Monopentaerythritol tetraesters and dipentaerythritol hexaesters of 2-ethylhexanoic and n-valeric acids

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Oral: OECD 422, rat, NOAEL systemic ≥ 1000 mg/kg bw/day

Read-across from structural analogue source substance Hexanoic acid, 2-ethyl-, 2,2-bis [ [(2-ethyl-1-oxohexyl)oxy] methyl] -1,3-propanediyl ester (CAS No. 7299-99-2)

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 Apr - 17 Jun 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc., Yokohama, Japan
- Age at study initiation: 10 weeks
- Weight at study initiation: males: 330.8 - 420.4 g (mean 374.0 g), females: 207.2 - 245.0 g (mean 228.3 g)
- Fasting period before study: no fasting period
- Housing: steel cage with wire floor
- Diet: CE-2 from CLEA Japan, Inc., Tokyo, Japan; ad libitum
- Water: tap water; ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23.0 - 24.5
- Humidity (%): 49.0 - 67.0
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
50% w/v solution was prepared by diluting the test substance in vehicle. This solution was diluted serially with vehicle to prepare 5 and 15% w/v solution. Gavage solution was prepared within 8 days before administration, since the stability was verified for 8 days. Prepared samples were stored at room temperature in the dark.

VEHICLE
- Justification for use and choice of vehicle (if other than water): test substance is insoluble in water, but not in corn oil
- Lot/batch no. (if required): V2P1825

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: 2 weeks
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as Day 0 of pregnancy
- Further matings after two unsuccessful attempts: no
- After successful mating, females were individually caged. After day 18 of pregnancy, they were cage in plastic breeding cages with paper and pulp chips (Paperclean, Japan SLC Inc.,) for nesting.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

0.5 mL of each prepared solution was mixed with hexane to be confirmed by GC. The resulting analytical concenrations were 107 - 110% of the nominal concentrations

Duration of treatment / exposure:

Males: 42 days
Females: from Day 14 before mating to Day 4 of lactation
Females(satellite): 42 days

Recovary period :
- Males: 14 days
- Females (satellite): 14 days

Frequency of treatment:

once daily

Dose / conc.:

100 mg/kg bw/day (nominal)

Dose / conc.:

300 mg/kg bw/day (nominal)

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

Control: 7 males, 100 mg/kg: 12 males, 300 mg/kg: 12 males, 1000 mg/kg: 7 males;
12 females per dose;
control and 1000 mg/kg for satellite group; 5 males and 5 females per dose

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dose were based on the results of a preliminary test, where groups of male and female rats received doses of 100, 300 and 1000 mg/kg bw, respectively. No effects on general condition, organ weights neither effects on liver, kidney or spleen was observed in any of the treated animals. Based on this, the maximum dose for the study was set to 1000 mg/kg bw, while the medium and low dose were set to 300 and 100 mg/kg bw, respectively.
- Post-exposure recovery period in satellite groups: 14 days

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice a day during treatment period and once a day during recovery period

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:
Males: Day 0, 7, 14, 21, 28, 35 and 42 of treatment
Males in satellite groups: Day 0, 7, 14, 21, 28, 35, 42 of treatment, Day 7 and 14 of recovery period
Females: Day 0, 7, 14, 21, 28, 35, 42 of treatment and once from Day 0 of lactation to Day 4 of lactation
Females in satellite groups: Day 0, 7, 14, 21, 28, 35, 42 of treatment, Day 7 and 14 of recovery period

BODY WEIGHT: Yes
- Time schedule for examinations:
Males: Day 1, 7, 14, 21, 28, 35, 42 of administration and before sacrifice
Males and females in satellite groups: Day 1, 7, 14, 21, 28, 35 , 42 of administration, Day 1, 7, 14 of recovery period and before sacrifice
Females: Day 1, 7, 14, 21 of treatment, Day 0, 7, 14, 20 of pregnancy, Day 0 and 4 of lactation, and before sacrifice

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Day 43 (on day after last treatment) for males, Day 15 of recovery period for satellite animals of both sexes, Day 4 of lactation for dams, Day 26 of pregnancy for non delivered dams
- Anesthetic used for blood collection: Yes (sodium pentobarbital)
- Animals fasted: Yes (18 - 24 hours)
- How many animals: 5 in each group
- Parameters checked: erythrocytes count (RBC), hemoglobin, hematocrit, Mean Cell Volume (MCV), Mean Corpuscular Hemoglobin (MCH), Mean Corpuscular Hemoglobin Con. (MCHC), leykocyte count (WBC), differential leukocyte count, platelet, Prothrombin time (PT), Activated partial thromboplastin time (APTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Day 43 (on day after last treatment) for males, Day 15 of recovery period for satellite animals of both sex, Day 4 of lactation for dams, Day 26 of pregnancy for non delivered dams
- Anaesthetic used for blood collection: Yes (pentobarbital sodium)
- Animals fasted: Yes (18 - 24 hours)
- Parameters checked: total protein, albumin, A/G, blood urea nitrogen, creatinine, glucose, total cholesterol, triglyceride, Alkaline phosphatase (ALP), Alanine transaminase (ALT (GPT)), Aspartate transaminase (AST (GOT)), γ-GTP, total bilirubin, inorganic phosphorous, calcium, Na, K, Cl

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: at the end of dosing period and at the end of recovery period
- Dose groups that were examined: all group
- Battery of functions tested: pupilary reflex, eyelid reflex, visual placing, withdrawal reflex, Payer's reaction, startle reaction, righting reflex

Oestrous cyclicity (parental animals):

Estrous cycles of all female except for satellite groups were observed before and after start of treatment. Vaginal smear samples were everyday prepared and observed until successful mating. Estrus cycles were divided into proestrus, estrus and anestrus, and frequencies of 4-day cycle, 4/5-day cycle and 5-day cycle were calculated.

Sperm parameters (parental animals):

Parameters examined in P male parental generations:
testis weight, epididymis weight

Litter observations:

Termination:
- Offspring: 4 days after birth

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was determined for pups born or found dead.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals at day 43 (one day after last treatment) and day 15 of recovery
- Maternal animals: All surviving animals at day 5 of lactation
- Females (satellite): All surviving animals at day 15 of recovery

GROSS NECROPSY
- Gross necropsy consisted of brain, the pituitary gland, spinal cord, heart, bronchotracheal, lungs (including bronchi), liver, kidney, thymus, spleen, adrenal, thyroid, parathyroid glands, stomach, duodenum, jejunum, ileum, cecum, colon, rectum, testis, epididymis, ventral prostate, seminal vesicles including coagulation glands, ovaries, uterus, vagina, bladder, lymph nodes under the jaw, mesenteric lymph nodes, sciatic nerve, femur

HISTOPATHOLOGY
Brain, the pituitary gland, spinal cord, heart, lung and bronchus, liver, kidney, thymus, spleen, adrenal gland, thyroid gland, mandibular lymph node, stomach, duodenum, jejunum, ileum, cecum, colon, rectum, testis, epididymis, prostate, seminal vesicles including coagulation glands, ovary, uterus, vagina, mesenteric lymph node, sciatic nerve, urinary bladder, bone marrow of femur

ORGAN WEIGHT
Brain, thymus, heart, liver, kidneys, spleen, adrenal glands, testes, epididymides

Postmortem examinations (offspring):

SACRIFICE
- All pups were subjected to postmortem macroscopic examination

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations

Statistics:

Fisher test, Mann-Whitney-U-Test, Student's t-test, Aspin-Welch test, Bartlett test, Kruskal-Wallis test, Dunnett-test

Reproductive indices:

Copulation index, fertility index, gestation index, implatation index, number of corpora lutea, number of implatation index

Offspring viability indices:

Number of newborns, number of dead pups, delivery index, birth index, live birth index, sex ration on day 0 and day 4, viability index

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

incidental effects which were not treatment-related (non adverse)

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

100 mg/kg bw: reduced body weight in males (non adverse)

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

100 mg/kg bw: reduced body weight in males (non adverse)

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

all changes were found distributed over all groups, thus they were regarded as spontanous incidences (non adverse)

Other effects:

no effects observed

Description (incidence and severity):

Test substance intake: 1000 mg/kg bw: significantly more food consumption during days 29 - 30 by satellite group (non adverse)

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

not examined

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

one animal in 1000 mg/kg was sterile (non adverse)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No mortality was observed in all groups.
An emaciation was observed at day 40 of treatment (Day 1 of lactation) in the female control group, but this was recovered after one day. Loss of the upper incisor was observed at day 25 - 32 in a female of the 300 mg/kg group but this was considered due to physical shock and were not affected by administration of the test substance.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Males of the 100 mg/kg group showed body weight loss compared to the control group, but this was not statistically significant. There was no difference in female groups and satellite groups compared with control groups.
Females of the satellite 1000 mg/kg group showed significantly more food consumption during days 29 - 30 compared to the control group. However, this change was considered as not compound-related as no difference in other groups was found, both within the treatment period as well as in the recovery period.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
Before mating, the estrous cycle of one animal in the 300 mg/kg group was 4/5-day and in one animal of the 100 mg/kg groups the estrous cycle was 5-day during the administration period. However, no significant difference was found.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Regarding coupling, all cases were successful but one animal in 1000 mg/kg was sterile. One dam in the control group was considered to show poor delivering behavior based on the dirty vagina. However, these changes were not compound-related, since no other significant change was found related to gestation rate, paring days until coupling, frequency of estrous, gestation length in days and delivery behaviour.

ORGAN WEIGHTS (PARENTAL ANIMALS)
At the end of dosing period, a significantly increase of absolute weight of brain was found in females (300 mg/kg), but the change of relative weight was not significant. No significant differences in males between treatment group and the control group were observed.
At the end of recovery period, a significantly increase of relative liver weight in females of the 1000 mg/kg group was noted, while no differences in males were found. The change was not considered as compound-related, since no corresponding abnormalities were observed in the livers.

GROSS PATHOLOGY (PARENTAL ANIMALS)
At the end of dosing period, no abnormalities were observed. At the end of recovery period, a tumor in right hind femur was observed in one male of the control group.

HISTOPATHOLOGY (PARENTAL ANIMALS)
At the end of dosing period, localized atrophy of the seminiferous tubules were observed in three males of the 100 mg/kg group and in one male of the 300 mg/kg group. Three of these animals had cell debris in lumen in epidedymis. Livers in males of the 1000 mg/kg and control groups exhibited fatty change in hepatocyte and microgranuloma. In all control group animals and three animals of 1000 mg/kg, eosinophilic bodies were found in cortex of the kidney, while slightly basophilic tubules were found in kidney cortex of two animals in the control group and in four animals of the 1000 mg/kg group. Extramedullary hematopoiesis and deposit of brown pigment was found in the spleen of all animals of the control and the 1000 mg/kg group. In addition, focal degeneration/fibrosis in the myocardium, accumulation of focal foam cell in the adveolus and mineralization of the arterial wall in lung, as well as lymphocytes and neutrophil cellular infiltration in prostate were observed both in control groups and 1000 mg/kg groups. However, these change were not significantly different but accidental.
In one female of the 1000 mg/kg group, follicular cyst, increased arterial follicle and decreased corpus luteum was found in ovary at the end of dosing period. In liver, fatty change in periportal hepatocytes and microgranuloma was observed in all animals of control and 1000 mg/kg groups, respectively. Basophilic tubule in the cortex of the kidney was observed in one animal at 1000 mg/ kg and slight mineralization was found in one control animal and one animal dosed with 1000 mg/kg, respectively. In the spleen, extramedullary hematopoiesis and brown deposit were found in all animals of the control and the 1000 mg/kg group. In addition, focal degeneration/fibrosis in myocardium, mineralization on arterial wall of lung and atrophy in thymus were also found in both control and 1000 mg/kg. However, these change were not significantly different but accidental.
At the end of recovery period, intramembranous ossification and periosteum proliferation were found in one male in control. These histopathological findings are due to fracture of the right femur. In one female of 1000 mg/kg, follicular cyst was found.

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects were observed in males and females of all dose groups

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

1000 mg/kg bw: one kinked tail (incidental)

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

VIABILITY (OFFSPRING)
Number of new borns, number of live newborns, birth index, live birth index, sex ration on Day 0, number of live pups on Day 4, and viability index on Day 4 and sex ratio on Day 4 showed no significant differences in all dose groups compared to the control group.

CLINICAL SIGNS (OFFSPRING)
A pup with kinked tail was observed in 1000 mg/kg. However, this external change was regarded as incidental, since no other significant external difference was found.

BODY WEIGHT (OFFSPRING)
No difference was found between treatment groups and control group.

GROSS PATHOLOGY (OFFSPRING)
No difference was found between treatment groups and control group.

Dose descriptor:

NOAEL

Remarks:

developmental

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 were observed in pups of all dose groups

Critical effects observed:

no

Reproductive effects observed:

no

Estrous cycle of dams:

Dose (mg/kg bw)	0*	100	300	1000
Number of dams examined	12	12	12	12
Pre-treatment period
Number of animals showing type of cycle
4-day cycle	12	12	12	12
Treatment period
Number of animals showing type of cycle
4-day cycle	12	11	11	12
4/5-day cycle	0	0	1	0
5-day cycle	0	1	0	0
Frequency of animals of which type of estrous cycle was changed after treatment	0/12	1/12	1/12	0/12
Mean times of estrous cycle during mating period (mean ± SD)	1 ± 0.0	1.1 ± 0.3	1.0 ± 0.0	1.0 ± 0.0

*vehicle control

Reproductive performance:

Dose (mg/kg bw)	0*	100	300	1000
Number of pairs examined	12	12	12	12
Number of pair copulated	12	12	12	12
Copulation index (%)	100	100	100	100
Number of pregnant females	12	12	12	11
Fertility index (%)	100	100	100	91.7
Pairing days until copulation (mean ± SD)	2.1 ± 1.0	2.9 ± 1.6	2.8 ± 1.1	3.4 ± 3.1

*vehicle control

Development of pups up to 4 days of lactation:

Dose (mg/kg bw)	0*	100	300	1000
Number of pregnant females	12	12	12	11
Number of pregnant females with live newborns	12	12	12	11
Gestation index (%)	100	100	100	100
Gestation length in days	22.5 ± 0.5	22.5 ± 0.5	22.3 ± 0.5	22.5 ± 0.5
Number of corpora lutea	16.4 ± 2.5	15.5 ± 1.7	15.8 ± 1.8	16.2 ± 1.0
Number if implantations	14.4 ± 2.8	15.3 ± 1.5	15.3 ± 1.7	15.5 ± 1.0
Implantation index	88.8 ± 18.7	98.5 ± 3.5	97.0 ± 4.7	95.6 ± 5.2
Day 0 of lactation (at birth)
Number of newborns	13.8 ± 2.7	13.5 ± 2.4	14.3 ± 2.1	14.4 ± 1.3
Delivery index	96.2 ± 5.0	88.4 ± 13.2	93.2 ± 6.8	93.0 ± 6.8
Number of live newborns	13.6 ± 2.8	13.2 ± 2.4	14.2 ± 2.2	14.0 ± 1.6
Males	6.5 ± 2.7	7.3 ± 2.5	7.0 ± 2.5	6.4 ± 1.8
Females	7.1 ± 1.8	5.9 ± 1.9	7.2 ± 1.6	7.6 ± 2.5
Birth index (%)	94.6 ± 8.1	86.3 ± 14.2	92.1 ± 6.7	90.5 ± 7.7
Live birth index (%)	98.2 ± 4.5	97.7 ± 6.3	98.9 ± 3.8	97.3 ± 5.0
Sex ratio on day 0	45.5 ± 18.2	53.9 ± 15.9	48.6 ± 12.5	46.0 ± 14.0
Day 4 of lactation
Number of live pups	13.4 ± 3.0	13.1 ± 2.5	14.1 ± 2.2	13.7 ± 1.6
Males	6.4 ± 2.8	7.2 ± 2.4	7.0 ± 2.5	6.3 ± 1.8
Females	7.0 ± 1.9	5.9 ± 1.9	7.1 ± 1.6	7.5 ± 2.4
Viability index	98.6 ± 4.8	99.4 ± 2.2	99.4 ± 2.2	98.2 ± 4.5
Sex ratio on day 4	45.4 ± 18.3	53.8 ± 15.6	48.8 ± 12.3	46.2 ± 14.1

*vehicle control

Mean body weights of pups up to day 4 of lactation (g):

Dose (mg/kg bw)	0*	100	300	1000
Day 0 of lactation (at birth)
Number of live newborns
Male	6.5 ± 2.7	7.3 ± 2.5	7.0 ± 2.5	6.4 ± 1.8
Female	7.1 ± 1.8	5.9 ± 1.9	7.2 ± 1.6	7.6 ± 2.5
Mean body weight (g)
Male	6.6 ± 0.5	7.1 ± 0.8	6.7 ± 0.9	6.7 ± 0.6
Female	6.4 ± 0.6	6.7 ± 0.8	6.3 ± 0.8	6.3 ± 0.5
Day 4 of lactation (at birth)
Number of live newborns
Male	6.4 ± 2.8	7.2 ± 2.4	7.0 ± 2.5	6.3 ± 1.8
Female	7.0 ± 1.9	5.9 ± 1.9	7.1 ± 1.6	7.5 ± 2.4
Mean body weight (g)
Male	10.6 ± 0.8	11.7 ± 2.0	10.6 ± 1.6	10.5 ± 1.2
Female	10.7 ± 1.4	11.2 ± 1.9	10.1 ± 1.6	10.0 ± 1.1

*vehicle control

Morphological observations of pups:

Dose (mg/kg bw)	0*	100	300	1000
Dead pups
Number of dead pups	5	5	3	7
Number of missing pups	0	0	1	3
Number of dead pups examined	5	5	2	4
Number of dead pups with external changes	0	0	0	0
Number of dead pups with visceral changes	0	0	0	0
Live pups
Number of newborns examined (at birth)	163	158	170	154
Number of newborns with external changes	0	0	0	1
Kinked tail	0	0	0	1
Number of dead pups examined at day 4 of lactation	161	157	169	151
Number of dead pups with external changes	0	0	0	0
Number of dead pups with visceral changes	0	0	0	0

*vehicle control

Conclusions:

The test material had no effect on reproductive performance and no effect on intrauterine development.

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 (Klimisch score 1) study from a source substance with similar structure and intrinsic properties. Read-across is justified based on common precursors and hydrolysis products and consistent trends in environmental fate, ecotoxicological and toxicological profile. The selected study is thus sufficient to fulfil the standard information requirements set out in Annexes VIII - X, 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

Justification for grouping of substances and read-across

The read-across from analogue source substances approach comprises aliphatic esters of poly-functional alcohols containing two to six reactive hydroxyl groups and one to six fatty acid chains. The analogue approach contains mono constituent, multi-constituent and UVCB substances with fatty acid carbon chain lengths ranging from C5 - C28, which are mainly saturated but also mono unsaturated C16 and C18, polyunsaturated C18, branched C5 and C9, branched C14 - C22 building mono-, di-, tri-, and tetra esters with an alcohol (i.e. the polyol).

The available data allows for an accurate hazard and risk assessment of the target substance and the read-across concept is applied for the assessment of environmental fate and environmental and human health hazards. Thus, where applicable, environmental and human health effects are predicted from adequate and reliable data for source substances within the group by interpolation to the target substance applying the group concept in accordance with Annex XI, Item 1.5, of Regulation (EC) No. 1907/2006. In particular, for each specific endpoint the source substance(s) structurally closest to the target substance is/are chosen for read-across, with due regard to the requirements of adequacy and reliability of the available data. Structural similarities and similarities in properties and/or activities of the source and target substance are the basis of read-across.

A detailed justification for the grouping of chemicals and read-across is provided in the technical dossier (see IUCLID Section 7.1 and 13).

Toxicity to reproduction

Toxicity to reproduction has been investigated using a structural analogue source substance in a reliable and adequate key study.

A combined repeated dose toxicity study with the reproduction/developmental toxicity screening test was performed according to OECD 422 and in compliance with GLP (MHWL, 2005b). 12 male and 12 female Crj: CD(SD) rats per dose were treated by gavage with 100, 300, and 1000 mg/kg bw/day of Hexanoic acid, 2-ethyl-, 2,2-bis [ [(2-ethyl-1-oxohexyl)oxy] methyl] -1,3-propanediyl ester (CAS No. 7299-99-2). Male rats and additional 5 female rats (recovery animals of control and 1000 mg/kg bw/day dose group) were dose once daily for 42 days. The other females were treated from day 14 before mating to day 4 of lactation. There was no substance-related mortality and no clinical signs that were related to treatment were observed during the study period. No effect on body weight gain was observed. No treatment-related effects were observed regarding organ weights, clinical chemistry and pathology. No substance-related effect on the estrus cycle was observed. No effect on the reproductive performance of the parental animals was observed. No effect on viability was observed in the offspring. In addition no clinical findings, effects on body weight, and no effects at necropsy were observed in the offspring. In conclusion, the NOAEL was ≥ 1000 mg/kg bw/day for maternal toxicity as well as reproductive toxicity.

Moreover, data taken from subchronic repeated dose toxicity studies are available for the analogue substances Fatty acids, C5-9, tetraesters with pentaerythritol (CAS No. 67762-53-2) and Pentaerythritol ester of pentanoic acids and isononanoic acid (CAS No. 146289-36-3). The studies also investigated effects on reproductive tissues and organs and indicate no potential for reproductive effects.

In a 13-week oral repeated-dose toxicity study performed comparable to OECD Guideline 408 with Pentaerythritol ester of pentanoic acids and isononanoic acid in rats (CAS No. 146289-36-3) reproductive organs were examined as well (Emery, 1998). Groups of 10 male and female Wistar rats each were exposed to the substance by gavage at 100, 300 and 1000 mg/kg bw for 90 consecutive days. Overall, there were no adverse effects found after oral application of the test substance for 90 days. With special regard to the reproductive organs (ovaries, epididymides, prostate, testes and uterus), the examination of organ weights as well as gross necropsy and histopathology revealed no substance-related findings. Based on the absence of effects up to the highest dose tested, the 90-day oral systemic NOAEL was found to exceed 1000 mg/kg bw/day.

A 90-day dermal toxicity study with Fatty acids, C5-9, tetraesters with pentaerythritol (CAS No. 67762-53-2) was performed comparable to OECD Guideline 411 (Exxon, 1988a).Groups of 10 male and female Sprague-Dawley rats each were once daily (5 days/week) exposed to the substance (purity not specified) at concentrations of 800 and 2000 mg/kg bw for 90 days (65 applications in total). Application to the skin was done open without coverage. Reproductive endpoints examined were sperm morphology and weights of reproductive organs. Overall, there were no adverse effects found after dermal application of the test substance for 90 days. Examination of reproductive organs (ovaries, epididymides, prostate and seminal vesicles, testes, uterus and vagina) revealed no substance-related findings. Additionally, cauda epididymal sperm of the control and high dose groups, examined at the end of the treatment period, did not show changes in morphology. Based on the absence of effects concerning weight of reproductive organs and sperm morphology up to the highest dose tested, the 90-day dermal systemic NOAEL was found to exceed 2000 mg/kg bw/day for the test substance in Sprague-Dawley rats.

A 90-day subchronic inhalation toxicity study was performed with Fatty acids, C5-9, tetraesters with pentaerythritol (CAS No. 67762-53-2) comparable to OECD guideline 413 in Sprague-Dawley rats (Exxon, 1992). 15 males and 15 females per group were whole body exposed to the test substance for 6 hours/day, 5 days/week at concentrations of 0.05, 0.15 and 0.5 mg/L air. The respective controls inhaled clean air under the same conditions. Reproductive parameters examined included reproductive organs and testicular spermatid count as well as epididymal sperm count and morphology. No changes in reproductive organs or effects on sperm count and morphology could be detected in the high dose group compared to the control animals. The NOAEC for systemic effects was therefore found to exceed 0.5 mg/L air.

Conclusion on reproductive toxicity / effects on fertility

Based on the results of all available studies, including a combined repeated dose toxicity study with the reproduction/developmental toxicity screening test and several subchronic studies investigating parameters of reproductive toxicity, all performed with adequate source substances and also taking into account toxicokinetic considerations, no adverse effects on reproductive organs and tissues as well as on intrauterine development are expected for the target substance Monopentaerythritol tetraesters and dipentaerythritol hexaesters of 2-ethylhexanoic and n-valeric acids.

Effects on developmental toxicity

Description of key information

Oral: OECD 422, rat, NOAEL developmental ≥ 1000 mg/kg bw/day

Read-across from structural analogue source substance Hexanoic acid, 2-ethyl-, 2,2-bis [ [(2-ethyl-1-oxohexyl)oxy] methyl] -1,3-propanediyl ester (CAS No. 7299-99-2)

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

(only 15 presumed pregnant females per group, exposure on day 0-19 of gestation, only 2 dose levels, nonstandard dermal exposure, limited details on exposure)

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Kingston, N.Y.
- Age at study initiation: approx. 9 weeks
- Mean weight at study initiation: 248 g
- Diet: Purina Certified Rodent Chow #5002 (Meal), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 22
- Humidity (%): 40 - 60
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

dermal

Vehicle:

unchanged (no vehicle)

Details on exposure:

TEST SITE
- Type of wrap if used: open exposure, no wrap
- Time intervals for shavings or clipplings: no data on frequency; clipped, intact skin
- Site: dorsal
Controls: The rats of the control group were clipped and collared. The intact dorsal skin of each rat was stroked with the tip of a syringe, but no test material was applied.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): The amount of the test material applied with a syringe was calculated based on the body weight of the animals and the density of the test substance.

USE OF RESTRAINERS FOR PREVENTING INGESTION: yes. To minimize ingestion of the test material, the rats were fitted with cardboard Elizabethan-style collars.

Analytical verification of doses or concentrations:

no

Details on mating procedure:

- Impregnation procedure: cohoused
- If cohoused: M/F ratio per cage: 1/1
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

gestation days 0 - 19

Frequency of treatment:

daily

Duration of test:

The animals were sacrificed on day 20 of gestation.

Dose / conc.:

800 mg/kg bw/day (nominal)

Dose / conc.:

2 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

15 presumed-pregnant females

Control animals:

yes, concurrent no treatment

Details on study design:

- Dose selection rationale: based on results of a 13-week dermal study previously conducted with the same material

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once daily
- Cage side observations checked: signs of pathosis, abortion, premature delivery, and death

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: day 0, 3, 6, 10, 13, 16, and 20 of gestation

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes
- Time schedule for calculation: days 0-3, 3-6, 6-10, 10-13, 13-16, and 16-20

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: The thoracic and abdominal cavities were exposed and all organs were examined grossly for evidence of pathosis.

OTHER:
- Clinical chemistry: alanine aminotransferase (ALT), albumin, albumin/globulin ration, alkaline phosphatase (ALP), aspartate aminotransferase (AST), bilirubin, calcium, chloride, cholesterol, creatinine, globuline, glucose, iron, lactate dehydrogenase (LDH), phosphorus, potassium, sodium, sorbitol dehydrogenase (SDH), total protein, triglycerides, urea nitrogen, and uric acid

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
- Number of live and dead fetuses: Yes
- Other: The ovaries of non-pregnant females were grossly examined and then discarded.

Fetal examinations:

- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: No

Statistics:

- Analysis of variances and group comparison using Fisher's Exact or Dunnett's test (maternal biophase and cesarean section data, and fetal data)
- ANOVA and Fisher's Exact test (fetal skeletal data)
- Fisher's Exact test (fetal visceral data)
- SAS procdures, Student-Newman-Keul's multiple comparison test (clinical chemistry data)
P < 0.05

Details on maternal toxic effects:

Maternal toxic effects:yes. Remark: slight local effects

Details on maternal toxic effects:
- General observations: neck lesions, red nasal exudate, and chromodacryorrhea in all groups (considered not to be test substance-related as these signs are common in animals that are collared)
- Local effects: mild dermal irritation including erythema and flaking of the skin in the treatment groups
- Body weight: similar to controls in both treatment groups
- Body weight gain: similar to controls in both treatment groups
- Uterine and net body weights: similar to controls in both treatment groups
- Food consumption: similar to control in both treatment groups; only difference (statistically significant) in high dose group on day 13-16: 31.5 g vs. 29.5 g (corresponding control data)
- Necropsy: no remarkable findings were observed
- Fetal status and uterine position: no parameter evaluated appeared to be adversly affected
- Clinical chemistry: no differences between treated and non-treated rats were observed

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

2 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: no adverse effects observed; slight local dermal irritation

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
- Fetal body measurements: Mean fetal body weights and crown-rump lengths, parameters of body growth and development were normal compared to controls.
- Fetal examination: No malformations or variations were observeed. Bruises were observed on the skin of 4 fetuses from the control group and 2 fetuses form the 800 mg/kg bw/day group (considered to be incidental). One fetus from one dam exposed to 800 mg/kg bw/day was pale in colour. No other remarkable findings were observed during external examination.
- Malformations (lumbar and sacral vertebrae, tail, sternebrae, and ribs) were observed. The incidence was low and there was no apparent dose-response relationship, these effects were considered to be not treatment-related. Comparable incidences of variant skeletal development were observed in both cotnrol and trated fetuses.
- Visceral examinations: A statistically significant (high dose) increase in the number of fetuses with levocardia was observed. The response appeared to be dose-related
Levocardia:
Litters: control: 0 (0%); 800 mg/kg bw/day: 2 (14.3%); 2000 mg/kg bw/day: 7 (50%); 14 litters per group examined
Fetuses: control: 0 (0%); 800 mg/kg bw/day: 3 (3.2%); 2000 mg/kg bw/day: 7 (10.1%); 94, 93, and 99 fetuses examined, respectively
Microphthalmia, anophtalmia and "apparent" hydronephrosis were also observed. Variant visceral development was observed in control and treated fetuses.

Dose descriptor:

LOAEL

Remarks:

developmental

Effect level:

800 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: increased number of fetuses with levocardia

Dose descriptor:

NOAEL

Remarks:

developmental

Effect level:

< 800 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: increased number of fetuses with levocardia

Abnormalities:

effects observed, treatment-related

Localisation:

visceral/soft tissue: cardiovascular

Description (incidence and severity):

An increased number of fetuses with levocardia was observed

Developmental effects observed:

yes

Lowest effective dose / conc.:

800 mg/kg bw/day (nominal)

Treatment related:

yes

Relation to maternal toxicity:

developmental effects in the absence of maternal toxicity effects

Dose response relationship:

yes

Relevant for humans:

no

Levocardia was the only parameter affected in fetuses of dams treated with the test substance during gestation. In other studies, levocardia was observed in control fetuses, too. However, the effect of the test substance on heart development should be focused on in further studies as well as the impact, this effect has on postnatal survival.

Conclusions:

In a developmental toxicity study, pregnant rats were dermally exposed to the test substance. No adverse effects were observed in any maternal or reproductive parameter nor on external and skeletal development of fetuses. Levocardia was observed in 3.2% and 10.1% of the fetuses exposed in utero to 800 and 2000 mg/kg bw /day, respectively. Thus, the developmental NOAEL was determined to be < 800 mg/kg bw.

Reference 2

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

22 Apr - 17 Jun 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 422

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc., Yokohama, Japan
- Age at study initiation: 10 weeks
- Weight at study initiation: males: 330.8 - 420.4 g (mean 374.0 g), females: 207.2 - 245.0 g (mean 228.3 g)
- Fasting period before study: no fasting period
- Housing: steel cage with wire floor
- Diet: CE-2 from CLEA Japan, Inc., Tokyo, Japan; ad libitum
- Water: tap water; ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23.0 - 24.5
- Humidity (%): 49.0 - 67.0
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
50 w/v% solution was prepared by diluting the test substance in vehicle. And this solution was diluted serially with vehicle to prepare 15 and 5 w / v% solution. Gavage solution was prepared within 8 days before administration, since the stability was verified for 8 days. Prepared samples were stored at room temperature in the dark.

VEHICLE
- Justification for use and choice of vehicle (if other than water): corn oil
- Lot/batch no. (if required): V2P1825

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

0.5 mL of each prepared solution was mixed with hexane to be confirmed by GC. The resulting analytical concenrations were 107 - 110% of the nominal concentrations

Details on mating procedure:

- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: 2 weeks
- Further matings after two unsuccessful attempts: no
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

Males: 42 days
Females: from Day 14 before mating to Day 4 of lactation
Females(satellite): 42 days

Recovary period :
- Males: 14 days
- Females (satellite): 14 days

Frequency of treatment:

once daily

Duration of test:

Males: 42 days
Females: from Day 14 before mating to Day 4 of lactation
Females(satellite): 42 days

Recovary period :
- Males: 14 days
- Females (satellite): 14 days

Dose / conc.:

100 mg/kg bw/day (nominal)

Dose / conc.:

300 mg/kg bw/day (nominal)

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

Males; control: 12 (5 for recovery), 100 mg/kg: 12, 300 mg/kg: 12, 1000 mg/kg: 12 (5 for recovery)
Females; control: 12, 100 mg/kg: 12, 300 mg/kg: 12, 1000 mg/kg: 12
Satellite females: control: 5, 1000 mg/kg: 5

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on a preliminary study, 14 days administration in females and males of 100 mg/kg, 300 mg/kg and 1000 mg/kg showed no effect of general condition, body weight gain and organ weights (liver, kidney and spleen).
- Post-exposure recovery period in satellite groups: 14 days

Maternal examinations:

Terminal killing:
- Females: Day 5 of lactation
- Females (satellite): Day 15 of recovery

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:
Males: Day 0, 7, 14, 21, 28, 35 and 42 of treatment
Males in satellite groups: Day 0, 7, 14, 21, 28, 35, 42 of treatment, Day 7 and 14 of recovery period
Females: Day 0, 7, 14, 21, 28, 35, 42 of treatment and once from Day 0 of lactation to Day 4 of lactation
Females in satellite groups: Day 0, 7, 14, 21, 28, 35, 42 of treatment, Day 7 and 14 of recovery period

BODY WEIGHT: Yes
- Time schedule for examinations:
Males: Day 1, 7, 14, 21, 28, 35, 42 of administration and before sacrifice
Males and females in satellite groups: Day 1, 7, 14, 21, 28, 35 , 42 of administration, Day 1, 7, 14 of recovery period and before sacrifice
Females: Day 1, 7, 14, 21 of treatment, Day 0, 7, 14, 20 of pregnancy, Day 0 and 4 of lactation, and before sacrifice

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation Day 26, if pups were not delivered.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: No
- Number of late resorptions: No

Fetal examinations:

- Terminal killing: 4 days after birth
- External examinations: Yes: all per litter
- Visceral examinations: Yes: all per litter
- Skeletal examinations: No
- Head examinations: No

Statistics:

Fisher test, Mann-Whitney-U-Test, Student's t-test, Aspin-Welch test, Bartlett test, Kruskal-Wallis test, Dunnett-test

Indices:

Reproductive indices: copulation index, fertility index, gestation index, implantation index, number of corpora lutea, number of implantation index
Offspring viability indices: number of newborns, number of dead pups, delivery index, birth index, live birth index, sex ration on day 0 and day 4, viability index

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No mortality was observed in all groups.
An emaciation was observed at day 40 of treatment (Day 1 of lactation) in the female control group, but this was recovered after one day. Loss of the upper incisor was observed at day 25 - 32 in a female of the 300 mg/kg group but this was considered due to physical shock and were not affected by administration of the test substance.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Males of the 100 mg/kg group showed body weight loss compared to the control group, but this was not statistically significant. There was no difference in female groups and satellite groups compared with control groups.
Females of the satellite 1000 mg/kg group showed significantly more food consumption during days 29 - 30 compared to the control group. However, this change was considered as not compound-related as no difference in other groups was found, both within the treatment period as well as in the recovery period.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
Before mating, the estrous cycle of one animal in the 300 mg/kg group was 4/5-day and in one animal of the 100 mg/kg groups the estrous cycle was 5-day during the administration period. However, no significant difference was found.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Regarding coupling, all cases were successful but one animal in 1000 mg/kg was sterile. One dam in the control group was considered to show poor delivering behavior based on the dirty vagina. However, these changes were not compound-related, since no other significant change was found related to gestation rate, paring days until coupling, frequency of estrous, gestation length in days and delivery behaviour.

ORGAN WEIGHTS (PARENTAL ANIMALS)
At the end of dosing period, a significantly increase of absolute weight of brain was found in females (300 mg/kg), but the change of relative weight was not significant. No significant differences in males between treatment group and the control group were observed.
At the end of recovery period, a significantly increase of relative liver weight in females of the 1000 mg/kg group was noted, while no differences in males were found. The change was not considered as compound-related, since no corresponding abnormalities were observed in the livers.

GROSS PATHOLOGY (PARENTAL ANIMALS)
At the end of dosing period, no abnormalities were observed. At the end of recovery period, a tumor in right hind femur was observed in one male of the control group.

HISTOPATHOLOGY (PARENTAL ANIMALS)At the end of dosing period, localized atrophy of the seminiferous tubules were observed in three males of the 100 mg/kg group and in one male of the 300 mg/kg group. Three of these animals had cell debris in lumen in epidedymis. Livers in males of the 1000 mg/kg and control groups exhibited fatty change in hepatocyte and microgranuloma. In all control group animals and three animals of 1000 mg/kg, eosinophilic bodies were found in cortex of the kidney, while slightly basophilic tubules were found in kidney cortex of two animals in the control group and in four animals of the 1000 mg/kg group. Extramedullary hematopoiesis and deposit of brown pigment was found in the spleen of all animals of the control and the 1000 mg/kg group. In addition, focal degeneration/fibrosis in the myocardium, accumulation of focal foam cell in the adveolus and mineralization of the arterial wall in lung, as well as lymphocytes and neutrophil cellular infiltration in prostate were observed both in control groups and 1000 mg/kg groups. However, these change were not significantly different but accidental.
In one female of the 1000 mg/kg group, follicular cyst, increased arterial follicle and decreased corpus luteum was found in ovary at the end of dosing period. In liver, fatty change in periportal hepatocytes and microgranuloma was observed in all animals of control and 1000 mg/kg groups, respectively. Basophilic tubule in the cortex of the kidney was observed in one animal at 1000 mg/ kg and slight mineralization was found in one control animal and one animal dosed with 1000 mg/kg, respectively. In the spleen, extramedullary hematopoiesis and brown deposit were found in all animals of the control and the 1000 mg/kg group. In addition, focal degeneration/fibrosis in myocardium, mineralization on arterial wall of lung and atrophy in thymus were also found in both control and 1000 mg/kg. However, these change were not significantly different but accidental.
At the end of recovery period, intramembranous ossification and periosteum proliferation were found in one male in control. These histopathological findings are due to fracture of the right femur. In one female of 1000 mg/kg, follicular cyst was found.

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: no adverse effects were observed in male and female offspring of all dose groups

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
A pup with kinked tail was observed at 1000 mg/kg. However, this external change was regarded as incidental since no other significant external difference was found.

Dose descriptor:

NOAEL

Remarks:

developmental

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: no adverse effects were observed in male and female offspring of all dose groups

Abnormalities:

no effects observed

Developmental effects observed:

no

Conclusions:

Pentaerythritol tetra(2-ethylhexanoate) 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 (Klimisch score 1) study from a source substance with similar structure and intrinsic properties. Read-across is justified based on common precursors and hydrolysis products and consistent trends in environmental fate, ecotoxicological and toxicological profile. The selected study is thus sufficient to fulfil the standard information requirements set out in Annexes VIII - X, 8.7, in accordance with Annex XI, 1.5, 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:

adverse effect observed

Dose descriptor:

LOAEL

800 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

The available information comprises an adequate and reliable (Klimisch score 2) study from a source substance with similar structure and intrinsic properties. Read-across is justified based on common precursors and hydrolysis products and consistent trends in environmental fate, ecotoxicological and toxicological profile. The selected study is thus sufficient to fulfil the standard information requirements set out in Annexes VIII - X, 8.7, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Additional information

Justification for grouping of substances and read-across

The read-across from analogue source substances approach comprises aliphatic esters of poly-functional alcohols containing two to six reactive hydroxyl groups and one to six fatty acid chains. The analogue approach contains mono constituent, multi-constituent and UVCB substances with fatty acid carbon chain lengths ranging from C5 - C28, which are mainly saturated but also mono unsaturated C16 and C18, polyunsaturated C18, branched C5 and C9, branched C14 - C22 building mono-, di-, tri-, and tetra esters with an alcohol (i.e. the polyol).

The available data allows for an accurate hazard and risk assessment of the target substance and the read-across concept is applied for the assessment of environmental fate and environmental and human health hazards. Thus, where applicable, environmental and human health effects are predicted from adequate and reliable data for source substances within the group by interpolation to the target substance applying the group concept in accordance with Annex XI, Item 1.5, of Regulation (EC) No. 1907/2006. In particular, for each specific endpoint the source substance(s) structurally closest to the target substance is/are chosen for read-across, with due regard to the requirements of adequacy and reliability of the available data. Structural similarities and similarities in properties and/or activities of the source and target substance are the basis of read-across.

A detailed justification for the grouping of chemicals and read-across is provided in the technical dossier (see IUCLID Section 7.1 and 13).

Developmental toxicity

Besides the combined repeated dose toxicity study with the reproduction/developmental toxicity screening test already discussed, there is a reliable and adequate study available for a structural analogue source substance investigating pre-natal developmental toxicity. While administration of the test substance in the first study was by oral gavage, the dermal route of exposure was utilised in the pre-natal developmental study.

In a developmental toxicity study Fatty acids, C5-9, tetraesters with pentaerythritol (CAS No. 67762-53-2) was investigated comparable to OECD Guideline 414 (prenatal developmental toxicity study) (Exxon, 1988b). Groups of 15 presumed pregnant female Sprague-Dawley rats received daily dermal doses of the test substance at concentrations of 800 and 2000 mg/kg bw/day during gestational days 0 to 19. Control animals remained untreated. On day 20 of gestation the animals were euthanized and examined for maternal and fetal parameters. There were no adverse effects found for all parameters examined in maternal animals. Based on the number of implantations, number of total litter losses by resorption, mortality, clinical signs, body weight, gross pathology and organ weights of maternal animals the NOAEL for maternal systemic toxicity was found to be 2000 mg/kg bw/day. Examination of fetus litter size and weights, offspring viability (number alive and number dead), sex ratio, grossly visible abnormalities, external, head, soft tissue and skeletal abnormalities revealed no differences to controls and thus no indication for teratogenic effects. Soft tissue examination revealed an increased number of fetuses with “levocardia” but the internal anatomy of the heart was normal and no accompanying heart or other malformations in the thoracic cavity were detected. It is important to note that the term levocardia as used in the study report describes the placement of the heart in the extreme left hemithorax without any reversal of organs or any other malformations or adverse effects. It is therefore concluded that the effect observed has no toxicological relevance for humans. However, since levocardia was observed in both treated groups, the NOAEL for embryo- / fetotoxicity and teratogenicity in rats for Fatty acids, C5-9, tetraesters with pentaerythritol under the experimental conditions of the study was determined to be < 800 mg/kg bw/day and the LOAEL = 800 mg/kg bw/day.

Conclusion on developmental toxicity

The available data on developmental toxicity from adequate source substances do not indicate any significant toxicological adverse effects on maternal and embryonal toxicity. Although in one of the two studies referred to “levocardia” in rats after dermal exposure was detected, it could be demonstrated that this effect has no toxicological relevance for humans as the effect described the positioning of the heart in the extreme left hemithorax without any further adverse implications and malformations. Therefore, based on the available data and following the analogue approach, no hazard for reproduction (development) was identified for the target substance Monopentaerythritol tetraesters and dipentaerythritol hexaesters of 2-ethylhexanoic and n-valeric acids and the NOAEL values for systemic and developmental toxicity of ≥ 1000 mg/kg bw/day are considered for hazard and risk assessment.

Justification for classification or non-classification

According to Article 13 of Regulation (EC) No. 1907/2006 information on intrinsic properties of substances may be generated by means other than tests, e.g. using information from structurally related substances (grouping or read-across), provided that conditions set out in Annex XI are met. Annex XI 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 read-across concept is applied to the target substance Monopentaerythritol tetraesters and dipentaerythritol hexaesters of 2-ethylhexanoic and n-valeric acids from representative structural analogue source substances to avoid unnecessary animal testing.

The read-across concept is also used to derive the classification of the target substance taking the properties of the source substances into account. Based on the read-across concept, all available data on reproductive and developmental toxicity do not meet the classification criteria according to Regulation (EC) No. 1272/2008 (CLP) and are therefore conclusive but not sufficient for classification.