Registration Dossier

Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
October 20 - November 20, 1997 to January 30, 1998
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Justification for type of information:
Data is from secondary literature

Data source

Reference
Reference Type:
other: NTRL
Title:
SUPPORT: FINAL REPORT, DEVELOPMENTAL TOXICITY EVALUATION OF TERGITOL NP-4 SURFACTANT ADMINISTERED BY GAVAGE TO CD(SPRAGUE-DAWLEY) RATS, WITH COVER LETTER DATED 4/23/1999
Author:
Rochelle W. Tyl, Ph.D., DABT Melissa C. Marr, B.A., LATG Christina B. Myers, M.S.
Year:
1999
Bibliographic source:
UNION CARBIDE CORP, NTRL: OTS0559310-1, 1999

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Principles of method if other than guideline:
Developmental Toxicity Evaluation of Tergitol NP-4 in rats
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study report): 4-Nonylphenol, branched, ethoxylated
- Molecular formula : C17H28O2
- Molecular weight : 264.409 g/mol
- Substance type:Organic
- Physical state: Liquid
Specific details on test material used for the study:
- Name of test material (as cited in study report): Tergitol NP-4
- Molecular formula (if other than submission substance): C23H40O5
- Molecular weight (if other than submission substance): 396.63 g/mol
- Substance type: Organic
- Physical state: Liquid
- Impurities (identity and concentrations): polyethylene glycol, <3%,dinonylphenol ethoxylate, <3%,

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
CO
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Inc., Raleigh, NC.
- Age at study initiation:
Female rats= 56 days
Male rats= 70 days
- Weight at study initiation: No data available
- Fasting period before study: No data available
- Housing: Males were housed singly in solid-bottom polycarbonate cages (8" x 19" x 10.5") with stainless steel wire lids. Non mated females were group housed (maximum three per cage), and mated females were singly housed in solid-bottom polycarbonate cages (8" x 19" x 10.5") with stainless steel wire lids (Laboratory Products, Rochelle Park, NJ). Sani-Chips® animal bedding (P. J. Murphy Forest Products, Montville, NJ) was used in all cages.
- Diet (e.g. ad libitum): Pelleted feed (No. 5002 Purina Certified Rodent Diet®; PMI Feeds, Inc., St. Louis, MO). ad libitum
- Water (e.g. ad libitum): tap water (Durham, NC water system, in plastic bottles with stainless steel sipper tubes) available ad libitum
- Acclimation period: 7 days quarantine periods for the females

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
target temp: 64-79°F (17.8 - 26.1°C)
maintained temp: 70.7-73.2°F (21.5 - 22.9 °C)

- Humidity (%):
target humidity: 30-70%
maintained humidity 35.7 - 63.8%

- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): 12:12 hour light: dark cycle

IN-LIFE DATES:
From: Dosing: November 3-15,1997 (gestation day 6 through 15)
To: Necropsy: November 17-20,1997 (gestation day 20)

Administration / exposure

Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The Tergitol NP-4 Surfactant for each dose was weighed out into a calibrated beaker (400 ml beaker to hold 200 ml, with adjustments for different volumes if needed). The appropriate volume of corn oil was added to each beaker, stirred with a stirring bar until the chemical was no longer visible, and then stirred for ten minutes more to give a dose range of 0, 50, 250 or 500 mg/Kg/day. The resulting formulation was sampled for archive and analytical samples and stored at 1-8°C in sealed, amber glass bottles protected from light. After storage and prior to use, the solutions were allowed to warm to room temperature and then stirred for ten minutes with a stirring bar. The formulations were then used for dosing.

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

VEHICLE
- Justification for use and choice of vehicle (if other than water): Corn oil
- Concentration in vehicle: 0, 50, 250 or 500 mg/Kg/day
- Amount of vehicle (if gavage): 1.0 ml/kg
- Lot/batch no. (if required): 325A7
- Purity: No data
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Prior to study dose formulation, formulations encompassing the range of dose employed (50 and 600 mg/mL) were assayed for homogeneity, stability, and dose level verification
Details on mating procedure:
- M/F ratio per cage: 1:1 (one male and one female)
- Length of cohabitation: No data available
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy Each morning the females were examined for the presence of vaginal sperm and/or copulation plug.
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility. No data available
- Further matings after two unsuccessful attempts: [no / yes (explain)] No data available
- After successful mating each pregnant female was caged (how): Sperm-positive females were individually housed until scheduled sacrifice on gestation day 20.
- Any other deviations from standard protocol: No data available
Duration of treatment / exposure:
10 consecutive days
Frequency of treatment:
Daily
Duration of test:
Gestation day 6 through gestation day 15
Doses / concentrations
Remarks:
0, 50, 250 and 500 mg/kg/day
No. of animals per sex per dose:
Total: 100
0 mg/kg/day: 25 females
50 mg/kg/day: 25 females
250 mg/kg/day: 25 females
500 mg/kg/day: 25 females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Doses was based on a range-finding study in pregnant rats performed at RTI. In range finding study, four group of eight sperm positive females were dosed once daily on gestation day 6 through 15 at concentartion 0, 500, 1500, and 3000 mg/kg/day at a dosing volume of 5.0 ml/kg. At 1500 and 3000 mg/kg/day, at least 50% of the dams died. At 500 mg/kg/day, all dams survived and were pregnant; seven of eight dams were pregnant at 0 mg/kg/day. Therefore, the highest dose level for the present study, 500 mg/kg/day, was chosen to induce overt maternal and developmental toxicity.

- Rationale for animal assignment (if not random): Sperm-positive female rats (dams) were assigned to treatment groups by a stratified randomization method, designed to provide uniform mean body weights across dose groups on gd 0 at the initiation ofthe study.
- Rationale for selecting satellite groups: No data available
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): No data available

Examinations

Maternal examinations:
Mortality, Clinical sign, Body weight and Food consumption and Gross pathology were examined.
Ovaries and uterine content:
Number of ovarian corpora lutea, total number of uterine implantation sites, on preimplantation loss, or on the incidence of resorptions were examined.
Fetal examinations:
number of live fetuses and fetal body weight, Gross pathology and Histopathology were examined.
Statistics:
No data available

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, treatment-related
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, non-treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
Mortality: One female rat were died at 500 mg/kg bw on day 6 as compared to control.

Clinical sign: Rust colored fur on the head, face, and back, a nonspecific indicator of stress (from hemoglobin breakdown products in the Harderian gland excreted via the tear ducts, so-called "bloody tears" or chromodacryorrhea, groomed from the eyes and nose) on gd 13 and 15, Rough coat (more severe than piloerection, indicative of more stress) on gd 12-15, Piloerection, another nonspecific sign of stress on gd 6 through 20, Lethargy on gd 6-15, Pica gd 6 and 7 and Prone positioning on gd 6-14 was observed at 500 mg/kg/day, and Piloerection, another nonspecific sign of stress on gd 6 through 20 and Pica on gd 6 and 8, at 250 mg/kg/day on gd 7-10 and 1315, and at 50 mg/kg/day on gd 14 in one dam were observed as compared to control.

Body weight:
Decreased in body weight were observed in ten dams at 250 mg/kg/day and 12 dams at 500 mg/kg/day on gd 9 (first scheduled weigh time point during dosing), in two dams at 500 mg/kg/day on gd 12 (next scheduled weigh timepoint), and in two dams at 500 mg/kg/day on gd 15 (next scheduled weigh time point), all during the dosing period as compared to control.

Food consumption: Statistically significantly decreased on at 250 and 500 mg/kg/day for gd 6-9, 9-12,12-15,6-1 and increased at 500 mg/kg/day for gd 18-20 and 15-20 (postdosing period), at 250 mg/kg/day for gd 18-20 and 15-20, and at 50 mg/kgfday for gd 18-20 and 15-20 as compared to control.

Organ weights:
Significantly increased absolute and relative liver weights were observed in 50 and 250 and 500 mg/kg/day and no effect were observed on gravid uterine weight of treated rats as compared to control.

Gross pathology: Alopecia (various areas), were observed unrelated to treatment and blood present in urogenital opening, urinary bladder, mouth and nose, and lung lobes hemorrhagic and edematous were observed in unscheduled deaths one at 500 mg/kg/day and area of extra hard tissue on left median liver lobe at 50 mg/kg/day in one female rats were observed as compared to control.

Maternal developmental toxicity

Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not specified
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified
Changes in number of pregnant:
not specified
Other effects:
not specified
Details on maternal toxic effects:
Reproductive function: estrous cycle: No significant effect were observed on number of ovarian corpora lutea, total number of uterine implantation sites, on preimplantation loss, or on the incidence of resorptions and dead fetuses per litter, of nonlive implants (resorbed plus dead) per litter, or of adversely affected implants (nonlive plus malformed) per litter were observed as compated to control.

Reproductive performance: No effect on number of live fetuses and sex ratio (% males) per litter were observed as compared to control.

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
50 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
organ weights and organ / body weight ratios
gross pathology
number of abortions
pre and post implantation loss
total litter losses by resorption
early or late resorptions
dead fetuses
other: No effect

Maternal abnormalities

Abnormalities:
not specified
Localisation:
not specified

Results (fetuses)

Fetal body weight changes:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified
Reduction in number of live offspring:
not specified
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Skeletal malformations:
effects observed, treatment-related
Visceral malformations:
no effects observed
Other effects:
not specified
Details on embryotoxic / teratogenic effects:
Mortality: No effects on number of live fetuses were observed as compared to control.

Body weight: No effect on fetal body weight was observed as compared to control.

Gross pathology: No effects on external malformations were observed as compared to control.

Skeletal malformations: Statistically significant skeletal variations such as cleft palate bilateral and unilateral rudimentary ribs on Lumbar I was observed in all fetuses or for male and female fetuses at 250 and 500 mg/kg/day as compared to control.

Visceral malformation: No effects on visceral malformations were observed as compared to control.

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
50 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
changes in sex ratio
fetal/pup body weight changes
changes in litter size and weights
external malformations
skeletal malformations
visceral malformations
other: No effect

Fetal abnormalities

Abnormalities:
not specified
Localisation:
other: not specified

Overall developmental toxicity

Developmental effects observed:
not specified
Treatment related:
not specified
Relation to maternal toxicity:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 50 mg/kg/day for P and F1 generation when Sprague-Dawley female rat by using Tergitol NP-4 orally by gavage from gd day 6 to 15.
Executive summary:

In a developmental toxicity study, Sprague-Dawley female rat by using Tergitol NP-4 in the concentration of 0, 50, 250 and 500 mg/kg/day in corn oil orally by gavage. One female rat were died at 500 mg/kg bw on day 6. Rust colored fur on the head, face, and back, a nonspecific indicator of stress (from hemoglobin breakdown products in the Harderian gland excreted via the tear ducts, so-called "bloody tears" or chromodacryorrhea, groomed from the eyes and nose) on gd 13 and 15, Rough coat (more severe than piloerection, indicative of more stress) on gd 12-15, Piloerection, another nonspecific sign of stress on gd 6 through 20, Lethargy on gd 6-15, Pica gd 6 and 7 and Prone positioning on gd 6-14 was observed at 500 mg/kg/day, and Piloerection, another nonspecific sign of stress on gd 6 through 20 and Pica on gd 6 and 8, at 250 mg/kg/day on gd 7-10 and 1315, and at 50 mglkg/day on gd 14 in one dam were observed. Decreased in body weight were observed in ten dams at 250 mg/kg/day and 12 dams at 500 mg/kg/day on gd 9 (first scheduled weigh time point during dosing), in two dams at 500 mg/kg/day on gd 12 (next scheduled weigh timepoint), and in two dams at 500 mg/kg/day on gd 15 (next scheduled weigh time point), all during the dosing period. Statistically significantly decreased in food consumption were observed at 250 and 500 mg/kg/day for gd 6-9, 9-12,12-15,6-1 and increased at 500 mg/kg/day for gd 18-20 and 15-20 (postdosing period), at 250 mg/kg/day for gd 18-20 and 15-20, and at 50 mg/kgfday for gd 18-20 and 15-20. No abortion or early delivery and no effect on numbers of litters (and fetuses) were observed in treated rats as compared to control. Significantly increased absolute and relative liver weights were observed in 50 and 250 and 500 mg/kg/day and no effect were observed on gravid uterine weight of treated rats as compared to control. Alopecia (various areas), were observed unrelated to treatment and blood present in urogenital opening, urinary bladder, mouth and nose, and lung lobes hemorrhagic and edematous were observed in unscheduled deaths one at 500 mg/kg/day and area of extra hard tissue on left median liver lobe at 50 mg/kg/day in one female rats were observed. In addition, no significant effect were observed on number of ovarian corpora lutea, total number of uterine implantation sites, on preimplantation loss, or on the incidence of resorptions and dead fetuses per litter, of nonlive implants (resorbed plus dead) per litter, or of adversely affected implants (nonlive plus malformed) per litter. No developmental effect were observed such as number of live fetuses, sex ratio (% males) per litter, and the average fetal body weight per litter (all fetuses or separately by sex) were also unaffected by treatment. There was no dose effect or dose x sex interaction for fetal body weight parameters were observed in treated female rats as compared to control. No effects on external and visceral malformations were observed in treated rats as compared to control. Statistically significant skeletal variations such as cleft palate bilateral and unilateral rudimentary ribs on Lumbar I was observed in all fetuses or for male and female fetuses at 250 and 500 mg/kg/day as compared to control. Therefore, NOAEL was considered to be 50 mg/kg/day for P and F1 generation when Sprague-Dawley female rat by using Tergitol NP-4 orally by gavage from gd day 6 to 15.