Amines, tri-C8-10-alkyl

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline and GLP compliant study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 22. March, 1996

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA OPPTS 870.3650, July 2000

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories, B.V., Horst, Netherlands (RccHanTM:WIST(SPF)
- Age at study initiation: 11 weeks
- Weight at study initiation: weight range: male 260 to 390 g; female 175 to 240 g
- Housing: in groups of three to four in Makrolon type-4 cages with wire mesh tops up to the day of randomization, afterwards individually in Makrolon type-3 cages
- Diet: Pelleted standard Harlan Teklad 2018C rodent maintenance diet, ad libitum
- Water: Community tap-water, ad libitum
- Acclimation period: minimum 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

The dose formulations will be prepared weekly using the test item as supplied by the Sponsor. Amines, tri-C8-10-alkyl will be weighed into a glass beaker on a tared precision balance and the vehicle will be added (w/v). Using an appropriate homogenizer, a homogeneous solution will be prepared. Separate formulations will be prepared for each concentration. Homogeneity of the test item in the vehicle will be maintained during the daily administration period using a magnetic stirrer.

Dose formulations will be stored at room temperature (15 - 25°C) in glass beakers. Based upon the results of stability analyses performed within the Harlan Laboratories study no. D71061 for at least 10 days at room temperature (15 - 25°C).

Animals will be treated orallay by gavage. The applicated volume is 4ml/kg body weight diluted in corn oil. Dosages are 0, 50, 150 and 400 mg/kg bw/d.

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation: 14 days
- Proof of pregnancy: vaginal smear sperm positive or a copulation plug was observed referred to as day 0 post coitum
- After successful mating each pregnant female was caged: individually

If a female does not mate during the 14-day pairing period, a second pairing of this female with a male in the same group, which has already mated successfully, will be considered. If mating is not recorded during this additional pairing period of a maximum of 14 days, the female will be sacrificed and, if indicated, the reproductive organs examined histopathologically in order to ascertain the reason for the infertility.
All dams were allowed to give birth and rear their litters (F1 pups) up to day 4 post partum. Day 0 was designated as the day on which a female has delivered all her pups.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

On the first dose formulation day samples from the control group as well as three samples (top, middle and bottom) of about 1 g of each concentration will be taken prior to dosing for analysis of concentration and homogeneity. Samples of about 1 g of each test item concentration will be taken from the middle to confirm the stability (10 days at room temperature 15-25°C). Towards the end of the study, samples will be taken from the middle to confirm concentration. The aliquots for analysis of dose formulations will be frozen (-20 ± 5 °C) and delivered on dry ice to Dr. B. Ludwig (Harlan Laboratories Ltd., Zelgliweg 1, 4452 Itingen / Switzerland) and stored there at -20 ± 5 °C until analysis.
The samples will be analyzed by HPLC-ELSD. The test item will be used as the analytical standard.

Duration of treatment / exposure:

Males: 28 days (14 days pre-pairing, pairing 14 days maximum, sacrifice after treatment of 28 days)
Females: 6-7 weeks (14 days pre-pairing, pairing 14 days maximum, 21 days gestation, 4 days lactation)

Frequency of treatment:

daily, 7 days/week

Remarks:

Doses / Concentrations:
50, 150, 400 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

12

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on the results of a 14-day range-finding study (BASF project number 01R0674/12X381) the dose levels of 50, 150, and 400 mg/kg bw per day were selected for the present study.

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily
Viability/Mortality: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once weekly
Once prior to the first administration of the test item (day 6 of acclimatization) and weekly thereafter (in the gestation period on day 0, 6, 13 and 20 post coitum), detailed clinical observations were performed outside the home cage in a standard arena. Animals were observed for the following: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies or bizarre behavior were also reported.

BODY WEIGHT: Yes
- Time schedule for examinations: daily, from treatment start to day of necropsy

FOOD CONSUMPTION:
- Food consumption for each animal determined: Males: Pre-pairing period days 1 - 4, 4 - 8, 8 - 11 and 11 – 14, after pairing period days 1 – 7 and 7-10. Females: Pre-pairing period days 1 - 4, 4 - 8, 8 - 11 and 11 - 14; gestation days 0 – 7, 7 14 and 14 – 21 and days 1 - 4 of the lactation. No food consumption was recorded during the pairing period.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: from males on day of necropsy and from females on day 5 post partum
- Anaesthetic used for blood collection: Yes (isofluorane)
- Animals fasted: Yes
- How many animals: 5 males and 5 females from each group
- Parameters checked: Complete Blood Cell Count: Erythrocyte count, Hemoglobin, Hematocrit, Mean corpuscular volume, Red cell volume distribution width, Mean corpuscular hemoglobin, Mean corpuscular hemoglobin concentration, Hemoglobin concentration distribution width, Leukocyte count (total), Differential leukocyte count: Platelet count & Reticulocytes; Coagulation: Prothrombin time (= Thromboplastin time), Activated partial Thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: from males of each group on day of necropsy and from females from each group on day 5 post partum
- Animals fasted: Yes
- How many animals: 5 males and 5 females from each group
- Parameters checked: Glucose, Urea, Creatinine, Bilirubin (total), Cholesterol (total), Triglycerides, Aspartate aminotransferase, Alanine aminotransferase, Alkaline phosphatase, Gamma-glutamyl-transferase, Bile acids, Sodium, Potassium, Chloride, Calcium, Phosphorus, Protein (total), Albumin, Globulin, Albumin/Globulin ratio

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: at one time during the study (males shortly before the scheduled sacrifice and in females of groups 1 and 2 on day 3 post partum and in females of groups 3 and 4 on day 23 or 24 post coitum)
- Dose groups that were examined: 5 males and 5 females randomly selected from each group
- Battery of functions tested:
• Cage-side observations: faeces-balls, urine and posture as well as resistance to removal.
• Hand-held observations: muscle tone, constitution, skin, pupil size, palpebral closure, lacrimation, salivation, reaction to handling and general abnormalities.
• Open field observations: level of ambulatory activity including rearing (one minute evaluation), unusual body movements (e.g. spasms, convulsions), gait evaluation, behavior, hair coat, respiration, quantity of faeces-balls and urine.
• Reflexes: blinking, palpebral closure, pinna reflex, extensor thrust response, paw pinch, responsiveness to sharp noise, righting reflex and hearing ability (Preyer’s reflex).
• Measurements / Counts: hind limb / fore limb grip strength, rectal temperature.

Oestrous cyclicity (parental animals):

examined

Sperm parameters (parental animals):

Parameters examined in male parental generation:
testis weight, epididymis weight

Litter observations:

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
litter size, live births, still births, sex ratio, individual weights on days (0) 1 and 4 post partum

GROSS EXAMINATION OF DEAD PUPS:
yes, for gross anomalies

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals after 28 days of treatment
- Maternal animals: All surviving animals on day 5 post partum

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera

HISTOPATHOLOGY
Testes, epididymides, prostate, seminal vesicles, ovaries, oviduct, vagina and uterus from all animals of the control and high dose group are examined. The same applies to all occurring gross lesions and to all animals, which die spontaneously or must be terminated in extremis. The remaining organs/tissues of 5 randomly selected males and females of the control and high dose group, respectively, are examined histopathologically.

The following tissues were preserved: Prostate, Seminal vesicles with coagulating gland, Testes, Epididymides (males); Ovaries, Uterus (females);
Gross lesions, Brain, Spinal chord (cervical, thoracic, lumbar), Small and large intestines2 (incl. Peyer’spatches), Stomach (forestomach and glandular stomach), Liver, Kidneys, Adrenals, Spleen, Lymph nodes (axillary and mesenteric), Aorta, Eyes with optic nerve and harderian gland, Lacrimal gland,
Heart, Thymus, Thyroids, and parathyroids if possible, Trachea and lungs (preserved by inflation with fixative and then immersion), Pituitary gland, Urinary bladder, Peripheral nerve (sciatic), Bone marrow (femur), Femur with knee joint, Mammary gland (male and female), Pancreas, Salivary glands – mandibular, sublingual, Skeletal muscle, Sternum with bone marrow, Pharynx, Larynx, Nasal cavity, Esophagus


ORGAN WEIGHTS:
At the scheduled sacrifice, the testes and epididymides of all parental males will be weighed separately.
In addition, from 5 males and 5 females killed at the end of the study which were selected for hematology and clinical chemistry examination from each group, the following organs will be trimmed from any adherent tissue, as appropriate, and their wet weight taken: Adrenal glands (weighed as pairs), Brain, Heart, Kidneys (weighed as pairs), Uterus (including cervix), Prostate, Liver, Thymus, Spleen, Thyroid (after fixation), Ovaries (weighed as pairs), Seminal vesicles (inclusive coagulating gland)

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring were sacrificed at day 4 post partum.

GROSS NECROPSY
- Gross necropsy consisted of examination for gross anomalies.

Statistics:

The following statistical methods will be used to analyze food consumption, body weights and reproduction data:
• Means and standard deviations of various data will be calculated and included in the report.
• The Dunnett-test [see References (2)] (many to one t-test) based on a pooled variance estimate will be applied if the variables can be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test [see References (3)] (many-one rank test) will be applied instead of the Dunnett-test when the data cannot be assumed to follow a normal distribution.
• Fisher's exact-test [see References (4)] will be applied if the variables can be dichotomized without loss of information.

Reproductive indices:

Percentage mated = (Females mated/Females paired)*100
Fertility index = (Females achieving a pregnancy/Females paired) *100
Conception rate = (Females achieving a pregnancy/Females mated)*100
Gestation index = (Number of females with living pups/Females pregnant)*100

Offspring viability indices:

Birth index = (pups born alive/number of implantations)*100
Viability index = (pups alive before cullig on day 4 post partum/pups born alive)*100

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

no unplanned deaths; for more informations see details on results.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

see details on results

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

see details on results

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

see details on results

Other effects:

not specified

Reproductive function: oestrous cycle:

effects observed, treatment-related

Description (incidence and severity):

see details on results

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
There were no unplanned deaths.
In two males at 400 mg/kg bw/day, slightly ruffled fur was noted during six consecutive days during pairing. This was accompanied by reddish nasal secretion in both animals for two consecutive days. One male at 50 mg/kg bw/day also showed one instance of reddish nasal secretion towards the middle of the after pairing. No further clinical signs were noted in males at these dose levels.
Due to the low number of animals affected, it could be deemed that this effect was not adverse although it cannot be excluded that it was a signs of discomfort associated with treatment with the test item.
Incidentally, in one male at 400 mg/kg bw/day a left eye slightly reduced in size was noted from the last two days of pre-pairing onwards.
No clinical signs were noted in males at 0 or 150 mg/kg bw/day.
At 400 mg/kg bw/day, one female showed bedding in mouth and salivation on days 5 to 7 of gestation, which was similar to the males and considered rather as sign of discomfort associated with treatment with the test item than an adverse effect of the test item.
One female at 150 mg/kg bw/day had vaginal bleeding on the last day of pairing and the first day of gestation period. This isolated finding and vaginal to this early time point was considered to be incidental.
In females, no clinical signs were noted at 0 and 50 mg/kg bw/day.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
In males treated at 400 mg/kg bw/day, body weights and body weight gain were lower than controls during the major part of the study after onset of treatment at the beginning of the pairing period. This effect reached statistical significance on several occasions during the study. This was consistent with the statistically significant reduction in food consumption noted in males during pre-pairing.
The overall differences in mean body weight gain at the dose levels of 0, 50, 150 and 400 mg/kg bw/day were: +11%, +12%, +11% and +6% during the pre-pairing period, +10%, +10%, +11% and +8% during the pairing period and +1%, +2%, +3% and +3% during the after pairing period (percentages refer to the body weight gain within the period).
Since no females at 150 and 400 mg/kg bw/day gave birth, they were excluded from the evaluation during gestation period and a comparison of these groups with controls was only possible during the pre-paring period.
In females at 400 mg/kg bw/day, reduced body weight gain was noted from the beginning of pre-pairing onwards. This was similar to the effect in males at this dose level, although in females no statistically significant reduction in body weights was noted.
In females at 50 mg/kg bw/day, there was a tendency towards higher body weights when compared with controls however mean body weight gain was similar in the gestation period and reduced in the gestation period (statistically significant on single days). Higher body weights were considered to be not an effect of treatment with the test item, since the difference was already noted during acclimatization and the start of pre-pairing.
The overall differences in mean body weight gain at the dose levels of 0, 50, 150 and 400 mg/kg bw/day were +6%, +7%, +8% and +4% during the pre-pairing period, +59% and +52% during gestation and +3% and +3% during the lactation period (percentages refer to the body weight gain within the period) at a dose level of 0 and 50 mg/kg bw/day.

In males treated at 400 mg/kg bw/day, mean food consumption was statistically significantly lower than controls during the major part of pre-pairing (up to -17% between days 4 to 8). However, towards the end of pre-pairing and during the remainder of the study, no statistically significant effect on food consumption was noted in males at 400 mg/kg bw/day. Therefore, this transient reduction in food consumption was deemed not to be adverse. No effect of treatment was noted in males at 150 and 50 mg/kg bw/day.
Since no females at 150 and 400 mg/kg bw/day gave birth, they were excluded from the evaluation during gestation period and a comparison of these groups with controls was only possible during the pre-paring period.
At 400 mg/kg bw/day during pre-pairing, a statistically significant reduction in food consumption (up to 20.5% between days 4 to 11) was noted. Since the reduction was less marked towards the end of pre-pairing and in analogy with the findings in males it can be deemed that this effect would have been reversible and not adverse. No effect of treatment was noted in females at 150 and 50 mg/kg bw/day.

REPRODUCTIVE FUNCTION: FEMALES (PARENTAL ANIMALS)
No effects on mating performance and fertility were observed at any dose level.
Two females each from the control group and at 50 mg/kg bw/day were not pregnant, as well as three females at 300 mg/kg bw/day and one female at 400 mg/kg bw/day. As a result the fertility index in was 83.3% in controls and at 50 mg/kg bw/day, 75.0% at 150 mg/kg bw/day and 91.7% at 400 mg/kg bw/day.

DURATION OF GESTATION
The mean duration of gestation was unaffected by exposure to the test item at 50 mg/kg bw/day.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
No effects on mating performance and fertility were observed at any dose level.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
No pairing was noted during the first pairing period for one female (no. 76) at 150 mg/kg bw/day and therefore these females were paired with a second male. A body weight gain of 32% indicating pregnancy was observed in this female on day 5 of the second pairing period and therefore pairing of this female was stopped. All other females were mated within the first pairing period.
Two females each from the control group and at 50 mg/kg bw/day were not pregnant, as well as three females at 300 mg/kg bw/day and one female at 400 mg/kg bw/day. As a result the fertility index in was 83.3% in controls and at 50 mg/kg bw/day, 75.0% at 150 mg/kg bw/day and 91.7% at 400 mg/kg bw/day.

ORGAN WEIGHTS (PARENTAL ANIMALS)
In males, statistically significantly reduced absolute testes and epididymides were noted at a dose level of 400 mg/kg bw/day. Calculated relative to body weights, the epididymides weights were also statistically significant reduced. The values for testes and epidymides were in the range of the historical control data (at the lower limit) but due to histopathological findings in testes and epididymides, these reductions were considered to be test item-related.
In females, no test item-related effects on organ weights were noted.

GROSS PATHOLOGY (PARENTAL ANIMALS)
At 150 and 400 mg/kg bw/d, testes were flaccid and/or reduced in size in single males, which was considered to be test item-related. One male at 400 mg/kg bw/d, had a pale discolored liver, which correlates to his high values of aspartate aminotransferase and alanine aminotransferase.
All other findings in males and females were typical of this strain and age of rat and were considered to be incidental.

HISTOPATHOLOGY (PARENTAL ANIMALS)
At 50, 150 and 400 mg/kg bw/day in the heart, a minimal to moderate multifocal dose-dependent cardiomyopathy (myocardial necrosis and/or degeneration with mononuclear inflammatory infiltration, and often accompanied by fibrosis/fibroplasias) was observed in the majority male and female rats. The males showed higher incidence and severity than the females.
Minimal to moderate foamy macrophages accumulation was observed at 50, 150 and 400 mg/kg bw/day in the lungs. The distribution of the lesion was focal to multifocal, and was mainly in the alveolar spaces around bronchioles and/or in terminal bronchioles. The males appeared to be more affected in incidence and severity.
Minimal to moderate increase in the formation of the residual bodies of abnormal shape and size was observed at 150 and 400 mg/kg bw/day. In the epididymides, the exfoliated residual bodies (cell debri) were noted at 400 mg/kg bw/day.

IMPLANTATION RATE AND POST-IMPLANTATION LOSS
The mean number of implantations per dam was in the range of the historical control data.
At a dose level of 150 and 400 mg/kg bw/day, no female gave birth. One and two females, respectively, had only implantation sites, all other pregnant females had only embryonic resorptions. The gestation index was therefore 0% at 150 and 400 mg/kg bw/day in contrast to 100% in the control group and at 50 mg/kg bw/day. This absolute post implantation loss was considered to be test item-related.
At 50 mg/kg bw/day a higher mean incidence of post-implantation loss was noted (19.1% compared to 11.3% in the control group; calculated as a percentage of total implantations; total 29 losses versus 16 losses). This higher post implantation loss did not reach statistical significance but was outside of the range of the historical control data. Due to total post implantation loss at 150 and 400 mg/kg bw/day, it cannot be excluded that this was a test item-related effect.

Dose descriptor:

LOAEL

Remarks:

developmental toxicity

Effect level:

<= 50 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: higher mean incidence of post-implantation loss

Dose descriptor:

LOAEL

Remarks:

systemic

Effect level:

<= 50 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Specific target organ toxicity: heart (cardiomyopathy)

Dose descriptor:

NOAEL

Remarks:

fertility

Effect level:

150 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: Reduced testis and epididymis weights

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

No Milk in the stomach one litter /5M (50mg/kg bw/d)

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

see details on results

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

see details on results

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

LITTER SIZE AND FIRST LITTER CHECK
No effects on litter size were observed at 50 mg/kg bw/day.
No dead pups were found at first litter check in either of those two groups.

POSTNATAL LOSS DAYS 0-4 POST PARTUM
There was no effect on postnatal loss at 50 mg/kg bw/day.

LITTER DATA
Only data from controls and females treated at 50 mg/kg bw/day were reported since females treated at 150 and 400 mg/kg bw/day did not give birth to any pups. No abnormal findings were noted at first litter check or during the first 4 days post partum in controls and at 50 mg/kg bw/day. Sex ratios at first litter check and on day 4 post partum were unaffected by exposure to the test item.
Mean pup weights on days 1 and 4 post partum were unaffected by treatment with the test item. The body weights of male pups from females treated at 50 mg/kg bw/day were slightly and statistically significantly lower than controls on day 1 post partum. However, when compared with the historical control data in Appendix VIII it can be noted that body weights of pups at 50 mg/kg bw/day are well within the range of historical data, whereas body weights of control pups are rather at the upper range of historical control range.
No test item-related findings were noted at macroscopic examination of F1 pups. The stomach of one control pup contained a watery fluid.

Dose descriptor:

LOAEL

Generation:

F1

Effect level:

<= 50 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

mortality

Remarks on result:

other: higher mean incidence of post-implantation loss

Reproductive effects observed:

not specified

Conclusions:

Based on the higher mean incidence of post-implantation loss a LOAEL of 50 mg/kg bw/d was determined.

Executive summary:

A study investigating the toxicological effects of the test item Amines, tri-C8-10-alkyl to rats according to OECD guideline 422 resulting from repeated oral-gavage administration is performed. Amines, tri-C8-10-alkyl was administered in corn oil as vehicle at dosages of 50, 150 and 400 mg/kg body weight/day, and controls received the vehicle only. Amines, tri-C8-10-alkyl was administered to male rats for 43 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum.

All animals survived until scheduled necropsy. There were no test item related clinical signs in both genders at 50 mg/kg bw/day. At 150 mg/kg bw/day, no signs of discomfort were observed; food consumption and body weights were similar to controls. A reduction in food consumption and body weight was noted in males and females at 400 mg/kg bw/day when compared with controls. Furthermore, higher values of aspartate aminotransferase, alanine aminotransferase and of alkaline phosphatase were noted in both genders at 400 mg/kg bw/day, without a histopathological correlate in the liver.

Mean precoital time, fertility index and conception rate were not affected by the treatment with the test item at any dose level. At a dose level of 150 and 400 mg/kg bw/day, no female gave birth. In both dose groups, a post implantation loss of 100% was noted, since all pregnant females had either only implantation sites or only embryonic resorptions. At 50 mg/kg bw/day, a higher mean incidence of post-implantation loss was noted.

In males, reduced testes and epididymides weights were noted at a dose level at 400 mg/kg bw/day.

In all treatment groups (50, 150 and 400 mg/kg bw/day), cardiomyopathy was noted in the heart, and foamy macrophages accumulation was observed in the lungs.

The mean number of pups at first litter check was not affected by the treatment with the test item. The sex ratio was also not affected. No abnormal pup was noted at any dose level. No effects on pup weight and pup weight gain were observed. At necropsy of pups, there were no abnormal findings.

Based on these results, no NOAEL (No Observed Adverse Effect Level) could be established for general toxicity and reproduction in this study. Nevertheless, accordingly to the applicant a LOAEL for systemic toxicity of 50 mg/kg bw/d, based on cardiomyopathy in males, and a LOAEL for reproduction/developmental toxicity of 50 mg/kg bw/day, based on the higher mean incidence of post-implantation loss, has to be established and used for calculation.

Effect on fertility: via oral route

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

150 mg/kg bw/day

Study duration:

subacute

Species:

rat

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Experimental data are available to assess the effects on fertillity Amines, tri-C8-10-alkyl. A study investigating the toxicological effects of the test item Amines, tri-C8-10-alkyl to rats according to OECD guideline 422 resulting from repeated oral-gavage administration is performed. Amines, tri-C8-10-alkyl was administered in corn oil as vehicle at dosages of 50, 150 and 400 mg/kg body weight/day, and controls received the vehicle only. Amines, tri-C8-10-alkyl was administered to male rats for 43 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum. All animals survived until scheduled necropsy. There were no test item related clinical signs in both genders at 50 mg/kg bw/day. At 150 mg/kg bw/day, no signs of discomfort were observed; food consumption and body weights were similar to controls. A reduction in food consumption and body weight was noted in males and females at 400 mg/kg bw/day when compared with controls. Furthermore, higher values of aspartate aminotransferase, alanine aminotransferase and of alkaline phosphatase were noted in both genders at 400 mg/kg bw/day, without a histopathological correlate in the liver. Mean precoital time, fertility index and conception rate were not affected by the treatment with the test item at any dose level. At a dose level of 150 and 400 mg/kg bw/day, no female gave birth. In both dose groups, a post implantation loss of 100% was noted, since all pregnant females had either only implantation sites or only embryonic resorptions. At 50 mg/kg bw/day, a higher mean incidence of post-implantation loss was noted. In males, reduced testes and epididymides weights were noted at a dose level at 400 mg/kg bw/day. In all treatment groups (50, 150 and 400 mg/kg bw/day), cardiomyopathy was noted in the heart, and foamy macrophages accumulation was observed in the lungs. The mean number of pups at first litter check was not affected by the treatment with the test item. The sex ratio was also not affected. No abnormal pup was noted at any dose level. No effects on pup weight and pup weight gain were observed. At necropsy of pups, there were no abnormal findings. Key study assignment: As there is only one reliable and relevant study investigating the reproductive toxicity of Amines, tri-C8-10-alkyl.this study is integrated as key study. Assessment of toxicity on reproduction: Based on these results, no NOAEL (No Observed Adverse Effect Level) could be established for general toxicity and reproduction in this study. Nevertheless, a dose response correlation could be observed showing that, for example, the post implantation losses decrease lowering the dosage. Unfortunatley no lower dose than 50mg/kg bw/d was tested but using the trend observed in effects an extrapolation may be suitable. Therefore the applicant corrected the LOAEL for systemic toxicity of 50 mg/kg bw/d, based on cardiomyopathy in males, by an additional safety factor of 10 to receive a NOAEL. The same is assumed for the effects above. The risk assessment and classification will be based on the assumed NOAEL of 5 mg/kg bw/d. The effects observed above are effects concerning the fertility/developmental. Based on these findings the substance will be classified for reproduction toxicity (fertility/developmental).  

Short description of key information:
- subacute toxicity / reproduction screening, rat, OECD 422 (BASF SE, 2013, 85R0674/12X380):
NOAEL reproduction (fertility) toxicity: 150 mg/kg bw/day (reduced testis and epididymis weights)
LOAEL reproduction (developmental) toxicity: 50 mg/kg bw/day (higher mean incidence of post-implantation loss)
LOAEL systemic: 50 mg/kg bw/day (Spefici target organ toxicity: heart (cardiomyopathy)

Justification for selection of Effect on fertility via oral route:
only avaialable result

Effects on developmental toxicity

Description of key information

- subacute toxicity / reproduction screening, rat, OECD 422 (BASF SE, 2013, 85R0674/12X380): 
NOAEL reproduction (fertility) toxicity: 150 mg/kg bw/day (reduced testis and epididymis weigh)
LOAEL reproduction (developmental) toxicity: 50 mg/kg bw/day (higher mean incidence of post-implantation loss)
LOAEL systemic: 50 mg/kg bw/day (Spefici target organ toxicity: heart (cardiomyopathy)

Effect on developmental toxicity: via oral route

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LOAEL

50 mg/kg bw/day

Study duration:

subacute

Species:

rat

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

Experimental data are available to assess the effects on developmental for Amines, tri-C8-10-alkyl.

A study investigating the toxicological effects of the test item Amines, tri-C8-10-alkyl to rats according to OECD guideline 422 resulting from repeated oral-gavage administration is performed. Amines, tri-C8-10-alkyl was administered in corn oil as vehicle at dosages of 50, 150 and 400 mg/kg body weight/day, and controls received the vehicle only. Amines, tri-C8-10-alkyl was administered to male rats for 43 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum. All animals survived until scheduled necropsy. There were no test item related clinical signs in both genders at 50 mg/kg bw/day. At 150 mg/kg bw/day, no signs of discomfort were observed; food consumption and body weights were similar to controls. A reduction in food consumption and body weight was noted in males and females at 400 mg/kg bw/day when compared with controls. Furthermore, higher values of aspartate aminotransferase, alanine aminotransferase and of alkaline phosphatase were noted in both genders at 400 mg/kg bw/day, without a histopathological correlate in the liver.Mean precoital time, fertility index and conception rate were not affected by the treatment with the test item at any dose level. At a dose level of 150 and 400 mg/kg bw/day, no female gave birth. In both dose groups, a post implantation loss of 100% was noted, since all pregnant females had either only implantation sites or only embryonic resorptions. At 50 mg/kg bw/day, a higher mean incidence of post-implantation loss was noted.In males, reduced testes and epididymides weights were noted at a dose level at 400 mg/kg bw/day.In all treatment groups (50, 150 and 400 mg/kg bw/day), cardiomyopathy was noted in the heart, and foamy macrophages accumulation was observed in the lungs. The mean number of pups at first litter check was not affected by the treatment with the test item. The sex ratio was also not affected. No abnormal pup was noted at any dose level. No effects on pup weight and pup weight gain were observed. At necropsy of pups, there were no abnormal findings.Key study assignment:As there is only one reliable and relevant study investigating the reproductive toxicity of Amines, tri-C8-10-alkyl.this study is integrated as key study.Assessment of toxicity on reproduction:Based on these results, no NOAEL (No Observed Adverse Effect Level) could be established for general toxicity and reproduction in this study. Nevertheless, a dose response correlation could be observed showing that, for example, the post implantation losses decrease lowering the dosage. Unfortunatley no lower dose than 50mg/kg bw/d was tested but using the trend observed in effects an extrapolation may be suitable. Therefore the applicant corrected the LOAEL for systemic toxicity of 50 mg/kg bw/d, based on cardiomyopathy in males, by an additional safety factor of 10 to receive a NOAEL. The same is assumed for the effects above. The risk assessment and classification will be based on the assumed NOAEL of 5 mg/kg bw/d. The effects observed above are effects concerning the fertility/developmental. Based on these findings the substance will be classified for reproduction toxicity (fertility/developmental). 

Justification for selection of Effect on developmental toxicity: via oral route:
see OECD 422 screening study

Justification for classification or non-classification

Based on these results, no NOAEL could be established for reproduction and developmental toxicity in this study. The LOAEL was 50mg/kg bw/kg, which was the lowest concentration tested. It cannot be excluded, that the test substance will cause adverse effects in concentrations lower than 50mg/kg bw/day. Nevertheless as the a trend in dose response relation concerning postimplantattions loses could be observed (decrease in loses with decrease dosage) an additional saftey factor correcting the LOAEL to a NOAEL of 10 is assumed to be suitable. As the effects seen are effects on fertility and developmental (postimplantation losses, reduced testis and epididymes weighs) the substance is labeled for reproduction and developmental toxicity as follows:

GHS: Repr. Cat. 1B, Reproductive toxicity, H360: May damage fertility or the unborn child,

DSD: Repr. Cat. 2, Reproductive toxicity, R60: May impair fertility; R61 May cause harm to the unborn child.