Registration Dossier

Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
other information
Study period:
191-04-23 to 1992-04-09
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report Date:
1993

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
; adopted 1981-05-12
Deviations:
yes
Remarks:
; exposure to test item from day 6 to 15 of pregnancy
Qualifier:
according to
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
; exposure to test item from day 6 to 15 of pregnancy
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): o-Chloraniline
- Physical state: yellow-orange, clear liquid
- Analytical purity: 99.6 % (IR spectroscopy)
- Impurities (identity and concentrations):
<0.01 % cyclohexylamine
<0.01 % N-isopropylaniline
<0.01 % 2-chloro-N-Isopropylaniline
0.08% aniline
<0.01% 2-Isopropylaniline
<0.01% o-nitrochlorobenzene
0.24% p-chloroaniline
<0.01% m-chloroaniline
<0.01% 2,4-dichloroaniline
<0.01% 2,5-dichloroaniline
<0.01% 2,3-dichloroaniline
<0.01% 2,3-dichloroaniline
<0.01% o-phenylendiamine
0.03 % unknown impurities
- Purity test date: 24 Apr 1991
- Lot/batch No.: Lagerkessel 15/ 594 488
- Stability under test conditions: stable and homogen throughout the test period
- Storage condition of test material: 3-7°C, dark

Test animals

Species:
rat
Strain:
other: Bor:WISW (SPF Cpb)
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Winkelmann, Borchen
- Age at study initiation: 2-3 months
- Weight at study initiation: male >300 g; nulliparous female 186-245 g
- Fasting period before study:
- Housing: groups of females per macrolon III cage before pairing and individual housing in macrolon cages II from day 0 p.c. on, male rats were housed individually in macrolon II cages, cages were changed three times per week, animals and cages marked individually
- Bedding: wood chips, Ssniff
- Diet : ad libitum; Altromin 1324
- Water: ad libitum; tap water
- Acclimation period: 7 d


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 33-90
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

0 mg/mL
2 mg/mL
10 mg/mL
50 mg/mL

Dosing solutions were tested for homgenicity and stability and kept in the dark at RT.


VEHICLE
- Justification for use and choice of vehicle (if other than water): polyethylene glycol enhances solubility and bioavailability of the test substance
- Amount of vehicle (if gavage): 5 mL/kg bw

- Source: BASF
Analytical verification of doses or concentrations:
yes
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/2 (not mor than 2 matings per male)
- Length of cohabitation: overnight
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
6 - 15 d
Frequency of treatment:
daily
Duration of test:
day 0 to day 20 of pregnancy
Doses / concentrations
Remarks:
Doses / Concentrations:
10, 50, 250 mg/kg body weight (in 5 ml/kg Polyethylen glycol)
Basis:
actual ingested
No. of animals per sex per dose:
control group and dose groups 10 mg/kg and 50 mg/kg: 25 dams
250 mg/kg dose group: 28 dams
Control animals:
yes, concurrent vehicle
Details on study design:
Sex: female
Duration of test: until 20 d

Examinations

Maternal examinations:
food consumption, body weight, spleen weight and histopathology, clinical signs of toxicity
Ovaries and uterine content:
number of corpora lutea, implantation sites, weight of uteri, weight and appearance of placenta, number of alive and dead fetuses, sex of alive fetuses
Fetal examinations:
visible morphological changes, visceral changes, (according to Wilson and Dawson)
Statistics:
rank sum test (Wilcoxon): body weight, number of fetuses with malformations per dam
Fisher-test: fertility and gravity rate
CHi square for: number of fetuses with different malformations, number of early and late resorptions, number of different sexes, preimplantation loss

F-test and t-test for the other parameters
Historical control data:
yes

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
no mortality; at 250 mg/kg: tremor following application, body weight gain and food consumption reduced; at 50 and 250 mg/kg: absolute spleen weight increased, increased hyperaemia, iron content and extramedullary haematopoesis;

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
10 mg/kg bw/day (actual dose received)
Basis for effect level:
other: maternal toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
teratogenic effects:
at 10, 50 mg/kg: no significant effects;                    
at 250 mg/kg: significant increase of late resorptions, number of viable pups/litter significantly decreased;                    
embryotoxic effects:
at 50, 250 mg/kg: necrotic placenta lining;no effects on development of skeletal system; 
at 250 mg/kg: increased number of spontaneous malformations.

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
50 mg/kg bw/day (actual dose received)
Basis for effect level:
other: teratogenicity

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Any other information on results incl. tables

no further data

Applicant's summary and conclusion

Executive summary:

Bartmann, K and Holzum, B (1993)

2 -Chloroaniline was tested by oral gavage for developmental toxicity study in rats according to OECD 414. the dams were treated daily with either 0, 10, 50 or 250 mg 2-chloroaniline in polyethylene glycol / kg body weight from day 6 to 15 of pregnancy. One day prior to delivery the uteri were removed and the fetuses as well as the uteri examinated. No mortality was observed in the dams. the 250 mg/ kg dose group displayed signs of toxicity like tremor, reduced food intake and reduced body weight gain. In the 50 and 250 mg/kg dose group the spleen weights were increased, most probably due to increased hematopoesis induced by methemoglobin formation. Beginning necrosis was more evident in these dose groups as well, probably caused by reduced oxygen supply. In the highest dose group the number of late resorptions and the number of spontaneous malformations increased and the number of fetuses decreased. Evidence for a specific teratogenic effect of 2-chloroaniline is lacking. The observed embryotoxicity

is in line with the observed maternal toxcicity. The maternal NOAEL was 10 mg 2-chloroaniline per kg body weight per day the fetal NOAEL was 50 mg 2-chloroaniline per kg body weight per day.