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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Remarks:
Reproductive / developmental toxicity screening study
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04 September 2013 - 28 August 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in Section 13.
Cross-reference
Reason / purpose:
read-across: supporting information
Reference
Endpoint:
short-term repeated dose toxicity: oral
Remarks:
Reproductive / developmental toxicity screening study
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
04 September 2013 - 28 August 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in Section 13.
Reason / purpose:
read-across source
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:
no effects observed
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Reproductive function: estrous cycle
no effects observed

Reproductive function: sperm measures
no effects observed

Reproductive performance
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No unscheduled mortality.

No treatment related clinical signs apparent for animals (f/m) treated with 3000, 7500 or 18000/15000 ppm.
One male treated with 3000 ppm had pilo-erection and hunched posture.

No treatment-related changes in:
-Behavioral Assessments
-Functional Performance Tests
-Sensory Reactivity Assessments

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
18000 pp: males showed a statistically significant reduction (p<0.01) in body weight gain during Week 1 and Week 5. Overall body weight gain for these males was reduced by 26% when compared to control males.
18000/15000 ppm: females showed a statistically significant (p<0.05-p<0.001) reduction in body weight on Day 20 of gestation and on Days 1 and 4 of lactation and a reduction in body weight gain during the final two weeks of gestation (no statistically significant).
7500 or 3000 ppm: No effects.

WATER CONSUMPTION (PARENTAL ANIMALS)
No effects

REPRODUCTIVE FUNCTION:
MATING
No treatment- related effects on mating performance

FERTILITY
No treatment- related effects on fertility

GESTATION LENGTH
No treatment- related effects on gestation length

LABORATORY INVESTIGATION
Hematology: No toxicologically significant effects were detected in the hematological parameters examined.
Blood Chemistry: 18000/15000 ppm: female showed a statistically significant increase (p<0.01) in bilirubin and males statistically significant reduction in albumin and albumin/globulin ratio (p<0.01 and p<0.05 respectively). Females and males showed a statistically significant reduction (p<0.01) in alkaline phosphatase. Females treated with 18000/15000 ppm showed a statistically significant reduction in alanine aminotransferase and aspartate amino transferase (p<0.05 and p<0.01 respectively).
7500 ppm: females showed a statistically significant reduction (p<0.05) in alanine aminotransferase.
Males showed a statistically significant reduction in alkaline phosphatase and albumi. Females also showed a statistically significant increase in bilirubin.

PATHOLOGY (PARENTAL ANIMALS)
18000/15000 ppm: pale lungs with white patches present were found in two females. Another female from this treatment group had brown discolored lungs with white patches present. All these females had alveolar macrophages evident at histopathological examination. No findings were detected in males treated with 18000 ppm or animals of either sex at 3000 and 7500 ppm

HISTOPATHOLOGY(PARENTAL ANIMALS)
Urinary Bladder: urothelial hyperplasia and vacuolation was evident in females treated with 18000/15000 ppm.
Lungs: increased incidence and severity (males only) of alveolar macrophages was evident in animals of either sex treated with 18000/15000 ppm.
Prostate: increased incidence and severity of inflammation and lymphoid infiltrates was evident in males treated with 18000 or 7500 ppm.
Key result
Dose descriptor:
NOEL
Effect level:
7 500 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Systemic Toxicity
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
18 000 other: 18000 ppm (males); 15000 ppm (females)
System:
respiratory system: lower respiratory tract
Organ:
lungs
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
The result of the study set the ‘No Observed Effect Level’ (NOEL) for systemic toxicity at 7500 ppm for either sex.
Executive summary:

This data is being read across from the source study that tested Resin acids and Rosin acids, fumarated, esters with glycerol based on category read across that is explained in the category justification document attached in Section 13 of the dossier.

The test material Resin acids and rosin acids, fumarated, esters with glycerol, CAS NO. 97489-11-7 was administrated to rats by gavage, at concentrations of 3000, 7500 or 18000/15000 ppm. The result of the study showed microscopic lung change in animals of either sex treated with 18000/15000 ppm, microscopic urinary bladder changes in females treated with 18000/15000 ppm, and microscopic prostate changes in males treated with 18000 or 7500 ppm.

 

The ‘No Observed Effect Level’ (NOEL) for systemic toxicity was considered to be 7500 ppm for either sex.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
Deviations did not affect the scientific integrity of the study
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Han™:RccHan™:WIST
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories U.K. Ltd., Blackthorn, Bicester, Oxon, UK
- Age at study initiation: approximately twelve weeks old
- Weight at study initiation: males: 302 to 352g, females: 186 to 215g
- Housing: Solid floor polypropylene cages with stainless steel mesh lids and softwood flake bedding
- Use of restrainers for preventing ingestion (if dermal): no
- Diet (e.g. ad libitum): A powdered diet (Rodent PMI 5002 (Certified), Harlan Laboratories U.K. Ltd., Oxon, UK.)
- Water (e.g. ad libitum): free access to water
- Acclimation period: six days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): e 22 ± 3 °C
- Humidity (%): 50 ± 20%
- Air changes (per hr): at least fifteen air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light, twelve hours darkness

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
DIET PREPARATION
- Rate of preparation of diet (frequency): prior to the first treatment, and approximately twice weekly for the next three mixes and then weekly thereafter
- Mixing appropriate amounts with (Type of food): A known amount of test item was mixed with a small amount of basal laboratory diet
- Storage temperature of food: eight days at room temperature -The diet was stored in labelled, double plastic bags in labelled, covered plastic bins at room temperature
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
About 6 weeks
Frequency of treatment:
Daily
Doses / concentrationsopen allclose all
Dose / conc.:
0 ppm
Remarks:
Control
Dose / conc.:
3 000 ppm
Remarks:
Low
Basis: nominal in diet
Dose / conc.:
7 500 ppm
Remarks:
Intermediate
Basis:nominal in diet
Dose / conc.:
18 000 other: 18000.0 ppm (males); 15000.0 ppm (females)
Remarks:
High
Basis: nominal in diet
No. of animals per sex per dose:
12
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale:
Dose levels were selected based on available toxicity data and including a fourteen day range finder performed at these Laboratories (Harlan Laboratories Ltd. Study Number 41302580). The range-finding did not show adverse effect at the highest doses of 20000 ppm. In view of the longer administration period, a high dosage of 18000 ppm (reduced to 15000 ppm for females during gestation and lactation) was chosen for the current study including a lower dosages of 7500 and and intermediate dose of 3000 ppm.

Examinations

Observations and examinations performed and frequency:
Parental animals: Observations and examinations
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: All animals were examined for overt signs of toxicity, ill-health and behavioral change immediately before dosing, soon after dosing, and one hour after throughout the treatment period (except for females during parturition where applicable).

FUNCTIONAL OBSERVATIONS
Prior to the start of treatment and at weekly intervals thereafter, all animals were observed for signs of functional/behavioral toxicity. Functional performance tests were also performed on five selected males and females from each dose level, prior to termination, together with an assessment of sensory reactivity to various stimuli.
- Behavioral Assessments
- Functional Performance Tests
- Sensory Reactivity

BODY WEIGHT: Yes
- Individual body weight were recordered on Day 1 (prior dosing) and then weekly for males until termination and weekly for females until mating was evident. Body weight were recorded for females on days 0, 7, 14, 20 post coitum, on days 1 and 4 post partum, and at terminal kill.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
During the pre-pairing period, weekly food consumption was recorded for each cage of adults. This was continued for males after the mating phase. For females showing evidence of mating, food consumption was recorded for the periods covering post coitum Days 0-7, 7-14 and 14-20. For females with live litters, food consumption was recorded on Days 1 and 4 post partum. Food efficiency (the ratio of body weight change/dietary intake) was calculated retrospectively for males throughout the study period (with the exception of the mating phase) and for females during the pre-pairing phase. Due to offspring growth and milk production, food efficiency could not be accurately calculated for females during gestation and lactation.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
Water intake was measured daily throughout the study (with the exception of the pairing phase).
- Time schedule for examinations:

REPRODUCTIVE PERFORMANCE
- Mating
- Pregnancy and Parturition
- Litter Data
- Physical Development

PHYSICAL DEVELOPMENT
All live offspring were assessed for surface righting reflex on Day 1 post partum.

Estrous cyclicity (parental animals)
Animals were paired on a 1 male: 1 female basis within each dose group, for a period of up to fourteen days. Cage tray-liners were checked each morning for the presence of ejected copulation plugs and each female was examined for the presence of a copulation plug in the vagina. A vaginal smear was prepared for each female and the stage of oestrus or the presence of sperm was recorded. The presence of sperm within the vaginal smear and/or vaginal plug in situ was taken as positive evidence of mating (Day 0 of gestation) and the males were returned to their original cages (unless requiredfor additional pairing). Mated females were housed individually during the period of gestation and
lactation.

Litter observations
Number of offspring born
Number of offspring alive recorded daily and reported on Days 1 and 4 post partum
Sex of offspring on Days 1 and 4 post partum
Clinical condition of offspring from birth to Day 5 post partum
Individual offspring weights on Days 1 and 4 post partum (litter weights were calculated retrospectively from this data)

ii.Number of offspring alive recorded daily and reported on Days 1 and 4 post partum
iii. Sex of offspring on Days 1 and 4 post partum
iv. Clinical condition of offspring from birth to Day 5 post partum
v. Individual offspring weights on Days 1 and 4 post partum (litter weights were calculated retrospectively from this data)

All live offspring were assessed for surface righting reflex on Day 1 post partum.
Sacrifice and pathology:
Postmortem examinations (parental animals)
Adult males were killed by intravenous overdose of sodium pentobarbitone followed by exsang
uination on Day 43.
Adult females: intravenous overdose of sodium pentobarbitone followed by exsanguination on Day 5
post partum.
Surviving offspring: intracardiac overdose of sodium pentobarbitone.
Any females which failed to achieve pregnancy or produce a litter were killed on or after Day 25 post
coitum.
One female which failed to mate was terminated on Day 57.
Organ weights ( 5 selected male and female animals):
Adrenals
Brain
Epididymides
Heart
Kidneys
Thym
Liver
Ovaries
Prostate
Seminal vesicles
Spleen
Testes
Thymus
Thyroid (weighed post-fixation with Parathyroid)
Uterus (weighed with Cervix)
Histopathology:
Samples of the following tissues were removed from five selected males and five selected:
Adrenals Muscle
Aorta
Ovaries Bone & bone marrow (femur including stifle joint)
Pancreas Bone & bone marrow (sternum)
Pituitary Brain (including cerebrum, cerebellum and pons)
Prostate Caecum Rectum
Coagulating gland Salivary glands (submaxillary)
Colon Sciatic nerve Duodenum Seminal vesicles Epididymides Skin (hind limb)
Esophagus Spinal cord (cervical, mid-thoracic and Eyes* lumbar)
Gross lesions Spleen Heart Stomach Ileum (including peyer’s patches)
Thyroid/parathyroid Jejunum
Trachea
Kidneys
Testes
Liver
Thymus Lungs (with bronchi)
Urinary bladder
Lymph nodes (mandibular and mesenteric)
Uterus/Cervix Mammary gland Vagina

Since there were indications of treatment-related liver, kidney, thyroid, pituitary and stomach changes, examination was subsequently extended to include similarly prepared sections of the liver, kidneys, thyroids, pituitary and stomach from animals in the low and intermediate groups.
Other examinations:
Reproductive indices
i. Pre-coital Interval
ii. Fertility Indices
i. Gestation Length
ii. Parturition Index

Offspring viability indices
i. Implantation Losses (%)
ii. Live Birth and Viability Indices
iii. Sex Ratio (% males)

Results and discussion

Results of examinations

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:
no effects observed
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Reproductive function: estrous cycle
no effects observed

Reproductive function: sperm measures
no effects observed

Reproductive performance
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No unscheduled mortality.

No treatment related clinical signs apparent for animals (f/m) treated with 3000, 7500 or 18000/15000 ppm.
One male treated with 3000 ppm had pilo-erection and hunched posture.

No treatment-related changes in:
-Behavioral Assessments
-Functional Performance Tests
-Sensory Reactivity Assessments

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
18000 pp: males showed a statistically significant reduction (p<0.01) in body weight gain during Week 1 and Week 5. Overall body weight gain for these males was reduced by 26% when compared to control males.
18000/15000 ppm: females showed a statistically significant (p<0.05-p<0.001) reduction in body weight on Day 20 of gestation and on Days 1 and 4 of lactation and a reduction in body weight gain during the final two weeks of gestation (no statistically significant).
7500 or 3000 ppm: No effects.

WATER CONSUMPTION (PARENTAL ANIMALS)
No effects

REPRODUCTIVE FUNCTION:
MATING
No treatment- related effects on mating performance

FERTILITY
No treatment- related effects on fertility

GESTATION LENGTH
No treatment- related effects on gestation length

LABORATORY INVESTIGATION
Hematology: No toxicologically significant effects were detected in the hematological parameters examined.
Blood Chemistry: 18000/15000 ppm: female showed a statistically significant increase (p<0.01) in bilirubin and males statistically significant reduction in albumin and albumin/globulin ratio (p<0.01 and p<0.05 respectively). Females and males showed a statistically significant reduction (p<0.01) in alkaline phosphatase. Females treated with 18000/15000 ppm showed a statistically significant reduction in alanine aminotransferase and aspartate amino transferase (p<0.05 and p<0.01 respectively).
7500 ppm: females showed a statistically significant reduction (p<0.05) in alanine aminotransferase.
Males showed a statistically significant reduction in alkaline phosphatase and albumi. Females also showed a statistically significant increase in bilirubin.

PATHOLOGY (PARENTAL ANIMALS)
18000/15000 ppm: pale lungs with white patches present were found in two females. Another female from this treatment group had brown discolored lungs with white patches present. All these females had alveolar macrophages evident at histopathological examination. No findings were detected in males treated with 18000 ppm or animals of either sex at 3000 and 7500 ppm

HISTOPATHOLOGY(PARENTAL ANIMALS)
Urinary Bladder: urothelial hyperplasia and vacuolation was evident in females treated with 18000/15000 ppm.
Lungs: increased incidence and severity (males only) of alveolar macrophages was evident in animals of either sex treated with 18000/15000 ppm.
Prostate: increased incidence and severity of inflammation and lymphoid infiltrates was evident in males treated with 18000 or 7500 ppm.

Effect levels

Key result
Dose descriptor:
NOEL
Effect level:
7 500 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Systemic Toxicity

Target system / organ toxicity

Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
18 000 other: 18000 ppm (males); 15000 ppm (females)
System:
respiratory system: lower respiratory tract
Organ:
lungs
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
not specified

Applicant's summary and conclusion

Conclusions:
The result of the study set the ‘No Observed Effect Level’ (NOEL) for systemic toxicity at 7500 ppm for either sex.
Executive summary:

The test material Resin acids and rosin acids, fumarated, esters with glycerol, CAS NO. 97489-11-7 was administrated to rats by gavage, at concentrations of 3000, 7500 or 18000/15000 ppm. The result of the study showed microscopic lung change in animals of either sex treated with 18000/15000 ppm, microscopic urinary bladder changes in females treated with 18000/15000 ppm, and microscopic prostate changes in males treated with 18000 or 7500 ppm.

 

The ‘No Observed Effect Level’ (NOEL) for systemic toxicity was considered to be 7500 ppm for either sex.