Fatty acids, C16-18, tetraesters with 3,3'-oxybis[1,2-propanediol]

Endpoint summary

Description of key information

Repeated dose toxicity: Oral NOAEL (rat, m/f): >1000 mg/kg bw/day (OECD 408, analogue approach)

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: 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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises adequate, reliable (Klimisch score 2) studies from reference substances with similar structure and intrinsic properties. Read-across is justified based on common origin, common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to endpoint discussion for further details). The selected studies are thus 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

Justification for grouping of substances and read-across

 

There are no data available for the repeated dose toxicity of Fatty acids, C16-18, tetraesters with 3,3'-oxybis[1,2-propanediol] (CAS# 91050-80-5).In order to fulfil the standard information requirements set out in Annex IX, 8.6.2, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, read-across from structurally related substances was conducted.

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

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

  

Overview of repeated dose toxicity, oral

CAS No.	NOAEL [mg/kg bw/day]
91050-80-5(a)	RA: deca-glycerol deca-oleate
Decaglycerol decaoleate(b)	10 % (m, f) (27780 (m) and 28985 (f) mg/kg bw/day)

 (a) Substances subject to the REACh  Phase-in registration deadline of 31 May 2013 are indicated in bold font. Only for this substance a full set of experimental results and/or read-across is given.

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

 

 

The above mentioned substances are considered to be similar on the basis of structural similarity resulting in similar properties and/or activities.The available endpoint information is used to predict the same endpoints for Fatty acids, C16-18, tetraesters with 3,3'-oxybis[1,2-propanediol] (CAS# 91050-80-5).

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

 

Discussion

Since no studies investigating the repeated dose toxicity, oral are available Fatty acids, C16-18, tetraesters with 3,3'-oxybis[1,2-propanediol] (CAS# 91050-80-5), in accordance to Regulation (EC) No. 1907/2006 Annex XI, 1.5 a read-across from the structurally related analogue decaglycerol decaoleate.

 

Decaglycerol decaoleate

A 90-day oral feeding study (King, 1971) was performed with the structurally related analogue substance decaglycerol decaoleate according a method equivalent to OECD Guideline 408. Test substance was administered via diet to 20 Sprague-Dawley rats per dose group at specified dose levels (2.5, 5, and 10% corresponding to approx. 6944, 13890 and 27780 mg/kg bw/day for males and 7246, 14493 and 28985 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 thatmales 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 SBO controls, 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 fetal 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 was determined to be 10% test substance, which refers to approximately 27780 mg/kg bw/day for males and 28985 mg/kg bw/day for females when administered by daily feeding to rats for 90 days.

 

There is no data available on the repeated dose toxicity after dermal application and inhalation of the category members.

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

Justification for classification or non-classification

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