Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

There is no study avalaible for this substance. Read across is made with hydrogenated tallow alkylamines inside the primary alkylamines category.

A read-across with tallow alkylamines is applied through anOECD TG 421 in which no reproductive toxicity effect is observed.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study according to GLP
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Italia
- Age at study initiation: (P) 8 wks
- Weight at study initiation: (P) Males: 225 - 250 g; Females: 200 - 225 g
- Fasting period before study: no
- Housing: macrolon cages
- Use of restrainers for preventing ingestion (if dermal): no
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2 °C
- Humidity (%): 55 +/- 5 %
- Air changes (per hr): 15 -20
- Photoperiod (hrs dark / hrs light): 12 hours periodically
Route of administration:
oral: gavage
Vehicle:
other: sesame oil
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: 16 h, 7 evenings/week for max. 14 evenings
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged: individually
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
Males: 14 days prior to mating, during mating; 28 days maximum
Females: 14 days prior to mating, during mating, during pregnancy, 3 days post-partum (lactation)
Females, positive smear, no offspring: 14 days prior to mating, during mating, 25 days post-mating
Females, negative smear: 14 days prior to mating, during mating, 25 days post-mating
Remarks:
Doses / Concentrations:
mg/kg body weight per day
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
12.5 mg/kg body weight per day
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
50 mg/kg body weight per day
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
150 mg/kg body weight per day
Basis:

No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on dose-range-finder in rats
- Rationale for animal assignment (if not random): random
- Other:
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data

OTHER:
Sperm parameters (parental animals):
Sections of testes were stained with PAS-hematoxylin to allow spermatogenesis to be classified into 14 stages, each stage based primariliy upon the changes of the acrosome and head morphology of the younger generation of spermatids.
Litter observations:
number and sex of pups
stillbirths
live births
postnatal mortality
presence of gross anomalies
weight gain (day 0, day 4)
gross examination of dead pups for external and internal abnormalities
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals on the day after the end of treatment
- Maternal animals: All surviving animals on day 4 of lactation with their pubs

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

HISTOPATHOLOGY / ORGAN WEIGHTS
The following tissues were prepared for microscopic examination and weighed, respectively:
- ovaries
- uteri (including horns, cervix, and vagina)
- testes (with special emphasis on stages of spermatogenesis and histopathology of interstitial cell structure)
- epididymides
- accessory sex organs
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed on day 4 of lactation and necropsied

GROSS NECROPSY
- Gross necropsy consisted of external examinations including the cervical, thoracic, and abdominal viscera.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
salivation, hunched posture at mid- and high dose animals, no changes at low-dose animals
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
decrease in mid- and high dose animals, no effect in low-dose animals
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
decrease in mid- and high dose animals, no effect in low-dose animals
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
effects observed, treatment-related
Description (incidence and severity):
Test substance intake: lower mean food consumption in mid- and high-dose animals, no effect in low-dose animals
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
decreased mating and fertility indices in high-dose animals
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
- 150 mg/kg: salivation, hunched posture, soft stools, piloerection; 6/10 male and 5/10 female animals died
- 50 mg/kg: salivation; 1/10 male and 1/10 female animals died
- 12.5 mg/kg: no changes observed
BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
- 150 mg/kg: decreased body weight gain (males and females)
- 50 mg/kg: decreased body weight gain during pre-mating period
- 12.5 mg/kg: slightly decreased body weight gain (males and females)
TEST SUBSTANCE INTAKE (PARENTAL ANIMALS)
- lower food intake in mid and high dose groups (males and females), slightly lower food intake in low dose group during lactation period in females
REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
- mating and fertility indices, mean pre-coital time and parturition were unaffected up to 50 mg/kg/day
ORGAN WEIGHTS (PARENTAL ANIMALS)
- lower absolute weight of epididymides and higher value of testis weight relative to body weight at 150 mg/kg/day
- no differences in ovary weights among the experimental groups
GROSS PATHOLOGY (PARENTAL ANIMALS)
- no pathological changes
HISTOPATHOLOGY (PARENTAL ANIMALS)
- histology of testes, epididymides and ovaries of high dose group animals did not show any compound-related changes
- no changes in testicular staging performed in the PAS-hematoxylin stained sections of control and high dose groups
OTHER FINDINGS (PARENTAL ANIMALS)
Dose descriptor:
NOEL
Effect level:
12.5 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: overall effects
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
no effects up to 50 mg/kg body weight
Mortality / viability:
mortality observed, treatment-related
Description (incidence and severity):
no litters present at highest dose
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
slightly lower pub weight at 50 mg/kg body weight
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
VIABILITY (OFFSPRING)
- no litters present at 150 mg/kg/day
- no effects on postnatal survival up to day 4 of lactation and no effects on sex ration at 50 mg/kg/day

CLINICAL SIGNS (OFFSPRING)
- no litters present at 150 mg/kg/day
- no significant effects up to 50 mg/kg/day

BODY WEIGHT (OFFSPRING)
- no litters present at 150 mg/kg/day
- slightly lower pup body weight seen in 50 mg/kg/day group
- no effects seen at 12.5 mg/kg/day

GROSS PATHOLOGY (OFFSPRING)
- no abnormalities observed in any pub either at birth or at autopsy on day 4 of lactation neither of the 50 or of the 12.5 mg/kg/day group
Dose descriptor:
NOEL
Generation:
F1
Effect level:
12.5 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: overall effects
Reproductive effects observed:
not specified
Conclusions:
Based on findings of general toxicity (death, clinical signs, reduced body weight gain) at daily dosages of 50 mg/kg bw/d a NOEL/systemic toxicity of 12.5 mg/kg bw/d can be derived from the study. Based on the findings of a lower fertility index and a lower conception rate at daily dosages of 50 mg/kg bw/d a NOEL/fertility of 12.5 mg/kg bw/d can be derived from the results of this screening study.

Based on the results of this study, 12.5 mg/kg/day is the NOEL for maternal toxicity and the NOEL for the offspring.
Executive summary:

Groups of 10 rats (Crl:CD (SD) BR) per sex were treated with dosages of 0, 12.5, 50, and 150 mg/kg bw/day of Genamin TA 100 by gavage (administration volume 10 ml/kg bw/day) using sesame oil as a vehicle. Males were treated daily from 14 days prior to mating until the end of the mating period for a maximum of 28 days. Females were treated daily for 14 days before start of the mating period, throughout the same, during pregnancy and until day 3 of lactation. The animals were mated one male with one female.

At daily doses of 150 mg/kg bw 6/10 males and 5/10 females died between day 9 and day 25 of treatment (during the premating and mating period). At daily doses of 50 mg/kg bw 1/10 males and 1/10 females died on day 13 respectively on day 24 of treatment. No animals died at 12.5 mg/kg bw/day, and in the control group 1 female died by accident.

Clinical observations at 150 mg/kg bw/day revealed salivation after treatment, hunched posture and in some cases soft stools and piloerection. The only clinical sign present at 50 mg/kg bw/day was salivation. No changes were seen at 12.5 mg/kg bw/day.

Body weight loss of about 22 g at the 150 mg/kg bw dose group and statistically significant (p > 0.01) lower body weight gain at the 50 mg/kg bw dose group during the premating period together with a lower mean food consumption was observed in the males. Also in the females body weight loss of about 17 g at the 150 mg/kg bw dose group and statistically significant lower body weight gain at the 50 mg/kg bw dose group together with a lower mean food consumption during the premating period was observed.

At sacrifice of the parental animals organ weights of ovaries, testes and epididymides were determined from all experimental groups. Histopathology was carried out on testes, epididymides and ovaries of the controls and of the animals of the 150 mg/kg bw dose group. A statistically significant(p> 0.01) lower absolute and relative weight of epididymides and a statistically significant(p> 0.01) higher value of relative testis weight was observed at 150 mg/kg bw/day. No significant differences were noted in ovary weights among the various experimental groups. Histopathology of testes and epididymides of high dose group animals did not show any compound-related changes. In particular, no changes were seen in the testicular staging performed in the PAS-hematoxylin stained sections of final sacrificed animals of control and of the high dose groups. In the ovaries, moderate increased frequency of atrophic corpora lutea was seen in the animals of the high dose group which died or were sacrificed, compared to the controls, this finding was considered a secondary effect to the decrease in body weight growth induced by treatment in this group.

At the dosages of 150 mg/kg bw/day only 3 females out of 7 mated females had positive vaginal smears and of these only one was pregnant, but with only implantation sites and no live pups. At this dosage level also the mean pre-coital interval was longer (13.4 days) than that of the control group (2.3 days). With regard to the mating index in the control, the 12.5 and the 50 mg/kg bw dose level groups 9/9 (100%), 9/10 (90%) and 9/9 (100%) mated females were sperm positive. With regard to the fertility index 9/9 (100%), 8/10 (80%) and 7/9 (78%) of the mated females became pregnant in the control, the 12.5 and the 50 mg/kg bw dose level groups. Mean pre-coital time and parturition were unaffected in the low and mid dose treated groups.

With regard to the conception rate in the control, the 12.5 and the 50 mg/kg bw dose level groups 9/9 (100%), 8/9 (89%) and 7/9 (78%) of the sperm positive females became pregnant and delivered live pups. Numbers of corpora lutea had not been determined during this study. Thus, any pre-implantation loss was not evaluated. Staining for the presence of implantation sites was only performed with the uteri of apparently non-pregnant females. The mean number of visible implantation sites per dam in the control, the 12.5 and the 50 mg/kg bw dose level groups were 17.6, 13.9 and 15.0. There were no stillborns or litters with only implantations in the control, low, and mid dose groups. The mean number of total pups born per litter was 16.9, 12.3 and 14.3 in the control, the 12.5 and the 50 mg/kg bw/day dose groups. Thus, the mean litter index for post-implantation loss was calculated to 3.4, 9.9 and 4.5% in the control, the low and the mid dose groups.

No effects were noted on the pup sex ratio. No abnormalities were observed in any pup either at birth or at autopsy on day 4 of lactation neither in the 12.5 nor in the 50 mg/kg bw/day treated groups. Lower mean values of live born per litter and of pups per litter alive at day 4 were found for both the low and the intermediate dose groups in comparison to those of the control group, however, the decreases were not dose dependent and the differences were statistically significant for the 12.5 mg/kg bw dose group only. Thus, these findings were considered unlikely to be related to the compound. A slightly lower pup body weight was observed in the 50 mg/kg bw/day treated group in comparison with the control group at birth and at day 4 of lactation but not for the pups of the 12.5 mg/kg bw/day treated group. Based on general toxicity findings a NOEL(systemic toxicity) of 12.5 mg/kg/day can be derived from stis study. Additionally, based on findings of a lower fertility index and a slightly lower conception rate at the mid dose level a NOEL(fertility) of 12 .5 mg/kg body weight per day is deducible.Further, during an oral 28-day study (OECD TG 407) with the same test substance Genamin TA 100 in rats (see section 7.5.1) also reproductive organ weights and reproductive organ histopathology of testes and epididymides, as well as organ weights of ovaries and uteri had been investigated. No substance-related findings had been observed during this study on these organs at oral doses up to and including 150 mg/kg bw/day.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
35.44 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The study is GLP compliant and the read-across is valid.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

The registered substance is an Alkyl phosphate and stearyl amine salts and is mainly composed by primary alkylamines, Monoalkyl dihydrogenophosphate and Dialkyl hydrogenophosphate salts. As the primary alkyl amine part is much more toxic than the alkyl di/hydrogenophosphates part, it is expected that the toxicological behaviour of the substance will not be far different from that of the primary alkylamine compounds or at least not more severe. Thus, assessment of the registered substance can be integrated in thecurrent approach of grouping of primary alkylamines substances. Regarding the reproductive toxicity with the registered substance on fertility, respectively on reproductive function, no experimental studies are available. However, data relevant for hazard assessment with regard to this endpoint, are available from an OECD TG 421 study as well as from repeated oral dose toxicity studies with histopathological investigation of the reproductive organs for related primary alkylamines, which are considered appropriate for extrapolation. In a study according to OECD TG 421 with tallow alkylamines, findings of general toxicity in close accordance to the results from the 28 -day repeated dose toxicity study with the same compound were obtained at identical dose levels. Since severe general toxicity due to the strong irritating / corrosive properties appears to be the predominating substance-related toxic effect, and since no histological compound-related changes were observed on testes, epididymides and ovaries, neither in the OECD 421 nor in the OECD 407 study, even at the highest dose of 150 mg/kg body weight per day tested, a specific substance-related effect on reproduction performance is not deducible. Signs of general toxicity without indications on the reproductive behaviour were also noted at the intermediate dose-level of 50 mg/kg body weight per day. No toxic effects were observed at the low-dose treatment with 12.5 mg/kg body weight per day which therefore can be considered to be a NOAEL (fertility). This conclusion is in line with the evaluation coming from the existing EU risk assessment on primary alkylamines. Additionally, according to the EU risk assessment further testing of primary alkylamines with regard to the endpoint fertility was not considered necessary.

As the registered substance is a Alkyl phosphate and stearyl amine salts with a molar ratio of 1:1, the NOAEL value should be converted as follows: Mw tallow alkylamine = 267, MW salt = 757 ; 12.5 x 757/267 = 35.44 mg/kg bw/day



Short description of key information:
No human data are available for any primary alkylamine indicative of potential toxicity to reproduction. Additionally, no animal data on this endpoint are available for hydrogenated tallow alkylamines. However, data from closely related primary alkylamines can be used as read-across tor assessment purposes. The results from an OECD TG 421 study with tallow alkylamines as well as from detailled histopathology of the reproductive organs of repeated-dose toxicity studies did not provide evidence for adverse effects on the gonads or any indications for a specific toxic potential adverse to fertility. Based on the results from these studies a NOAEL for fertility of 12.5 mg/kg body weight per day is derived on a read-across basis for hydrogenated tallow alkylamines, which is in line with the conclusion from the existing EU risk assessment on primary alkylamines in general.

Effects on developmental toxicity

Description of key information

Developmental Toxicity studies on rabbits by oral route (OECD 414, GLP) based on read-across from tallow alkyl amine (values converted on mw basis from tallow alkyl amine to the registered salt ): NOAEL maternal toxicity = 85 mg/kg bw/day, NOAEL developmental toxicity > 85 mg/kg bw/day. OECD 414 with tallow alkyl amine in rat is also available with higher NOAELs.

Effect on developmental toxicity: via oral route
Dose descriptor:
NOAEL
30 mg/kg bw/day
Additional information

The registered substance is an Alkyl phosphate and stearyl amine salts and is mainly composed by primary alkylamines, Monoalkyl dihydrogenophosphate and Dialkyl hydrogenophosphate salts. As the primary alkyl amine part is much more toxic than thealkyl di/hydrogenophosphates part, it is expectedthat the toxicological behaviour of the substance will not be far different from that of the primary alkylamine compounds or at least not more severe. This has been demonstrated by results obtained with the registered substance in a short-term repeated toxicity study (e.g. 14 days). Thus, assessment of the registered substance can be integrated in thecurrent approach of grouping of primary alkylamines substances. Experimental studies with the registered substance with respect to the endpoint developmental toxicity are not available. However, data from guideline according prenatal developmental toxicity testing in two species (rats, rabbits) is available for the closely related primary alkylamine (Z)-octacec-9 -enylamine (CAS No. 112 -90 -3). From the rat study a NOAEL (maternal toxicity) of 10 mg/kg body weight per day and a NOAEL (developmental toxicity) of greater 80 mg/kg body weight was derived. In addition, a guideline conform teratology study in rabbits with (Z)-octadec-9 -enylamine revealed general findings of irritation in the gastrointestinal tract and dose-dependent body weight loss or reduced weight gain during the treatment in the 30 mg/kg dose group. Caesarean section data did not reveal any significant differences in reproductive parameters and no indications of an embryotoxic, fetotoxic or teratogenic effect at any tested dose-level. A NOAEL (developmental toxicity) of greater 30 mg/kg body weight per day was retained.

When converted to the corresponding values for the registered salt, the levels become:

- NOAEL maternal toxicity = 85 mg/kg bw/day 

- NOAEL embryo-fetal toxicity = 85 mg/kg bw/day

 (mw tallow alkyl amine = 267, mw registered substance = 757; Correction factor: 757/267 = 2.83)  

GLP study with the lowest NOAELs 

Justification for selection of Effect on developmental toxicity: via inhalation route: The registered substance is a solid with no expected inhalable particles and a neglectable vapour pressure (<<10 -6) at 20°C . Also the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be unlikely to occur. In addition, considering the apparently low systemic toxicity of the primary alkylamines, no need for further inhalative testing was seen. 

Justification for selection of Effect on developmental toxicity: via dermal route: The registered substance as a solid and large salt is not expected to pass the skin. Therefore, the dermal route is not an adapted route of testing.

Justification for classification or non-classification

Reproductive toxicity studies on hydrogenated tallow alkylamines are not available. However, data relevant for hazard assessment with respect to the endpoints fertility and developmental toxicity are available for the closely related primary alkylamines (Z)-octadec-9 -enylamine (CAS No. 112 -90 -3) and tallow alkylamines (CAS No. 61790 -33 -8). In line with the existing EU risk assessment on primary alkylamines it is considered approriate to extrapolate from these data and therefore read-across is considered adequate for hazard assessment. Also in line with the existing EU risk assessment, additional testing is not regarded to be necessary. Thus, according to the results from the available studies classification and labelling of hydrogenated tallow alkylamines with regard to reproductive toxicity is not warranted as it is not for all other primary alkylamines of this category.

Additional information