Trisodium 4-[[1-hydroxy-6-[[[[5-hydroxy-6-(phenylazo)-7-sulphonato-2-naphthyl]amino]carbonyl]amino]-3-sulphonato-2-naphthyl]azo]benzoate

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data from secondary source

Data source

Reference

Reference Type:

secondary source

Title:

Reproductive and developmental toxicity in rats for CAS no: 114959-46-5

Author:

US EPA

Year:

2017

Bibliographic source:

High Production Volume Information System (HPVIS)| OPPT | US EPA, 2017

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Principles of method if other than guideline:

Reproductive and developmental toxicity study of Benzoic acid, 2-hydroxy-, mono-C14-18-alkyl derivs., calcium salts (2:1) was performed on rats according OECD guideline 421

GLP compliance:

not specified

Limit test:

no

Justification for study design:

No data available

Test material

Test material information

Constituent 1

Reference substance name:

Benzoic acid, 2-hydroxy-, mono-C14-18-alkyl derivs., calcium salts (2:1)

EC Number:

601-337-1

Cas Number:

114959-46-5

IUPAC Name:

Benzoic acid, 2-hydroxy-, mono-C14-18-alkyl derivs., calcium salts (2:1)

Details on test material:

- Name of test material (as cited in study report):Benzoic acid, 2-hydroxy-, mono-C14-18-alkyl derivs., calcium salts (2:1)
- Molecular formula :Unspecified
- Molecular weight : 388.6014g/mol
- Substance type: Organic
- Physical state:Solid

Specific details on test material used for the study:

- Name of test material (as cited in study report):Benzoic acid, 2-hydroxy-, mono-C14-18-alkyl derivs., calcium salts (2:1)
- Molecular formula (if other than submission substance):Unspecified
- Molecular weight (if other than submission substance): 388.6014g/mol
- Substance type: Organic
- Physical state:Solid

Test animals

Species:

rat

Strain:

Crj: CD(SD)

Details on species / strain selection:

No data available

Sex:

male/female

Details on test animals or test system and environmental conditions:

Details on test animals and env. conditions
TEST ANIMALS
- Source: No data available
- Age at study initiation: approximately 62 days
- Weight at study initiation: No data available
- Fasting period before study: No data available
- Housing: No data available
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum): No data available
- Water (e.g. ad libitum): No data available
- Acclimation period: No data available
ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%):No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available
IN-LIFE DATES: From

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material dissolved in corn oil
DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food )
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water): Test material soluble in corn oil
- Concentration in vehicle: 0, 50, 150 and 500 mg/kg/day
- Amount of vehicle (if gavage): 5ml/kg
- Lot/batch no. (if required): No data available
- Purity: No data available

Details on mating procedure:

- M/F ratio per cage:1/1
- Length of cohabitation:For up to 1 4 days or until positive evidence of mating was observed
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy :No data available
- 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):No data available
- Any other deviations from standard protocol:No data available

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analysis of dosing solutions confirmed that the preparations were homogeneous and that they were at the appropriate concentrations. The stability of the dosing solutions was previously determined.

Duration of treatment / exposure:

F0 males- study days 0-27, 14 day premating period through lactation day 3.

Frequency of treatment:

once/day, 7 days/week

Details on study schedule:

No data available

Doses / concentrations

Remarks:

0, 50, 150 and 500 mg/kg/day

No. of animals per sex per dose:

Total:96
0 mg/kg bw/day:24
50mg/kg bw/day:24
150mg/kg bw/day:24
500mg/kg bw/day:24

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Positive control:

No data available

Examinations

Parental animals: Observations and examinations:

Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes
DETAILED CLINICAL OBSERVATIONS: Yes
Time schedule: Twice daily throughout the study. Detailed individual examinations conducted once weekly

BODY WEIGHT: Yes
Time schedule for examinations: F0 males : pretest, weekly throug h termination.
F0 females: pretest, weekly during premating and mating; gesta tion days0, 4, 7, 11, 14, 17 and 20; females with litters weighed on lactation days 1 and 4.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Pretest and weekly during treatment period. F0 males: pretest, weekly throughout the study except during mating.
F0 females: pretest, weekly during premating, days 0, 4, 7, 11, 14, 17 and 20 of gestation and days 1- 4 of lactation for females with litters.

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 / No / No data: No data available


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
Time schedule for examinations:

OTHER:

Oestrous cyclicity (parental animals):

No data available

Sperm parameters (parental animals):

No data available

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: [yes/no]No data available
- If yes, maximum of [...] pups/litter ([...]/sex/litter as nearly as possible); excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in [F1 offspring:
Litter Size: Recorded daily.
Individual Pup Body Weights: Pup weights recorded on days 1 and 4 of lactation.
Sex Determination: Days 0 and 4 of lactation.


GROSS EXAMINATION OF DEAD PUPS:Litters observed as soon as possible after Delivery for number of live and dead pups and pup abnormalities. Thereafter litters observed daily for dead pups and/or obvious irregularities.

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY:No data available

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY:No data available

Postmortem examinations (parental animals):

Postmortem examinations (Parent Animal)
SACRIFICE : Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]: No data
Maternal animals: yes
All surviving animals [describe when, e.g. after the last litter of each generation was weaned :

GROSS NECROPSY: Organ Weights:
Performed on all animals. Microscopic Examinations: Special emphasis was placed on the stages of spermatogenesis and histopathology of interstitial testicular cell structure. Microscopic examination was
performed on the cervix, coagulating glands, mammary gland, thyroids with parathyroids, testes with epididymides and vas deferens, seminal
vesicles, ovaries and oviducts, pituitary gland, uterus with vagina, prostate gland and all gross lesions for all animals in the control and 500 mg/kg/day groups at the scheduled necropsies; gross lesions from all dosage groups were also examined.
HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.:
macroscopic examination was performed

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows: Pups found dead and pups sacrificed on day 4

GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]No data available

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.No data available

Statistics:

Parental mating, fertility, conception and copulation indices were analysed using the Chi-square test with Yates' correction factor. Mean parental body weights (weekly, gestation and lactation), body weight changes and food consumption, offspring body weights and body weight changes, gestation length, numbers of corpora lutea, implantation sites, number of pups born, live litter size on PND 0, unaccounted-for sites, absolute and relative organ weights and pre-coital intervals were subjected to a parametric one-way analysis of variance (ANOVA) to determine intergroup differences. If the ANOVA revealed statistically significant (p<0.05) intergroup variance, Dunnett's test was used to compare the test article-treated groups to the control group. Mean litter proportions (percent per litter) of males at birth and postnatal survival were subjected to the Kruskal-Wallis nonparametric ANOVA to determine intergroup differences. If the ANOVA revealed statistically significant (p<0.05) intergroup variance, Dunn's test was used to compare the test article-treated groups to the control group. Histopathological findings in the test article-treated groups w ere compared to the control group using a two-tailed Fisher's Exact test.

Reproductive indices:

No data available

Offspring viability indices:

No data available

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Clinical findings of excessive pawing at the cage floor and/or walls and wiping of the mouth on the cage floor and/or walls were noted for all 12 females in the 150 and 500 mg/kg/day groups at the time of dose administration, and for 1 and 8 females in these same groups 1-2 hours following dose administration. Salivation and related findings (consisting of clear material on various body surfaces) were noted for females in the 150 and 500 mg/kg/day groups just prior to, at the time of and/or 1-2 hours following dose administration. Excessive pawing at the cage floor and/or walls and salivation-related findings were noted to a lesser degree for males in the 500 mg/kg/day group at the time of dose administration. Salivation-related findings were also noted for males in this group just prior to and 1-2 hours following dose administration. The behavioural findings of excessive pawing of the cage floor and/or walls (males and females) and wiping of the mouth on the cage floor and/or walls (females) were likely due to test article taste aversion. Therefore, salivation-related findings noted for males and females were attributed to the test article, but were not considered adverse.

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Body weight was unaffected by test article administration at 50, 150 and 500 mg/kg/day.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

food consumption was unaffected by test article administration at 50, 150 and 500 mg/kg/day.

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

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Description (incidence and severity):

Reproductive parameters (e.g., mating, fertility and copulation/conception indices) were unaffected by test article administration at 50, 150 and 500 mg/kg/day

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

500 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

mortality

body weight and weight gain

food efficiency

organ weights and organ / body weight ratios

gross pathology

reproductive performance

Remarks on result:

other: No effects were observed on reproductive performance

Target system / organ toxicity (P0)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

no mortality observed

Description (incidence and severity):

postnatal survival from birth to PND 4 were unaffected at 50, 150 and 500 mg/kg/day.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

pup body weights were unaffected at 50, 150 and 500 mg/kg/day.

Food consumption and compound intake (if feeding study):

not specified

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

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

Other effects:

no effects observed

Description (incidence and severity):

F1 pups were unaffected by maternal test article administration. The mean number of pups born, live litter size on postnatal day (PND) 0, general physical condition, and postnatal survival from birth to PND 4 were unaffected at 50, 150 and 500 mg/kg/day.

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not specified

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

500 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

other: The mean number of pups born, live litter size on postnatal day (PND) 0, general physical condition, pup body weights and postnatal survival from birth to PND 4 were unaffected

Remarks on result:

other: No developmental effects were observed

Target system / organ toxicity (F1)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Overall reproductive toxicity

Reproductive effects observed:

not specified

Treatment related:

not specified

Relation to other toxic effects:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Applicant's summary and conclusion

Conclusions:

No Observed Adverse Effect Level (NOAEL) forreproductive and developmental toxicity in F0 and for F1 generationon the basis of reproductive parameter and overall developmental effects was considered to be 500 mg/kg/day.When femaleCrl:CD(SD)rats were treated with Benzoic acid, 2-hydroxy-, mono-C14-18-alkyl derivs., calcium salts (2:1)(114959-46-5)orally.

Executive summary:

The reproductive and developmental toxicity study of Benzoic acid, 2-hydroxy-, mono-C14-18-alkyl derivs., calcium salts (2:1)(114959-46-5) was performed according to OECD guideline 421 on male and female Crl:CD(SD)rats. 96 animals i.e 24/dose group were used in the study. The test material dissolved in corn oil in dose concentration0, 50, 150 and 500 mg/kg/day and administered by oral gavage route in dose volume 5ml/kg for once/day, 7 days/week. Analysis of dosing solutions confirmed that the preparations were homogeneous and that they were at the appropriate concentrations. The stability of the dosing solutions was previously determined. Doses were selected based on prior studies conducted with this test article.F0 males were exposed to test material on 0-27, 14 day premating period through lactation day 3 females were exposed. Mating ratio was One male to one female for up to 1 4 days or until positive evidence of mating was observed.All the animals were observed for clinical signs and toxicity Twice daily throughout the study. Detailed Individual examinations conducted once weekly. Body Weights in F0 males pretest, weekly through termination. Body weight of F0 females were observed weekly during premating and mating; gestation days 0, 4, 7, 11, 14, 17 and 20; females with litters weighed on lactation days 1 and 4.Food Consumption was Pre-test and weekly during treatment period. In F0 males it pre-test, weekly throughout the study except during mating while in F0 females it pre-test, weekly during premating, days 0, 4, 7, 11, 14, 17 and 20 of gestation and days 1- 4 of lactation for females with litters.

 

Macroscopic Examinations were performed on all animals. Organ Weights also noted in all animals. In microscopic Examinations Special emphasis was placed on the stages of spermatogenesis and histopathology of interstitial testicular cell structure. Microscopic examination was performed on the cervix, coagulating glands, mammary gland, thyroids with parathyroids, testes with epididymides and vas deferens, seminal vesicles, ovaries and oviducts, pituitary gland, uterus with vagina, prostate gland and all gross lesions for all animals in the control and 500 mg/kg/day groups at the scheduled necropsies; gross lesions from all dosage groups were also examined.

Litters observed as soon as possible after delivery for number of live and dead pups and pup abnormalities. Thereafter litters observed daily for dead pups and/or obvious irregularities. Litter Size was recorded daily. Individual Pup Body Weights recorded on days 1 and 4 of lactation. While sex determination performed on Days 0 and 4 of lactation. Macroscopic Examination carried out to on pups found dead and pups sacrificed on day 4 post-partum were carefully examined externally for gross external abnormalities, and a macroscopic examination was performed. Gross lesions and malformations were retained.

Clinical findings of excessive pawing at the cage floor and/or walls and wiping of the mouth on the cage floor and/or walls were noted for all 12 females in the 150 and 500 mg/kg/day groups at the time of dose administration, and for 1 and 8 females in these same groups 1-2 hours following dose administration. Salivation and related findings (consisting of clear material on various body surfaces) were noted for females in the 150 and 500 mg/kg/day groups just prior to, at the time of and/or 1-2 hours following dose administration. Excessive pawing at the cage floor and/or walls and salivation-related findings were noted to a lesser degree for males in the 500 mg/kg/day group at the time of dose administration. Salivation-related findings were also noted for males in this group just prior to and 1-2 hours following dose administration. The behavioural findings of excessive pawing of the cage floor and/or walls (males and females) and wiping of the mouth on the cage floor and/or walls (females) were likely due to test article taste aversion. Therefore, salivation-related findings noted for males and females were attributed to the test article, but were not considered adverse. Body weight, food consumption were unaffected by test article administration at 50, 150 and 500 mg/kg/day. Higher mean liver weights were noted for males (↑ 5.7%) and females (↑ 14.6%) in the 500 mg/kg/day group, but were not considered adverse, as these differences are probably consistent with hepatic enzyme induction that was adaptive in nature. Similar effects on liver weights with no morphologic correlates were reported for this test article in a previous study. No test article-related macroscopic or microscopic findings and no adverse effects on mean organ weights were observed for males and females at any dosage level. Reproductive parameters (e.g., mating, fertility and copulation/conception indices) were unaffected by test article administration at 50, 150 and 500 mg/kg/day. F1 pups were unaffected by maternal test article administration.

The mean number of pups born, live litter size on postnatal day (PND) 0, general physical condition, pup body weights and postnatal survival from birth to PND 4 were unaffected at 50, 150 and 500 mg/kg/day. Hence No Observed Adverse Effect Level (NOAEL) forreproductive and developmental toxicity in F0 and for F1 generationon the basis of reproductive parameter and overall developmental effects was considered to be 500 mg/kg/day.When femaleCrl:CD(SD)rats were treated with Benzoic acid, 2-hydroxy-, mono-C14-18-alkyl derivs., calcium salts (2:1)(114959-46-5)orally.