Tall oil, reaction products with diethylenetriamine

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

No data are available for the submission substance. Data requirements are addressed based on read across to available data for the major components (AAI_DETA and Rosin) of the submission substance, DTO_DETA.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

It is considered appropriate to address the data requirements for DTO_DETA by read-across to the available studies on the main components of DTO_DETA: AAI_DETA and Rosin.
DTO_DETA contains comparatively lower levels of imidazolines and higher levels of resin acids than AAI_DETA and therefore consideration of data for resin acids is also considered necessary. The main resin acid in DTO_DETA is abietic acid, but abietic acid derivatives and other acids, such as pimaric acid, are also found in notable quantities, and the resin acids collectively are known as ‘rosin’. DTO_DETA contains up to 25% unreacted rosin, and taking into account the compositional information available for the rosin in DTO_DETA and Rosin (CAS# 8050-09-07, EC# 232-475-7), the latter was considered appropriate for read-across to DTO_DETA.
A combined repeated dose oral toxicity study with reproduction/developmental toxicity (OECD 422) conducted with AAI_DETA is available (NOTOX, 2010). The NOAEL for effects on fertility was 100 mg/kg bw/d (the highest dose tested). In addition, a screening study in the rat (OECD 421) conducted with Gum Rosin is available. The NOAEL was 3000 ppm (equivalent to 248 and 309 mg/kg bw/d in males and females, respectively) based on parental toxicity; there was no evidence for a direct reproductive/developmental toxic effect.
Read across to these studies is considered appropriate since AAI_DETA and Rosin are the main components in the target substance, DTO_DETA. No concerns are raised from the read across data and it can therefore be concluded that DTO_DETA is unlikely to be a reproductive/developmental toxicant.

Reason / purpose for cross-reference:

read-across source

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

No toxicologically significant changes in body weight (gain) occurred.

Males at 100 mg/kg/day showed a slight, but statistically significant, lower mean body weight and body weight gain during the treatment period, which recovered to control levels during the recovery phase. Some of these males showed (notable) weight loss during the premating period, which (largely) recovered during the mating/repro period. Overall, these changes were slight and reversible in nature. The statistically significant lower body weight gain on Day 4 of Lactation occurred in the absence of a dose- and time related trend, and was very slight in nature. Hence, these variations in body weight (gain) were not considered to be of toxicological relevance.

Body weight and body weight gain of males at 10 and 30 mg/kg/day was similar to control levels throughout the observation period.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Food consumption before or after allowance for body weight of males at 100 mg/kg/day appeared slightly lower than controls during the premating and mating/repro phase. However during the recovery phase food intake levels were similar to controls. Therefore, these changes were not considered to be of toxicological relevance.

Males at 10 and 30 mg/kg/day, and females at 10, 30 and 100 mg/kg/day showed normal food consumption levels, before or after allowance for body weight. The statistically significant lower food consumption before or after allowance for body weight of females at 100 mg/kg/day over Days 4-7 of the post coitum phase was of a very slight nature and within the range considered normal for rats of this age and strain. Food intake of these females remained similar to control levels during the remainder of the post coitum and lactation phase.

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

The following microscopic findings were considered to be related to treatment:
- Ileum: foamy macrophage foci in 5/5 males (minimal to slight) and in 5/5 females (minimal to slight) at 100 mg/kg/day.
- Mesenteric lymph node: slightly increased incidence and degree of macrophage foci in males and females at 100 mg/kg/day.
- Thymus: increased incidence of lymphoid atrophy in females at 100 mg/kg/day.
At the end of the 14-day recovery period for males, foamy macrophage foci in the ileum had completely resolved, whilst macrophage foci in the mesenteric lymph node persisted at slightly increased incidence and severity, with the mean severity grade being higher than encountered at the end of treatment.

All other microscopic findings recorded were considered to be within the normal range of background pathology encountered in Wistar rats of this age and strain.

No abnormalities were seen in the reproductive organs of non-fertile animals which could account for infertility.

The assessment of the integrity of the spermatogenetic cycle did not provide any evidence of impaired spermatogenesis.

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

Description (incidence and severity):

No toxicologically significant effects on reproductive parameters, fertility index and conception rate were noted.

Precoital time and number of corpora lutea and implantation sites were unaffected by treatment.

No deficiencies in maternal care were observed. Examination of cage debris of pregnant females revealed no signs of abortion or premature birth. No signs of difficult or prolonged parturition were noted among the pregnant females.

No toxicologically significant effects on number of females with live pups, gestation index, duration of gestation were noted.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 30 - < 100 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

histopathology: non-neoplastic

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

100 mg/kg bw/day (nominal)

System:

gastrointestinal tract

Organ:

ileum

mesenteric lymph node

thymus

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

yes

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

DEVELOPMENTAL DATA
No toxicologically significant effects on early postnatal pup development (body weight, clinical signs, viability index and external macroscopy) were noted.

The incidence of dead/missing pups among the dose groups was within the range considered normal for this type of study. Incidental macroscopic findings among these pups included absence of milk in the stomach and autolysis. No relationship with treatment was established for these deaths and they were considered to be of no toxicological significance.

Incidental clinical symptoms and macroscopic findings among surviving pups consisted of a small size and absence of milk in the stomach. No relationship with treatment was established for these observations and they were considered to be within the normal biological variation for rats of this age and strain.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

>= 100 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no significant effects observed at highest dose level tested

Key result

Critical effects observed:

no

Key result

Reproductive effects observed:

no

Conclusions:

No reproductive/developmental toxicity was observed at any dose level. A reproductive/developmental NOAEL of 100 mg/kg/day was derived.

Executive summary:

A guideline (OECD 422) Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test has been conducted with the read across (source) substance AAI_DETA (NOTOX, 2010). AAI_DETA was administered to rats by daily gavage at dose levels of 0, 10, 30 and 100 mg/kg bw/d; males were exposed for 28 days starting 2 weeks prior to mating, females were exposed from 2 weeks prior to mating until day 4 of lactation. There were no effects of treatment on reproductive/developmental toxicity at any dose level, and the NOAEL was therefore 100 mg/kg bw/d (the highest dose tested). Based on the increased incidence/severity of macrophage foci in the mesenteric lymph node at both the end of treatment and recovery period in males, a parental NOAEL of 30 mg/kg/day was derived. AAI_DETA is a major component of DTO_DETA therefore read across to the NOAEL obtained with AAI_DETA is considered justified. A NOAEL of 3000 ppm (equivalent to 248 and 309 mg/kg bw/d in males and females, respectively) was identified in a screening study with the read across substance Rosin, based on parental toxicity; there was no evidence for a direct reproductive/developmental toxic effect (Inveresk, 2003). The study on Rosin is presented as supporting information. It can be concluded, based on read across to AAI_DETA (supported by the Rosin dataset) that DTO_DETA is unlikely to be a reproductive/developmental toxicant and no concerns are raised with respect to developmental/reproductive toxicity.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

100 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

OECD 422 screening studies are available for the component substances.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

A guideline (OECD 422) Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test has been conducted with the read across (source) substance AAI_DETA (NOTOX, 2010). There were no effects of treatment on reproductive/developmental toxicity at any dose level, and the NOAEL was therefore 100 mg/kg bw/d (the highest dose tested). Based on the increased incidence/severity of macrophage foci in the mesenteric lymph node at both the end of treatment and recovery period in males, a parental NOAEL of 30 mg/kg/day was derived. In addition, a screening study in the rat (OECD 421) conducted with Gum Rosin is available as supporting information (Inveresk, 2003). The NOAEL was 3000 ppm (equivalent to 248 and 309 mg/kg bw/d in males and females, respectively) based on parental toxicity; there was no evidence for a direct reproductive/developmental toxic effect. Based on the available data, it is concluded that DTO_DETA is unlikely to be a reproductive/developmental toxicant.

Effects on developmental toxicity

Description of key information

This endpoint is addressed based on read across to available data for the major component (AAI_DETA) of the target substance, DTO_DETA.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

It is considered appropriate to address the data requirements for DTO_DETA by read-across to the available studies on the main components of DTO_DETA: AAI_DETA and Rosin.
DTO_DETA contains comparatively lower levels of imidazolines and higher levels of resin acids than AAI_DETA and therefore consideration of data for resin acids is also considered necessary. The main resin acid in DTO_DETA is abietic acid, but abietic acid derivatives and other acids, such as pimaric acid, are also found in notable quantities, and the resin acids collectively are known as ‘rosin’. DTO_DETA contains up to 25% unreacted rosin, and taking into account the compositional information available for the rosin in DTO_DETA and Rosin (CAS# 8050-09-07, EC# 232-475-7), the latter was considered appropriate for read-across to DTO_DETA.
A prenatal developmental toxicity study (OECD 414) conducted with the read across substance AAI_DETA (WIL Research, 2014) is available. The developmental NOAEL was 150 mg/kg bw/d (the highest dose tested). Read across to AAI_DETA is appropriate since this is a major component of the target substance, DTO_DETA. Two EPA developmental toxicity studies (Chun, 1993; range-finding and Neeper-Bradley, 1992) conducted with Imidazolium compounds, 4,5-dihydro-1-methyl-2-nortallow alkyl-1-(2-tallow amidoethyl) Me sulfate) (CAS No 68122-86-1) were submitted in support of the AAI_DETA 2013 REACH registration. These studies are provided to support the conclusion that AAI_DETA, and therefore DTO_DETA, is not teratogenic

Reason / purpose for cross-reference:

read-across source

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

At 150 mg/kg, one female (no. 85) showed hunched posture, piloerection and reduced faeces production during the last two days of treatment. This animal also showed affected body weight, food consumption and macroscopic findings, and therefore these clinical signs were regarded treatment related. At 150 mg/kg, eight additional animals also showed clinical signs (rales, piloerection and/or pale faeces). However, as these signs were only noted for 1-3 days, these signs were not considered toxicologically relevant. Incidental findings that were noted for the other groups consisted of black or red discolouration of the tail, rales, alopecia, and pale faeces. These findings occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study. At the incidence observed, these were considered signs of no toxicological relevance.

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Reduced body weights and body weight gain from Day 12 post-coitum onwards, and lower body weight gain corrected for gravid uterus weight were noted for approximately half of the animals at 150 mg/kg. Two animals (nos. 85 and 86) were most severely affected; female no. 86 gained only 2% and female no. 85 lost 18% by the end of the treatment period. At 15 and 50 mg/kg, body weights and body weight gain (also body weight gain corrected for uterine weight) remained in the same range as controls over the treatment period.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

At 150 mg/kg, food consumption was statistically significantly reduced during the complete treatment period (Days 6-20 post-coitum). Food consumption of female nos. 85 and 86 was severely reduced from Day 12 post-coitum onwards. Food consumption before or after allowance for body weight was similar between controls and animals treated at 15 and 50 mg/kg.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

At 150 mg/kg, two females (nos. 85 and 86) had the gastro-intestinal tract distended with gas and small thymus and spleen. In addition, one of these also showed irregular surface of the stomach and was emaciated. The incidence of other incidental findings among control and treated animals was within the background range of findings that are encountered among rats of this age and strain, and did not show a dose-related incidence trend. These necropsy findings were therefore not considered to be toxicologically relevant, and included several reddish foci on the thymus, uterus containing fluid, alopecia, and pelvic dilation of the kidneys.

Details on maternal toxic effects:

There were 22, 21, 21, and 18 pregnant females in the control, 15, 50 and 150 mg/kg groups, respectively, with 22, 21, 21, and 18 litters available for evaluation. No significant differences were observed between control and treated groups regarding the number of corpora lutea, implantation sites, viable or dead fetuses, early or late resorptions, or pre- and post-implantation loss.

Key result

Dose descriptor:

NOAEL

Effect level:

> 50 - < 150 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

body weight and weight gain

clinical signs

food consumption and compound intake

gross pathology

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): effects observed, non-treatment-related
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): No toxicologically relevant change was noted for fetal body weights up to 150 mg/kg. One litter at 150 mg/kg showed a very low mean fetal body weight (i.e. 2.0 gram). This was considered secondary to the health status of this dam. Body weights of fetuses (sexes combined) were 3.5, 3.4, 3.5 and 3.3 grams for the control, 15, 50 and 150 mg/kg groups, respectively.

Changes in litter size and weights:

no effects observed

Description (incidence and severity):

No treatment related effect on litter size was noted up to 150 mg/kg. The mean number of viable fetuses per litter was 11.3, 11.5, 11.9 and 11.3 in the control, 15, 50 and 150 mg/kg groups, respectively.

External malformations:

effects observed, non-treatment-related

Description (incidence and severity):

There were no treatment related effects on fetal external morphology up to 150 mg/kg. Two external malformations were noted in this study. One fetus of the control group had a cleft palate (over the entire length) and one fetus at 50 mg/kg had exencephaly (absence of part of the cranial vault, with the brain protruding outside the skull). One external variation (i.e. subcutaneous edema in the neck) was noted for one fetus at 150 mg/kg. At this single occurrence in the control, mid and high dose group, these findings were not considered to be treatment related.

Skeletal malformations:

effects observed, non-treatment-related

Description (incidence and severity):

There were no treatment related effects on fetal skeletal morphology up to 150 mg/kg. Skeletal malformations were observed in four fetuses (three litters) of the control group and five fetuses (four litters) at 150 mg/kg. One control fetus showed fusion of the sternebrae (nos. 4 and 5 with thread-like ossification bridge) with severely malaligned sternebrae (no. 4; slight no. 3). As this concerned a control fetus, it was not treatment related. Bent limb bones were noted for three control fetuses (two litters) and five high dose fetuses (four litters). One of these high dose fetuses also showed sternoschisis (nos. 1-6). Bent limb bones have been noted in 17 historical control fetuses (15 litters) with a maximum incidence of 2.0%. The litter incidence of the concurrent control group is 1.2% and 3.6% at 150 mg/kg. Even though the incidence at 150 mg/kg is outside the historical control range, these findings were not considered treatment related as the absolute number of affected fetuses at 150 mg/kg was only slightly above the concurrent control group (5 fetuses at 150 mg/kg versus 3 control fetuses). Moreover, bent limb bones are considered reversible (Ref. 12) and are therefore not adverse. In addition, as sternoschisis was noted for a single fetus, and as it was seen previously in the historical control data (2 fetuses), it was considered a chance finding and not toxicologically relevant. Skeletal variations noted for fetuses in the control and/or high dose group were unossified or reduced ossification of several bones (skull, entire sternum, vertebral centra, vertebral arches, pubis, ischium, ribs, sternebrae nos. 1, 2, 3, 4 5 and/or 6, vertebral centra, hyoid, metacarpals and/or metatarsals), 14th full or rudimentary ribs, bent ribs, 7th cervical full or rudimentary ribs, cervical centrum no. 1 ossified, slight or moderate malaligned sternebrae, caudal shift of pelvic girdle, and branched sternebrae. These variations were not considered to be treatment related, because they occurred at similar or higher frequencies in the control group, occurred infrequently and/or occurred at frequencies within the historical control range.

Visceral malformations:

effects observed, non-treatment-related

Description (incidence and severity):

There were no treatment related effects on fetal visceral morphology up to 150 mg/kg. Visceral malformations were observed in four fetuses (three litters). Internal hydrocephaly was noted for one fetus of the control group and one fetus at 50 mg/kg. At 50 mg/kg, one fetus had a small eye and one fetus showed persistent truncus arteriosus and absent eyes. At this low incidence and without a dose response relationship, these malformations were not considered treatment related. In addition, all findings (except persistent truncus arteriosus) were noted before in historical control fetuses. As persistent truncus arteriousus was noted for only one fetus at the mid dose group, it was considered a chance finding and not related to treatment. Visceral variations noted for fetuses in the control and/or treated group(s) were small supernumerary liver lobe, dilated ureter, liver appendix, partially undescended thymus horns, and convoluted ureter. These variations were not considered to be treatment related, because they occurred infrequently, in the absence of a dose related trend and/or at frequencies within the historical control range.

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No developmental effects observed up to 150 mg/kg bw/d (highest dose tested).

Key result

Abnormalities:

no effects observed

Key result

Developmental effects observed:

no

Conclusions:

The prenatal developmental toxicity study (OECD 414) on AAI-DETA resulted to a maternal NOAEL of 50 mg/kg. The developmental NOAEL was at least 150 mg/kg. Based on read across to AAI_DETA, DTO_DETA is not considered to cause developmental toxicity.

Executive summary:

A guideline (OECD 414) Prenatal Developmental Toxicity Study has been conducted with the read across (source) substance AAI_DETA (WIL Research, 2014). AAI_DETA was administered to female rats by daily gavage at dose levels of 0, 15, 50 and 150 mg/kg bw/d from Days 6 to 19 of gestation. There were no effects of treatment on development at any dose level, and the NOAEL was therefore 150 mg/kg bw/d (the highest dose tested). Based on reduced body weight gain and food consumption, a maternal NOAEL of 50 mg/kg/day was derived. AAI_DETA is a major component of DTO_DETA therefore read across to the NOAEL obtained with AAI_DETA is considered justified. Two EPA developmental toxicity studies (Chun, 1993; range-finding and Neeper-Bradley, 1992) conducted with Imidazolium compounds, 4,5-dihydro-1-methyl-2-nortallow alkyl-1-(2-tallow amidoethyl) Me sulfate) (CAS No 68122-86-1) were submitted in support of the AAI_DETA 2013 REACH registration. These studies are provided to support the conclusion that AAI_DETA, and therefore DTO_DETA, is not teratogenic.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

150 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Read across to a guideline and GLP-compliant study

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

A guideline (OECD 414) Prenatal Developmental Toxicity Study has been conducted with the read across (source) substance AAI_DETA (WIL Research, 2014). There were no effects of treatment on development at any dose level, and the NOAEL was therefore 150 mg/kg bw/d (the highest dose tested). Based on reduced body weight gain and food consumption, a maternal NOAEL of 50 mg/kg/day was derived. Two EPA developmental toxicity studies (Chun, 1993; range-finding and Neeper-Bradley, 1992) conducted with Imidazolium compounds were submitted in support of the AAI_DETA 2013 REACH registration. These studies are provided to support the conclusion that AAI_DETA, and therefore DTO_DETA, is not teratogenic.

Justification for classification or non-classification

Based on the results of the available studies, DTO_DETA is not a developmental or reproductive toxin and classification according to Regulation (EC) No. 1272/2008 is not required.