Reaction mass of ethoxylated N-oleyl-1,3-propanediamine, hydrofluorides of and ethoxylated N-palmityl-1,3-propanediamine, hydrofluorides of and ethoxylated N-stearyl-1,3-propanediamine, hydrofluorides of and olaflur

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

In a Segment I, II and III approach, which in general cover all aspects investigated in developmental and one generation studies, the NOAEL for developmental toxicity, as well as reproductive and fertility performance of Olaflur was established to be at 25 mg/kg bw/day. There were no adverse effects attributable to Olaflur, only some slight toxic effects occurred. Thus, the NOAEL for reproductive toxicity by far exceeds the NOAEL for systemic toxicity derived from a chronic toxicity study and clearly indicates that Olaflur does not represent a specific concern with regard to reproductive or developmental toxicity.

Effect on fertility: via oral route

Dose descriptor:

NOAEL

25 mg/kg bw/day

Additional information

Key Studies:

Segment I rat fertility study:

In a reproduction / fertility toxicity study Amine fluorides 297/242 (mixture of 1.315 % Olaflur as leading substance and 0.347 % Hetaflur) were administered orally to sexually mature male and female Long-Evans rats (10 males, 20 females per dose) from prior to mating to the time of sacrifice at dose levels of 0, 1, 5, and 25 mg/kg bw/day at a volume of 10 mL /kg bw/day in a vehicle of 0.25% methylcellulose. The control group received the vehicle only (Menley & James Laboratories, 1973).

The NOEL is considered to be 25 mg/kg bw/day for both males and females.

There were no adverse effects which could be attributed to amine fluoride 297/242 as evidenced in evaluations of female body weights and weight gains, adult mortality, mating performances, oestrous cycles, pregnancy rates, lengths of gestation, implantation efficiencies, numbers of implantations or litter size. Evaluations of postnatal offspring viabilities, litter survivals, offspring weights, sex ratios and incidences of malformations showed no adverse effects attributable to amine fluoride 297/242 administration. There were statistically significant (p < 0.05) higher incidences of apparent intrauterine deaths in the mid and high-dose groups. These findings were adjudged to be the results of experimental sampling and of little toxicological significance.

Seg III, rat

In a Seg III developmental toxicity study (Menley & James 1973) Amine fluorides 297/242 were administered orally to mated Long-Evans rats from day 15 of gestation through day 21 of lactation. Dose levels of 1.0, 5.0, and 25.0 mg/kg bw/day were administered at a volume of 10 mL/kg bw/day in a vehicle of 0.25 % methylcellulose. The control group received the vehicle only.

There were no adverse effects upon pregnancy rate, maternal mortality, prenatal or postnatal offspring viability, percent of litters weaned, mean offspring weights., sex ratios, or soft-tissue malformations. Some slight effects concerning intrauterine deaths and postnatal viability were observed, however, these effects are considered to be not dose dependent or test item related.

Developmental, rat (2 generations)

In a developmental toxicity study (LTP, 2005) Olaflur (Amine fluoride 297 -103) was administered to pregnant CD/Crl:CD rats (20 /group, F0 -generation) by oral gavage at dose levels of 0, 6, 20 or 60 mg/kg bw/day from implantation (6th day of gestation) until weaning (the 21st day of lactation). After spontaneous delivery the F1-generation animals were examined and their development was observed employing parameters such as body weight gain, morphological landmarks and functional tests. Dams which did not deliver spontaneously were laparotomised on the 3rd day after the calculated day of delivery. The uterus was examined macroscopically and the implantation sites were determined. One male and one female rat of each litter and group (F1-generation; with a body weight nearest to the mean litter weight) were raised to maturity and mated at the age of 13 weeks. The males were sacrificed after the mating period. After spontaneous delivery, the pups (F2-generation) were weaned and their development was observed employing body weight gain. Dams which did not deliver spontaneously were laparotomised on the 3rd day after the calculated day of delivery. The uterus was examined macroscopically and the implantation sites were determined. The dams and pups were sacrificed after 3 lactation weeks.

The NOEL for maternal (F0 -dams) and developmental toxicity (F1 -pups) was considered to be 20 mg/kg bw/day and 60 mg/kg bw/d, respectively. At 60 mg/kg bw/day, reduced food intake, reduced body weight and body weight gain were noted in the dams. The variation in the F1-pup development at 60 mg/kg bw/d are not considered to be of toxicological relevance as no such effects were seen in the F2 generation, receiving the substance over a significant longer period of time and over 2 generations.

Since no test item related changes were noted both in F1 male and female parental animals as well as no findings in F2 -pups were discovered the NOEL for both is considered to be > 60 mg/kg bw/d.

Effects on developmental toxicity

Description of key information

Olaflur neither possessed teratogenic properties nor caused developmental toxicity in the two tested species rat and rabbit, even tested at systemic toxic doses.

Effect on developmental toxicity: via oral route

Dose descriptor:

NOAEL

60 mg/kg bw/day

Additional information

Key studies:

1) Developmental, rat

In a developmental toxicity study (LTP, 2004), Olaflur (Amine fluoride 297) was administered to female CD/Crl:CD rats (25 mated animals/group) by oral gavage at dose levels of 0, 6, 20 or 60 mg/kg bw/day from days 6 through 17 of gestation. The NOAEL for maternal and prenatal developmental toxicity was considered to be 60 mg/kg bw/day. At 60 mg/kg bw/day, reduced food intake was noted in the dams. The test item Olaflur possessed no teratogenic properties, no increase in malformations and retardations was noted, not even at the materno-toxic dose. Marginal effects were noted at the materno-toxic dose (60 mg/kg bw/day) only in form of an increased incidence of soft tissue variations which however were well within the historic control range and not considered to indicate a embryotoxic effect.

2) Developmental, rat

In a developmental toxicity study (LTP, 2005) Olaflur (Amine fluoride 297 -103) was administered to pregnant CD/Crl:CD rats (20 /group, F0 -generation) by oral gavage at dose levels of 0, 6, 20 or 60 mg/kg bw/day from implantation (6th day of gestation) until weaning (the 21st day of lactation). After spontaneous delivery the F1-generation animals were examined and their development was observed employing parameters such as body weight gain, morphological landmarks and functional tests. Dams which did not deliver spontaneously were laparotomised on the 3rd day after the calculated day of delivery. The uterus was examined macroscopically and the implantation sites were determined. One male and one female rat of each litter and group (F1-generation; with a body weight nearest to the mean litter weight) were raised to maturity and mated at the age of 13 weeks. The males were sacrificed after the mating period. After spontaneous delivery, the pups (F2-generation) were weaned and their development was observed employing body weight gain. Dams which did not deliver spontaneously were laparotomised on the 3rd day after the calculated day of delivery. The uterus was examined macroscopically and the implantation sites were determined. The dams and pups were sacrificed after 3 lactation weeks.

The NOEL for maternal (F0 -dams) and developmental toxicity (F1 -pups) was considered to be 20 mg/kg bw/day and 60 mg/kg bw/d, respectively. At 60 mg/kg bw/day, reduced food intake, reduced body weight and body weight gain were noted in the dams. The variation in the F1-pup development at 60 mg/kg bw/d are not considered to be of toxicological relevance as no such effects were seen in the F2 generation, receiving the substance over a significant longer period of time and over 2 generations.

Since no test item related changes were noted both in F1 male and female parental animals as well as no findings in F2 -pups were discovered the NOEL for both is considered to be > 60 mg/kg bw/d.

 

3) Developmental, rabbit

In a developmental toxicity study (LTP, 2004), Dectaflur (Amine fluoride 335) was administered to female Himalayan rabbits (24 mated animals/group) by oral gavage at dose levels of 0, 6, 20 or 60 mg/kg bw/day from days 6 through 20 of gestation.

The NOAEL for maternal and prenatal developmental toxicity was considered to be 20 and 60 mg/kg bw/day, respectively.

At 60 mg/kg bw/day, pilo-erection and diarrhoea were noted in 2 of 21 dams. In addition, reductions were noted for the body weight, body weight gain and food Intake. Necropsy revealed a dilated gall bladder in 5 of 21 dams and a dilated Vena cava caudalis in 2 of 21 dams. The changes are considered to be test item related as these changes were only observed in the high dose. Moreover, in fetuses at 60 mg/kg bw/day, slight but statistically significant increases were noted for skeletal variations (incidence of sternebra(e) fused and total variations). However, these findings were well within the normal background range. No test item-related increase was noted in the incidence of malformations, variations and retardations at any of the tested doses.

In conclusion, the test item Dectaflur possessed no teratogenic properties, even tested at systemic toxic doses.

Supportive studies:

Seg II rat

In a Seg II developmental toxicity study (Menley & James 1973) Amine fluorides 297/242 were administered orally to mated Long-Evans rats from day 6 through day 15 of gestation. Dose levels of 1.0, 5.0, and 25.0 mg/kg bw/day were administered at a volume of 10 mL/kg bw/day in a vehicle of 0.25 % methylcellulose. The control group received the vehicle only. There were no adverse effects upon maternal body weight, pregnancy rate, mortality, early deliveries or abortions, implantation efficiency, fetal size, fetal sex, or malformations. There was no biologic or toxicologic significance attached to the incidence of ossification variations observed in this study.

There was a statistically significant Increase (p<0.05) from control in the incidence of resorptions in the high-dose group, but this did not effect any other parameter linked to reproduction (e.g. total fetal size, abortions or early deliveries) and was not confirmed in developmental studies with Olaflur in rats and rabbits.

Seg II, rabbit

In a Seg II developmental toxicity study (Menley & James 1973) Amine fluorides 297/242 were administered orally to mated sexually mature New Zealand White female rabbits from day 7 through day 19 of gestation (day 0 = day of mating). Dose levels of 1, 5, and 25 mg/kg bw/day were administered at a volume of 1 mL/kg bw/day in a vehicle of 0.25 % methylcellulose. The control group received the vehicle only. There were no adverse effects upon pregnancy rate, incidences of early deliveries and abortions, fetal size, fetal sex, or ossification variations which could be attributed to the administration of Amine fluoride 297/242. Four fetuses in the treatment groups (two low, one mid, and one high) were observed to be malformed. This low incidence was not considered to express evidence of a teratogenic effect resulting from the administration of the compound. Maternal toxicity was expressed as reduced body weight gain and weight loss in the high-dose group, and increased maternal and fetal mortality in the mid and high-dose groups. These effects were attributed to a secondary toxemia resulting from the antibacterial properties of the amine fluorides against gram-positive bacteria followed by an overgrowth of gram-negative bacteria.

Seg III, rat

In a Seg III developmental toxicity study (Menley & James 1973) Amine fluorides 297/242 were administered orally to mated Long-Evans rats from day 15 of gestation through day 21 of lactation. Dose levels of 1.0, 5.0, and 25.0 mg/kg bw/day were administered at a volume of 10 mL/kg bw/day in a vehicle of 0.25 % methylcellulose. The control group received the vehicle only.

There were no adverse effects upon pregnancy rate, maternal mortality, prenatal or postnatal offspring viability, percent of litters weaned, mean offspring weights., sex ratios, or soft-tissue malformations. Some slight effects concerning intrauterine deaths and postnatal viability were observed, however, these effects were not dose dependent and not considered test item related, respectively.

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available data on toxicity to reproduction, the test item is not classified according to Regulation (EC) No 1272/2008 (CLP), as amended for the eighth time in Regulation (EU) No 2016/918.