Trimethylenediamine

Toxicity to reproduction

Administrative data

Endpoint:

two-generation reproductive toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

pre-GLP Trimethylenediamine is structurally very similar to ethylenediamine (difference: chain length / one CH2-group) or ethylenediammonium dichloride (doses of ammonium and chloride not toxicological relevant), respectively. Thus, the test result is justified for read across to assess the toxic potential to reproduction of trimethylenediamine.

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1984

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Ethylenediammmoniumdichloride

IUPAC Name:

Ethylenediammmoniumdichloride

Constituent 2

Reference substance name:

Ethylenediammonium dichloride

EC Number:

206-369-6

EC Name:

Ethylenediammonium dichloride

Cas Number:

333-18-6

IUPAC Name:

ethane-1,2-diaminium dichloride

Details on test material:

High purity Ethylenediamine (99,9% ) was used to synthesize ethylenediammoniumchloride (EDA*2HCl). Analysis by chromatographic and spectrophotometric procedures showed the salt to be free from impurities.

Test animals

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc., Portage, Michigan.
- Age at study initiation: (P) 43 wks;
- Weight at study initiation: (P) Males: 112-141 g; Females: 85-111 g;
- Housing: Two or three males or three or five females were housed per cage prior to mating;
- Diet : ad libitum, Purina Certified Rodent Chow 5002.
- Water :ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22
- Humidity (%): 55
- Photoperiod (hrs dark / hrs light): 12 hr light cycle

Administration / exposure

Route of administration:

oral: feed

Details on exposure:

Fresh diet was prepared biweekly with the percentage of EDA*2HCl in the diet adjusted to maintain a constant dosage level in grams per kilogram
for each sex according to the average animal body weight gain and diet consumption. At monthly intervals two diet samples were analysed.

Details on mating procedure:

The F0 parents were mated afier having received dietary EDA * 2HCl for 100 days. Twenty-six females and 13 males (randomly selected from 25 males) were mated per treated group while 52 females and 26 males (randomly selected from 50 males) were mated for the control group. During mating, two females were placed in a cage with one male; at this time all rats received EDA * 2HCl at the concentration in the diet for the respective female groups. Fifteen days after cohabitation, mating was discontinued and the females were placed in individual shoebox cages fitted with wire rod metal tops.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

At monthly intervals two diet samples were analysed.

Duration of treatment / exposure:

for two generations

Frequency of treatment:

daily

Details on study schedule:

At each dose level 13 male and 26 female rats were mated in both F0 and F1 generation (control group: 26 male and 52 female rats each); continuous treatment starting 100 days prior to cohabitation of F0 rats until weaning of F2 rats.

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
50 mg/kg bw/day
Basis:
actual ingested

Doses / concentrations 2

Remarks:

Doses / Concentrations:
150 mg/kg bw/day
Basis:
actual ingested

Doses / concentrations 3

Remarks:

Doses / Concentrations:
500 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

13 male and 26 female. Control group: 26 male and 52 female rats

Control animals:

yes, concurrent no treatment

Examinations

Parental animals: Observations and examinations:

BODY WEIGHT: Yes, measured weekly
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Food consumption determined on each cage of animals every 2 week. Dosage adjusted to keep a constant dosage level.

Results and discussion

Results: P0 (first parental generation)

Effect levels (P0)

1

Dose descriptor:

NOAEL

Effect level:

50 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Body weight; food consumption; organ weights;

2

Dose descriptor:

NOAEL

Effect level:

22.6 mg/kg bw/day

Based on:

test mat.

Remarks:

recalculated to EDA

Sex:

male/female

Basis for effect level:

other: Body weight; food consumption; organ weights; (Reclaculated to EDA base)

3

Dose descriptor:

NOAEL

Effect level:

27.9 mg/kg bw/day

Based on:

test mat.

Remarks:

calculated from EDA to trimethylenediamine

Sex:

male/female

Basis for effect level:

other: Study result generated with EDA*2HCl as test material was recalculated to EDA base and then - for read across - calculated for trimethylenediamine.

Results: F1 generation

Effect levels (F1)

1

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

500 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Body weight; food consumption; organ weights; histopathology: Hepatocellular pleomorphism

2

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

225.9 mg/kg bw/day

Based on:

test mat.

Remarks:

recalculated to EDA

Sex:

male/female

Basis for effect level:

other: Body weight; food consumption; organ weights; histopathology: Hepatocellular pleomorphism; (Recalculated to EDA base)

3

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

278.6 mg/kg bw/day

Based on:

test mat.

Remarks:

calculated from EDA to trimethylenediamine

Sex:

male/female

Basis for effect level:

other: Study result generated with EDA*2HCl as test material was recalculated to EDA base and then - for read across - calculated for trimethylenediamine.

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

Significant reduction in parental body weight gain of female rats in the intermediate and high dose group of the F0 generation, in the high dose group of the F1 generation and of male rats in the high dose group of both F0 and F1 generations. No substance-related parental deaths in the F0 or F1 generation. Significant decrease of absolute liver weight in male rats of the high dose F1 generation.

No macroscopic or histopathologic findings except a significant higher incidence of hepatocellular pleomorphism in both sexes of the high dose group of the F1 generation (6/10 male, 10/10 female; control: 0/20 each) and a significant decreased prevalence of kidney tubular mineralization in female rats of the high dose group of the F1 generation (0/10 female; control: 10/20).

In conclusion there was no evidence of fertility impairment or embryotoxic effect at dose levels that show maternal or paternal toxicity.

REPRODUCTIVE INDICES
	Dosage (g/kg,lday)
	0.50	0.15	0.05	0
	F0 → F1
Fertility index (%)
Male	92	100	92	92
Female	81	77	73	77
Gestation index (%)	100	100	100	100
Gestation survival index (%)	99.0 ± 3.4	100.0 ± 0.0	97.2 ± 11.8	99.2 ± 5.2
0- to 4-Day survival index (%)	90.0 ± 30.0	100.0 ± 0.0	94.4 ± 23.6	93.2 ± 22.4
4- to 14-day survival indexb (%)	99.4 ± 2.5	100.0 ± 0.0	94.1 ± 24.3	100.0 ± 0.0
4- to 2I-day survival indexb (%)	99.4 ± 2.5	100.0 ± 0.0	94.1 ± 24.3	99.7 ± 2.0
Pups born alive/litter	6.8 ± 3.5	8.6 ± 2.8	6.2 ± 3.9	8.4 ± 3.0
Days from first mating to parturition	28.3 ± 3.8	29.6 ± 2.0	29.8 ± 3.0	28.1 ± 3.0
	F1 → F2
Fertility index (%)
Male	100	92	92	100
Female	92	88	73	88
Gestation index (%)	100	100	100	100
Gestation survival index (%)	98.6 ± 6.7	99.1 ± 3.1	99.4 ± 2.5	99.5 ± 2.0
0- to 4-Day survival index (%)	98.0 ± 4.1	99.3 ± 3.5	98.6 ± 3.3	98.5 ± 4.1
4- to 14-day survival index (%)	100.0 ± 0.0	98.7 ± 6.3	100.0 ± 0.0	99.6 ± 3.0
4- to 21-day survival index (%)	100.0 ± 0.0	98.3 ± 8.3	100.0 ± 0.0	99.6 ± 3.0
Pups born alive/litter	8.5 ± 3.0	8.8 ± 2.2	9.4 ± 2.8	10.0 ± 2.5
Days from first mating to parturition	28.5 ± 3.2	28.8 ± 3.3	27.3 ± 3.6	28.4 ± 3.8

Conclusion: There was no indication of reproductive toxicity in Fischer 344 rats, following exposure to dietary EDA for two generations. A read across approach is justified as trimethylenediamine is structurally very similar to ethylenediamine (difference: chain length / one CH2-group) or ethylenediammonium dichloride (applied doses of ammonium and chloride of no toxicological relevance), respectively. Thus, there is no evidence that trimethylenediamine has reproductive potential.