Registration Dossier

Reference

Endpoint:: screening for reproductive / developmental toxicity
Type of information:: migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:: key study
Reliability:: 2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:: other: see 'Remark'
Remarks:: GLP - Guideline study. According to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
Qualifier:: according to guideline
Guideline:: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Principles of method if other than guideline:: Method: other: OECD Combined Repeat Dose and Reproductive/Developmental Toxicity Screening Test
GLP compliance:: yes
Limit test:: no
Species:: rat
Strain:: other: Slc:SD
Sex:: male/female
Details on test animals or test system and environmental conditions:: TEST ANIMALS
- Weight at study initiation: male: 304-343 g; femalws: 196-226 g
- Housing: pregnant females should be caged individually
- Diet ad libitum
- Water ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-24
- Humidity (%): 50-60
- Photoperiod 12 hrs dark / 12 hrs light
Route of administration:: oral: gavage
Vehicle:: water
Details on exposure:: dosing of both sexes should begin 2 weeks prior to matingm continued through mating period
males: dosing continued up to the day when females are killed
females: dosing continued throughout pregnancy and up to day 4 of lactation
Details on mating procedure:: one female to one male until pregnancy occurs:
day 0 of pregnancy is defined as the day sperm is found
Analytical verification of doses or concentrations:: yes
Details on analytical verification of doses or concentrations:: details not given
Duration of treatment / exposure:: Exposure period: male, 45 days; female, from 14 days before mating to day 3 of lactation
Duration of test: terminal kill: male, day 46; female and pups, day 4 of lactation
Frequency of treatment:: daily
Details on study schedule:: -Age at mating of the mated animals 10 weeks
Remarks:: Doses / Concentrations:
0, 12.5, 50, 200, 800 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:: 12
Control animals:: yes, concurrent vehicle
Details on study design:: as requested by OECD TG 422
Positive control:: no
Parental animals: Observations and examinations:: at least once per day:
--behavioural changes, signs of difficult or prolonged parturition, mortality and all signs of toxicity
cage side observations:
changes in skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system fucnction
--food consumption, males and females should be weighed
--duration of gestation, examination of the litter as soon as possible, number and sex of pups, stillbirth, live birth, pup weight, and the presence of gross anomalies
--clinical examinations: hematologym clinical chemistry, urinalysis
--pathology: gross necropsy, histopathology
Oestrous cyclicity (parental animals):: no data,
Sperm parameters (parental animals):: no data
Litter observations:: PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities,

GROSS EXAMINATION OF DEAD PUPS:
yes, for external abnormalities;
Postmortem examinations (parental animals):: pathology: gross necropsy, histopathology
Postmortem examinations (offspring):: external malformation
Statistics:: yes but method not mentioned
Reproductive indices:: number of mated pairs
number of copulated pairs
copulation index
number of pregnant animals
fertility index
pairing days until copulation
implantation index
delivery index
Offspring viability indices:: number of pups born,
number of pups alive
birth index
live birth index
sex ratio
number of pups alive on day 4
viability index
body weight of F1 pups on day 4
Clinical signs:: no effects observed
Body weight and weight changes:: effects observed, treatment-related
Food consumption and compound intake (if feeding study):: effects observed, treatment-related
Organ weight findings including organ / body weight ratios:: effects observed, treatment-related
Histopathological findings: non-neoplastic:: effects observed, treatment-related
Other effects:: no effects observed
Reproductive function: oestrous cycle:: no effects observed
Reproductive function: sperm measures:: no effects observed
Reproductive performance:: no effects observed
Dose descriptor:: NOAEL
Remarks:: systemic
Effect level:: 800 mg/kg bw/day
Based on:: test mat.
Sex:: male/female
Basis for effect level:: other: see 'Remark'
Dose descriptor:: NOAEL
Remarks:: fertility
Effect level:: 800 mg/kg bw/day
Based on:: test mat.
Sex:: male/female
Basis for effect level:: other: based on no signs indicative for reproductive / developmental toxicity up to the highest test dose.
Clinical signs:: no effects observed
Mortality / viability:: no mortality observed
Body weight and weight changes:: not examined
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
Dose descriptor:: NOAEL
Remarks:: developmental
Generation:: F1
Effect level:: 800 mg/kg bw/day
Based on:: test mat.
Sex:: male/female
Basis for effect level:: other: based on no signs indicative of developmental toxicity up to the highest test dose
Reproductive effects observed:: not specified
Executive summary:: Trimethylolpropane was studied for oral toxicity in an OECD TG 422 (combined repeat dose and reproductive/ developmental toxicity screening test) at doses of 0. 12.5, 50, 200, and 800 mg/kg bw/day given by gavage.No signs indicative of reproductive/developmental toxicity were observed . The NOAEL for reproductive performance and offspring development were both 800 mg/kg bw/day. The NOAEL (general toxicity) was 200 mg/kg bw/day (MHLW 1994).

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises an adequate, reliable study (Klimisch score 2) from a reference substances with similar structure and intrinsic properties. Read-across is justified based on the structural similarity between the source and target substance, as the source substance comprises a hydrolysis product (CAS 77-99-6) of the target substance. Refer to endpoint discussion for further details. The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.7, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Endpoint conclusion:

no study available

Endpoint conclusion:

no study available

Justification for grouping of substances and read-across

There are no data available for the assessment of reproduction toxicity of Fatty acids, C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane (CAS 147256-33-5).

In order to fulfil the standard information requirements set out in Annex IX, 8.7, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, read-across from structurally related substances was conducted.

In accordance with Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met.” In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across).

Having regard to the general rules for grouping of substances and read-across approach laid down in Annex XI, Item 1.5, of Regulation (EC) No 1907/2006 whereby substances may be predicted as similar provided that their physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity.

Overview of toxicity to reproduction:

CAS	147256-33-5 (a)	77-99-6 (b)
Chemical Name	Fatty acids,C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane	Propylidynetrimethanol
MW	624.97 1115.78; 1606.58; 677.05; 1219.92; 1762.80	134.18
Toxicity to reproduction – screening studies	RA: CAS 77-99-6	Experimental result: NOAEL (fertility) ≥ 800 mg/kg bw/day NOAEL (developmental toxicity) ≥ 800 mg/kg bw/day
Toxicity to reproduction – (pre-natal) development	Waiving	--

(a) Substances subject to the REACh Phase-in registration deadline of 31 May 2013 are indicated in bold font.

(b) Substances that are either already registered under REACh or not subject to the REACh Phase-in registration deadline of 31 May 2013 are indicated in normal font. Lack of data for a given endpoint is indicated by “--“.

The above mentioned substances are considered to be similar on the basis of the structural similar properties and/or activities. The available endpoint information is used to predict the same endpoints for Fatty acids, C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane (CAS 147256-33-5).

A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

Toxicity to reproduction – screening studies

CAS 77-99-6

In a GLP-compliant reprotoxicity/developmental toxicity screening study according to OECD Guideline 422, male and female Slc:SD rats were exposed to propylidynetrimethanol at dose levels of 12.5, 50, 200 and 800 mg/kg bw/day (MHLW, 1994). The test substance dissolved in distilled water was orally administered to 12 animals per sex and dose via gavage. Males were treated for a period of 45 days, whereas females were exposed to the test substance 14 days prior to mating, during gestation and until Day 3 of lactation. A similar constituted group of animals received the vehicle and served as control. No mortality and no clinical signs were observed during the study period. During the 14-day pre-mating period, the absolute body weights in males and the body weight gain in females was statistically significantly decreased at 800 mg/kg bw/day compared to the controls. However, no effects on the body weights were observed in treated animals during the remainder of the study. No effect on food consumption was observed in male and females of any treatment groups compared to controls. A significant increase in absolute and relative liver weight in males was observed at 800 mg/kg bw/day. In females of the same dose group, only a slight and not statistically significant increase in absolute and relative liver weight was noted. At necropsy, enlargement of the liver was found in 3/10 males, which correlated with the observed increase in liver weight in this gender. In females, the increase in the incidence of macroscopic findings in the lungs (red patch/zone) at all dose levels was not considered to be of toxicological relevance, since they were not unequivocally attributable to the test substance administration due to a lacking dose-response relationship. At histopathological examination, a slight basophilic alteration of the renal tubular epithelial cells was observed in 1/11 females receiving 50 mg/kg bw/day, in 2/10 females receiving 200 mg/kg bw/day and in 5/10 females (stat. significant) receiving 800 mg/kg bw/day. However, these changes were not unequivocally attributable to the test substance administration, because of their limited distribution and limited degree, and because similar lesions were observed in male rats of all groups including the controls. Although necropsy revealed liver enlargement in 3/10 male rats receiving 800 mg/kg bw/day, histopathological examination did not show any definite morphological lesions in the liver. All other histopathological alterations in treated animals were also observed to a similar degree in the corresponding control animals, and were thus of no toxicological significance.

No effects of the test substance were observed on copulation, fertility or oestrus cycle of parental rats, as well as on delivery and on dams during the lactation period. External examination of pups revealed no increase in the incidence of abnormalities. Body weight gain of pups was normal up to Day 4 of the lactation period. Stillborn, dead pups and pups sacrificed at Day 4 of the lactation period showed no abnormal gross lesions, which were attributable to treatment with the test substance.

Based on these results, the NOAEL for systemic toxicity and fertility in parental rats and the NOAEL for developmental toxicity in pubs was ≥ 800 mg/kg bw/day, which corresponded to the highest dose level administered.

Toxicity to reproduction – Two-generation studies

This information is not available

Conclusions for toxicity to reproduction (fertility)

There are no data available on the toxicity to reproduction (fertility) of Fatty acids, C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane. However, oral, dermal or inhalative absorption of the registered substance is deemed unlikely due to its very high MW and physico-chemical properties. As hydrolysis is possible to a limited extend, it will most likely be the cleavage products that are absorbed, thus a worst case approach considering the reproduction toxicity of potential hydrolysis products was performed. Hazard assessment is conducted by means of read-across from the available data on the hydrolysis product propylidynetrimethanol (CAS 77-99-6), which did not show any adverse effect on reproductive parameters and post-natal offspring development up to and including the highest dose in a combined repeated dose and reproductive/developmental toxicity screening test in rats. Thus, the NOAEL for fertility and offspring development was considered to be ≥ 800 mg/kg bw/day.

No two-generation reprotoxicity study is available, neither for the target substance nor the surrogate substances. However, data on the oral repeated dose toxicity of hydrolysis products of the target substance, propylidynetrimethanol (CAS 77-99-6) and Fatty acids, C18-unsatd., dimers (61788-89-4), demonstrate no adverse effects on reproduction organs and tissues after subchronic exposure up to dose levels that are near the conventional limit dose of 1000 mg/kg bw/day. Therefore, based on the weight of evidence, a two-generation reprotoxicity study by any route of exposure is considered scientifically unjustified and not necessary in terms of animal welfare.

The NOAEL of 800 mg/kg bw/day for propylidynetrimethanol (molecular weight = 134.18 g/mol) is selected for hazard assessment of Fatty acids, C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane. In order to account for differences in molecular weight, full enzymatic hydrolysis of Fatty acids, C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane into Fatty acids, C18-unsatd., dimers, oleic acid and trimethylolpropane is assumed. A resulting estimated NOAEL for fertility and developmental toxicity ranging from 3726.2-10510.1 mg/kg bw/day was established for Fatty acids, C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane.

However, the NOAEL for fertility of 800 mg/kg bw/day used for hazard assessment represents an overestimation, as the registered substance is not anticipated to be absorbed and hydrolysed in any significant amounts. Having regard to the fact that the NOAEL was near to the conventional limit dose of 1000 mg/kg bw/day, no adverse effects for humans are expected and no hazard was identified.

In summary, the available data provide sufficient evidence to conclude that the substance Fatty acids, C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane is not toxic to reproduction.

Effects on fertility:

A waiver for the requirement to perform an extended one-generation reproduction toxicity study (standard configuration or with additional modules) was included, as the study is not scientifically necessary and, considering concerns regarding the use of vertebrate animals for experimental purposes, unjustified.

Short description of key information:
Based on read-across from Propylidynetrimethanol (CAS 77-99-6), which is a hydrolysis product of the target substance, no hazard was identified.
NOAEL fertility and developmental toxicity (rat, m/f) = 3726.2-10510.1 mg/kg bw/day (based on read-across from Propylidynetrimethanol and after correction for differences in molecular weight)

Justification for selection of Effect on fertility via oral route:
Hazard assessment is conducted by means of read-across from a structural surrogate. The selected study is the most adequate and reliable study based on the identified similarities in structure and intrinsic properties between source and target substance and overall assessment of quality, duration and dose descriptor level (refer to the endpoint discussion for further details).

Toxicity to reproduction – (pre-natal) development: no study available

Endpoint conclusion:

no study available

Endpoint conclusion:

no study available

Endpoint conclusion:

no study available

Justification for grouping of substances and read-across

There are no data available for the assessment of reproduction toxicity of Fatty acids, C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane (CAS 147256-33-5).

In order to fulfil the standard information requirements set out in Annex IX, 8.7, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, read-across from structurally related substances was conducted.

In accordance with Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met.” In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across).

Having regard to the general rules for grouping of substances and read-across approach laid down in Annex XI, Item 1.5, of Regulation (EC) No 1907/2006 whereby substances may be predicted as similar provided that their physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity.

Overview of toxicity to reproduction:

CAS	147256-33-5 (a)	77-99-6 (b)
Chemical Name	Fatty acids,C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane	Propylidynetrimethanol
MW	624.97 1115.78; 1606.58; 677.05; 1219.92; 1762.80	134.18
Toxicity to reproduction – screening studies	RA: CAS 77-99-6	Experimental result: NOAEL (fertility) ≥ 800 mg/kg bw/day NOAEL (developmental toxicity) ≥ 800 mg/kg bw/day
Toxicity to reproduction – (pre-natal) development	Waiving	--

(a) Category members subjected to the REACh Phase-in registration deadline of 31 May 2013 are indicated in bold font

(b) Substances that are either already registered under REACh, or not subject to the REACh Phase-in registration deadline of 31 May 2013, are indicated in normal font. Lack of data for a given endpoint is indicated by “--“.

The above mentioned substances are considered to be similar on the basis of the structural similar properties and/or activities. The available endpoint information is used to predict the same endpoints for Fatty acids, C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane (CAS 147256-33-5).

A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

Toxicity to reproduction – (pre-natal) development

This information is not available.

Conclusions for toxicity to reproduction (development)

There are no data available for the prenatal developmental toxicity of Fatty acids, C18 unsatd., dimers, mixed esters with oleic acid and trimethylolpropane. However, data from a reproduction/developmental screening test with the hydrolysis product propylidynetrimethanol (CAS 77-99-6) do not indicate any adverse effects on intrauterine development up to the highest dose tested (NOAEL developmental toxicity ≥ 800 mg/kg bw/day). Furthermore, all available data on Fatty acids, C18 unsatd., dimers, mixed esters with oleic acid and trimethylolpropane and structurally related substances show that the substance is of low acute toxicity after oral and inhalation exposure. In addition, data on the oral repeated dose toxicity of the hydrolysis products Fatty acids, C18-unsatd., dimers (CAS 61788-89-4) and propylidynetrimethanol (CAS 77-99-6) demonstrate no adverse systemic effects after oral exposure. As oral, dermal or inhalative absorption of the registered substance is deemed unlikely due to its very high MW and physico-chemical properties and hydrolysis is not anticipated to occur in significant amounts, a very low bioavailability in humans is anticipated for Fatty acids, C18 unsatd., dimers, mixed esters with oleic acid and trimethylolpropane and its hydrolysis after any route of exposure. Based on the physicochemical (high molecular weight, low water solubility and high lipophilicity) and toxicological properties (low acute and repeated systemic toxicity), there is sufficient weight of evidence provided leading to the assumption/conclusion that the substance is not toxic to reproduction. Therefore, further testing on vertebrate animals for (pre-natal) development shall be omitted and further testing not involving vertebrate animals may be omitted.

Based on read-across from the surrogate substances, the available data on the toxicity to reproduction (fertility) do not meet the classification criteria according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.