[No public or meaningful name is available]

Endpoint summary

Description of key information

Repeated dose toxicity: Oral NOAEL (rat, m/f): = 1000 mg/kg bw/day

Read-across from source substances 1,2,3 propanetriol, homopolymer, diisooctadecanoate (CAS 63705-03-3), decaglycerol decaoleate (no CAS No. available) and2,3-dihydroxypropyl oleate

(CAS 111-03-5)

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

sub-chronic toxicity: oral

Type of information:

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

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented publication which meets basic scientific principles (lack of details on test material, no ophthalmologic or neurobehavioral examinations).

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

yes

Remarks:

lack of details on test material, no ophthalmologic or neurobehavioral examinations

GLP compliance:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: mean body weight - 72 g (males), 69 g (females)
- Housing: in individual cages
- Diet: basal diet consisting of ground pellets (Purina Lab Chow), inorganic salts, vitamins, casein and fat, ad libitum, except during periods of urine collection
- Water: ad libitum, except during periods of urine collection

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: for exact composition of dosing solutions see Table 1 under "Any other information on materials and methods including tables"

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

daily, 7days/week (except during urine collection)

Remarks:

Doses / Concentrations:
2.5 % (Diet No.1), 5 % (Diet No.2), 10 % (Diet No.3)
Basis:
nominal in diet

No. of animals per sex per dose:

10 males and females

Control animals:

other: Diet No.4: free oleic acid + glycerol corresponding to total fatty acid and glycerol content of Diet No.2 (to determine if free fatty acids and glycer... (see attached file)

Details on study design:

- Dose selection rationale: substance is used as a food additive and is therefore tested at the specified dose levels

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: regular observations were done

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: regular observations were done

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined: Yes

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes

HAEMATOLOGY: Yes
- Time schedule for collection of blood: during fifth and eleventh week and from severed neck vessels at necropsy
- Anaesthetic used for blood collection: No data
- Animals fasted: No
- How many animals: all animals
- Parameters checked in table [No.6] were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: during third and ninth week of the study
- Metabolism cages used for collection of urine: No data
- Animals fasted: Yes
- Parameters examined: total nitrogen, specific gravity and pH

NEUROBEHAVIOURAL EXAMINATION: Yes
- Dose groups that were examined: regular observations of behavior of animals was done


OTHER:
- collection of feces during fourth and tenth week for determination of total fatty acid (TFA) absorption

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (see Table 2)
HISTOPATHOLOGY: Yes (see Table 2)

Statistics:

Statistic testing for significance compared to soybean control groups was done but method used is not specified.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

effects observed, treatment-related

Description (incidence and severity):

higher nitrogen excretion in females fed 10 % PGE but no changes during the histologic examination of the urinary tract were found

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
Throughout the study all animals appeared to be in excellent health

BODY WEIGHT GAIN/FOOD CONSUMPTION AND FOOD EFFICIENCY
Males fed PGE at the high dietary level (10 %) consumed more food and had a poorer feed efficiency value than did male control animals, however despite decreased feed efficiency those animals consumed enough food to maintain normal growth.

HAEMATOLOGY/CLINICAL CHEMISTRY
Values fell within normal ranges, and there were no indications of any blood disorder. Some values were significantly different from SBO controls, however differences were usually quite small and in no case established any trend or pattern indicative of a dose-related effect.

URINALYSIS
Urine collected from each animal appeared to be normal in regard to color clarity, sediment, specific gravity and pH, total nitrogen excretion during the third and ninth week by females fed PGE at the 10 % level was significantly greater than the control value and appears to be related to dietary treatment, the highest nitrogen excretion (male and female) was from the animals fed the highest dose PGE, the reason for this difference is not understood but it should be recognized that other parameters, including three derived from histologic examination of the urinary tract were normal

ORGAN WEIGHTS
There were no statistical differences for all organs examined.

GROSS PATHOLOGY
The only gross observation of significance was what appeared to be very mild, chronic murine pneumonia in 18% of the animals, but the affected animals were scattered throughout all the groups and the effect was not related to the feeding of PGE.

HISTOPATHOLOGY: NON-NEOPLASTIC
Microscopic examination confirmed the pneumonitis but did not reveal any other changes as evidence of toxicity

OTHER FINDINGS
The percentage of dietary fatty acids absorbed decreased as the level of PGE in the diet was increased. In all cases fat absorption by animals fed PGE at the 5 and 10 % dietary levels was significantly less than corresponding SBO control values. Absorption in animals fed the lowest dose was also less but not significantly different. Values from the group fed oleic acid and glycerol fell between those of the 5 % PGE and SBO control.
Gas-liquid Chromatography analyses of faecal fatty acids showed that excretion of oleic acid increased in a dose-related fashion: the oleic acid content of faecal fatty acids from animals fed the SBO control diet was 23 % compared to 32, 41 and 51 % when PGE was fed at levels of 2.5, 5, and 10 %. The increased excretion of fatty acids in general and oleic acid in particular shows that absorption of dietary PGE was not complete. The faecal oleic acid may have resulted from the excretion of intact PGE or from hydrolyzed or partially hydrolyzed but unabsorbed material. The oleic acid content of faecal fatty acids from animals fed free oleic and glycerol was 41 % corresponding exactly to the 5 % PGE group.

Dose descriptor:

NOAEL

Effect level:

10 other: %

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: decreased total fat absorption

Critical effects observed:

not specified

Table3: Food Consumption, Body Weight Gain and Feed Efficiency

	Cumulative Food consumption (g)		Cumulative gain in body weight (g)		Cumulative feed efficienies^a
Dietary Group	Male	Female	Male	Female	Male	Female
2.5 % PGE	1585	1262	393	195	24.8	15.6
5 % PGE	1589	1365	384	191	24.2	14.4
10 % PGE	1753	1293	397	205	22.9^b	15.9
Free Acid	1527	1140	382	191	25.0	16.8
Soybean Oil control (SBO)	1615	1237	404	193	25.0	15.8

^a - Weigth gained g/100g diet consumed

^b - Significantly different from the SBO control (P < 0.05)

Table4: Urine data from the 90 -day Feeding study

	Total Nitrogen Excretion^a
	Third week		Ninth week
Dietary Group	Males	Females	Males	Females
2.5 % PGE	97.3	93.6	145.4	131.8
5 % PGE	94.1	93.0	142.0	106.2
10 % PGE	115.3	114.8^b	169.6	138.7^b
Free Acid	99.9	95.4	143.0	120.8
Soybean Oil Control (SBO)	106.3	91.0	143.7	104.6

^a - Nitrogen (mg) excreted during the 16 hour collection period

^b - Significantly different from the SBO control (P < 0.05)

Table5: Total Fatty Acid (TFA) Absorption data

	TFA absorption (% of fed)
	Fourth week		Tenth week
Dietary Group	Males	Females	Males	Females
2.5 % PGE	88.3	90.5^a	90.1^a	91.8^a
5 % PGE	83.6^a	86.7^a	85.2^a	89.5^a
10 % PGE	73.1^a	73.8^a	70.1^a	68.8^a
Free Acid	87.0^a	89.5^a	88.8^a	91.5
Soybean Oil Control (SBO)	89.3	92.9	91.9	93.9

^a - Significant different from the SBO control (P<0.05)

Table6: Blood values from the 90 -day feeding study

Dietary group	Hemoglobin^a	Hematocrit^b	RBC count^c	WBC count^d	Lymphocytes^e	Neutrophils^e	Monocytes^e	Eosinophils^e
Males
2.5 % PGE	15.7	47	8.18	6700	92	6	1	1
5 % PGE	15.7	48	8.36	7700	91	7	1	1
10 % PGE	15.9	48	8.66	8750	94	4	1	1
Free Acid	16.1	48	8.32	7250	0	8	1	1
Soybean Oil control (SBO)	15.5	47	8.48	8250	91	7	1	1
Females
2.5 % PGE	15.2	46	8.47*	4500	91	6	2	1
5 % PGE	16.0	47	8.04*	5625	93	6	1	0*
10 % PGE	15.3	46	7.06	4675	92	7	1	0*
Free Acid	16.1	48	7.70	5925	94	5	1	0*
Soybean Oil control (SBO)	15.9	46	7.34	5575	95	3	1	1

a - g/100mL

b - Packed Cell Volume (%)

c - RBC = Millions of red blood cells per cubic millimeter of blood

d - WBC = Number of white blood cells per cubic millimeter of blood

e - Differential count of white blood cells, expressed as a percentage of total white cells

*Significantly different from the SBO control (P<0.05)

Conclusions:

All animals fed PGE appeared to be in excellent health throughout the study and no adverse effects were found upon survival, growth, organ weights, organ, body weight ratios and hematological values. There were no significant gross or microscopic tissue changes which would be attributed to dietary treatment. Total fat absorption decreased in a dose-related response, showing that absorption of PGE was not complete. Additionally, excretion of nitrogen in the urine by females fed 10 % PGE was significantly greater than the control but this difference is not understood.

Reference 2

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP - Guideline 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:

Sprague-Dawley

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: 352-426 g (males; mean: 372 g), 192-249 g (females; mean: 224 g)
- Housing: individual in stainless steel cages
- Diet: ad libtum
- Water: ad libitum
- Acclimation period: 11 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.1 - 23.2
- Humidity (%): 48 - 61
- Air changes (per hr): more than 10
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
The test substance was diluted in appropritate amounts of corn oil for each dose level. Aliquots of the dosing solution corresponding to the amount of daily administration were stored in the dark at 2 - 6 °C. The stability of the dosing solution was 7 days in a refrigerator and 1 day at room temperature. Therefore, the dosing solution was used within 7 days.

VEHICLE
- Justification for use and choice of vehicle (if other than water): the test substance showed low solubility in water.
- Amount of vehicle (if gavage): 5 mL/kg
- Lot/batch no. (if required): V4N3566

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

No details reported.

Duration of treatment / exposure:

males: 42 days (14 days prior to mating and 28 days thereafter)
females: 42-52 days (from 14 days before mating to day 4 of lactation)
satellite males and females: 42 days and 14 days post-exposure observation period

Frequency of treatment:

once daily, 7 days/week

Remarks:

Doses / Concentrations:
100, 300 and 1000 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

12 females in control and test groups
7 males in control and 1000 mg/kg bw groups
12 males in 100 and 300 mg/kg bw groups
5 animals per sex in satellite control and 1000 mg/kg bw/day groups (in addition to number listed above)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
Two dose range finding studies were performed. 2000 mg/kg bw of test substance was administered for 3 days in male and female rats. No abnormalities were found in general condition and body weight. The second study was performed at dose levels of 0, 330, 100, 300 and 1000 mg/kg bw/day for 14 days. No abnormalities of general condition, body weight, food consumption, hematological findings, blood biochemical findings, gross pathology and organ weight were found. Therefore, 1000 mg/kg bw/day was selected as the highest dose.
- Post-exposure recovery period in satellite groups: 14 days

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once prior to the first exposure and weekly thereafter

BODY WEIGHT: Yes
- Time schedule for examinations:
Males: Day 1 (before administration), 7, 14, 21, 28, 35 and 42 (before sacrifice)
Females: Day 1 (before administration), 7, 14, during pregnancy on Day 0, 7 14 and 21, during lactation on Days 0 and 4 (before sacrifice)
Satellite males and females: Day 1 (before administration), 7, 14, 21, 28, 35, 42, 49 (Day 7 of recovery period) and 56 (Day 14 of recovery period, 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: one day after last application
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes
- How many animals: 5 per group
- Parameters checked: RBC, Hemoglobin, Hematocrit, MCV, MCH, MCHC, Platelets, Reticulocytes, PT, APTT, WBC, Differential leukocytes: Lymphocytes, Neutrophils, Eosinophils, Basophils, Monocytes

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: one day after last application
- Animals fasted: Yes
- How many animals: 5 per group
- Parameters checked: AST, ALT, ALP, γ-GTP, T-protein, Albumin, A/G, T-bilirubin, BUN, Creatinine, Glucose, T-cholesterol, Triglyceride, Na, K, Cl, Ca, P, LDH, choline esterase

URINALYSIS: Yes (males)
- Time schedule for collection of urine: Day 37 during administration period or Day 9 during recovery period
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked: Colour, cloudiness, water consumed, volume, specific gravity, Na, K, pH, Protein, Glucose, Ketone body, Bilirubin, Occult blood, Urobilinogen, Epithelial cells, Erythrocytes, Leukocytes, Casts, Crystals, Fat

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations:
Males: on Week 6 of the administration period
Females: on Week 6 of the administration period
Males and females of satellite groups: on Week 6 of the administration period and on Week 2 of the recovery period
- Dose groups that were examined: all
- Battery of functions tested: hearing reaction, eye sight reaction, sense of touch reaction, pain reaction, pupil reflex, pinna reflex, ipsilateral flexor reaction, eyelid reflex, righting reflex

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Body surface, mucous membranes and internal organs

HISTOPATHOLOGY: Yes
Brain, pituitary, thyroid, thymus, lung trachea (after liquid immersion fixation), stomach, intestines, heart, liver, spleen, kidney, adrenal gland, bladder, testis, epididymis, prostate, seminal vesicles, ovaries, uterus, spinal cord (cervical, thoracic, lumbar), sciatic nerve, bone marrow (femur), lymph nodes (cervical lymph node, mesenteric lymph nodes), mammary gland, and other gross abnormalities
Spermatogenic cycle (Stage II, III, V, VII, and XII) was also investigated.

SACRIFICE
- Males: on Day 43
- Females: on Day 5 of lactation
- Females (unsuccessful mating): on Day 53
- Females (mated but non-pregnant): on Day 5 after scheduled delivery
- Satellite males and females: on Day 15 of the recovery period

Other examinations:

Organ weight: brain, liver, kidney, spleen, heart, thymus, thyroid, pituitary, adrenal, testis, sminal vesicle, epididymis
Estrous cycle
Number of ovarian corpora lutea
Number of uterine implantation
Observation of pups

Statistics:

ANOVA, Barlettm Kruskal-Willis, Dunnett, F test, Studen t-test, Aspin-Welch t-test, Mann-Whitney U-test, Fisher's exact test

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

1000 mg/kg bw/day (satellite group, males): significant increase was observed (non adverse)

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

1000 mg/kg bw/day (satellite group, females): significant increase on Day 14 of administration (non adverse)

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

100 and 300 mg/kg bw/day (males): decrease in APTT without dose-dependency after administration period (non adverse)

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

300 and 1000 mg/kg bw/day (females): significant increase in organic P in females after administration period without dose-dependency (non adverse); 1000 mg/kg bw/day (satellite group, females): significant increase in choline esterase (non adverse)

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

100 mg/kg bw/day: significant decrease in absolute weight of seminal vesicles in male and significant decrease in relative weight of spleen in females after administration period (non adverse)

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

300 mg/kg bw/day: mass in subcutis in one female after administration period (not treatment-related)

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

control and 1000 mg/kg bw/day: after administration period, low incidence of commonly found microscopic changes, evenly distributed between groups (not treatment-related)

Histopathological findings: neoplastic:

effects observed, treatment-related

Description (incidence and severity):

300 mg/kg bw/day: after administration period, benign fibroadenoma of the mammary gland in one female (not treatment-related)

Details on results:

CLINICAL SIGNS AND MORTALITY
No abnormalities and no mortality were observed.

BODY WEIGHT AND WEIGHT GAIN
No change of body weight and weight gain was observed during the administration period. During the recovery period, a significant increase in body weight was noted in males at 1000 mg/kg bw/day. This was caused by a tendency of the control group animals to lose weight. One male in control group showed a significant decrease in body weight during the recovery period. However, no other abnormalities were observed in this male.

FOOD CONSUMPTION
No change was observed during the administration period in the test groups. A significant increase in food consumption was found in satellite females of the 1000 mg/kg bw/day group on Day 14 of administration. However, it was regarded as an incidental finding since the food consumption of the corresponding control group was relatively small on that day. Therefore, this change was not regarded as a compound-related effect.

HAEMATOLOGY
No changes were observed. After administration, a significant decrease in APTT was observed in males of the 100 and 300 mg/kg bw/day groups in comparison with control group. However, this change was not regarded as compound-related, because no dose-dependency was observed and this was within reference range.

CLINICAL CHEMISTRY
A significant decrease in inorganic P was observed in females of the 300 and 1000 mg/kg bw/day groups after administration. However, this change was not regarded as compound-related, since a dose-dependency was not observed and almost all the individual data were within the reference ranges except for one female of 300 mg/kg bw/day.
Choline esterase was increased in females of 1000 mg/kg bw/day after the recovery period but this was within the reference range. This change was not compound-related effect since this change was slightly and no change was observed after the administration period.

URINALYSIS
No significant changes were found during the administration and recovery periods.

NEUROBEHAVIOUR
No abnormality was observed for the sensory/reflex function, landing foot, grip strength and motor activity during the administration period. During the recovery period, motor activity increased from 0 to 60 min but not from 0 to 30 min in females dosed 1000 mg/kg bw/day. This effect was not compound-related since the observed change was slight and no change was observed during and after the administration period.

ORGAN WEIGHTS
After the administration period, a significant decrease in the absolute weight of seminal vesicles in males given and a significant decrease in relative weight of spleen in females were observed in the 100 mg/kg bw/day group. However, this change was not compound-related because no abnormalities were found at histopathological examination and there was no dose-dependency.
After the recovery period, a significant decrease in relative weight of pituitary in males of the 1000 mg/kg bw/day group and relative weight of thyroid in females of the 1000 mg/kg group bw/day was observed. Nevertheless, this effect was not compound-related because no abnormality was reported regarding these organs after administration period.

GROSS PATHOLOGY
No test substance-related changes were found. No abnormalities were found in breeding pairs which were not successful at mating and in those which were successful at mating but not pregnant.
After administration period, reddish thymus was noted in one male of the control group and subcutis mass was found in one female of 300 mg/kg by/day group (see also HISTOPATHOLOGY: NEOPLASTIC). After the recovery period, larger spleen and capsular thickening in spleen was found in one male and reddish area in thymus was observed in one female at 1000 mg/kg bw/day.

HISTOPATHOLOGY: NON-NEOPLASTIC
No test substance-related changes were observed. There were also no changes regarding spermatogenic cycle.
Myocardial degeneration/fibrosis, foam cell accumulation in lung, mineralization in artery in lung, fatty degeneration of hepatocyte, microgranuloma in liver, solitary cyst in kidney, hyaline cast in kidney, lymphocyte infiltration in cortex of kidney, mineralization of cortico-medullary junction in kidney, fibrosis of cortex in kidney and haemorrhage in thymus were observed both in the control and 1000 mg/kg bw/day groups or only in the control group with low incidence. Hyaline droplet of proximal tubular epithelium in kidney was found in all males of the control and 1000 mg/kg bw/day groups. Brown deposit pigment and extramedullary haematopoiesis in spleen were found in all males and females of the control and 1000 mg/kg bw/day groups. However, there were no differences between control and test group. In the 1000 mg/kg bw/group, lymphocyte interstitium infiltration in prostate was found in one male, interstitial focal inflammation in lung and focal necrosis in liver was observed in one female. These changes occur naturally and were not compound-related.
No abnormalities were found in uterus and ovary in the non-pregnant females and the unsuccessful copulation females of the control and 1000 mg/kg bw/day groups, respectively. Degeneration of seminiferous tubules of testis, decrease in sperm and atrophy in prostate was observed in one control male.

HISTOPATHOLOGY: NEOPLASTIC
Mass on abdomen was found in a female after 40 days of administration in the 300 mg/kg bw/day group. However, this subcutaneous tumour of the mammary gland was a benign fibroadenoma and generated naturally.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects

Critical effects observed:

not specified

Reference 3

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP - guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA OPPTS 870.3650 “Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test”

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories, B.V., Kreuzelweg 53, 5961 NM Horst / Netherlands
- Age at study initiation: 11 weeks
- Weight at study initiation: 302 - 380 g (males),180 - 247 g (females)
- Housing: In groups of three to five in Makrolon type-4 cages with wire mesh tops up to the day of randomization and afterwards individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding (‘Lignocel’ J. Rettenmaier & Söhne GmbH & CoKG, 73494 Rosenberg / Germany, imported by Provimi Kliba SA, 4303 Kaiseraugst / Switzerland) with paper enrichment (Enviro-dri from Lillico, Biotechnology, Surrey, UK, batch/lot nos. 72 and 79 and ISO-BLOX from Harlan Laboratories B.V., Netherlands, batch/lot nos. T.6960C.CS-100099, 100267). During the pre-pairing period, cages with males were interspersed amongst those holding females to promote the development of regular estrus
cycles.
- Diet (ad libitum): pelleted standard Harlan Teklad 2018C (batch nos. 43/12 and 56/12) rodent maintenance diet (Provimi Kliba SA, 4303 Kaiseraugst / Switzerland)
- Water (ad libitum): community tap-water
- Acclimation period: at least seven days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): Based on an observation on 01-Feb-2013, the automatic light cycle of 12 h on / 12 h off was not functional within a block of rooms including room numbers 015A and 011 B. Due to the breeding performance from November 2012 onwards during this and other studies conducted in rooms which belong to the affected block, it was considered probable that the animals had been maintained under constant lighting throughout the study period for the first delivery animals. From 01-Feb-2013 onwards, the automatic light cycle was restored.

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: daily. The test substance was weighed into a glass beaker on a tared precision balance and approximately 80% of the vehicle was added (w/v). Using an appropriate homogenizer, a homogeneous suspension was prepared. Having obtained a homogeneous mixture, the remaining vehicle was added. Separate formulations were prepared for each concentration.
Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.

VEHICLE
- Amount of vehicle: 4 mL/kg bw (dose volume)

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

On the first dose formulation day samples from the control group as well as three samples (top, middle and bottom) of about 1 g of each concentration were taken prior to dosing for analysis of concentration and homogeneity. Samples of about 1 g of each test item concentration were taken from the middle to confirm the stability (4 hours and 8 days at room temperature (20 ± 5°C)).
Towards the end of the study of the first and third delivery of animals, samples were taken from the middle to confirm concentration. Additional samples were taken on 21-Feb-2013 (first application of the second delivery of animals) from the middle to confirm concentration. The aliquots for analysis of dose formulations were frozen (-20 ± 5 °C) and delivered on dry ice or at room temperature to Dr. G. Heinemann (Harlan Laboratories Ltd., Zelgliweg 1, 4452 Itingen /Switzerland) and stored there at -20 ± 5 °C until analysis or were analyzed directly.

As the results obtained were out of the accepted limits, a new analytical method was implemented, which necessitated additional dose formulation samples mainly from the low dose levels (for details please refer the analytical report). For the implementation of the method additional samples were taken during the study, which were not reported.

The samples were analyzed by derivatisation with MSTFA/BSTFA and analysis by GC-FID or analyzed by HPLC using a method which was implemented within the present study. Analyzed samples were not discarded without written consent from the study director.

Duration of treatment / exposure:

Males: Minimum 28 days
Females: Approximately 6 weeks

Frequency of treatment:

once daily (within four hours)

Remarks:

Doses / Concentrations:
100, 300, 1000 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

at least 12 (see any other information on materials and methods)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on single dose toxicity data of the test compound and repeated dose toxicity data of chemically similar compounds, the results of a 14-day range-finding study the dose levels of 100, 300, and 1000 mg/kg bw per day were selected for the present study.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once prior to the first administration of the test item and weekly thereafter (in the gestation period on day 0, 6, 13 and 20 post coitum).
- detailed clinical observations were performed outside the home cage in a standard arena and the animals were observed for the following changes in: skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies or bizarre behavior were also reported.

BODY WEIGHT: Yes
- Time schedule for examinations: daily from treatment start to day of necropsy.

FOOD CONSUMPTION: Yes
- Males: Pre-pairing period days 1 - 4, 4 - 8, 8 - 11 and 11 - 14; after pairing period weekly.
- Females: Pre-pairing period days 1 - 4, 4 - 8, 8 - 11 and 11 - 14; gestation days 0 – 7, 7 14 and 14 – 21 and days 1 - 4 of the lactation
No food consumption was recorded during the pairing period.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Blood samples were obtained on the day of the scheduled necropsy from 5 males randomly selected from each group. Blood samples from 5 lactating females from each group will be obtained on day 5 postpartum.
- Anaesthetic used for blood collection: Yes (light isoflurane anesthesia)
- Animals fasted: Yes, for approximately 18 hours
- How many animals: 5 males randomly selected from each group and from 5 lactating females from each group
- Parameters checked: erythrocyte count, haemoglobin, haematocrit, mean corpuscular volume, red cell volume distribution width, mean corpuscular haemoglobin, mean corpuscular hemoglobin concentration, haemoglobin concentration distribution width, leukocyte count (total), differential leukocyte count: platelet count, reticulocytes; prothrombin time (= thromboplastin time), activated partial thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: see haematology
- Animals fasted: Yes, for approximately 18 hours
- How many animals: see haematology
- Parameters checked:
Glucose
Urea
Creatinine
Bilirubin, total
Cholesterol, total
Triglycerides
Aspartate aminotransferase
Alanine aminotransferase
Alkaline phosphatase
Gamma-glutamyl-transferase
Bile acids
Sodium
Potassium
Chloride
Calcium
Phosphorus
Protein, total
Albumin
Globulin
Albumin/Globulin ratio

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: males towards the end of the first pairing period and females on day 3 post partum
- Dose groups that were examined: five P generation males and five P generation females randomly selected from each group
- Battery of functions tested:
• Cage-side observations: faeces-balls, urine and posture as well as resistance to removal.
• Hand-held observations: muscle tone, constituation, skin, pupile size, palpebral closure, lacrimation, salivation, reaction to handling and general abnormalities.
• Open field observations: level of ambulatory activity including rearing (one minute evaluation), unusual body movements (e.g. spasms, convulsions), gait evaluation, behavior, hair coat, respiration, quantity of faeces-balls and urine.
• Reflexes: blinking, palpebral closure, pinna reflex, extensor thrust response, paw pinch, responsiveness to sharp noise, righting reflex and hearing ability (Preyer’s reflex).
• Measurements / Counts: hind limb / fore limb grip strength, rectal temperature.
Additionally, locomotor activity was measured quantitatively for the same animals. Activity was measured with an Activity Monitor AMS-0151 (FMI, Germany). Activity of the animals (based on beam count) was recorded for 6-minute intervals over a period of 30 minutes. These data and the total activity over 30 minutes were reported.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
At the scheduled sacrifice, the testes and epididymides of all parental males were weighed separately. In addition, from 5 males and 5 females killed at the end of the study which were selected for hematology and clinical chemistry examination from each group, the following organs were trimmed from any adherent tissue, as appropriate, and their wet weight taken.
Adrenal glands (weighed as pairs)
Brain
Heart
Kidneys (weighed as pairs)
Uterus (including cervix)
Prostate
Liver
Thymus
Spleen
Thyroid (after fixation)
Ovaries (weighed as pairs)
Seminal vesicles (inclusive coagulating gland)

From all animals of the third delivery, following organs were weighed:
Testes
Epididymides
Heart
Liver
Kidneys (weighed as pairs)
Adrenal glands (weighed as pairs)

HISTOPATHOLOGY: Yes
The following tissues from all parental males were preserved in neutral phosphate buffered 4% formaldehyde solution:
Prostate
Seminal vesicles with coagulating gland
Testes (in Bouin’s fixative)
Epididymides (in Bouin’s fixative)
The following tissues from all parental females were preserved in neutral phosphate buffered 4%
formaldehyde solution:
Ovaries (with oviduct) Uterus (with vagina)

In addition, from all males and females the following tissues were preserved in neutral phosphate
buffered 4% formaldehyde solution:
Gross lesions
Brain
Spinal chord (cervical, thoracic, lumbar)
Small and large intestines (incl. Peyer’s patches)
Stomach (forestomach and glandular stomach)
Liver
Kidneys
Adrenals
Lymph nodes (axillary and mesenteric)
Urinary bladder
Aorta
Eyes with optic nerve and harderian gland
Lacrimal gland
Larynx
Nasal cavity
Esophagus
Heart
Thymus
Thyroids and parathyroids
Trachea and lungs (preserved by inflation
with fixative and then immersion)
Pituitary gland
Spleen
Peripheral nerve (sciatic)
Bone marrow (femur)
Femur with knee joint
Mammary gland (male and female)
Pancreas
Salivary glands – mandibular, sublingual
Skeletal muscle
Sternum with bone marrow
Pharynx

All organ and tissue samples to be examined by the study pathologist were processed, embedded and cut at an approximate thickness of 2 - 4 micrometers and stained with hematoxylin and eosin. Additionally, the testis was stained by PAS-hematoxylin.
Testes, epididymides, prostate, seminal vesicles, ovaries, oviduct, vagina and uterus from all animals of all three animal deliveries of the control and high-dose group were examined. The same applied to all occurring gross lesions. The remaining organs/tissues of 5 randomly selected males and females of the control and high-dose group, respectively, were examined histopathologically. From the 3th delivery of animals no additional organs/tissues of 5 randomly selected males and females of the control and high dose group were examined.
Special emphasis was made on the stages of spermatogenesis and histopathology of interstitial cell structure.
Histological examination of ovaries was carried out on the females that did not give birth. In addition, microscopic examination of the reproductive organs of males that did not successfully mate with the allocated female was performed.

Statistics:

The following statistical methods were used to analyze food consumption, body weights and reproduction data:
• The Dunnett-test (many to one t-test) based on a pooled variance estimate was applied if the variables were assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
• Fisher's exact-test will be applied if the variables could be dichotomized without loss of information

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

see details on results

Mortality:

mortality observed, treatment-related

Description (incidence):

see details on results

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

see details on results

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

see details on results

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

see details on results

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

see details on results

Urinalysis findings:

not examined

Behaviour (functional findings):

effects observed, treatment-related

Description (incidence and severity):

see details on results

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

see details on results

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

see details on results

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

see details on results

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY
At a dose level of 1000 mg/kg bw/day, one male (no. 171, third animal delivery) died spontaneously on day 1 of the after pairing period. No clinical signs were observed and a normal body weight gain was recorded until day 13 of the paring period. One day before he died he showed a body weight loss of 3%. During necropsy, a dark red discolored thymus was recorded.

BODY WEIGHT AND WEIGHT GAIN
Males:
Mean body weight gain was statistically significantly lower at 1000 mg/kg bw/day during the pre-pairing period, resulting in slightly reduced mean body weights (-3% at the end of this period compared to the controls). This was a result of the minor body weight loss (1%) between days 1 to 2. During the pairing period, body weight gain at the high dose level was similar to the control value. Due to this fact, the reduction in mean body weight gain and body weights were considered to be not adverse. The overall differences in mean body weight gain at the dose levels of 0, 100, 300 and
1000 mg/kg bw/day were: +13%, +12%, +11% and +11% during the pre-pairing period, +5%, +5%, +3% and +4% during the pairing period and +7%, +9%, +7% and +8% during the after pairing period (percentages refer to the body weight gain within the period).

Females:
No effect of the test item on mean body weights and mean body weight gain was noted for females at any dose level. Towards the end of the gestation period, body weights and body weight gain were reduced in females at 1000 mg/kg bw/day (statistically significantly different body weight gains from day 17 post coitum onwards). These differences were considered to be a result of the increased post implantation loss and low number of pups, which were noted for dams at this dose level. The overall differences in mean body weight gain at the dose levels of 0, 100, 300 and 1000 mg/kg bw/day were: +8%, +9%, +8% and +7% during the pre-pairing period, +63%, +57%, +57% and +51% during the gestation period and +4%, +7%, +4% and +3% during days 1 to 4 of the lactation period (percentages refer to the body weight gain within the period).

FOOD CONSUMPTION AND COMPOUND INTAKE
Males:
At 1000 mg/kg bw/day, mean food consumption was statistically significantly reduced during the first 11 days of treatment in males of the first and second delivery (-9.3% on day 11 of prepairing when compared with controls). Thereafter, the food consumption remained slightly lower than in controls, but without reaching statistical significance. Food consumption was also lower in males of the third delivery, but not to a statistically significant degree. Since this reduction in food consumption occurred only transiently, the reduction was considered to be not adverse.

Females:
There were no effects on mean food consumption at any dose level and in any study phase.

HAEMATOLOGY
The assessment of the hematology data did not reveal any test item-related effects in males and females at any dose level.

CLINICAL CHEMISTRY
The assessment of the clinical biochemistry data did not reveal any test item-related effects in males and females at any dose level.
Slightly higher urea levels were noted in males at 300 mg/kg bw/day and in females at 1000 mg/kg bw/day. Additionally, lower alkaline phosphatase levels were noted in females at 300 mg/kg bw/day, higher creatine levels in males and lower glucose levels in females at 1000 mg/kg bw/day, respectively. However, in the absence of dose-dependency and/or since values were additionally within the historical control range, a test item-related effect can be excluded.

NEUROBEHAVIOUR
No clinical signs were noted in males during the course of this study.
Incidentally, one control female (no. 45) showed a slightly decreased activity, ruffled fur and vaginal bleeding on day 25 post coitum. Necropsy of this female revealed masses in the uterus and vagina. Furthermore, one female (no. 66) treated at 100 mg/kg bw/day that did not give birth around day 21 post coitum either showed a wound on the anterior dorsum during the last three days prior to necropsy on day 25 post coitum. During necropsy reddish discolored ovaries and enlarged axillary lymph nodes were noted. Both females were pregnant.
These isolated findings were not related to treatment with the test item.

No findings were noted at detailed weekly clinical observation during the whole course of the
study.

There were no treatment-related changes noted at functional observational battery. One male at 100 mg/kg bw/day had increased faeces-balls and one female at 300 mg/kg bw/day showed decreased rearings. These isolated findings showing no dose relationship were deemed unrelated to treatment with the test item. A statistically significantly lower mean body temperature was noted in males of the first and second delivery at 1000 mg/kg bw/day (37.8 °C compared to 38.5 °C in the control group). No similar effect was noted for males treated at 100 and 300 mg/kg bw/day or in females. Since the value of 37.8°C was borderline to the range of the historical control data (38.0°C-38.6°C), and no change was noted in females, this finding was considered to be incidental.

ORGAN WEIGHTS
Treatment with the test item at 1000 mg/kg bw/day resulted in statistically significantly higher absolute and relative liver and kidney weights in animals of both genders, which was confirmed in animals of the third delivery except for kidney weights in females. Since there was no evidence for an impairment of organ function by clinical pathology and histopathology, these findings were not considered to be adverse. Additionally, in females, absolute and relative heart weights were also increased. This finding was without histopathological correlate, and was therefore considered to be incidental.
There were no effects of treatment at 100 and 300 mg/kg bw/day.

GROSS PATHOLOGY
All findings were typical of this strain and age of rat and were considered to be incidental.

HISTOPATHOLOGY: NON-NEOPLASTIC
There were no treatment-related findings.
In particular, qualitative examination of the stages of spermatogenesis in the testis did not reveal any treatment-related abnormalities in the integrity of the various cell types present within the different stages of the sperm cycle. No treatment-related microscopic abnormalities were observed
in the evaluation of the ovarian follicles and corpora lutea of the ovaries or the evaluation of the uterus.
All findings were typical of this strain and age of rat and were considered to be incidental.

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects

Key result

Dose descriptor:

NOEL

Remarks:

systemic

Effect level:

>= 300 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects

Critical effects observed:

not specified

Conclusions:

Based on these results, the NOAEL for general toxicity was considered to be 1000 mg/kg bw/day. The NOEL for general toxicity was established at 300 mg/kg bw/day.

Reference 4

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

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

Remarks:

Summary of available data used for the endpoint assesssment of the target substance

Adequacy of study:

key study

Justification for type of information:

refer to the analogue justification provided in IUCLID section 13

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Description (incidence and severity):

see details on results

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects

Remarks on result:

other:

Remarks:

Source CAS 63705-03-3

Key result

Dose descriptor:

NOEL

Remarks:

systemic

Effect level:

>= 300 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects

Remarks on result:

other:

Remarks:

Source CAS 63705-03-3

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects

Remarks on result:

other:

Remarks:

Source CAS 111-03-5

Critical effects observed:

not specified

Conclusions:

As detailed in the analogue justification, it is considered that the target and the source substances are unlikely to lead to differences in repeated dose systemic toxicity potential. With both source substances, the NOAEL for general systemic toxicity was determined to be 1000 mg/kg bw/day. Therefore, the NOAEL for general systemic toxicity for the target substance is also expected to be 1000 mg/kg body weight/day.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

The available information comprises adequate, reliable studies from reference substances with similar structure and intrinsic properties. Read-across is justified based on common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to the endpoint discussion for further details). Taken together, the information from these independent sources is consistent and provides sufficient weight of evidence for hazard assessment leading to an endpoint conclusion in accordance with Annex XI, 1.2, of Regulation (EC) No 1907/2006. Therefore, the available information as a whole is sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

No repeated dose toxicity data are available on Fatty acids, C16-18 (even numbered), esters with glycerol oligomers. However, in EFSA’s scientific opinion on the re-evaluation of polyglycerol esters of fatty acids (PEPA, E 475), as a food additive, it is stated that the safety of polyglycerols and specific fatty acids has recently been assessed and no adverse effects were identified in the available studies. No adverse effects of PEFA at any dose have been observed in short-term, subchronic or chronic toxicity studies. A no observed adverse effect level (NOAEL) of 9,000 mg/kg bw per day was identified from subchronic studies and of 2,500 mg/kg bw per day from chronic studies, the highest doses tested (EFSA, 2017).

The registrants are currently trying to get hold of the repeated dose toxicity studies mentioned in the aforementioned EFSA report.  Therefore, as an interim measure, the repeated dose toxicity hazard of the registered substance was assessed based on the available data from the source substances 1,2,3 propanetriol, homopolymer, diisooctadecanoate (CAS 63705-03-3), deca-glycerol deca-oleate and 2,3-dihydroxypropyl octadec-9-enoate (CAS 111-03-5). In accordance with Regulation (EC) No. 1907/2006 Annex XI, 1.5 “Grouping of substances and read across” and following the Read across assessment framework (RAAF, ECHA 2017) read across from analogue substances have been applied to support the human health hazard assessment of Fatty acids, C16-18 (even numbered), esters with glycerol oligomers.

 

CAS 63705-03-3

 

A study investigating the toxicological effects of the test item 1,2,3 -Propanetriol, homopolymer, diidoctadecanoate (CAS 63705-03-3) to rats according to OECD guideline 422 is available. 1,2,3 -Propanetriol, homopolymer, diidoctadecanoate was administered to the rats by gavage, using corn oil as vehicle, at doses of 100, 300 and 1000 mg/kg bw/day; the control group received the vehicle only.

To meet guideline requirements concerning number of pregnant females per group, male and female rats were added in a second delivery for breeding due to reduced fertility observed in all groups of the first delivery. It is believed, that a disturbance of the light / dark cycle was the reason for the reduced mating rate of the females of the first delivery. The additional animals ran through the whole study as the animals from the first delivery with the exception that the males went to necropsy on day 24 after mating period and not on day 16 as the males of the first delivery (logistical reasons). Therefore, 1,2,3-Propanetriol, homopolymer, diisooctadecanoate was administered to male rats for up to 28 days (first delivery) and up to 41 days (second delivery) and to female rats for 14 days prior to mating, through the mating and gestation periods until the F1 generation reached day 4 post partum.

Because an impact of the light/dark cycle disturbance during the study on these results (a prolonged duration of gestation and an increased post-implantation loss was observed at the high dose) could not be excluded, the study was repeated with a third animal delivery of control and high-dose (1000 mg/kg bw/day) groups under proper light conditions. Test item was administered to 12 male rats per group for up to 33 days and to 12 female rats per group for 14 days prior to mating, through the mating and gestation periods until the F1 generation reached day 4 post partum. All animals survived until scheduled necropsy. There were no treatment-related clinical signs or effects on fuctional observation battery or locomotor activity. No test-titem related findings were noted during the clinical laboratory investigations.

At necropsy, liver and kidney weights were slightly increased at 1000 mg/kg bw/day in both sexes of the first and second delivery, which was confirmed in animals of the third delivery except for kidney weights in females. These changes were considered compound-related, but not adverse, since there was no evidence for an impairment of organ functional by clinical pathology and histopathology. There were no test item-related gross pathological or histopathological abnormalities.

Overall at 1000 mg/kg bw/day, food consumption and body weight gain were transiently reduced in males only at the beginning of the treatment period which was considered to be not adverse.

Based on these results, the NOAEL (No Observed Adverse Effect Level) for general parental toxicity was considered to be 1000 mg/kg body weight/day.

 

CAS 111-03-5

 

A GLP-compliant oral toxicity study according to OECD 422 was performed with 2,3-dihydroxypropyl oleate at doses of 100, 300 and 1000 mg/kg bw/day (Yamaguchi, 2005). Male and female Sprague Dawley rats (12 per sex and group, except for 1000 mg/kg bw/day: only 7 males) received the test substance in corn oil once daily via gavage. A control group, consisting of 7 males and 12 females, was treated with the vehicle alone. The duration of treatment was 42 days (14 days prior to mating and 28 days thereafter) in males and 42-52 days (from 14 days before mating to day 4 of lactation) in females, respectively. Satellite groups of 5 animals per sex, each for the control and test groups, were used to investigate reversibility of effects during a 14-day post-exposure recovery period. No mortality and no clinical signs and effects on neurobehaviour were observed during the whole study period. No adverse effects on body weight development were observed during treatment and recovery period. The analysis of clinical, haematological and urinary parameters did not reveal any dose-dependent and toxicologically relevant changes in treated animals compared to controls. At 100 mg/kg bw/day, a significant decrease in absolute weight of seminal vesicles in males and a significant decrease in relative weight of spleen in females were observed at necropsy. Since these effects did not follow a dose-dependent relationship and were not accompanied by any histopathological changes in the respective organs, they were not considered to be toxicologically relevant. A decrease in relative weights of pituitary in males and thyroid in females was observed at 1000 mg/kg bw/day at the end of the recovery period. Since no abnormalities were reported for these organs after the administration period, the findings were considered not compound-related. No test substance-related changes were found at gross pathology. A low incidence of commonly found microscopic changes was observed, which was evenly distributed between all groups and therefore considered as incidental findings. Although one female showed a subcutaneous tumour of the mammary gland after 40 days exposure to 300 mg/kg bw/day, this benign fibroadenoma was considered to be generated naturally, since no effects were observed at the higher dose levels. Based on the overall effects observed in the present study, a NOAEL of 1000 mg/kg bw/day for male and female Sprague Dawley rats was derived.

 

Decaglycerol decaoleate

 

A 90-day oral feeding rat study (King, 1971) was performed with the structurally related analogue substance decaglycerol decaoleate according a method equivalent to OECD Guideline 408. The test substance was administered via diet to 20 Sprague-Dawley rats (10 per sex) per dose group at specified dose levels (2.5, 5, and 10% corresponding to approx. 6944, 13,890 and 27,780 mg/kg bw/day for males and 7246, 14,493 and 28,985 mg/kg bw/day for females). Soyabean oil was used as the control fat. All animals appeared to be in excellent health during the duration of the study, and no adverse effects were observed with regard to survival, growth, absolute and relative organ weights, and histopathology. With regard to food consumption, it was found that males fed 10% PGE consumed more food and had a poorer feed efficiency value than male control animals. However, despite decreased feed efficiency those animals consumed enough food to maintain normal growth.

Concerning hematology and clinical chemistry the values fell within normal ranges, and there were no indications of any blood disorder. Some values were significantly different from Soybean Oil control (SBO), however differences were usually quite small and in no case established any trend or pattern indicative of a dose-related effect. Urine collected from each animal appeared to be normal in regard to color clarity, sediment, specific gravity and pH, total nitrogen excretion during the third and ninth week by females fed PGE at the 10% level was significantly greater than the control value and appears to be related to dietary treatment, the highest nitrogen excretion (male and female) was from the animals fed the highest dose PGE, the reason for this difference is not understood but it should be recognized that other parameters, including three derived from histologic examination of the urinary tract were normal.

The only gross observation of significance was what appeared to be very mild, chronic murine pneumonia in 18% of the animals, but the affected animals were scattered throughout all the groups and the effect was not related to the feeding of PGE.

The percentage of dietary fatty acids absorbed decreased as the level of PGE in the diet was increased. In all cases fat absorption by animals fed PGE at the 5 and 10% dietary levels was significantly less than corresponding SBO control values. Absorption in animals fed the lowest dose was also less but not significantly different. Values from the group fed oleic acid and glycerol fell between those of the 5% PGE and SBO control.

Gas-liquid Chromatography analyses of fecal fatty acids showed that excretion of oleic acid increased in a dose-related fashion: the oleic acid content of fecal fatty acids from animals fed the SBO control diet was 23% compared to 32, 41 and 51% when PGE was fed at levels of 2.5, 5, and 10%. The increased excretion of fatty acids in general and oleic acid in particular shows that absorption of dietary PGE was not complete. The fecal oleic acid may have resulted from the excretion of intact PGE or from hydrolyzed or partially hydrolyzed but unabsorbed material. The oleic acid content of fecal fatty acids from animals fed free oleic and glycerol was 41% corresponding exactly to the 5% PGE group.

 The 90-day oral NOAEL in diet was determined to be 10% of the test substance, which refers to approximately 27,780 mg/kg bw/day for males and 28,985 mg/kg bw/day for females when administered by daily feeding to rats for 90 days.

 

Conclusions

 

In conclusion, the available data on repeated dose toxicity via the oral route with all three source substances showed no adverse effects indicative of systemic toxicity resulting from repeated dose exposure. Thus, a similar result is expected for the target substance, based on read-across.

Justification for classification or non-classification

Based on read-across from structurally similar substances, the available data on repeated dose toxicity do not meet the criteria for classification according to Regulation (EC) No. 1272/2008, and are therefore conclusive but not sufficient for classification.