Fatty acids, C16-18 and C18-unsatd., esters with propylene glycol

Endpoint summary

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Effects on fertility

Link to relevant study records

Reference

Endpoint:

extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the extended one-generation reproductive toxicity study does not need to be conducted because there are no results from available repeated dose toxicity studies that indicate adverse effects on reproductive organs or tissues, or reveal other concerns in relation with reproductive toxicity

Reproductive effects observed:

not specified

Effect on fertility: via oral route

Endpoint conclusion:

no study available

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

In accordance with Regulation (EC) No 1907/2006, Annex IX, 8.7.3, column 1, a two-generation toxicity study is required if the repeated dose studies (28- or 90-day exposure) indicate adverse effects on reproductive organs and tissues. In regard to the available data on repeated dose toxicity, sufficient and reliable data (Klimisch score 2 to point to the fact that the studies were conducted on a read-across analogue substance) are available after long-term exposure to the read-across analogue substances Fatty acids, C18 and C18 unsatd. epoxidized, ester with ethylene glycol (CAS 151661-88-0) and Decanoic acid, mixed diesters with octanoic acid and propylene glycol (CAS 68583-51-7) which did not reveal adverse effects on reproductive organs and tissues in rats in either sex. Therefore, performance of a two-generation study is scientifically unjustified and, in accordance with Annex XI, Section 1.2 of Regulation (EC) 1907/2006 further testing on vertebrate animals for that property shall be omitted.

Moreover, according to Regulation (EC) No 1907/2006, Annex IX, 8.7, Column 2, ”reproductive toxicity studies do not need to be conducted if the substance is of low toxicological activity and it can be proven from toxicokinetic data that no systemic absorption occurs via relevant routes of exposure”. As target substance is considered to be hydrolysed before absorption occurs as discussed in the toxicokinetic statement (please refer to chapter 5 in the technical dossier), distribution of the intact glycol diester is not relevant but rather the absorption of the breakdown products of intestinal hydrolysis, which are considered to be either rapidly absorbed through the GI tract (propylene glycol; ATDSR, 1997, 2010; ICPS, 2001) or into the walls of the intestine villi (oleate; Lehninger, 1970). However, in accordance with the general rules set out in Regulation (EC) No 1907/2006, Annex XI, section 1.2, a weight-of-evidence approach based on the available toxicity data clearly demonstrates that the target substance exhibits no toxicological potency although absorption of the breakdown products occurs.

In conclusion, performance of a two-generation study is scientifically unjustified and, according to Annex XI, Section 1.2 of Regulation (EC) No 1907/2006 further testing on vertebrate animals for that property shall be omitted.

Justification for selection of Effect on fertility via oral route: In accordance with Regulation (EC) No 1907/2006, Annex IX, 8.7.3 column 1, no two-generation study is required since no adverse effects on reproductive organs were seen in subchronic toxicity studies with structural analogues.

Effects on developmental toxicity

Description of key information

Oral (OECD 414, rat): NOAEL developmental toxicity: >900 mg/kg bw/day 
Oral (OECD 414, rat): NOAEL maternal toxicity: >9000 mg/kg bw/day 

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 Jul 1994 - 15 Mai 1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

GLP-Guideline study with acceptable restrictions. Administration of test article only during organogenesis.

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

only organogenesis covered

Qualifier:

according to guideline

Guideline:

EU Method B.31 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

only organogenesis covered

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Sprague-Dawley, CD

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles Rover, Sulzfeld, Germany
- Age at study initiation: 8 weeks
- Weight at study initiation: 198 g
- Housing: animals were housed individually in Makrolon M3 cages with standard softwood bedding.
- Diet: pelleted Altromin Maintenance Diet 1324, Lot No. 170994/1340 (Altromin GmbH, Lage, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-24
- Humidity (%): 47-82
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

other: 0.5% sodium carboxymethylcellulose and 0.25% Cremophor in aqua dest.

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: The test article was prepared daily before administration.

Analytical verification of doses or concentrations:

yes

Details on mating procedure:

- Impregnation procedure: purchased timed pregnant
- They were received at the testing facility on Day 0 of gestation.

Duration of treatment / exposure:

Day 6-15 of gestation

Frequency of treatment:

daily, 7 days/week

Duration of test:

Day 20 of gestation

No. of animals per sex per dose:

24 P females

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: dose levels were based on the results of toxicological examinations (no further information, Henkel Report TBD 710070).

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least twice daily (working days) for mortality

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least twice daily (working days) for signs of reaction to treatment and/or symptoms of illness

BODY WEIGHT: Yes
- Time schedule for examinations: on Day 0, 6, 16 and 20 post coitum

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation Day 20
- Organs examined: Gross macroscopic examination of all maternal organs with emphasis on the uterus, uterine contents, and position of foetuses in the uterus.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

- External examinations: Yes: all per litter
- Visceral examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter

Statistics:

If the variables could be assumed to follow a normal distribution, the Dunnett-test, based on a pooled variance, was applied for the comparison between the treated groups and the control group. The Steel-test was applied when the data could not be assumed to follow a normal distribution. Fisher’s Exact test for 2x2 tables was applied if the variables could be dichotomized without loss of information (Bonferroni-Holm-corrected).

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
No mortalities occurred during the study period. No compound-related symptoms were observed in the treatment groups in comparison to the control group. Body weight, body weight gains and corrected body weight were within expected ranges. No compound related differences were noted between the mean reproduction data of the test groups in comparison to the control group. At scheduled necropsy no macroscopic changes were noted in the dams of the treatment groups.

Dose descriptor:

NOAEL

Effect level:

>= 900 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No substance-related symptoms were observed in the treatment groups. Pre-implantation loss, post-implantation loss, mean number of resorptions, embryonic deaths and total foetuses were not affected by treatment. Only one dead foetus was observed in the middle- and one dead foetus in the high-dose group from 345 and 306 live foetuses, respectively. Mean foetal placental and uterus weights were not affected by treatment. Foetal sex ratio was comparable in all groups. No treatment-related foetal abnormalities were found at necropsy. The examined foetuses showed no treatment-related malformations. The figures of visceral variations in the test groups were considered to be similar to the control group.
The mean weight of live male and female foetuses in the mid-dose group was significantly increased (see Table 1 under "Any other information on results incl. tables"). The weights of live foetuses of the other treatment groups exhibited no significant differences on a litter and individual basis e.g. mean weight in comparison to the control group.
The figures of skeletal ossifications and variations showed no treatment-related deviations, significant evidences between treated groups and the control group are listed in Table 2 under "Any other information on results incl. tables". The statistically significant differences concerning increased or decreased figures of various findings were considered to be incidental. Furthermore, no dose-relationship in any finding was observed.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 900 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: teratogenicity

Key result

Developmental effects observed:

no

Table 1. Results of study.

Parameter	Group 1 0 mg/kg bw	Group 2 100 mg/kg bw	Group 3 300 mg/kg bw	Group 4 900 mg/kg bw
Number of corpora lutea [total/number of dams ± SD]	16.7 ± 1.9	17.1 ± 2.0	16.7 ± 1.9	16.5 ± 1.4
Implantations [total/number of dams ± SD]	15.3 ± 2.2	15.4 ± 2.7	14.9 ± 2.3	14.8 ± 1.5
Pre-implantation loss	34	39	44	38
Post-implantation loss	15	18	13	20
Embryonic death [total]	15	18	12	19
Embryonic resorptions [total/number of dams ± SD]	0.6 ± 1.1	0.7 ± 1.1	0.5 ± 0.7	0.7 ± 0.9
Fetal resorptions [total/number of dams ± SD]	0.0 ± 0.2	0.0 ± 0.2	0.0 ± 0.0	0.1 ± 0.4
Live fetuses	336	337	345	306
Dead fetuses	0	0	1	1
Malformed fetuses [total/number of dams ± SD]	0.0 ± 0.2	0.0 ± 0.0	0.0 ± 0.0	0.0 ± 0.2
Sex of fetuses (%males : % females)	47.3 : 52.7	51.6 : 48.4	50.3 : 49.4	52.1 : 47.6
Weights of live fetuses (mean ± SD) Males Females	4.3 ± 0.9 4.0 ± 0.8	4.2 ± 0.8 4.0 ± 0.7	5.0 ± 0.9* 4.6 ± 0.8*	4.5 ± 0.7 4.3 ± 0.7
Weights of placenta (mean ± SD)	0.6 ± 0.1	0.6 ± 0.1	0.6 ± 0.1	0.6 ± 0.1
Weights of uteri (mean ± SD)	88.7 ± 14.9	89.3 ± 19.2	97.9 ± 18.9	90.9 ± 11.7

*: Dunnett-test based on pooled variance, significant at level 5%

Table 2. Skeletal examination of fetuses (stage of development).

Parameter (%)	Group 1 0 mg/kg bw	Group 2 100 mg/kg bw	Group 3 300 mg/kg bw	Group 4 900 mg/kg bw
Fetal skeleton No abnormal findings	4.0	9.8	16.1##	10.0
Skull bones, single incompletely ossified	4.5	11.6#	10.0	5.0
Sternebrae incompletely ossified abnormally ossified	66.5 24.4	84.4## 17.9	51.1## 9.4##	66.3 17.5
Coccygeal vertebrae, 4 and more ossified	32.4	22.5	56.1##	47.5#
Pelvis, pubis: incompletely ossified	6.2	0.6#	0.6##	1.3

#/##: Fishers exact test (two-sided) significant at level 5% (#) or 1% (##), Bonferroni-Holm-corrected

Conclusions:

The test substance had no effect on intrauterine development.

Reference 2

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

key study

Study period:

05 Jul 1994 - 15 Mai 1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

GLP-Guideline study with acceptable restrictions. Administration of test article only during organogenesis.

Justification for type of information:

Please refer Section 13 for read across justification.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

only organogenesis covered

Qualifier:

according to guideline

Guideline:

EU Method B.31 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

only organogenesis covered

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Sprague-Dawley, CD

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles Rover, Sulzfeld, Germany
- Age at study initiation: 8 weeks
- Weight at study initiation: 198 g
- Housing: animals were housed individually in Makrolon M3 cages with standard softwood bedding.
- Diet: pelleted Altromin Maintenance Diet 1324, Lot No. 170994/1340 (Altromin GmbH, Lage, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-24
- Humidity (%): 47-82
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

other: 0.5% sodium carboxymethylcellulose and 0.25% Cremophor in aqua dest.

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: The test article was prepared daily before administration.

Analytical verification of doses or concentrations:

yes

Details on mating procedure:

- Impregnation procedure: purchased timed pregnant
- They were received at the testing facility on Day 0 of gestation.

Duration of treatment / exposure:

Day 6-15 of gestation

Frequency of treatment:

daily, 7 days/week

Duration of test:

Day 20 of gestation

No. of animals per sex per dose:

24 P females

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: dose levels were based on the results of toxicological examinations (no further information, Henkel Report TBD 710070).

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least twice daily (working days) for mortality

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least twice daily (working days) for signs of reaction to treatment and/or symptoms of illness

BODY WEIGHT: Yes
- Time schedule for examinations: on Day 0, 6, 16 and 20 post coitum

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation Day 20
- Organs examined: Gross macroscopic examination of all maternal organs with emphasis on the uterus, uterine contents, and position of foetuses in the uterus.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

- External examinations: Yes: all per litter
- Visceral examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter

Statistics:

If the variables could be assumed to follow a normal distribution, the Dunnett-test, based on a pooled variance, was applied for the comparison between the treated groups and the control group. The Steel-test was applied when the data could not be assumed to follow a normal distribution. Fisher’s Exact test for 2x2 tables was applied if the variables could be dichotomized without loss of information (Bonferroni-Holm-corrected).

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
No mortalities occurred during the study period. No compound-related symptoms were observed in the treatment groups in comparison to the control group. Body weight, body weight gains and corrected body weight were within expected ranges. No compound related differences were noted between the mean reproduction data of the test groups in comparison to the control group. At scheduled necropsy no macroscopic changes were noted in the dams of the treatment groups.

Dose descriptor:

NOAEL

Effect level:

>= 900 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No substance-related symptoms were observed in the treatment groups. Pre-implantation loss, post-implantation loss, mean number of resorptions, embryonic deaths and total foetuses were not affected by treatment. Only one dead foetus was observed in the middle- and one dead foetus in the high-dose group from 345 and 306 live foetuses, respectively. Mean foetal placental and uterus weights were not affected by treatment. Foetal sex ratio was comparable in all groups. No treatment-related foetal abnormalities were found at necropsy. The examined foetuses showed no treatment-related malformations. The figures of visceral variations in the test groups were considered to be similar to the control group.
The mean weight of live male and female foetuses in the mid-dose group was significantly increased (see Table 1 under "Any other information on results incl. tables"). The weights of live foetuses of the other treatment groups exhibited no significant differences on a litter and individual basis e.g. mean weight in comparison to the control group.
The figures of skeletal ossifications and variations showed no treatment-related deviations, significant evidences between treated groups and the control group are listed in Table 2 under "Any other information on results incl. tables". The statistically significant differences concerning increased or decreased figures of various findings were considered to be incidental. Furthermore, no dose-relationship in any finding was observed.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 900 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: teratogenicity

Key result

Developmental effects observed:

no

Table 1. Results of study.

Parameter	Group 1 0 mg/kg bw	Group 2 100 mg/kg bw	Group 3 300 mg/kg bw	Group 4 900 mg/kg bw
Number of corpora lutea [total/number of dams ± SD]	16.7 ± 1.9	17.1 ± 2.0	16.7 ± 1.9	16.5 ± 1.4
Implantations [total/number of dams ± SD]	15.3 ± 2.2	15.4 ± 2.7	14.9 ± 2.3	14.8 ± 1.5
Pre-implantation loss	34	39	44	38
Post-implantation loss	15	18	13	20
Embryonic death [total]	15	18	12	19
Embryonic resorptions [total/number of dams ± SD]	0.6 ± 1.1	0.7 ± 1.1	0.5 ± 0.7	0.7 ± 0.9
Fetal resorptions [total/number of dams ± SD]	0.0 ± 0.2	0.0 ± 0.2	0.0 ± 0.0	0.1 ± 0.4
Live fetuses	336	337	345	306
Dead fetuses	0	0	1	1
Malformed fetuses [total/number of dams ± SD]	0.0 ± 0.2	0.0 ± 0.0	0.0 ± 0.0	0.0 ± 0.2
Sex of fetuses (%males : % females)	47.3 : 52.7	51.6 : 48.4	50.3 : 49.4	52.1 : 47.6
Weights of live fetuses (mean ± SD) Males Females	4.3 ± 0.9 4.0 ± 0.8	4.2 ± 0.8 4.0 ± 0.7	5.0 ± 0.9* 4.6 ± 0.8*	4.5 ± 0.7 4.3 ± 0.7
Weights of placenta (mean ± SD)	0.6 ± 0.1	0.6 ± 0.1	0.6 ± 0.1	0.6 ± 0.1
Weights of uteri (mean ± SD)	88.7 ± 14.9	89.3 ± 19.2	97.9 ± 18.9	90.9 ± 11.7

*: Dunnett-test based on pooled variance, significant at level 5%

Table 2. Skeletal examination of fetuses (stage of development).

Parameter (%)	Group 1 0 mg/kg bw	Group 2 100 mg/kg bw	Group 3 300 mg/kg bw	Group 4 900 mg/kg bw
Fetal skeleton No abnormal findings	4.0	9.8	16.1##	10.0
Skull bones, single incompletely ossified	4.5	11.6#	10.0	5.0
Sternebrae incompletely ossified abnormally ossified	66.5 24.4	84.4## 17.9	51.1## 9.4##	66.3 17.5
Coccygeal vertebrae, 4 and more ossified	32.4	22.5	56.1##	47.5#
Pelvis, pubis: incompletely ossified	6.2	0.6#	0.6##	1.3

#/##: Fishers exact test (two-sided) significant at level 5% (#) or 1% (##), Bonferroni-Holm-corrected

Conclusions:

The test substance had no effect on intrauterine development.

Reference 3

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

GLP-Guideline study, tested with the source substance Decanoic acid, mixed diesters with octanoic acid and propylene glycol (CAS 68583-51-7). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (Dec 2012)”, 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 414 (Prenatal Developmental Toxicity Study)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.31 (Prenatal Developmental Toxicity Study)

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Sprague-Dawley, CD

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga, Sulzfeld, Germany
- Age at study initiation: 8-10 weeks
- Weight at study initiation: mean 216 g
- Housing: individual in Makrolon cages
- Diet: pelleted Altromin Maintenance Diet 1324 (Lot No. 090792/0826), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 41-65
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12 (lux values 20-430)

Route of administration:

oral: gavage

Vehicle:

arachis oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Dosing solutions were prepared daily by dissolving of the test material in arachidis oil yielding a final concentration of 20-200 mg/mL.

VEHICLE
- Concentration in vehicle: 2, 6, and 20% (w/v)
- Amount of vehicle (if gavage): 5 mL/kg bw

Analytical verification of doses or concentrations:

no

Details on mating procedure:

- Impregnation procedure: the females were mated at the supplier with an accurate day of mating. The animals were reveived at the testing facility on day 0 of gestation.

Duration of treatment / exposure:

Day 6-15 of gestation

Frequency of treatment:

daily, 7 days/week

Duration of test:

10 days

No. of animals per sex per dose:

24 P females

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Dose levels were based on the results of toxicological examinations (Potokar, 1988).

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least twice daily
- Cage side observations which were included: clinical signs

BODY WEIGHT: Yes
- Time schedule for examinations: Day 0, 6, 16 and 20 of gestation

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: all maternal organs, with emphasis on the uterus

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

- External examinations: Yes: [all per litter]
- Soft tissue examinations: Yes: [half per litter]
- Skeletal examinations: Yes: [half per litter]
- Head examinations: Yes: [half per litter]

Statistics:

In case of a normal distribution, the Dunnett-Test, based on a pooled variance, was applied for the comparison between the treated groups and the control group. The Stell-Test was applied when the data could not be assumed to follow a normal distribution. Fisher´s Exact test for 2x2 tables was applied if the variable could be dichotomized without loss of information (Bonferroni-Holm-corrected).

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
During the study:
- no maternal mortality occurred,
- no compound-related symptoms were observed,
- body weight profiles were similar in all groups,
- no compound-related differences between the mean reproduction data of the test groups in comparison to the control group occurred,
- and no macroscopic changes were noted.

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:yes. Remark: No adverse or treatment-related effects

Details on embryotoxic / teratogenic effects:
The body weights of live foetus/weights of placenta and uterus exhibited no significant differences between treatment and control groups. The sex ratio was not affected by treatment (see Tables 1, 2 and 3 under ‘Any other information on results incl. tables’).

External Examination:
Only in the control group, 6 dead foetuses of dam no. 15 were observed. All 6 foetuses had malformations as hydrocephalus, exencephaly, agenesis of the mandibula and maxilla, in 3/6 exophthalmus and in 1/6 spina bifida, as well. In group 2 (100 mg/kg bw) one foetus with hydrocephalus was noted.

Visceral Examination:
In the control group 157 foetuses were examined with 17/157 showing hydronephrosis, 2/157 ureter dilatation, 3/157 ureter waved and 1/157 thorax blood coagulum. In the dose group 100 mg/kg bw, 26/150 examined foetuses had hydronephrosis, 6/150 ureter dilatation and 8/150 ureter waved. In the dose group 300 mg/kg bw, 21/150 examined foetuses had hydronephrosis, 7/150 ureter dilatation and 7/150 ureter waved. One runt was observed with normal organs and 1 foetus with hydrocephalus internus. In dose group 1000 mg/kg bw, 31/150 examined foetuses had hydronephrosis, 6/150 ureter dilatation and 16/150 ureter waved. According to the author the observed abnormalities were not treatment-related.

Skeletal Examination:
The observation revealed in the control group 12/168 foetuses with incomplete ossified skull bones and 6/168 with non ossified skull bones. In the first treatment group only 1/166 examined foetuses showed incomplete ossified skull bones and there were no findings in group 3 (300 mg/kg bw). In dose group 1000 mg/kg bw, 1/173 foetuses had incomplete ossified skull bones. The findings were considered to be identical in all groups and therefore not treatment-related.

Table 1. Maternal effects

Parameter	Group 1 0 mg/kg bw	Group 2 100 mg/kg bw	Group 3 300 mg/kg bw	Group 4 1000 mg/kg bw
Number of dams examined	24	24	24	24
Clinical findings	No clinical signs were observed during the study period in all groups.
Mortality of dams [%]	No death occurred in the dams of all groups.
Body weight gain [g] Day 6-20	131.9	128.0	131.2	137.8
Uterus weight (mean) [g]	84.9	82.1	86.0	86.8
Pregnancies [%]	96	96	92	96

 

Table 2. Litter response (Caesarean section data)

Parameter	Group 1 0 mg/kg bw	Group 2 100 mg/kg bw	Group 3 300 mg/kg bw	Group 4 1000 mg/kg bw
Corpora lutea [total number]	399	381	374	402
Corpora lutea [total/ no of dams with implantations ± SD]	17.3 ± 2.1	16.6 ± 1.6	17.0 ±2.5	17.5 ±2.2
Implantations[total number]	337	328	327	347
Implantations[total/number of dams ± SD]	14.7 ± 2.3	14.3 ± 2.6	14.9 ± 2.7	15.1 ± 2.5
Total number of live foetuses	319	316	315	333
Total number of dead foetuses	6	0	0	0
Pre-implantation loss [total/no. of dams with implantations]	2.7	2.3	2.1	2.4
Post-implantation loss [total/no. of dams with implantations]	0.8	0.5	0.5	0.6
Foetal sex ration[% male/female]	48.9/49.2	50.6/49.4	52.1/47.9	51.1/48.9
Foetus weight(mean) [g]	4.0	4.1	4.0	4.0
Placenta weight(mean) [g]	0.6	0.6	0.6	0.6
Total number of litters	23	23	22	23

Table 3: Examination of the foetuses

Parameter	Group 1 0 mg/kg bw	Group 2 100 mg/kg bw	Group 3 300 mg/kg bw	Group 4 1000 mg/kg bw
Number of foetuses per group	325	316	315	333
Total number of foetuses with malformations	6	1	0	0
% of foetuses with malformations	1.8	0.3	0	0

Conclusions:

The test substance had no effect on intrauterine development.
CLP: not classified

Reference 4

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

GLP-Guideline study, tested with the source substance Decanoic acid, mixed diesters with octanoic acid and propylene glycol (CAS 68583-51-7). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (Dec 2012)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Justification for type of information:

Please refer Section 13 for read across justification.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.31 (Prenatal Developmental Toxicity Study)

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Sprague-Dawley, CD

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga, Sulzfeld, Germany
- Age at study initiation: 8-10 weeks
- Weight at study initiation: mean 216 g
- Housing: individual in Makrolon cages
- Diet: pelleted Altromin Maintenance Diet 1324 (Lot No. 090792/0826), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 41-65
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12 (lux values 20-430)

Route of administration:

oral: gavage

Vehicle:

arachis oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Dosing solutions were prepared daily by dissolving of the test material in arachidis oil yielding a final concentration of 20-200 mg/mL.

VEHICLE
- Concentration in vehicle: 2, 6, and 20% (w/v)
- Amount of vehicle (if gavage): 5 mL/kg bw

Analytical verification of doses or concentrations:

no

Details on mating procedure:

- Impregnation procedure: the females were mated at the supplier with an accurate day of mating. The animals were reveived at the testing facility on day 0 of gestation.

Duration of treatment / exposure:

Day 6-15 of gestation

Frequency of treatment:

daily, 7 days/week

Duration of test:

10 days

No. of animals per sex per dose:

24 P females

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Dose levels were based on the results of toxicological examinations (Potokar, 1988).

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least twice daily
- Cage side observations which were included: clinical signs

BODY WEIGHT: Yes
- Time schedule for examinations: Day 0, 6, 16 and 20 of gestation

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: all maternal organs, with emphasis on the uterus

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

- External examinations: Yes: [all per litter]
- Soft tissue examinations: Yes: [half per litter]
- Skeletal examinations: Yes: [half per litter]
- Head examinations: Yes: [half per litter]

Statistics:

In case of a normal distribution, the Dunnett-Test, based on a pooled variance, was applied for the comparison between the treated groups and the control group. The Stell-Test was applied when the data could not be assumed to follow a normal distribution. Fisher´s Exact test for 2x2 tables was applied if the variable could be dichotomized without loss of information (Bonferroni-Holm-corrected).

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
During the study:
- no maternal mortality occurred,
- no compound-related symptoms were observed,
- body weight profiles were similar in all groups,
- no compound-related differences between the mean reproduction data of the test groups in comparison to the control group occurred,
- and no macroscopic changes were noted.

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:yes. Remark: No adverse or treatment-related effects

Details on embryotoxic / teratogenic effects:
The body weights of live foetus/weights of placenta and uterus exhibited no significant differences between treatment and control groups. The sex ratio was not affected by treatment (see Tables 1, 2 and 3 under ‘Any other information on results incl. tables’).

External Examination:
Only in the control group, 6 dead foetuses of dam no. 15 were observed. All 6 foetuses had malformations as hydrocephalus, exencephaly, agenesis of the mandibula and maxilla, in 3/6 exophthalmus and in 1/6 spina bifida, as well. In group 2 (100 mg/kg bw) one foetus with hydrocephalus was noted.

Visceral Examination:
In the control group 157 foetuses were examined with 17/157 showing hydronephrosis, 2/157 ureter dilatation, 3/157 ureter waved and 1/157 thorax blood coagulum. In the dose group 100 mg/kg bw, 26/150 examined foetuses had hydronephrosis, 6/150 ureter dilatation and 8/150 ureter waved. In the dose group 300 mg/kg bw, 21/150 examined foetuses had hydronephrosis, 7/150 ureter dilatation and 7/150 ureter waved. One runt was observed with normal organs and 1 foetus with hydrocephalus internus. In dose group 1000 mg/kg bw, 31/150 examined foetuses had hydronephrosis, 6/150 ureter dilatation and 16/150 ureter waved. According to the author the observed abnormalities were not treatment-related.

Skeletal Examination:
The observation revealed in the control group 12/168 foetuses with incomplete ossified skull bones and 6/168 with non ossified skull bones. In the first treatment group only 1/166 examined foetuses showed incomplete ossified skull bones and there were no findings in group 3 (300 mg/kg bw). In dose group 1000 mg/kg bw, 1/173 foetuses had incomplete ossified skull bones. The findings were considered to be identical in all groups and therefore not treatment-related.

Table 1. Maternal effects

Parameter	Group 1 0 mg/kg bw	Group 2 100 mg/kg bw	Group 3 300 mg/kg bw	Group 4 1000 mg/kg bw
Number of dams examined	24	24	24	24
Clinical findings	No clinical signs were observed during the study period in all groups.
Mortality of dams [%]	No death occurred in the dams of all groups.
Body weight gain [g] Day 6-20	131.9	128.0	131.2	137.8
Uterus weight (mean) [g]	84.9	82.1	86.0	86.8
Pregnancies [%]	96	96	92	96

 

Table 2. Litter response (Caesarean section data)

Parameter	Group 1 0 mg/kg bw	Group 2 100 mg/kg bw	Group 3 300 mg/kg bw	Group 4 1000 mg/kg bw
Corpora lutea [total number]	399	381	374	402
Corpora lutea [total/ no of dams with implantations ± SD]	17.3 ± 2.1	16.6 ± 1.6	17.0 ±2.5	17.5 ±2.2
Implantations[total number]	337	328	327	347
Implantations[total/number of dams ± SD]	14.7 ± 2.3	14.3 ± 2.6	14.9 ± 2.7	15.1 ± 2.5
Total number of live foetuses	319	316	315	333
Total number of dead foetuses	6	0	0	0
Pre-implantation loss [total/no. of dams with implantations]	2.7	2.3	2.1	2.4
Post-implantation loss [total/no. of dams with implantations]	0.8	0.5	0.5	0.6
Foetal sex ration[% male/female]	48.9/49.2	50.6/49.4	52.1/47.9	51.1/48.9
Foetus weight(mean) [g]	4.0	4.1	4.0	4.0
Placenta weight(mean) [g]	0.6	0.6	0.6	0.6
Total number of litters	23	23	22	23

Table 3: Examination of the foetuses

Parameter	Group 1 0 mg/kg bw	Group 2 100 mg/kg bw	Group 3 300 mg/kg bw	Group 4 1000 mg/kg bw
Number of foetuses per group	325	316	315	333
Total number of foetuses with malformations	6	1	0	0
% of foetuses with malformations	1.8	0.3	0	0

Conclusions:

The test substance had no effect on intrauterine development.
CLP: not classified

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises adequate, reliable (Klimisch score 2, partially due to read-across) studies from reference substances with similar structure and intrinsic properties. Read-across is justified based on common functional group(s), common precursors/breakdown products and similarities in PC/ECO/TOX properties (refer to endpoint discussion for further details). Taken together, the information from these independent sources is consistent and provides sufficient weight of evidence for hazard assessment leading to an endpoint conclusion in accordance with Annex XI, 1.2, of Regulation (EC) No 1907/2006.
Therefore, the available information as a whole is sufficient to fulfil the standard information requirements set out in Annex IX, 8.7.2.

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

There are no data on developmental toxicity available for the target substance. In accordance with Regulation (EC) No 1907/2006, Annex XI, 1.5 read-across from appropriate substances is conducted to fulfill the standard information requirements set out in Regulation (EC) No 1907/2006, Annex IX, 8.7.2.

According to 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) “to avoid the need to test every substance for every endpoint”. 

The target substance Fatty acids, C16-18 and C18-unsatd., esters with propylene glycol represents a UVCB substance predominantly comprised of diesters of an aliphatic diol (1,2-propyleneglycol (PG)) chemically linked to mainly oleic acid (C18:1) but as well to palmitic acid (C16), palmitoleic acid (C16:1), stearic acid (C18) and/or linoleic acid (C18:2). Glycol esters are in general known to be stepwise hydrolysed by gastrointestinal enzymes into the free fatty acid component and the respective alcohol (Long, 1958; Lehninger, 1970; Mattson and Volpenhein, 1972).

Based on the common metabolic fate of glycol esters, the read-across approach is based on the presence of common functional groups, common precursors and the likelihood of common breakdown products via biological processes, which result in structurally similar chemicals, common functional groups, structural similarities and similar physico-chemical, toxicological and toxicokinetic behaviour. For further details on the read-across approach, please refer to the analogue justification in section 13 of the technical dossier.

As no data are available on pre-natal developmental toxicity after exposure to Propyleneglycol dioleate, read-across to reliable data on the analogue substances Decanoic acid, mixed diesters with octanoic acid and propylene glycol (CAS 68583-51-7) and Fatty acids, C16-18, esters with ethylene glycol (CAS 91031-31-1) was conducted.

 

CAS 68583-51-7

Prenatal developmental toxicity of Decanoic acid, mixed diesters with octanoic acid and propylene glycol was characterised according to OECD guideline 414 under GLP conditions (Pittermann, 1994).

Groups of 24 female Sprague Dawley rats per dose level were orally dosed with 100, 300 and 1000 mg/kg bw/day by gavage from Day 6-15 of gestation. A concurrent negative control group receiving the vehicle (arachis oil) only was included in the testing as well. No maternal mortality occurred and no substance-related clinical signs of toxicity were observed. In addition, maternal body weight profiles were similar in all groups. Furthermore, at scheduled necropsy no macroscopic changes were noted in maternal animals.

No compound-related differences were observed in pre- and post-implantation loss, embryonic deaths, mean numbers of resorptions and total fetuses of the test groups in comparison to the control group. Mean fetal placental and uterus weights and body weights of live fetus exhibited no significant differences between treatment and control groups. In addition, the fetal sex ratio was not affected by treatment. No treatment-related fetal abnormalities or malformations were found at necropsy. The figures of skeletal and visceral variations in the treatment and control groups were considered to be comparable or of incidental origin due to missing dose-dependencies. Therefore, due to the lack of adverse effects in this study, the NOAEL for maternal toxicity and developmental toxicity for rats was considered to be ≥ 1000 mg/kg bw/day.

CAS 91031-31-1

Fatty acids, C16-18, esters with ethylene glycol was tested in an oral prenatal developmental toxicity study according to OECD guideline 414 in compliance with GLP (Pittermann, 1997).

Groups of 24 female Sprague-Dawley rats were dosed with 100, 300 and 900 mg/kg bw/day of Fatty acids, C16-18, esters with ethylene glycol via gavage from Day 6-15 of gestation. Concurrent negative control groups receiving the vehicle alone were included in all testings.

No mortalities in maternal animals and no compound-related symptoms were observed in the treatment groups. In addition, body weight and body weight gains were within the expected ranges. No compound-related differences were noted between the mean reproduction data of the test groups in comparison to the control group. At scheduled necropsy no macroscopic changes were noted in the dams of the treatment groups. Furthermore, pre-implantation loss, post-implantation loss, mean number of resorptions, embryonic deaths and total fetuses were not affected by treatment with the test substance. In addition, mean fetal placental and uterus weights were not affected. The fetal sex ratio was comparable in all groups and no treatment-related fetal abnormalities were found at necropsy. The examined fetuses showed no treatment-related malformations and the figures of visceral variations in the test groups were considered to be similar to the control group. The mean weight of live male and female fetuses in the mid-dose group was significantly increased, whereas the weights of live fetuses of the other treatment groups exhibited no significant differences. The figures of skeletal ossifications and variations showed no treatment-related deviations; thus various findings, all without dose-relationship, were considered to be incidental.

Based on the lack of adverse effects in this study, the NOAEL for maternal toxicity and developmental toxicity for rats was considered to be ≥ 900 mg/kg bw/day.

 

Conclusion for developmental toxicity

The available data on developmental toxicity of the read-across analogue substances Fatty acids, C16-18, esters with ethylene glycol (CAS 91031-31-1) and Decanoic acid, mixed diesters with octanoic acid and propylene (CAS 68583-51-7 did not show treatment-related effects on maternal animals or the offspring up to the highest tested dose level. Thus, no hazard for developmental toxicity was identified and hence, the target substance is not considered as hazardous for intrauterine development.

 

References

Agency for Toxic Substances and Disease Registry (ATSDR) (1997). Toxicological Profile for Propylene Glycol. US Department of Health and Human Services. Atlanta, US.

Agency for Toxic Substances and Disease Registry (ATSDR) (2010). Toxicological Profile for Ethylene Glycol. US Department of Health and Human Services. Atlanta, US.

Lehninger, A.L. (1970). Biochemistry. Worth Publishers, Inc.Long, C.L. et al. (1958). Studies on absorption and metabolism of propylene glycol distearate. Arch Biochem Biophys, 77(2):428-439.

Mattson, F.H. and Volpenhein, R.A. (1972). Hydrolysis of fully esterified alcohols containing from one to eight hydroxyl groups by the lipolytic enzymes of the rat pancreatic juice. Journal of Lipid Research 13: 325-328

Miller, O.N., Bazzano, G. (1965).Propanediol metabolism and its relation to lactic acid -metabolism. Annals of the New York Academy of Sciences 119:957-973.

Ritchie, A.D. (1927). Lactic acid in fish and crustacean muscle. Journal of Experimental Biology 4:327-332.

Justification for selection of Effect on developmental toxicity: via oral route: Hazard assessment is conducted by means of read-across from structural analogues. All available studies are adequate and reliable based on the identified similarities in structure and intrinsic properties between source and target substances and overall quality assessment (refer to the endpoint discussion for further details). 

Justification for selection of Effect on developmental toxicity: via inhalation route: In accordance with Regulation (EC) No 1907/2006, Annex IX, no study is required to fulfil the data requirements. 

Justification for selection of Effect on developmental toxicity: via dermal route: In accordance with Regulation (EC) No 1907/2006, Annex IX, no study is required to fulfil the data requirements.

Justification for classification or non-classification

Based on the analogue read-across approach, the available data on developmental toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008, and are therefore conclusive but not sufficient for classification.