Reaction mass of 4-tert-butylphenol and 1,3- phenylenedimethanamine and 2-({[3-(aminomethyl) benzyl]amino}methyl)-4-tert-butylphenol

Toxicity to reproduction

Administrative data

Endpoint:

toxicity to reproduction

Remarks:

other: 90 d repeated dose toxicity study

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Follows GLP and OECD 408

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EPA OPPTS 870.3100 (90-Day Oral Toxicity in Rodents)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 4-tert-butylphenol and 1,3- phenylenedimethanamine and 2-({[3-(aminomethyl) benzyl]amino}methyl)-4-tert-butylphenol

EC Number:

939-071-6

Molecular formula:

(C10 H14 O . C8 H12 N2 . C H2 O)x

IUPAC Name:

Reaction mass of 4-tert-butylphenol and 1,3- phenylenedimethanamine and 2-({[3-(aminomethyl) benzyl]amino}methyl)-4-tert-butylphenol

Details on test material:

- Name of test material: Paraformaldehyde, oligomeric reaction products with 4-tert-butylphenol mphenylenebis(methylamine)

Test animals

Species:

rat

Strain:

other: F344/DUCrl

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (Kingston, New York)
- Age at study initiation: Approximately 7 weeks
- Housing: After assignment, animals were housed two per cage in stainless steel cages. Cages had solid floors with corncob bedding and shredded aspen for enrichment. Cages contained a feed crock and a pressure activated lixit valve-type watering system. The following environmental conditions were maintained in the animal room.
- Diet: Animals were provided LabDiet ad libitum.
- Water: ad libitum
- Acclimation period: Upon arrival, the animals were housed two to three per cage in stainless steel cages.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C with a range of 20°C-26°C
- Humidity (%): 50% with a range of 30-70%
- Air changes (per hr): 10-15 times/hour (average)
- Photoperiod (hrs dark / hrs light): 12-hour light/dark (on at 6:00 a.m. and off at 6:00 p.m.)

IN-LIFE DATES: From: November 5,2014 To: February 3 and 4,2015

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

DOSE SOLUTION PREPARATION
All dosing solutions were prepared by mixing the test material in propylene glycol (PG) at concentrations of 1.67, 5, or 16.7 mg/ml, and administered at a dose volume of 6 ml/kg body weight to achieve the targeted dose levels.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Mannich base PTBP-MXDA was determined to be soluble in PG at a concentration of 250 mg/ml
- Concentration in vehicle: 1.67, 5, or 16.7 mg/ml
- Lot/batch no. (if required): MKBS5987V.

Details on mating procedure:

animals were not mated

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

90 or 91 days

Frequency of treatment:

once daily seven days/week

Details on study schedule:

animals were not mated

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 10, 30, 100 mg/kg bw/d
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: A cage-side examination was conducted at least once a day, approximately at the same time each day (usually in the morning).


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Detailed clinical observations (DCO) were conducted on all animals pre-exposure and once per week throughout the study. The DCO was conducted on all animals, at approximately the same time each day according to an established format.

BODY WEIGHT: Yes
- Time schedule for examinations: All rats were weighed pre-exposure, twice during the first week, and weekly thereafter during the dosing period. Body weight gains were calculated relative to day 1.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

OTHER: more details in section "repeated dose toxicity"

Oestrous cyclicity (parental animals):

not examined

Sperm parameters (parental animals):

not examined

Litter observations:

n.a.

Postmortem examinations (offspring):

n.a.

Reproductive indices:

n.a.

Offspring viability indices:

n.a.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Other effects:

not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

not examined

Details on results (P0)

ORGAN WEIGHTS (PARENTAL ANIMALS)
Males given 100 mg/kg/day had higher mean relative testes and epididymides weights, that were statistically significant and interpreted to be reflective of the lower body weights of males at this dose level.
Females given 100 mg/kg/day had treatment-related statistically significant lower mean absolute and relative uterus weights, which corresponded to the histopathologic observation of very slight or slight diffuse atrophy of the uterus in 9/10 females at this dose level. Females given 100 mg/kg/day had lower absolute ovaries weights that were statistically significant and interpreted to be reflective of the lower body weights of females at this dose level.

GROSS PATHOLOGY (PARENTAL ANIMALS)
There were no treatment-related gross pathologic observations. All gross pathologic observations were considered to be spontaneous alterations, unassociated with oral gavage administration of Mannich base PTBP-MXDA.

HISTOPATHOLOGY (PARENTAL ANIMALS)
Nine females given 100 mg/kg/day had treatment-related very slight or slight diffuse atrophy of the cervix, uterus, and vagina. The atrophy of the uterus was characterized by diffuse decreased thickness of the endometrial mucosa, endometrial stroma, and tunica muscularis, and corresponded to the lower mean absolute and relative uterine weights at this dose level. The atrophy of the cervix and vagina was characterized by diffuse decreased thickness of the mucosa and fibromuscular wall of these tissues.

More details are given in section "repeated dose toxicity"

Effect levels (P0)

Dose descriptor:

NOAEL

Remarks:

fertility parameters

Effect level:

30 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: see 'Remark'

Results: F1 generation

General toxicity (F1)

Clinical signs:

not examined

Mortality / viability:

not examined

Body weight and weight changes:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Histopathological findings:

not examined

Overall reproductive toxicity

Reproductive effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

In this 90 d repeated dose toxicity study the follwing effects on reproductive organs were noted:
Females given 100 mg/kg/day had treatment-related statistically significant lower mean absolute and relative uterus weights, which corresponded to the histopathologic observation of very slight or slight diffuse atrophy of the uterus in 9/10 females at this dose level. 9/10 females given 100 mg/kg/day had treatment-related very slight or slight diffuse atrophy of the cervix, uterus, and vagina.
The NOAEL for fertility parameters is 30 mg/kg bw/d.