Phenol, paraalkylation products with C10-15 branched olefins ( C12 rich) derived from propene oligomerization, calcium salts, sulfurized, including distillates (petroleum), hydrotreated, solvent-refined, solvent-dewaxed, or catalyc dewaxed, light or heavy paraffinic C15-C50

Toxicity to reproduction

Administrative data

Endpoint:

one-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study following GLP.

Justification for type of information:

This is the key study to determine the reproductive toxicity of alkyl phenate sulfides without residual EC 310-154-3.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]

Version / remarks:

OECD 415, One-Generation Reproduction Toxicity Study, 26 May 1983

Deviations:

yes

Remarks:

These deviations did not negatively impact the quality or integrity of the data nor the outcome of the study.

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Phenol, paraalkylation products with C10-15 branched olefins ( C12 rich) derived from propene oligomerization, calcium salts, sulfurized, including distillates (petroleum), hydrotreated, solvent-refined, solvent-dewaxed, or catalyc dewaxed, light or heavy paraffinic C15-C50

EC Number:

701-249-4

Molecular formula:

A molecular formula for this substance does not exist because it is a UVCB. The molecular formula for a theoretical representative structure is C36H58Ca2O4Sx where x = 1-3.

IUPAC Name:

Phenol, paraalkylation products with C10-15 branched olefins ( C12 rich) derived from propene oligomerization, calcium salts, sulfurized, including distillates (petroleum), hydrotreated, solvent-refined, solvent-dewaxed, or catalyc dewaxed, light or heavy paraffinic C15-C50

Details on test material:

Phenol, dodecyl-, sulfurized, calcium salts
Testing was performed on a commercial sample of this material. Typical purity of this material as distributed in commerce is 60% alkyl phenol sulfide and 40% highly refined lubricant base oil.

Specific details on test material used for the study:

The test material was a calcium alkyl phenate sulfide purifed of residual EC 310-154-3. This material did not have carbonation in order to ensure the most bioavailble molecule was tested. See the read across justification in section 13 for more information.

Test animals

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Diet: conventional laboratory diet provided ad libitum.
- Water: Reverse osmosis-purified drinking water provided ad libitum.
- 6 weeks old at the initiation of test item administration

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Remarks:

corn oil (lot nos. ZT1301 and 2AE0415, exp. dates: 1 July 2012 and 4 May 2013, respectively, received from Spectrum Chemical Manufacturing Corporation, New Brunswick, NJ).

Details on exposure:

Both sexes of the parental generation were treated with concentrations of 0 (corn oil vehicle), 250, 500 or 1000 mg/kg/day by oral gavage for approximately 70 consecutive days prior to mating. Males were dosed daily until euthanasia, 84-85 doses. Female rats were dosed through mating, gestation, lactation and until euthanasia, 127-128 doses.

Details on mating procedure:

Vaginal lavages were performed daily and the slides were evaluated microscopically to determine the stage of estrus of each F0 female for 21 days prior to cohabitation and continuing until evidence of mating was observed or until the end of the mating period.

The F0 animals were paired on a 1:1 basis within each treatment group after a minimum of 70 days of treatment. All animals were randomly selected for cohabitation, avoiding sibling matings. A breeding record containing the male and female identification numbers and the start date of cohabitation was maintained. Each female was housed in the home cage of the male. Positive evidence of mating was confirmed by the presence of a vaginal copulatory plug or the presence of sperm in a vaginal lavage and verified by a second biologist.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analyzed dosing formulations were within WIL Research’s SOP range for suspensions (85% to 115%), were homogeneous. Based on these results, the protocolspecified dosages of test item were administered to the animals. The test item was not detected in the vehicle formulation that was administered to the control group (Group 1).

Duration of treatment / exposure:

Males were dosed daily until euthanasia, 84-85 doses. Female rats were dosed through mating, gestation, lactation and until euthanasia, 127-128 doses.

Frequency of treatment:

daily, 7 days each week

Details on study schedule:

See other sections

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 250, 500 or 1000 mg/kg/day
Basis:
other: analytically measured values

No. of animals per sex per dose:

30 Males and 30 Females per dose group
(4) dose groups total (including vehicle control)

Control animals:

yes, concurrent vehicle

Details on study design:

Estrous cyclicity was evaluated 21 days prior to mating. Developmental landmarks (balanopreputial separation and vaginal patency) were evaluated for the F1 rats selected to constitute the F1 generation. Nonselected F1 pups were necropsied on PND 21. Selected F1 pups were necropsied on PND 60.

Positive control:

Not applicable

Examinations

Parental animals: Observations and examinations:

Mean body weights and body weight gains in the 250, 500, and 1000 mg/kg/day groups were generally similar to the control group throughout the entire generation (males) and throughout pre-mating, gestation, and lactation (females). Test item-related, higher mean food consumption was noted throughout the entire treatment period in the 500 and 1000 mg/kg/day group males and sporadically during the pre-mating treatment period in the 1000 mg/kg/day group females. The increased food consumption noted in these groups was considered non-adverse because it did not affect mean body weights or body weight gains. Mean food consumption in the 250 mg/kg/day group males throughout the entire generation, in the 250 and 500 mg/kg/day group females during the pre-mating period, and in the 250, 500, and 1000 mg/kg/day group females during gestation and lactation was unaffected by test item administration.

Increased occurrences of red material around the nose and/or red, yellow and/or clear material around the mouth were noted in the 500 mg/kg/day group F0 males and the 1000 mg/kg/day group F0 males and females intermittently throughout the treatment period at approximately 1 hour following dose administration; these findings were more frequently noted in males than in females. These material findings were attributed to the test item. There were no test item-related clinical findings noted at the detailed physical examinations for the F0 generation.

Oestrous cyclicity (parental animals):

Estrous cyclicity was evaluated 21 days prior to mating.

Sperm parameters (parental animals):

Hypospermia

Litter observations:

Pup clinical observations, body weights, and body weight gains prior to weaning were unaffected by parental test item administration. No internal findings attributed to parental administration of the test item were noted at the necropsies of pups that were found dead or at the PND 21 necropsy.

There were no test item-related effects on survival, clinical signs, body weights, body weight gains, or food consumption for F1 males and females following weaning at any dosage level. There were no test item-related macroscopic findings noted in the F1 males or females at the scheduled necropsy.

Postmortem examinations (parental animals):

See table

Postmortem examinations (offspring):

See table

Statistics:

All statistical tests were performed using WTDMS™ unless otherwise noted. Analyses were conducted using two-tailed tests (except as noted otherwise) for minimum significance levels of 1% and 5%, comparing each test item-treated group to the control group by sex.

Reproductive indices:

F0 reproductive performance values (fertility, mating, and conception/copulation indices, and number of days between pairing and coitus) and estrous cycle length in the test item-treated groups were similar to the control group.

Offspring viability indices:

See table

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

see below

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

see below

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

see below

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No treatment-related effects were observed.

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

No treatment-related effects were observed.

Details on results (P0)

Mean body weights and body weight gains in the 250, 500, and 1000 mg/kg/day groups were generally similar to the control group throughout the entire generation (males) and throughout pre-mating, gestation, and lactation (females). Test item-related, higher mean food consumption was noted throughout the entire treatment period in the 500 and 1000 mg/kg/day group males and sporadically during the pre-mating treatment period in the 1000 mg/kg/day group females. The increased food consumption noted in these groups was considered non-adverse because it did not affect mean body weights or body weight gains. Mean food consumption in the 250 mg/kg/day group males throughout the entire generation, in the 250 and 500 mg/kg/day group females during the pre-mating period, and in the 250, 500, and 1000 mg/kg/day group females during gestation and lactation was unaffected by test item administration.

Increased occurrences of red material around the nose and/or red, yellow and/or clear material around the mouth were noted in the 500 mg/kg/day group F0 males and the 1000 mg/kg/day group F0 males and females intermittently throughout the treatment period at approximately 1 hour following dose administration; these findings were more frequently noted in males than in females. These material findings were attributed to the test item. There were no test item-related clinical findings noted at the detailed physical examinations for the F0 generation.
F0 reproductive performance values (fertility, mating, and conception/copulation indices, and number of days between pairing and coitus) and estrous cycle length in the test item-treated groups were similar to the control group. No test item-related effects were noted on mean gestation lengths or the process of parturition at any dosage level. The number of former implantation sites and the number of unaccounted-for sites were unaffected by test item administration.

Effect levels (P0)

1

Key result

Dose descriptor:

NOAEL

Remarks:

NOAEL (Reproductive Toxicity)

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: NOAEL = highest dose tested

Remarks on result:

other: Generation: F0 and F1 (migrated information)

2

Key result

Dose descriptor:

NOAEL

Remarks:

NOAEL (Systemic Toxicity)

Effect level:

1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: NOAEL = highest dose tested

Remarks on result:

other: Generation: F0 & F1 (migrated information)

Results: F1 generation

General toxicity (F1)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

see below

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

see below

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

see below

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

Details on results (F1)

Pup clinical observations, body weights, and body weight gains prior to weaning were unaffected by parental test item administration. No internal findings attributed to parental administration of the test item were noted at the necropsies of pups that were found dead or at the PND 21 necropsy.

There were no test item-related effects on survival, clinical signs, body weights, body weight gains, or food consumption for F1 males and females following weaning at any dosage level. There were no test item-related macroscopic findings noted in the F1 males or females at the scheduled necropsy.

Mean ages of attainment of balanopreputial separation and vaginal patency and mean body weights at the age of attainment for F1 animals were unaffected by parental test item administration.

Lower postnatal survival was noted from birth to PND 4 (pre-selection) in the 500 and 1000 mg/kg/day groups compared to the control group. The lower values were attributed to 1 and 3 females in the 500 and 1000 mg/kg/day groups, respectively, with higher numbers of pup deaths or total litter losses; these losses were attributed to lower body weight pups and/or smaller litter size for the effected dams and were not considered test item-related. The mean number of pups born, live litter size, and percentage of males at birth were unaffected by parental test item administration.

Effect levels (F1)

Dose descriptor:

NOAEL

Remarks:

NOAEL (Neonatal Toxicity

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: NOAEL = highest dose tested

Overall reproductive toxicity

Reproductive effects observed:

not specified

Applicant's summary and conclusion

Executive summary:

There was no F0 and F1 test item-related mortality at any dosage level. No adverse effects on mean F0 and F1 male and female body weights, body weight gains, and food consumption were noted at any dosage level. In addition, no adverse effects were noted on macroscopic findings for the F0 and F1 males and females or microscopic findings and organ weights for the F0 males and females at any dosage level. Therefore, the no-observed-adverse-effect level (NOAEL) for F0 and F1 male and female systemic toxicity was considered to be 1000 mg/kg/day (the highest dosage level evaluated). In the absence of effects on F0 male and female reproductive performance, a dosage level of 1000 mg/kg/day was considered to be the NOAEL for F0 reproductive toxicity of the test material administered orally to male and female Crl:CD(SD) rats. In the absence of neonatal toxicity at all dosage levels, a dosage level of 1000 mg/kg/day was considered to be the NOAEL for neonatal toxicity.

This study demonstrates that alkyl phenate sulfides are not reproductive hazards but, instead, the reproductive toxicity is attributed to the residual starting material, EC 310 -154 -3.