1-Butanamine, N-butyl-, reaction products with polyethylene-polypropylene glycol ether with trimethylolpropane (3:1) acrylate

Endpoint summary

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2012/2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

adopted 1996

Qualifier:

according to guideline

Guideline:

other: EPA OPPTS 870.3650, adopted July 2000

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:Charles River Laboratories, Research Models and Services, Germany GmbH, Sulzfeld, Germany
- Age at study initiation: 10-11 wks;
- Weight at study initiation: Males: 294g; Females: 197g, on average
- Fasting period before study: no
- Housing: individually (except during overnight mating and lactation) in Makrolon type M III cages
- Diet (e.g. ad libitum): Kliba maintenance diet mouse-rat “GLP” meal, ad lib.
- Water (e.g. ad libitum): tap water ad lib.
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24°C
- Humidity (%): 30-70%
- Air changes (per hr):15

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:The test substance was applied as suspension, which was prepared at least once a week. An appropriate amount of the test substance was weighed, and corn oil was added up to the desired volume. The suspension was kept homogenous during administration by stirring with a magnetic stirrer.

The administration volume was 4ml/kg b.w.

VEHICLE
- Justification for use and choice of vehicle (if other than water): The test substance is poorly soluble in water.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The stability of the test substance in corn oil for a period of 7 days at room temperature was proven before the start of the study (BASF Project No. 11L00386).
For homogeneity and concentration control analyses, each 6 samples of all concentrations was drawn at the start of the study and towards the end of the administration period. Homogeneity was confirmed, all concentrations measured were within 90-110% of the target concentration.

Duration of treatment / exposure:

males: 36 days
females: 51 days

Frequency of treatment:

daily, except to animals being in labor

Remarks:

Doses / Concentrations:
100, 300, 1000mg/kg (high dose was reduced to 600mg/kg on day 19)
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: based on results of a range-finding study - no severe signs of toxicity were observed in animals treated with 1000mg/kg of the test substance for 14 days.

Positive control:

no

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily on workdays, daily on weekends and public holidays
- Cage side observations: check for moribund animals, pertinent behavioral changes, signs of overt toxicity, parturation, littering and lactation behavior of the dams

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: prior to the first administration, weekly thereafter
- The following parameters were examined: abnormal behavior during “handling”, fur, skin, posture, salivation, respiration, activity/arousal level, tremors, convulsions, abnormal movements, impairment of gait, lacrimation, palpebral closure, exophthalmus, feces (appearance/consistency), urine, pupil size

BODY WEIGHT: Yes
- Time schedule for examinations: day 0, weekly thereafter with the following exceptions for females: During the mating period the parental females were weighed on the day of positive evidence of sperm (GD 0) and on GD 7, 14 and 20. Females with litter were weighed on the day of parturition (PND 0) and on PND 4.

FOOD CONSUMPTION:
- Food consumption for each animal determined: Yes, except during mating

WATER CONSUMPTION: Monitored by daily visual inspection

HAEMATOLOGY: Yes
- Time schedule for collection of blood: on the day of necropsy
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes (16-20h)
- How many animals: 5 per sex and group
- The following parameters were examined: Leukocyte count, Erythrocyte count, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelet count, differential blood count, reticulocytes, prothrombin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on the day of necropsy
- Animals fasted: Yes (16-20h)
- How many animals: 5 per sex and group
- The following parameters were examined: ALT, AST, ALP, GGT, sodium, potassium, chlorid, inorganic phosphate, calcium, urea, creatinine, glucose, total bilirubin, total protein, albumin, globulins, triglycerides, cholesterol

URINALYSIS: Yes
- Time schedule for collection of urine: day 30 (males) and day 50 (females)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: yes, during collection
- How many animals: 5 per sex and group
- The following parameters were examined: pH, protein, glucose, ketones, urobilinogen, bilirubin, blood, specific gravity, sediment, color, turbity, volume

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: 3 days prior to necropsy (day 32 males, day 49 females)
- Dose groups that were examined: 5 animals per sex and group (only females with litter)
- Battery of functions tested: home cage and open field observation, motor activity, sensory motor test / reflexes (including: Reaction to an object being moved towards the face, touch sensitivity, vision (Visual placing response), pupillary reflex, pinna reflex, auditory startle response, righting response, behavior during handling, vocalization, pain perception (Tail pinch), grip strength of forelimbs and hindlimbs, landing foot-splay test)

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
The following tissues were weighed
- in all animals: epididymides, testes
- in 5 males and females of each group: adrenal glands, brain, heart, kidneys, liver, spleen, thymus
The following tissues were examined histotechnically in at least 5 animals per sex of the control and high dose group unless noted otherwise
adrenal glands, gross lesions (all affected animals in all dosage groups), bone marrow (femur), brain, cecum, cervix, coagulation glands, colon, duodenum, epididymides, heart, ileum, jejunum, kidneys, liver, lungs, lymph nodes (auxillary and mesenteric), ovaries, oviducts, prostate gland, peyer's patches, rectum, sciatic nerve, seminal vesicles, spinal cord, spleen, forestomach (all animals from all dose groups), glandular stomach, testes, thymus, thyroid glands, trachea, urinary bladder, uterus, vagina

Other examinations:

Reproductive performance: see entry in chapter 7.8.1

Details on results:

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
In test group 3 (1000 and 600 mg/kg bw/d) female animal No. 80 was found dead on study day 17 (Mating Day 3), female animal No. 74 was sacrificed moribund on study day 19 (GD 2), and male animal No. 32 was found dead on study day 35. These premature deaths were assessed as being related to the test substance. One female animal of test group 2 (300 mg/kg bw/d) was also found dead on study day 44. The animal died because of a gavage error. Therefore, a relation to treatment was excluded.

Almost all animals of both sexes of the mid and high dose group showed mostly slight, sometimes moderate salivation within 2 hours after the test substance administration on several days of the study. The same was true for individual low dose animals. From the temporary, short appearance immediately after dosing it could be assumed that salivation was induced by a bad taste of the test substance or local affection of the upper digestive tract. The effect was related to the test substance but assessed as being non-adverse.

Animal No. 74 of test group 3 (1000 and 600 mg/kg bw/d) showed poor general condition, piloerection and respiration sounds on GD 1 and GD 2. Therefore, the animal was sacrificed in a moribund condition on GD 19. Animal No. 75 of the same group showed semiclosed eyelids, poor general condition, hunched posture, smeared fur, piloerection (up to GD 7) and gasping on GD 2 and respiration sounds on GD 3. The same animal showed respiration gasping and sounds on LD 1. These findings were assessed as being related to treatment.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
No treatment-related changes in body weight data were observed in animals of all test groups.

FOOD CONSUMPTION
No treatment-related changes in food consumption were observed.

HAEMATOLOGY
No treatment-related changes among hematological parameters were observed.

CLINICAL CHEMISTRY
No treatment-related changes were measured.

URINALYSIS
No test substance-related effects were observed.

NEUROBEHAVIOUR
No test substance-related effects were observed.

ORGAN WEIGHTS
Absolute thymus weights were significantly decreased in high dose male animals when compared to control group animals. This finding was not considered to be treatment related, since no relative organ weights were significantly altered.

GROSS PATHOLOGY
Erosions, ulcers, and in one case a thickened forstomach were observed in all high dose male and females (with the exception of one female) and 5 mid dose females and 4 mid dose males.

HISTOPATHOLOGY:
Five male and 6 female animals of test group 3 (1000 and 600 mg/kg bw/d) as well as each 2 males and females of test group 2 (300 mg/kg bw/d) revealed multifocal erosions/ulcers in the forestomach. Accompanying these erosions/ulcers a mild to extreme diffuse or focal squamous hyperplasia with hyperkeratosis and a mild to severe submucosal edema was present. When the hyperplasia of the squamous epithelium was focal, it was located at the cardia region (stomach entrance). Within the edema a higher number of inflammatory cells were observed. Not all animals with a macroscopically diagnosed erosion/ulcer showed this finding. But the consistency of the accompanying findings (hyperplasia with hyperkeratosis and edema) which were observed in animals with and without erosion/ulcer led to the conclusion that this all belongs to the same complex. As the erosions/ulcer that could be seen microscopically were very small it is possible that they were not on the level of section. But taken all these findings together they point towards an irritating effect of the substance on the squamous epithelium of the forestomach. Especially the cardia region (entrance of the stomach) was affected.
The three animals of test group 3 (1000 and 600 mg/kg bw/d) which were found dead or were sacrificed in a moribund (animal Nos. 32, 74 and 80) did not show any other findings that could explain the death beside the ones in the stomach. Two of them revealed a moderate to severe thymus atrophy which was regarded to be a consequence to the stress caused by the findings in the forestomach. But it is not clear whether these observed findings were the cause of the death although death was regarded to be a consequence to the treatment with the test substance.

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

>= 300 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: deaths and poor general state in high dose animals

Dose descriptor:

NOAEL

Remarks:

local

Effect level:

>= 100 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: irritation in the stomach

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

There are no studys available.

The test item is Propylidynetrimethanol, ethoxylated, propoxylated, esters with acrylicacid, reaction products with 1-butanamine, n-butyl (Cas:173011-06-8). The concentration of EO/PO is >1 <6.5. The chosen read-across substances is Propylidynetrimethanol, ethoxylated, esters with acrylicacid, reaction products with 1-butanamine, n-butyl (Cas: 195008-76-5) concentration of. EO>1 <6.5.

In general basic physicochemical properties such as water solubility, vapour pressure and the partition coefficient (LogPow) are quite similar. The water solubility was 0.9 g/l the vapour pressure 2.3 hPa and the logPow 4.2 and 0.8 g/l, 1.6 hPa and the logPow 3.5 for the read-across substance.

Both molecules show no acute toxicity after oral or dermal exposure and did not induce gene mutations in bacteria. Both showed no skin irritation and Cas: 195008-76-5 was slightly eye irritating. Due to the very similar structure, physicochemical properties and almost identical results in the toxicological studies mentioned above, comparable results are also expected for the endpoints for which read across was performed.

Readacross

Propylidynetrimethanol, ethoxylated, esters with acrylic acid, reaction products with 1-Butanamine, N-butyl- was administered daily in corn oil via gavage to groups of 10 male and 10 female Wistar rats at dose levels of 100, 300, and 1000mg/kg. Because of the premature death of 2 female animals, the high dose was reduced from 1000 to 600 mg/kg bw/d from study day 19 onwards. The duration of treatment covered 36 days in males and 51 days in females, including premating, mating, gestation and 4 days of lactation.

A detailed clinical observation was performed in all animals before initial test substance administration and at weekly intervals thereafter. Food consumption and body weight of the animals were determined in app. weekly intervals. Towards the end of the administration period a functional observational battery was performed and motor activity was measured in 5 animals per sex and test group. Clinicochemical and hematological examinations as well as urinalyses were also performed towards the end of the administration period in 5 animals per sex and test group. All animals were sacrificed by decapitation, under isoflurane anesthesia, and were assessed by gross pathology. Weights of selected organs were recorded and a histopathological examination was performed.

Of the high dose animals, one male and one female were found dead, and one female was sacrificed in moribund condition. Piloerection, smeared fur, respiratory sounds, poor general state, hunched posture, and semiclosed eyelids were observed in single animals of this group at different time points. Pathology revealed erosion/ulcer in the forestomach of all male and 9/10 female high dose animals, mild to severe diffuse or focal hyperplasia with hyperkeratosis in the forestomach of all animals, and mild to severe submucosal edema in the forestomach of 8 male and 3 female animals. Animals receiving 300mg/kg b.w. showed comparable signs of local irritation in the forestomach, though the incidence was clearly reduced. No clinical signs were noted in this group and in low dose animals, which also showed no local irritation in the stomach. Reproductive performance was unaffected in all groups. Though the clinical signs and deaths of high dose animals are most likely secondary to the severe irritation in the stomach, systemic toxicity cannot be completely excluded. Following the precautionary principle, the NOAEL was thus set to 300mg/kg b.w. for systemic effects, and to 100mg/kg for local irritation.

Justification for classification or non-classification

No specific target organ toxicity was oberseved; only local irritation occured in the repeated dose study. Thus Propylidynetrimethanol, ethoxylated, propoxylated esters with acrylic acid, reaction products with 1-Butanamine, N-butyl- does not have to be classified according to 67/548/EEC or CLP/EU-GHS.