Decanoic acid, mixed diesters with octanoic acid and propylene glycol

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Two-generation toxicity study (OECD 416, rat, m/f): NOAEL (fertility) = 1000 mg/kg bw/day

The hazard assessment is based on the data currently available. New studies with the registered substance and/or other member substances of the glycol esters category will be conducted in the future. The finalised studies will be included in the technical dossier as soon as they become available and the hazard assessment will be re-evaluated accordingly.

For further details, please refer to the category concept document attached to the category object (linked under IUCLID section 0.2) showing an overview of the strategy for all substances within the glycol esters category.

Link to relevant study records

Reference

Endpoint:

two-generation reproductive toxicity

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

17 Nov 2003 - 18 Jul 2004

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Lack of test material details.

Qualifier:

according to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Deviations:

yes

Remarks:

lack of test material details.

GLP compliance:

yes (incl. QA statement)

Remarks:

Freie und Hansestadt Hamburg, Behörde für Wissenschaft und Gesundheit, Hamburg, Germany

Limit test:

no

Species:

rat

Strain:

other: CD® / Crl:CD®

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH, Sulzfeld, Germany
- Age at study initiation: (P) 40-47 days (males/females)
- Weight at study initiation: (P) 164.0 - 206.3 g (males) and 139.9 - 176.1 g (females)
- Housing: pre-mating animals and the pregnant females were housed individually, the rearing females were housed together with their pups and the mating animals were kept as pairs in Makrolon type III cages. Coarse granulated wood (Brandenburg, Goldenstedt-Arkeburg, Germany) was used as bedding material.
- Diet: ssniff R-Z V1324 (ssniff Spezialdiäten GmbH, Soest, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: once daily, 7 days/week

VEHICLE
- Amount of vehicle: 2 mL/kg bw
- Lot/batch no.: 31107453

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: until copulation occurred or 2 weeks had elapsed
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as Day 0 of pregnancy
- After 14 days of unsuccessful pairing the animals were separated without further opportunity for mating.
- After successful mating each pregnant female was caged: individually

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The determination of the test substance in corn oil mixtures was performed applying a GC method with FID detection. The test substance was quantified applying a reference standard. The following parameters were determined: linearity, accuracy, precision, sensitivity, specificity and stability. The results of the analysis showed that the test item vehicle mixtures were correctly prepared and the concentration and stability found were in good agreement to those expected. The actual concentrations of the test item-carrier mixtures were within the measured range of 97.4-104.7% of the nominal test item concentrations.

Duration of treatment / exposure:

(F0) Males: 10 weeks before mating, 2 weeks during mating.
(F0) Females: 10 weeks before mating, up to 2 weeks during mating, approximately 3 weeks during pregnancy, 3 weeks during lactation.
(F1) Males: 10 weeks before mating, 2 weeks during mating.
(F1) Females: 10 weeks before mating, up to 2 weeks during mating, approximately 3 weeks during pregnancy, 3 weeks during lactation.

Frequency of treatment:

daily, 7 days/week

Details on study schedule:

- F1 parental animals not mated until at least 10 weeks after selected from the F1 litters.
- Selection of parents from F1 generation when pups were 3 weeks of age.

Remarks:

Doses / Concentrations:
100, 300 and 1000 mg/kg bw/day
Basis:
nominal conc.

No. of animals per sex per dose:

20

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dose levels for this study were selected in agreement with the sponsor based on available toxicity data.

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: Parental animals (F0 and F1) were weighed on the first day of dosing and at least weekly thereafter. Parental females (F0 and F1) were weighed daily during gestation and during lactation on the same days as the weighing of litters and on the day the animals were sacrificed.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal was determined weekly during the pre-mating period and daily during the gestation periods. After parturition and during lactation, food consumption measurements were performed on the same day as the weighing of the litters.

Oestrous cyclicity (parental animals):

Oestrous cycle length and normality were evaluated in F0 and F1 females by vaginal smear 3 weeks prior to mating until evidence of mating was found. When obtaining vaginal/cervical cells, care was taken to avoid disturbance of mucosa and subsequently, the induction of pseudopregnancy.

Sperm parameters (parental animals):

Parameters examined in [F0/F1] male parental generations: testis weight, epididymis weight and one of each paired organ was reserved for histopathological examination. Of a subset of at least 10 males of each group, the remaining testes and epididymides were used for enumeration of ultra-sound resistant (homogenisation-resistant) spermatids and cauda epididymides sperm reserves, respectively. From the same groups, sperm motility and sperm morphology was determined.

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 ] offspring: number and sex of pups, weight, presence of milk in the stomach, externally visible abnormalities, stillbirths.
- Functional development: mid-air righting reflex, auditory startle reflex, pupillary reflex, open field, passive avoidance learning and memory.
- Physical development: pinna detachment, ear and eye opening, cleavage of the balanopreputial gland, vaginal opening, upper incisor eruption.

GROSS EXAMINATION OF DEAD PUPS:
yes, all pups with external abnormalities or clinical signs and at least one randomly selected pup/sex/litter from F1 and F2 generation, were examined macroscopically for any structural abnormalities or pathological changes.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals at the end of the mating period
- Maternal animals: All surviving animals after 3 lactation weeks

GROSS NECROPSY
- Gross necropsy consisted of external surfaces, orifices, cranila cavity, carcass and all organs. Special attention was paid to the organs of the reproductive system.

HISTOPATHOLOGY / ORGAN WEIGHTS
The following tissues were prepared for microscopic examination: adrenal glands, coagulation gland, epididymides, grossly abnormal tissue, ovary, pituitary, prostate, seminal vesicle, testicle, uterus (with oviducts and cervix) and vagina.
The following organs were weighed: adrenal glands, brain, epididymides, kidney, liver, ovary, pituitary, prostate, seminal vesicle, spleen, testicle, thyroid and uterus (with oviducts and cervix).

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring not selected as parental animals or selected for open field and passive avoidance testes and all F2 offspring were sacrificed after 3 lactation weeks.
- These animals were subjected to postmortem examinations macroscopic and microscopic examination as follows: full histopathology of the organs were performed for all high dose and control F1 parental animals per sex.

GROSS NECROPSY
- Gross necropsy consisted of grossly abnormal tissue.

HISTOPATHOLOGY
- Grossly abnormal tissue from all pups with external abnormalities or clinical signs, as well as from at least one randomly selected pup/sex/litter from F1 and F2 generation were fixed for possible histopathological examinations. In F1 females evaluation of Corpora lutea of lactation in ovaries was performed.

Statistics:

For all numerical values homogeneity of variances was tested by using the BARTLETT chi-square test. If the variances were homogenous, the DUNNETT test (p ≤ 0.01) was used to compare the experimental groups with the control group.
In case of heterogeneity of variances, the STUDENT´s t-test was carried out, limit of significance was p ≤ 0.01.
For the comparison of classification measurements the FISHER´s exact test (≤ 0.05), n < 100 or chi²-test with Yates´correction for continuity, n ≥ 100 (p ≤ 0.01) was employed.

Reproductive indices:

For each group the following calculations were performed:
- Fertility female index: (number of pregnancies/number of matings) x 100
- Gestation index: (number of litters with live pups/number of pregnancies) x 100

For each litter and group the following calculations were performed:
- Live-born index: (number of pups born alive/total number of pups born) x 100
- Lactation index: (number of pups alive on Day 21/number of pups alive on Day 4) x 100
- Sex ratio: number of males/number of females
- Percentage by Sex: number of males (females)/total number of animals x 100
- Viability index: (number of pups alive on Day 4/number of pups alive and kept on Day 0) x 100

Offspring viability indices:

For each litter and group the following calculations were performed:
- Live-born index: (number of pups born alive/total number of pups born) x 100
- Lactation index: (number of pups alive on Day 21/number of pups alive on Day 4) x 100
- Sex ratio: number of males/number of females
- Percentage by Sex: number of males (females)/total number of animals x 100
- Viability index: (number of pups alive on Day 4/number of pups alive and kept on Day 0) x 100

Clinical signs:

no effects observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

F0: Low-/mid-dose (f): increased body weight gain; high-dose (f): decreased body weight gain, low-dose (f): increased food consumption, non-adverse; F1: mid-dose (f): decreased body weight; high-dose (f): increased body weight, non-adverse

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

F0: Low-/mid-dose (f): increased body weight gain; high-dose (f): decreased body weight gain, low-dose (f): increased food consumption, non-adverse; F1: mid-dose (f): decreased body weight; high-dose (f): increased body weight, non-adverse

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

F0: mid- and high dose: reduced live-born indices; high-dose: increased post-implantation loss and prolonged gestation length, non-adverse; F1: high-dose (f): increased fertility index, non-adverse

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
F0: No mortality and no signs of systemic toxicity were observed.
F1: No mortality and no signs of systemic toxicity were observed.

BODY WEIGHT (PARENTAL ANIMALS)
F0: No influence was noted on the body weight and the body weight gain of the male rats during the pre-mating and mating period. Statistically significant increase in body weight gain during the pre-mating and mating period of the low- mid-dose female group were observed. Additionally, decrease of body weight gain during the gestation period was observed in the mid-dose group. The differences were not considered to be test item-related (Table 3 under "Any other information on results incl. tables").
F1: No influence was noted on the body weight and the body weight gain of the male rats during the pre-mating and mating period. In the mid-dose female group a decreased body weight gain and in the high and low-dose female group an increased body weight gain was apparent but not considered to be test-item related. A statistically significant increase in food consumption in the low- and high-dose female group was determined and was not considered to be test item-related.

FOOD CONSUMPTION (PARENTAL ANIMALS)
F0: No test item-related influence was observed on the food consumption or the food efficiency of the male animals treated with 100, 300 or 1000 mg/kg bw/day during the pre-mating period. A statistically significant increase in food consumption in the low dose female group was determined and was not considered to be test item-related.
F1: No test item-related influence was observed on the food consumption or the food efficiency of the male animals treated with 100, 300 or 1000 mg/kg bw/day during the pre-mating period.
No test item-related influence was observed on the food consumption or the food efficiency of the female animals treated with 100, 300 or 1000 mg/kg bw/day during the pre-mating, gestation or lactation period. Statistically significant increases in food consumption were observed in the high and low dose group in week 8 and week 20, respectively and were not considered to be test item-related.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
F0: No test item-related influence on the number or the length of the oestrous cycles was observed at any of the tested dose levels during the pre-mating and the mating period. No test item-related difference was noted for the number of abnormal cycles between the dose level groups and the control group.
F1: No test item-related influence on the number or the length of the oestrous cycles was observed at any of the tested dose levels during the pre-mating and the mating period. No test item-related difference was noted tor the number of abnormal cycles between the dose level groups compared to the control group.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
F0: No test item-related influence was noted on the male fertility of the animals.
The number of ultrasound-resistant spermatids per gram testicular tissue was not influenced at any of the tested dose levels. No test item related changes were noted in the percentage of motile spermatozoa in the epididymal cauda for the males treated with either 100, 300 or 1000 mg/kg bw/day. The mean percentage of morphologically normal spermatids was not different in the dosed males as compared to the control animals.
F1: No test item-related influence was noted on the male fertility of the animals. The number of ultrasound-resistant spermatids per gram testicular tissue was not influenced. No test item related changes were noted in the percentage of motile spermatozoa in the epididymal cauda for the males treated with either 100, 300 or 1000 mg/kg bw/day. The mean percentage of morphologically normal spermatids was not different in the dosed males as compared to the control animals.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
F0: In the high dose group an increased postimplantation loss (statistically significant at p ≤ 0.05) and a statistically (at p ≤ 0.01) significantly prolonged gestation length was observed. The value of post-implantation loss was slightly out of the range of the historical control data and was not considered to be of toxicological relevance. The marginally and not dose-related prolonged gestation length in the high dose group was 21.8 days compared to 21.2 days of the control group.The slight but statistically (at p ≤ 0.05) significantly reduced live-born indices at 300 or 1000 mg/kg bw/day were regarded to be spontaneous; the values were still within the background data of the testing laboratory. Furthermore, the reduced values in each group are based on one litter each only with no viable pups. In the high dose dam no. 171 revealed only one pup which was dead, in the intermediate dose dam no. 126 revealed a litter with 9 dead pups. If both litters are excluded from calculation, the mean live-born index is 98.8% for the intermediate dose group and 98.6% tor the high dose group. These values are within the range of the control group. No test item-related influence was noted for the number of implantation sites, pups at birth (alive and dead), runts or malformed pups, in the number of dams with runts or malformed pups as well as in the value calculated for the gestation index. No female aborted. No malformed pups ware noted. No test item-related influence was noted on the sex ratio, the percentage by sex and the survival indices (viability indices and lactation index) in any of the tested dose groups (see Table 1 and 5 under "Any other information on results incl. tables").
F1: No test substance-related influence on post-implantation loss and gestation length was observed in any animal.

SEXUAL MATURATION
F1: No test item-related influence was noted on the female fertility index of the animals treated with 100, 300 or 1000 mg/kg bw/day. The statistically significantly (at p ≤ 0.01) increased fertility index of the high dosed females is due to the relatively low fertility index of the control females and, therefore, of no biological relevance (see Table 1). No test item-related influence was noted on the pre-coital time of the females.
There were no test item-related differences between the control group and the treated animals in the number of implantation sites, pups at birth (alive and dead), stillbirths, live born pups, in the number of dams with stillborn pups, runts and malformed pups as well as in the values calculated for the gestation index, the gestation length, the live-born Index and the post-implantation loss. No female aborted.
No test item-related influence was noted on the sex ratio, the percentage by sex and the survival indices (viability indices and lactation index) in any of the tested dose groups.

ORGAN WEIGHTS (PARENTAL ANIMALS)
F0: In the high-dose females a statistically significant increase in relative organs weights of brain, spleen and pituitary were observed. These slight alterations in comparison to the control animals were without biological relevance. In the low dose female group, increased pituitary and uterus weight was observed and not considered to be test item-related. The respective absolute organ weights remained unchanged (see Table 2 under "Any other information on results incl. tables").
F1: No test item-related influence was noted tor the relative and absolute organ weights of male and female animals treated with 100, 300 or 1000 mg/kg bw/day. A slight but statistically significantly (at p ≤ 0.01) reduced relative organ weight of the left adrenal is regarded as spontaneous as the effect was observed for the left adrenal only.

GROSS PATHOLOGY (PARENTAL ANIMALS)
F0: Macroscopic inspections at necropsy revealed no test item-related changes in the organs or tissues of the animals treated with either 100, 300 or 1000 mg/kg bw/day (see Table 2 under "Any other information on results incl. tables").
F1: No test item-related changes were observed during necropsy.

HISTOPATHOLOGY (PARENTAL ANIMALS)
F0: The histomorphological examination of rats treated with the high-dose did not reveal any microscopical morphological changes which are considered to be related to the treatment with the test item. Type, incidence and severity of the lesions recorded were not increased in the treated animals as compared to the control animals. All lesions observed were interpreted as spontaneous organ changes and were thus not test item-related.
F1: The histopathological examination of the high-dose group did not reveal any morphological changes which are considered to be related to the test item. In F1 females unaffected Corpora lutea of lactation were present in ovaries of all females. No statistically significant difference was noted on the number of primordial and small growing follicles during examination of the ovaries.

Dose descriptor:

NOEL

Effect level:

300 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: overall effects

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: Increased post-implantation loss and increased gestation length.

Critical effects observed:

not specified

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

F1 (until weaning) high-dose (m, f): increased body weight

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

F1 (until weaning) low- and high-dose (f, m): decreased relative brain weight, non-adverse

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

F1 (until weaning) low- and high-dose: delayed ear opening; high dose (m, f): delayed pinna detachement and eye opening; mid- and high-dose (f): delayed vaginal opening, non-adverse

Histopathological findings:

no effects observed

CLINICAL SIGNS AND MORTALITY (OFFSPRING)
F1/F2: No clinical signs and no mortality were reported.

DEVELOPMENTAL LANDMARKS (OFFSPRING)
F1: No test item-related influence was noted on the physical development. No delayed development was noted for the examined pinna detachment, upper incisor eruption, ear and eye opening.
Statistically significant differences in morphological landmarks which are not considered to be test item-related are listed in Table 4 under "Any other information on results incl. tables".

BODY WEIGHT (OFFSPRING)
F1: No test item-related influence was noted on the mean and total litter weight at any of the tested dose levels. Statistically significant differences in mean body weight where apparent for the following groups but were not considered to be test item-related: increase in mean body weight of the high dose females and males.
F2: No test item-related influence was noted on the mean and total litter weight at any of the tested dose levels.

ORGAN WEIGHTS (OFFSPRING)
F1: No test item-related influence was noted for the relative and absolute organ weights. A statistically significant (at p ≤ 0.01) decreased relative brain weight of the male and female pups of the low and high dose group is regarded to be due to the slightly increased body weight at autopsy observed for these dose groups.
F2: No test item-related influence was noted for the relative and absolute organ weights of the selected male and female F2 pups at any of the tested dose levels.

GROSS PATHOLOGY (OFFSPRING)
F1: No test item-related changes were observed in the organs or tissues of the F1 pups selected for necropsy.
F2: No test item-related changes were observed in the organs or tissues of the F2 pups selected for necropsy.

SEXUAL MATURATION (OFFSPRING)
F1: No delayed development was noted for the cleavage of balanopreputial gland and vaginal opening.

HISTOPATHOLOGY (OFFSPRING)
F1: No test item-related changes in the organs or tissues of the F1 pubs were observed at any of the tested dose levels.
F2: No test item-related changes in the organs or tissues of the F1 pubs were observed at any of the tested dose levels.

FUNCTIONAL TESTS (OFFSPRING)
F1: No test item-related influence on the functional development of the F1 pups was observed. The number of pups showed no auditory startle reflex, midair righting reflex and pupillary reflex as well as the number of pups failed the passive avoidance test revealed no difference between the test item-treated animals and the control group. No test item-related influence in the open field test was apparent. The statistically significant (at p ≤ 0.01) decreased number of pups failed the passive avoidance learning test observed in the intermediate and high dose group is regarded to be a spontaneous finding. Under the present test conditions a decreased number of pups tailed the passive avoidance learning test compared to the control is of no biological relevance.

MATERNAL CARE (OFFSPRING)
F1: No deficiencies in maternal care were noted for the females treated with the test substance. No inadequate lactation or cannibalism of alive and healthy pups was observed during the 3-week lactation period.

ANOGENITAL DISTANCE (OFFSPRING)
F2: No test item-related influence was noted on the anogenital distance at any of the tested dose levels. The statistically significantly (at p ≤ 0.01) decreased anogenital distance noted for male pups of the intermediate dose group is regarded to be a spontaneous finding as no dose response relationship was observed.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed

Critical effects observed:

no

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects

Reproductive effects observed:

not specified

Table 1. Table for reproductive toxicity study.

Group /Dose level	Fertility Index in % F0         F1		No. of males evaluated for fertility F0                F1		No. of fertile males F0          F1
Group 1 (control)	83	79	24	24	22	23
Group 2 (100 mg/kg bw/day)	92	79	24	24	23	24
Group 3 (300 mg/kg bw/day)	92	96*	24	24	24	23
Group 4 (1000 mg/kg bw/day)	96*	88	24	24	24	23

*: statistically significant at p ≤ 0.01 (Fisher test)

Table 2. Organ weights of F0 females.

Organ	Group	Increase ↑ Decrease ↓
Brain	Group 4 (1000 mg/kg bw/day)	↑*
Pituitary	Group 2 (100 mg/kg bw/day) Group 4 (1000 mg/kg bw/day)	↑* ↑*
Spleen	Group 4 (1000 mg/kg bw/day)	↑*
Uterus	Group 2 (100 mg/kg bw/day)	↑*

*: statistically significant at p ≤ 0.01 (Fisher test)

Table 3. Body weight gain.

Period / Day of age	Group	Increase ↑ Decrease ↓
Pre-mating and mating	Group 2 (100 mg/kg bw/day); F0 females Group 3 (300 mg/kg bw/day); F0 females	↑* ↑*
Gestation	Group 3 (300 mg/kg bw/day); F0 females	↓*
Day 7, 14, 21	Group 4 (1000 mg/kg bw/day); F1 (until weaning) males and females	↑*
Week 1-2	Group 3 (300 mg/kg bw/day); F1 (parental animals) females	↓*
Week 9-10	Group 4 (1000 mg/kg bw/day); F1 (parental animals) females	↑*

*: statistically significant at p ≤ 0.01 (Fisher test)

Table 4. Examination of morphological landmarks F1 (until weaning).

Parameter	Group	Accelerated ↑ Delayed ↓	Reason	Time point
Pinna detachment	Group 4 (1000 mg/kg bw/day); females and males	↓*	A	Day of life
Ear opening	Group 2 (100 mg/kg bw/day); females and males Group 4 (1000 mg/kg bw/day); females and males	↓* ↓*	A, B A	Day of life
Eye opening	Group 4 (1000 mg/kg bw/day); females and males	↓*	A	Day of life
Vaginal opening	Group 3 (300 mg/kg bw/day); females Group 4 (1000 mg/kg bw/day); females	↓* ↓*	A, B A	Day after conception Day of life

*: statistically significant at p ≤ 0.01 (Fisher test)

A: the slight alternation in comparison to control animals is without biological relevance

B: lacking dose dependence

C: effect is due to the relative low or high value observed for the control group

D: effect observed before start of treatment

Table 5. Table for reproductive performance.

	Group 1 (control)	Group 2 (100 mg/kg bw/day)	Group 3 (300 mg/kg bw/day)	Group 4 (1000 mg/kg bw/day)
Fertility Index in % F0 F1	83 79	92 79	92 96*	96* 88
No. of males evaluated for fertility F0 F1	24 24	24 24	24 24	24 24
No. of fertile males F0 F1	22 23	23 24	24 23	24 23
Gestation length (days) F0 F1	21.2 21.2	21.5 21.5	21.5 21.2	21.8## 21.5
Live-Born Index (%) F0 F1	99.7 100	96.5 100	93.9* 100	93.7* 99.5
Post-implantation loss (%) F0 F1	8.0 7.6	15.9 6.1	14.8 7.6	18.9* 6.7
Gestation Index (%) F0 F1	100 100	100 100	100 100	100 100
Implantation sites (%) F0 F1	339 325	334 312	299 324	310 341

*: statistically significant at p ≤ 0.05 (Fisher test)

##: statistically significant at p ≤ 0.01 (Dunnett test or Student´s t-test)

Table 6. Maternal effects.

Parameter	Group 1 Control	Group 2 100 mg/kg bw/day	Group 3 300 mg/kg bw/day	Group 4 1000 mg/kg bw/day
Total treated F0/F1	20	20	20	20
No. (%) dead F0/F1	0 (0)	0 (0)	0 (0)	0 (0)
Body weight gain [g] mean (Day 1-21) F0 F1	53.3 ± 6.4 56 ± 8.1	53.1 ± 6.4 54 ± 6.9	51.1 ± 7.9 51.5 ± 5.4	50.0 ± 11.7 53.1 ± 6.7
Gravid uterine weight [g/kg bw] F0 F1	1.744 ± 0.277 1.488± 0.544	2.281 ± 0.649** 1.354± 0.243	1.759 ± 0.464 1.557± 1.592	2.167 ± 0.710 1.496± 0.461
No. (%) pregnant F0 F1	83 79	92 79	92 96*	96* 88

* p ≤ 0.05 Fisher test

** p ≤ 0.01 Dunnett test or Student´s t-test

Table 7. Litter response.

Parameter	Group 1 0 mg/kg bw/day	Group 2 100 mg/kg bw/day	Group 3 300 mg/kg bw/day	Group 4 1000 mg/kg bw/day
Implantations [total number] F0 F1	339 325	334 312	299 324	310 341
Implantations F0 [total/number of dams ± SD]	17 ± 2.3	16.7 ± 2.0	15 ± 3.0	15.5 ± 3.2
Total number of live and dead foetuses at birth from F0 F1	313 300	286 293	269 299	265 320
Total number of dead foetuses F0 F1	1 0	8 0	12 0	5 1
Total number of live born pups F0 F1	312 300	278 293	257 299	260 319
Post-implantation loss(%) F0 F1	8.0 7.6	15.9 6.1	14.8 7.6	18.9* 6.7
Foetal sex rationat birth (mean %, m:f) F1 F2	44.7/55.3 50/50	52.8/47.2 52/48	50.9/49.1 49/51	51.4/48.6 51/49
Foetus weight (mean at Day of Age 1) [g] F1 F2	6.4 6.5	6.7 6.7	6.7 6.6	6.8 6.8
Number of malformed pups F1 F2	0 0	0 0	0 0	0 1#
Number of runts F1 F2	1 1	2 0	0 1	0 0

*: statistically significant at p≤ 0.05 (Fisher test)

#: short tail was observed for pup no. 487-5 m

Conclusions:

The test substance had no effect on reproductive performance.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The available information comprises an adequate and reliable study (Klimisch score 2) from a reference substance with similar structure and intrinsic properties. Read-across is justified based on common functional group(s), common precursors and breakdown products and similarities in PC/ECO/TOX properties (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.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

The hazard assessment is based on the data currently available. New studies with the registered substance and/or other member substances of the glycol esters category will be conducted in the future. The finalised studies will be included in the technical dossier as soon as they become available and the hazard assessment will be re-evaluated accordingly.

For further details, please refer to the category concept document attached to the category object (linked under IUCLID section 0.2) showing an overview of the strategy for all substances within the glycol esters category.

CAS 627-83-3, CAS 68583-51-7, CAS 84988-75-0 and CAS 624-03-3

Within the category of Glycol Esters, one study with Butylene glycol dicaprylate / dicaprate (CAS 853947-59-8) is available. The study of the category member was considered for assessment and read-across was conducted based on a category approach.

Butylene glycol dicaprylate / dicaprate was tested for toxicity to reproduction in a two-generation reproduction toxicity study according to OECD 416 in compliance with GLP (LPT Laboratory, 2005). Groups of 20 CD® / Crl:CD® rats per sex and dose were given 100, 300 and 1000 mg/kg bw/day of the test material by gavage. Males were given the test material daily for 10 weeks until mating and 2 weeks during mating. Females were treated with the test material in the same way and additionally approximately 3 weeks during pregnancy and 3 weeks during lactation. A concurrent negative control group receiving the vehicle corn oil only was included in the testing as well. Examination of the parental animals revealed no clinical signs of toxicity or mortality in relation to the test substance. Furthermore, no influence was noted on the body weight and the body weight gain of the male rats during the pre-mating and mating period. Statistically significant differences in body weight gain in females were observed but were not considered to be test item-related. No influence on food consumption or food efficiency was noted in parental males during the premating period and in females during the premating, gestation and lactation period at any of the tested dose levels. In male animals of the parental F0 and F1 generation, examination showed no test-item related influence on the male fertility period, sperm number, viability and morphology within the treated groups. In female animals, no test item-related influence on female fertility indices was noted. In addition, no influence on the number or the length of the oestrous cycles was observed at any of the tested dose levels during the pre-mating and the mating period and no test item-related difference was noted for the number of abnormal cycles between the treated groups and the control group. However, in females of the high-dose group (F0) an increased but not dose-related post-implantation loss was observed. The value of post-implantation loss was slightly out of the range of the historical control data and was not considered to be of toxicological relevance. Furthermore, in the high-dose group a marginally and not dose-related increased prolonged gestation length was observed (21.8 days compared to 21.2 days of the control group). Post-implantation loss and gestation length in F1 females were not influenced as in the F0 females. No further effects on reproductive performance of F0 and F1 animals were observed and evaluation of the pre-coital time showed no test-item related influence in parental animals, as well. Moreover, no deficiencies in maternal care of the parental animals were noted. The macroscopic examination at necropsy revealed no substance-related differences in the organs and tissues of the parental animals. No changes in absolute and relative organ weights of the parental animals were observed, as well. Histopathological examination revealed no morphological changes which were considered to be test item-related.

Examinations of the F1 and F2 offspring showed no effects on the mean and total litter weight. Furthermore, no effects on the physical development of the F1 offspring were noted in any treated group. The anogenital distance in the F2 pups was not influenced by the test substance. At necropsy, no substance-related differences were noted between control and treated animals in the organs or tissues of the selected animals. Furthermore, the absolute and relative organ weights of male and female F1 and F2 pups was not influenced. Functional tests showed no test item-related differences in the functional development of the F1 offspring.

In summary, under the conditions of the study, the substance had no effect on reproductive performance and the NOAEL for reproduction toxicity of the parental and the F1 and F2 generations is considered to be above 1000 mg/kg bw/day (m, f).

Based on the availability of a two-generation reproductive toxicity study performed in 2005, performance of an extended one-generation reproduction toxicity study is not considered as necessary to fulfil the standard data requirements according to Regulation (EC) No. 1907/2006, Annex X, 8.7.3 Column 2, which states that two-generation reproductive toxicity studies (OECD TG 416) initiated before 13 March 2015 shall be considered appropriate to address the standard information requirement.

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.

Gubicza, L., Kabiri-Badr, A., Keoves, E., Belafi-Bako, K. (2000): Large-scale enzymatic production of natural flavour esters in organic solvent with continuous water removal. Journal of Biotechnology 84(2): 193-196.

Heymann, E. (1980): Carboxylesterases and amidases. In: Jakoby, W.B., Bend, J.R. & Caldwell, J., eds., Enzymatic Basis of Detoxication, 2nd Ed., New York: Academic Press, pp. 291-323.Gubicza, L. et al. (2000). Large-scale enzymatic production of natural flavour esters in organic solvent with continuous water removal. Journal of Biotechnology 84(2): 193-196.

International Programme on Chemical Safety (IPCS) (2001): Ethylene Glycol. Poisons Information Monograph. PIM 227.

Lilja, J. et al. (2005). Esterification of propanoic acid with ethanol, 1-propanol and butanol over a heterogeneous fiber catalyst. Chemical Engineering Journal, 115(1-2): 1-12.

Liu, Y. et al. (2006). A comparison of the esterification of acetic acid with methanol using heterogeneous versus homogeneous acid catalysis. Journal of Catalysis 242: 278-286.

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.

Radzi, S.M. et al. (2005). High performance enzymatic synthesis of oleyl oleate using immobilised lipase from Candida antartica. Electronic Journal of Biotechnology 8: 292-298.

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

Stryer, L. (1994): Biochemie. 2nd revised reprint, Heidelberg; Berlin; Oxford: Spektrum Akad. Verlag.

Tocher, D.R. (2003): Metabolism and Functions of Lipids and Fatty Acids in Teleost Fish. Reviews in Fisheries Science 11(2), 107-184.

WHO (2002): Ethylene Glycol: Human Health Aspects. Concise International Chemical Assessment Document 45.

Zhao, Z. (2000). Synthesis of butyl propionate using novel aluminophosphate molecular sieve as catalyst. Journal of Molecular Catalysis 154(1-2): 131-135.

Justification for selection of Effect on fertility via oral route:
Hazard assessment is conducted by means of read-across from a structural analogue. 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).

Effects on developmental toxicity

Description of key information

NOAEL oral (developmental): 1000 mg/kg bw/day (OECD 414, GLP)

The hazard assessment is based on the data currently available. New studies with the registered substance and/or other member substances of the glycol esters category will be conducted in the future. The finalised studies will be included in the technical dossier as soon as they become available and the hazard assessment will be re-evaluated accordingly.

For further details, please refer to the category concept document attached to the category object (linked under IUCLID section 0.2) showing an overview of the strategy for all substances within the glycol esters category.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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

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

Remarks:

Doses / Concentrations:
100, 300 and 1000 mg/kg bw/day
Basis:
other: nominal dose

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

Remarks:

maternal toxicity

Effect level:

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

Based on:

test mat.

Basis for effect level:

other: no adverse effects observed

Abnormalities:

no effects observed

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.

Dose descriptor:

NOAEL

Remarks:

developmental toxicity

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed

Abnormalities:

no effects observed

Developmental effects observed:

no

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.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The available information comprises an adequate and reliable study (Klimisch score 1), and is thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.7, of Regulation (EC) No 1907/2006.

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

The hazard assessment is based on the data currently available. New studies with the registered substance and/or other member substances of the glycol esters category will be conducted in the future. The finalised studies will be included in the technical dossier as soon as they become available and the hazard assessment will be re-evaluated accordingly.

For further details, please refer to the category concept document attached to the category object (linked under IUCLID section 0.2) showing an overview of the strategy for all substances within the glycol esters category.

CAS 68583-51-7

One study with Decanoic acid, mixed diesters with octanoic acid and propylene glycol is available. The oral prenatal developmental toxicity study was conducted according to OECD 414 under GLP conditions (Henkel, 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. Skeletal ossification showed in the low- and high-dose treatment groups one fetus each with incomplete ossified skull bones and additionally one fetus in the high-dose group with non-ossified skull bones. In the control group, 12 fetuses showed incomplete ossified skull bones and 6 fetuses showed non-ossified skull bones as well. Thus, the number of incomplete- and non-ossified skull bones was decreased in the treatment groups in comparison to the control group and the findings were considered to be identical and therefore not treatment-related.

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 624-03-3, CAS 627-83-8 and CAS 84988-75-0

Altogether, four studies investigating the developmental toxicity are available within the Glycol Ester category. The studies from the category members Fatty acids, C16-18, esters with ethylene glycol (CAS 91031-31-1), Decanoic acid, mixed diesters with octanoic acid and propylene (CAS 68583-51-7), Butylene glycol dicaprylate / dicaprate (CAS 853947-59-8) and Fatty acids, C6-12, ester with propylene glycol (CAS 85883-73-4) were considered for assessment and read-across was conducted based on a category and weight of evidence approach.

The results of the study with Decanoic acid, mixed diesters with octanoic acid and propylene are already described above.

Fatty acids, C16-18, esters with ethylene glycol, Butylene glycol dicaprylate / dicaprate and

Fatty acids, C6-12, ester with propylene glycol were tested in oral prenatal developmental toxicity studies according to OECD 414 in compliance with GLP (Henkel, 1997; Research Toxicology Centre, 2007; WIL Research Laboratories, 2007).

Groups of 24 or 25 female rats per dose were dosed with the respective test compound via gavage from Day 6-15, Day 6-17 or Day 6-19 post mating. Concurrent negative control groups receiving the vehicle alone were included in all testings.

Animals were dosed via gavage with 100, 300 and 900 mg/kg bw/day of Fatty acids, C16-18, esters with ethylene glycol. 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 above 900 mg/kg bw/day.

 

In the study with Butylene glycol dicaprylate / dicaprate, animals were dosed with the limit dose of 1000 mg/kg bw/day. No mortality or treatment-related signs of toxicity in maternal animals occurred during the study period. Body weight, body weight gain and food consumption were unaffected by treatment. No treatment-related effects in uterus weight or macroscopic changes were detected in treated females. No compound-related differences were observed in pre-and post-implantation loss, embryonic deaths and mean numbers of resorptions of the test groups in comparison to the control group. In fetuses of the treatment group, a slight but statistically significant lower mean fetal weight was observed when compared to the control group but within the historical control data for this rat strain. This slight difference was attributed to the higher presence of fetuses in the treated group compared to controls in which mean fetal weight was also unusually higher when compared to the internal historical control data of the laboratory. Three small fetuses were observed in the control and treatment group and one control fetus and one fetus of the treated group showed multiple anomalies. These changes were considered incidental. Furthermore, spontaneous changes at skeletal examination of the fetuses were noted between the treated and the control group. Visceral examination of fetuses showed unilateral cryptorchidism in two fetuses from two different litters of the treated group. The identification of the same stock male being mated with these two females gave rise to the hypothesis that the male parent could genetically transmit the finding. In addition, considering also the high presence of displaced testes in the control group, the findings described were considered evidence of spontaneous pathology, often seen in this species under the experimental conditions.

Thus, based on 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.

 

Fatty acids, C6-12, ester with propylene glycol was administered to groups of female rats at dose levels of 500, 1500 and 2500 mg/kg bw/day. No mortality occurred in maternal animals during the study period. In the mid- and high-dose group, test article-related salivation was noted in 8 and 6 females, respectively. Further findings noted in the treated groups included hair loss, scabbing and red material on various body surfaces as well as rales and soft stool. These findings occurred infrequently and were not dose-related. Mean maternal body weights, body weight gains, net body weights, net body weight gains and gravid uterine weights were unaffected by test article administration. A slightly lower food consumption in the high-dose group was not considered to be adverse based on the lack of an effect on mean body weights. Post-implantation loss, live litter size, mean fetal body weights, fetal sex ratios, mean numbers of corpora lutea and implantation sites and the mean litter proportions of pre-implantation loss were similar across all groups. There were no external developmental variations for any fetuses and no visceral and no skeletal malformations were noted for fetuses in this study which were considered to be test substance-related. Fetal malformations and developmental variations, when observed in treatment groups, occurred infrequently or at a frequency similar to that in the control group and did not occur in a dose-related manner or were within the historical control data ranges.

Thus, no adverse effects in the maternal animals and the offspring were observed and a NOAEL of 1000 mg/kg bw/day for maternal and developmental toxicity was considered.

 

Conclusion for developmental toxicity

Four studies investigating the developmental toxicity are available within the Glycol Ester category. The studies from the category members Fatty acids, C16-18, esters with ethylene glycol (CAS 91031-31-1), Decanoic acid, mixed diesters with octanoic acid and propylene (CAS 68583-51-7), Butylene glycol dicaprylate / dicaprate (CAS 853947-59-8) and Fatty acids, C6-12, ester with propylene glycol (CAS 85883-73-4) did not show treatment-related effects up to the highest tested dose level. Thus, no hazard for developmental toxicity was identified.

A waiver for the requirement to perform a prenatal developmental toxicity study in a 2nd species was included, as this requirement is considered not to add new information for hazard assessment and therefore is scientifically and, considering concerns regarding the use of vertebrate animals for experimental purposes, unjustified.

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.

Gubicza, L., Kabiri-Badr, A., Keoves, E., Belafi-Bako, K. (2000): Large-scale enzymatic production of natural flavour esters in organic solvent with continuous water removal. Journal of Biotechnology 84(2): 193-196.

Heymann, E. (1980): Carboxylesterases and amidases. In: Jakoby, W.B., Bend, J.R. & Caldwell, J., eds., Enzymatic Basis of Detoxication, 2nd Ed., New York: Academic Press, pp. 291-323.Gubicza, L. et al. (2000). Large-scale enzymatic production of natural flavour esters in organic solvent with continuous water removal. Journal of Biotechnology 84(2): 193-196.

International Programme on Chemical Safety (IPCS) (2001): Ethylene Glycol. Poisons Information Monograph. PIM 227.

Lilja, J. et al. (2005). Esterification of propanoic acid with ethanol, 1-propanol and butanol over a heterogeneous fiber catalyst. Chemical Engineering Journal, 115(1-2): 1-12.

Liu, Y. et al. (2006). A comparison of the esterification of acetic acid with methanol using heterogeneous versus homogeneous acid catalysis. Journal of Catalysis 242: 278-286.

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.

Radzi, S.M. et al. (2005). High performance enzymatic synthesis of oleyl oleate using immobilised lipase from Candida antartica. Electronic Journal of Biotechnology 8: 292-298.

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

Stryer, L. (1994): Biochemie. 2nd revised reprint, Heidelberg; Berlin; Oxford: Spektrum Akad. Verlag.

Tocher, D.R. (2003): Metabolism and Functions of Lipids and Fatty Acids in Teleost Fish. Reviews in Fisheries Science 11(2), 107-184.

WHO (2002): Ethylene Glycol: Human Health Aspects. Concise International Chemical Assessment Document 45.

Zhao, Z. (2000). Synthesis of butyl propionate using novel aluminophosphate molecular sieve as catalyst. Journal of Molecular Catalysis 154(1-2): 131-135.

Justification for selection of Effect on developmental toxicity: via oral route:
There is only one study available.

Justification for classification or non-classification

According to Article 13 of Regulation (EC) No. 1907/2006 "General Requirements for Generation of Information on Intrinsic Properties of substances", information on intrinsic properties of substances may be generated by means other than tests e.g. from information from structurally related substances (grouping or read-across), provided that conditions set out in Annex XI are met. Annex XI, "General rules for adaptation of this standard testing regime set out in Annexes VII to X” states that “substances whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity may be considered as a group, or ‘category’ of substances. This avoids the need to test every substance for every endpoint". Since the group concept is applied to the members of the Glycol Ester Category, data will be generated from representative reference substance(s) within the category to avoid unnecessary animal testing. Additionally, once the group concept is applied, substances will be classified and labeled on this basis.

Therefore, based on the group concept, all available data on toxicity to reproduction 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.