addressed:

developmental toxicity / teratogenicity

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-diphenyl-p-phenylenediamine

EC Number:

200-806-4

EC Name:

N,N'-diphenyl-p-phenylenediamine

Cas Number:

74-31-7

Molecular formula:

C18H16N2

IUPAC Name:

N,N'-diphenyl-p-phenylenediamine

Test material form:

solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source of test material: Oy Nokia Ab, Nokia
- Purity test date: technical grade

Test animals

Species:

other: chicken

Strain:

other: White Leghorn chicken egg

Details on test animals or test system and environmental conditions:

- Source: hatchery of Siipikarjanhoitajain Liito ry, Hämeenlinna, Finland

Administration / exposure

Route of administration:

other: injection

Vehicle:

acetone

Details on exposure:

VEHICLE
- Purity: p.a. grade, from Merck

Doses / concentrations

Dose / conc.:

3 other: µmol per egg

Remarks:

Equals highest feasible dose based on solubility or suspendibility in 5 µL of acetone

Control animals:

yes, concurrent vehicle

Details on study design:

CONTROLS
- The solution containing test compound was injected into the egg in a total volume of 5 µL. Ten control eggs, injected with 5 µL of acetone, were incubated with each batch of eggs. Results from solvent controls were not subtracted from experimental values. The background level of 1-2% was insignificant against the variation of the method.

EXPERIMENTAL DETAILS
- Three-day (72-76 hours) embryos were selected by candling.
- The test compound is injected on the inner shell membrane, focusing the solution drop exactly on the embryo.
- Two days after the injection, the eggs were candled again. Eggs containing dead embryos were counted and discarded.
- The remaining eggs were then candled every second or third day. Those containing dead embryos were opened and the embryos checked for external malformations and the developmental stage.
- The incubation was terminated 11 days after the injection, after a total incubation of 14 days. The remaining eggs were opened and the embryos inspected for survival and for external malformations.
- The temperature was kept at 37.7 “C and the humidity between 66% and 71% throughout the incubation period. The eggs were turned two to four times per day.

CLASSIFICATION
The affected embryos were classified into the following categories:
- Early deaths, embryos that died before day 5 of the incubation, within two days of the treatment.
- Late deaths, non-malformed, externally normal embryos that died between days 5 and 14.
- Late deaths, malformed, externally malformed embryos that died between days 5 and 14.
- Malformed survivors, externally malformed embryos alive on day 14 of the incubation.
LD50 and ED50 values were calculated using a Wang table computer.

Results and discussion

Details on results:

The test compound dit not produce effects above the vehicle background.