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EC number: 604-608-2 | CAS number: 147853-32-5
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Description of key information
Based on read-across from Fatty acids, C18-unsaturated, dimers (CAS No. 61788-89-4):
Oral: NOAEL (rat, subchronic) = 741 (males) and 855 (females) mg/kg bw/day (similar to OECD 408, GLP)
Key value for chemical safety assessment
Repeated dose toxicity: via oral route - systemic effects
Link to relevant study records
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- 30 Nov Nov 1992 - 05 Mar 1993
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: GLP-compliant, comparable to guideline study with acceptable restrictions. No functional observations were conducted; most but not all haematological/clinical chemistry, organ weight and histopathological examinations were conducted.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
- Deviations:
- yes
- Remarks:
- no functional observations were conducted; most but not all haematological/clinical chemistry, organ weight and histopathological examinations were conducted
- 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 (UK) Ltd.
- Age at study initiation: 4-5 weeks
- Weight at study initiation: 114.5-155.5 g (males); 104.6-142.0 g (females)
- Housing: groups of 5 animals per cage
- Diet: ESL modified AIN-76A (MODAIN) purified diet, ad libitum
- Water: tap water, ad libitum
- Acclimation period: 1 week
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2
- Humidity (%): 55 ± 10
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 30 Nov 1992 To: 01-05 Mar 1993 - Route of administration:
- oral: feed
- Vehicle:
- unchanged (no vehicle)
- Details on oral exposure:
- DIET PREPARATION
- Rate of preparation of diet (frequency): the experimental diets were initially prepared on a weekly basis from 26 Nov - 10 Dec 1992. After data was obtained confirming stability of the test material in diet for 14 days, diets were prepared biweekly from 17 Dec 1992 to the end of the study.
- Mixing appropriate amounts with (Type of food): ESL modified AIN-76A (MODAIN) purified diet
- Storage temperature of food: diets in sealed bags were UV-sterilised for a minimum of 10 h at room temperature before entry in the SPF unit, where they were stored at ca. 4 °C before use. - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- -Stability and homogeneity in diet
Stability of the test substance at 0.1%, 1% and 5% in diet was determined over periods of 7 and 14 days when stored at 4°C or in the animal room.
Diets were analysed in order to confirm homogeneous test substance distribution in the diet. After diet mixing, 5 samples were taken for analysis from the top, the middle and bottom centre, and left and right centre of the mixing bowl.
Diet samples were extracted into propan-2-ol and centrifuged to remove particulate matter. An aliquot was concentrated to dryness, then redissolved and analysed by HPLC on a 5µ Lichrosorb Diol column, detection by a light scattering detector. Quantitation was achieved by comparison of peak areas with external standards of the test substance.
Separation on the HPLC system was based on the interaction of the carboxylic acid of the test substance with free hydroxyl groups at the surface of the diol phase. Thus, interaction increased with the number of carboxylic acid groups. The test substance contains mixtures of mono, di and polyacids. In the assay preparation based on two peaks, di-acid denoted dimer and tri or greater (poly) acid denoted trimer.
The test substance was shown to be stable in diet over 14 days. The results for 0.1% (w/w) test substance in diet showed more variation that would normally be accepted, but the level was very close to the limits of detection of the analytical method.
Using the methods described, the test substance was shown to be mixed homogeneously in the diet at a concentration of 0.1%, 1% and 5% (w/w).
-Confirmation of achieved concentration
Diets containing 5%, 1% and 0.1% test substance prepared on 26 Nov 1992, 07 Jan 1993 and 18 Feb 1993 were analysed for the achieved concentration of the test substance after first passing the UV lock. The methodology was the same as that used for the determination of stability.
Analysis of the diets prepared on Week 1 and 13 confirmed the nominal concentration had been achieved within the expected experimental error of the analytical method. The results for 0.1% (w/w) test substance in diet showed more variation that would normally be accepted, but the level was very close to the limits of detection of the analytical method. - Duration of treatment / exposure:
- 13 weeks
- Frequency of treatment:
- daily, 7 days/week
- Remarks:
- Doses / Concentrations:
0.1, 1, 5% (w/w)
Basis:
nominal in diet - Remarks:
- Doses / Concentrations:
74.1, 740.9, 3591.2 mg/kg bw/day (males)
Basis:
other: as calculated from the reported mean body weight and food intake data - Remarks:
- Doses / Concentrations:
90.5, 854.9, 4085.5 mg/kg bw/day (females)
Basis:
other: as calculated from the reported mean body weight and food intake data - No. of animals per sex per dose:
- 20
- Control animals:
- yes, plain diet
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice per day (once on Saturdays and Sundays)
- Cage side observations included: signs of ill-health or reaction to treatment
BODY WEIGHT: Yes
- Time schedule for examinations: on the first day of the study and once per week thereafter
FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No. Food intake values were determined two times per week and weekly values were calculated for each cage of 5 animals,
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No
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
WATER CONSUMPTION: Yes
- Time schedule for examinations: water consumption values were determined two times per week and weekly values were calculated for each cage of 5 animals,
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: during the week before study start and end, respectively
- Dose groups that were examined: all
HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the end of the treatment period
- Anaesthetic used for blood collection: Yes (halothane)
- Animals fasted: no data
- How many animals: all
- Parameters examined: red blood cell count, platelets, haemoglobin, total white blood cells, differential white cell count (lymphocytes, neutrophils, monocytes, eosinophils, basophils, large unstained cells), mean red cell volume (derived from the red cell volume histogram), haematocrit, mean red cell haemoglobin, mean red cell haemoglobin concentration, reticulocytes, prothrombin time (PT), activated partial thromboplastin time (APTT).
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of the treatment period
- Animals fasted: no data
- How many animals: all
- Parameters examined: sodium, potassium, calcium, magnesium, chloride, inorganic phosphate, creatinine, urea, glucose, triglyceride, total cholesterol, total bilirubin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), hydroxybutyrate dehydrogenase (HBD), alkaline phosphatase (AP), pseudocholinesterase, creatine kinase, 5’-nucleotidase, gamma-glutamyltransferase, total protein, albumin, alpha1-globulin, alpha2-globulin, β-globulin, gamma-globulin, albumin:globulin ratio. - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes. All surviving animals received a detailed necropsy. All macroscopic abnormalities were recorded and a score was allocated as an assessment of the level of intra-abdominal fat deposition. The following organs were weighed prior to fixation: brain, heart, liver, kidneys, spleen, testes and adrenal glands.
HISTOPATHOLOGY: Yes. The following tissues were taken from each rat and preserved in 10% buffered formalin: adrenal glands, jejunum, prostate, aorta, kidneys, rectum, bladder, larynx, sciatic nerve, brain, liver, spinal cord, caecum, lungs, spleen, cervix, lymph nodes (cervical and mesenteric), sternum, colon, mammary glands, stomach, duodenum, muscle, thymus, femur and stifle joint, oesophagus, thyroid and parathyroids, head, ovaries and fallopian tubes, tongue, heart, pancreas, trachea, ileum, pituitary, uterus.
The following tissues were taken from each rat and preserved in Bouin's fixative: epididymides, salivary glands, seminal vesicles, skin, testes, vagina.
The eyes/harderian glands were taken from each rat and fixed in Davidson's fluid.
Bone marrow smears were taken and stained with May-Grunwald Giemsa.
Microscopic examination was carried out on all of the tissues from all animals in the control and hig-dose groups, and on all tissues showing macroscopic abnormalities which were designated as lesions at necropsy. In addition, livers, adrenals, mesenteric lymph nodes, spleens and thyroids (females only) from all animals fed 1.0 and 0.1% test substance were examined microscopically following identification of treatment-related lesions in the high-dose group.
During the examination of tissues, histopathological changes were recorded as present or absent, or graded according to their morphology, using a numerical scale of 0.0–5.0 in order to assess degrees of severity or activity. This procedure provided a means of ranking degrees of change which can assist in the interpretation of biological differences. - Statistics:
- Statistical analysis was conducted for data on body weight, food and water intake, food conversion efficiency, clinical pathology and organ weights. Initially the data were examined to see if parametric or non-parametric analysis was appropriate.
For parametric data, one way analysis of variance was used to assess differences. A t-test was used to show any significant differences between control and test groups at the 5, 1 and 0.1% probability levels. A multiple T test was used for pairwise comparisons between groups. - 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):
- effects observed, treatment-related
- Description (incidence and severity):
- refer to Details on results
- Food efficiency:
- effects observed, treatment-related
- Description (incidence and severity):
- refer to Details on results
- Water consumption and compound intake (if drinking water study):
- no effects observed
- Ophthalmological findings:
- no effects observed
- Haematological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- refer to Details on results
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- refer to Details on results
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- refer to Details on results
- Gross pathological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- refer to Details on results
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- refer to Details on results
- Histopathological findings: neoplastic:
- no effects observed
- Details on results:
- CLINICAL SIGNS AND MORTALITY
There were no unscheduled deaths during the study period.
Clinical signs generally involved scabs, alopecia, excoriation, nasal discharge and ocular discharge, and were considered to be not treatment-related, since they were observed in all groups (control and treatment).
BODY WEIGHT AND WEIGHT GAIN
There were no treatment-related effects on body weight (gain) during the study. Statistically significant changes in body weight were few, minor, randomly distributed and of no biological significance. The mean body weight of females fed 1% test material was slightly (3.7%) increased in Week 0. In the same treatment group, body weight gain was slightly (7.6%) increased during Week 0-4.
FOOD CONSUMPTION AND COMPOUND INTAKE
There was a significant decrease in food consumption during the first 4 weeks of the study in males (-5.4%) and females (-8.4%) in the 5.0% group, which may reflect an initial reluctance of the animals to eat the diet.
FOOD EFFICIENCY
There was a significant increase in food conversion efficiency (9.5%) during the first four weeks of the study in females of the 5.0% group.
WATER CONSUMPTION
There were statistically significant changes (increases and decreases) in water consumption but without a clear treatment-related pattern. Accumulated water consumption was not significantly different between groups.
OPHTHALMOSCOPIC EXAMINATION
A persistent pupillary membrane was recorded for one male in the 0.1% group and ocular opacity was recorded for one female in the 5.0% group at the end of the study. These findings were considered to be not treatment-related.
HAEMATOLOGY (Table 2)
In the 5.0% group, mean cell haemoglobin was slightly (2.3%) but statistically significantly increased in females; prothrombin time was likewise slightly but significantly increased in males (3%) and females (4.3%).
In the 1.0% group, prothrombin time was slightly but significantly increased in females (2.6%).
The observed changes were slight and unlikely to be toxicologically relevant.
A number of other changes (e.g. decreased neutrophil counts in females of the 1 .0 and 5.0% groups) were statistically significant by Student's t-test but did not trigger the multiple t-test as significant.
CLINICAL CHEMISTRY (Table 3)
The following statistically significant changes were observed but not considered to be toxicologically relevant, since they were minor (< 10%) and/or no dose-relationship was noted:
Decrease in plasma calcium (5.0% male group and all female groups), glucose (1.0% female group), total protein (5.0% male and female groups) and increase in albumin/globulin ratio (1.0 and 5.0% male groups).
Aspartate aminotransferase was increased in females fed 0.1 and 5.0% test item and serum albumin was increased in males and females given 5.0% test item, but there was no dose-relationship.
Likewise, there was no clear evidence for a dose dependency for the decrease in 5’-nucleotidase (0.1% male groups and 5.0% male and female groups) and the increase in alanine aminotransferase (males and females of the 5.0% group).
Bilirubin was statistically significantly increased in males fed 1.0 and 5.0% test item in diet. However, the levels measured were below the sensitivity of the method used and must therefore be viewed with caution.
Total cholesterol and triglycerides were significantly and dose-dependently decreased at 1.0 and 5.0% in males and females. There was a decrease in beta-globulin in males of the 5.0% group.
The statistically significant and dose-dependent increase in alkaline phosphatase observed in males and females of the 1.0 and 5.0% groups was marked. The determined level was more than double that of the control group in animals of the 5.0% group.
ORGAN WEIGHTS (Table 4)
There was a statistically significant reduction in absolute and relative spleen weights in males of the 1.0 and 5.0% groups. There was a slight but statistically significant reduction in kidney weights in females fed 1.0% (only relative weight) and 5.0% (absolute and relative weights) test item in diet. In females of the 0.1% group, absolute liver weight was significantly reduced, while relative weights were significantly reduced in all female groups, but without showing any dose-relationship. Absolute and relative liver weights were slightly but significantly reduced in the 1.0 and 5.0% male groups.
The observed effects had no relation to any effect which might have been expected on the basis of the histopathology findings.
GROSS PATHOLOGY (Table 5)
There were no treatment-related effects on bodily condition as assessed by estimation of abdominal fat reserves at necropsy.
Slightly enlarged mesenteric lymph nodes were noted in 1/20 males and 2/20 females of the 0.1% groups, in 13/20 males and 14/20 females of the 1.0% groups and in 13/20 males and 16/20 females of the 5.0% groups. Moderate enlargement was observed in 1/20 males at 1.0% and in 7/20 males and 2/20 females at 5.0%.
The colour of caecal contents was affected by feeding with the test substance. Caecal contents were predominantly yellow in animals of the 5.0% group, predominantly yellow-green in animals of the 1.0% group, and green or gray-green in animals of the 0.1% or the control group, respectively. It should be note that the test substance was a yellow liquid.
There was a slightly increased incidence of uterine fluid distension in animals fed 5.0% test item (15/20 animals) compared with the control group (8/20 animals).
All other macroscopic findings were considered to be incidental and within the range expected for this age and strain of rats.
HISTOPATHOLOGY: NON-NEOPLASTIC (Table 6)
Microscopic examination revealed treatment-related findings in mesenteric lymph nodes, spleen, liver, adrenal glands and thyroid glands (in females). Only those effects seen in the mesenteric lymph nodes and spleen extended down to the group fed 0.1% test substance.
-Mesenteric lymph nodes:
Macrophage aggregation(s) (some containing a golden brown pigment) were noted in the paracortex and in the medullary cords in all animals of the 5.0% group, in 11/19 male and 17/19 females of the 1.0% group and in 3/18 male and 8/19 female animals of the 0.1% group. The incidence and number of aggregations were dose-related, and there were only a few aggregations present in animals of the 0.1% group. There was a correlation between histological findings in the mesenteric lymph nodes and lymph node enlargement noted at necropsy.
-Spleen:
Golden/dark brown pigmented macrophages were seen in the red and white pulp of all males and females of the 5.0% groups, 16/20 males and 19/20 females of the 1.0% groups and 5/20 females of the 0.1% group. Incidence and amount of pigmented macrophages was dose-related in male and female rats. The effect was more pronounced in females.
-Liver:
In male animals of the 5.0% group, there was an increased incidence of bile duct proliferation (18/20 animals vs. 5/20 animals in the control group) and sclerotic bile ducts (13/20 animals vs. 3/20 animals in the control group). Sclerosis was associated with minimal infiltration of mixed inflammatory cells. A very slight increase in the incidence of bile duct proliferation was seen in females of the 5.0% group (5/20 treated animals vs. 2/20 control animals). Periportal cytoplasmic vacuolation was decreased in males and females fed 1.0% and 5.0% test substance in diet.
-Adrenals:
In female animals of the 5.0% or 1.0% groups, cortical vacuolation was seen in the adrenal glands (13/20 and 20/20 rats, respectively). Trace levels of vacuolation were noted in one female of the 0.1% group. This finding was considered to be not toxicologically important, since vacuolation may occasionally be seen in control female animals. Cytoplasmic rarefaction was decreased in females fed 5.0% test item in diet (0/20 treated animals vs. 19/20 control rats).
Cortical extramedullary haemopoiesis was not present in females of the 5.0% group. A slightly reduced incidence of extramedullary haemopoiesis was noted in females given 1.0% or 0.1% test item in diet (6/20 and 5/20 animals, respectively, vs. 10/20 control animals). However, this was considered to be of no toxicological importance since the incidence of this finding generally varies considerably among groups of untreated animals.
-Thyroids:
A slight increase in follicular epithelial hypertrophy was noted in females of the 5.0% group (15/20 animals vs. 5/20 control animals).
-Spontaneous pathology:
Microscopic examination of the uteri from rats showing macroscopic fluid distension showed that this finding was due to a variety of different reasons: luminal dilatation or dilated/cystic endometrial glands. This finding was not thought to be of any toxicological importance, in view of this, and in view of the variation in uterine size with different phases of the estrous cycle.
The incidence of retinal folding/atrophy was higher in treated rats. However, given that the overall incidence was very low and there was no dose relationship, these lesions were not considered to be toxicologically relevant.
A slight increase in the incidence of lesions in the nasal passage was observed in animals of the 5.0% group when compared with the control groups. Lesions were minor in nature and included focal epithelial hypertrophy or hyperplasia associated with mucosal inflammatory cells or a luminal inflammatory exudate. Lesions of this nature are a common finding in control rats, and, while it is possible that they could be exacerbated by inhalation of diet containing irritant test material, the incidence in treated animals was still within the normal range.
A variety of spontaneous changes was noted in animals of all treatment groups without evidence of a treatment-related distribution. Findings were within the spectrum of spontaneous lesions commonly seen in laboratory rats of this age and strain and were therefore considered to be not substance-related. - Dose descriptor:
- NOAEL
- Effect level:
- 1 other: % in diet
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: adaptive effects on clinical chemistry (increase in alkaline phosphatase, slight decrease in total cholesterol and triglycerides) and histopathology (pigmented macrophage aggregation in lymph nodes and spleen)
- Dose descriptor:
- NOAEL
- Effect level:
- 741 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male
- Basis for effect level:
- other: corresponding to 1% in diet based on reported mean body weight and food intake data
- Dose descriptor:
- NOAEL
- Effect level:
- 855 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- female
- Basis for effect level:
- other: corresponding to 1% in diet based on reported body weight and food intake data
- Critical effects observed:
- not specified
- Conclusions:
- 1.0 % (w/w) test material in diet can be considered a NOAEL based on clinical chemistry parameters and histopathological findings, this corresponds to a dose level of approximately 741 and 855 mg/kg bw/day for male and female rats, respectively.
Reference
Table 1. Achieved dose levels.
Treatment |
Body weight (g) |
Mean body weight (g) |
Food intake (g) |
Mean food intake (g/day) |
Mean compound intake (g/day) |
Mean dose level (mg/kg bw/day) |
|
Week 0 |
Week 13 |
Week 0-13 |
Week 0-13 |
Week 0-13 |
Week 0-13 |
Week 0-13 |
|
Males |
|||||||
5.0% |
133.7 |
541.5 |
337.6 |
2182.3 |
24.2 |
1.21 |
3591.2 |
1.0% |
135.1 |
533.5 |
334.3 |
2229.1 |
24.8 |
0.25 |
740.9 |
0.1% |
135.8 |
560.8 |
348.3 |
2321.6 |
25.8 |
0.03 |
74.1 |
Control |
133.3 |
545.5 |
339.4 |
2215.3 |
24.6 |
0.00 |
0.0 |
Females |
|||||||
5.0% |
120.2 |
317.5 |
218.85 |
1609.4 |
17.9 |
0.89 |
4085.5 |
1.0% |
123.6 |
335.9 |
229.75 |
1767.7 |
19.6 |
0.20 |
854.9 |
0.1% |
120.2 |
317.7 |
218.95 |
1782.7 |
19.8 |
0.02 |
90.5 |
Control |
119.2 |
317.5 |
218.35 |
1693.8 |
18.8 |
0.00 |
0.0 |
Table 2. Haematology
Treatment |
Mean cell haemoglobin (pg) |
Prothrombin time (s) |
Males |
||
5.0% |
18.16* |
11.90** |
1.0% |
17.74 |
11.65 |
0.1% |
17.58 |
11.54 |
Control |
17.75 |
11.55 |
Females |
||
5.0% |
18.45 |
11.92*** |
1.0% |
18.57 |
11.73* |
0.1% |
18.51 |
11.58 |
Control |
18.41 |
11.43 |
* Statistically significantly different versus control (P = 0.5)
** Statistically significantly different versus control (P = 0.1)
*** Statistically significantly different versus control (P = 0.01)
Table 3. Clinical chemistry.
Treatment |
Calcium (mmol/L) |
5’-Nucleotidase (U/L) |
Alkaline phosphatase (U/L) |
Alanine aminotransferase (U/L) |
Aspartate aminotransferase (U/L) |
Triglycerides (mmol/L) |
Total cholestertol (mmol/L) |
Males |
|||||||
5.0% |
2.502*** |
28.4*** |
810.6*** |
65.1*** |
104.5 |
0.618*** |
1.901*** |
1.0% |
2.559 |
32.7 |
606.3*** |
52.8 |
107.3 |
1.247*** |
2.380* |
0.1% |
2.598 |
32.2* |
424.7 |
40.4 |
95.1 |
1.701 |
2.517 |
Control |
2.604 |
36.3 |
389.5 |
41.0 |
93.8 |
1.820 |
2.803 |
Females |
|||||||
5.0% |
2.567*** |
47.0** |
512.6*** |
43.1*** |
93.1*** |
0.475*** |
1.766*** |
1.0% |
2.626** |
63.4 |
393.3*** |
30.5 |
75.9 |
0.876 |
2.181* |
0.1% |
2.612*** |
74.3 |
250.4 |
29.1 |
81.5** |
0.965 |
2.362 |
Control |
2.694 |
76.3 |
216.4 |
29.6 |
71.1 |
1.120 |
2.441 |
Treatment |
Glucose (mmol/L) |
Bilirubin (µmol/L) |
Total protein (g/L) |
Albumin (g/L) |
Albumin/globulin ratio |
beta-globulin (g/L) |
Males |
||||||
5.0% |
9.353 |
3.1*** |
62.07*** |
29.23* |
0.905** |
10.64*** |
1.0% |
9.815 |
2.7* |
64.09* |
30.44 |
0.905** |
11.89 |
0.1% |
10.258 |
2.5 |
65.44 |
30.16 |
0.860 |
12.29 |
Control |
10.053 |
2.3 |
66.58 |
30.52 |
0.845 |
12.36 |
Females |
||||||
5.0% |
9.420 |
2.7 |
67.29** |
33.76*** |
1.015 |
10.65 |
1.0% |
9.373* |
2.8 |
70.59 |
36.24 |
1.055 |
10.70 |
0.1% |
9.993 |
2.6 |
71.10 |
35.97 |
1.025 |
11.15 |
Control |
9.852 |
2.6 |
72.35 |
37.12 |
1.050 |
11.43 |
* Statistically significantly different versus control (P = 0.5)
** Statistically significantly different versus control (P = 0.1)
*** Statistically significantly different versus control (P = 0.01)
Table 4. Organ weights.
Treatment |
Spleen (g) |
Spleen (g/100 g bw) |
Kidney (g) |
Kidney (g/100 g bw) |
Liver (g) |
Liver (g/100 bw) |
Males |
|
|
|
|
|
|
5.0% |
0.7520*** |
0.1381*** |
3.429 |
0.631 |
18.169* |
3.343*** |
1.0% |
0.8436* |
0.1566* |
3.441 |
0.641 |
18.739* |
3.472** |
0.1% |
0.9571 |
0.1691 |
3.565 |
0.633 |
20.756 |
3.660 |
Control |
0.9454 |
0.1723 |
3.635 |
0.663 |
20.610 |
3.737 |
Females |
|
|
|
|
|
|
5.0% |
0.5573 |
0.1765 |
2.028* |
0.640** |
10.781 |
3.395** |
1.0% |
0.5904 |
0.1764 |
2.178 |
0.650* |
10.804 |
3.210*** |
0.1% |
0.5808 |
0.1851 |
2.064 |
0.657 |
9.862*** |
3.125*** |
Control |
0.6079 |
0.1924 |
2.167 |
0.688 |
11.520 |
3.640 |
* Statistically significantly different versus control (P = 0.5)
** Statistically significantly different versus control (P = 0.1)
*** Statistically significantly different versus control (P = 0.01)
Table 5. Incidence of macroscopic findings.
Macroscopic observation |
Treatment |
||||
Control |
0.1% |
1.0% |
5.0% |
||
Males |
|||||
No. of animals/group |
20 |
20 |
20 |
20 |
|
Caecum |
Contents green |
5 |
6 |
4 |
0 |
Contents gray-green |
15 |
13 |
0 |
0 |
|
Contents yellow |
0 |
0 |
0 |
19 |
|
Contents yellow-green |
0 |
1 |
16 |
1 |
|
Mesenteric lymph node |
Enlarged (moderate) |
0 |
0 |
1 |
7 |
Enlarged (slight) |
0 |
1 |
13 |
13 |
|
Females |
|||||
No. of animals/group |
20 |
20 |
20 |
20 |
|
Caecum |
Contents green |
7 |
8 |
3 |
0 |
Contents gray-green |
13 |
12 |
1 |
0 |
|
Contents yellow |
0 |
0 |
0 |
20 |
|
Contents yellow-green |
0 |
0 |
16 |
0 |
|
Mesenteric lymph node |
Enlarged (moderate) |
0 |
0 |
0 |
2 |
Enlarged (slight) |
0 |
2 |
14 |
16 |
|
Uterus |
Moderate fluid distention |
1 |
4 |
6 |
3 |
Severe fluid distention |
1 |
0 |
0 |
0 |
|
Slight distention |
0 |
0 |
1 |
0 |
|
Slight fluid distention |
8 |
7 |
6 |
15 |
Table 6. Incidence of microscopic findings.
Microscopic observation |
Treatment |
||||
Control |
0.1% |
1.0% |
5.0% |
||
Males |
|||||
Adrenals |
No. examined |
20 |
20 |
20 |
20 |
|
Without abnormalities |
3 |
0 |
0 |
1 |
Adrenal cortex |
Vacuolation |
17 |
20 |
20 |
19 |
|
Extramedullary haemopoiesis |
0 |
2 |
0 |
0 |
Thyroids |
No. examined |
20 |
0 |
0 |
20 |
|
Without abnormalities |
10 |
0 |
0 |
8 |
|
Follicular epithelium hypertrophy |
10 |
0 |
0 |
12 |
Mesenteric lymph node |
No tissue available |
0 |
2 |
1 |
0 |
|
No. examined |
20 |
18 |
19 |
20 |
|
Without abnormalities |
12 |
12 |
1 |
0 |
|
Pigmented macrophage aggregation(s) |
0 |
1 |
11 |
20 |
|
Macrophage aggregation(s) |
1 |
3 |
10 |
20 |
Spleen |
No. examined |
20 |
20 |
20 |
20 |
|
Without abnormalities |
20 |
19 |
4 |
0 |
|
Pigmented macrophages |
0 |
0 |
16 |
20 |
|
Extramedullary haemopoiesis |
0 |
1 |
2 |
0 |
Liver |
No. examined |
20 |
20 |
20 |
20 |
|
Without abnormalities |
0 |
0 |
1 |
0 |
|
Cytoplasmic vacuolation (periportal) |
7 |
6 |
1 |
0 |
|
Bile duct proliferation |
5 |
4 |
6 |
18 |
|
Sclerotic bile duct(s) |
3 |
4 |
7 |
12 |
|
Mixed inflammatory cell infiltration (periportal) |
1 |
1 |
1 |
11 |
Females |
|||||
Adrenal cortex |
No. examined |
20 |
20 |
20 |
20 |
|
Without abnormalities |
0 |
0 |
1 |
0 |
|
Vacuolation |
0 |
1 |
13 |
20 |
|
Extramedullary haemopoiesis |
10 |
6 |
5 |
0 |
|
Cytoplasmic rarefaction |
19 |
16 |
14 |
0 |
Thyroids |
Follicular epithelium hypertrophy |
5 |
5 |
6 |
15 |
Mesenteric lymph node |
No tissue available |
0 |
1 |
1 |
0 |
|
No. examined |
20 |
19 |
19 |
20 |
|
Without abnormalities |
17 |
5 |
1 |
0 |
|
Pigmented macrophage aggregation(s) |
0 |
3 |
9 |
19 |
|
Macrophage aggregation(s) |
0 |
8 |
17 |
20 |
|
Focus(i) foamy macrophages |
0 |
1 |
0 |
0 |
Spleen |
No. examined |
20 |
20 |
20 |
20 |
|
Without abnormalities |
20 |
15 |
1 |
0 |
|
Pigmented macrophages |
0 |
5 |
19 |
20 |
Liver |
No. examined |
20 |
20 |
20 |
20 |
|
Without abnormalities |
0 |
1 |
5 |
5 |
|
Cytoplasmic vacuolation (periportal) |
10 |
13 |
6 |
2 |
|
Bile duct proliferation |
2 |
0 |
0 |
5 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed
- Quality of whole database:
- The available information comprises an adequate and reliable study (Klimisch score 2) from a reference substance with similar structure and intrinsic properties. Read-across is justified based on common functional group(s), common precursors/breakdown products and similarities in physicochemical and (eco)toxicological properties (refer to endpoint discussion for further details).
The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6.2, 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 on the repeated dose toxicity of Fatty acids, C18-unsatd., dimers, di-Me esters, hydrogenated (CAS 147853-32-5). 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 a structurally related substance is 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 physicochemical, toxicological and ecotoxicological properties may be predicted from data for reference substance(s) by interpolation to other substances on the basis of structural similarity, Fatty acids, C18-unsatd., dimers (CAS 61788-89-4) is selected as reference substance for assessment of repeated dose toxicity.
The read-across is based on structural similarity between the source and target substances, which share a common origin and metabolic fate. A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).
Overview of repeated dose toxicity
|
Target substance |
Source substance |
CAS No. |
147853-32-5 |
61788-89-4 |
Chemical name |
Fatty acids, C18-unsatd., dimers, di-Me esters, hydrogenated |
Fatty acids, C18-unsatd., dimers |
Repeated dose toxicity: oral (subacute) |
RA: CAS 61788-89-4 |
Experimental result: NOAEL (rat, m) ≥ 1450 mg/kg bw/day NOAEL (rat, f) ≥ 1692 mg/kg bw/day (OECD 421) |
Repeated dose toxicity: oral (subchronic) |
RA: CAS 61788-89-4 |
Experimental result: NOAEL (rat, m) = 741 mg/kg bw/day NOAEL (rat, f) = 855 mg/kg bw/day (similar to OECD 408) |
Repeated dose toxicity: oral
Subacute
CAS 61788-89-4
A Reproduction/Developmental Toxicity Screening Test was conducted with Fatty acids, C18-unsatd., dimers following OECD guideline 421 and under GLP conditions (Clubb and Sutherland, 2004). This study is discussed in detail under Toxicity to reproduction.
Based on the lack of toxicologically relevant effects up to the highest dose level tested, the systemic toxicity as well as the reproductive toxicity/fertility NOAEL values were ≥ 1450 mg/kg bw/day and ≥ 1692 mg/kg bw/day for males and females, respectively.
Subchronic
CAS 61788-89-4
Fatty acids, C18-unsaturated, dimers were tested in a subchronic oral toxicity study in rats according to a method similar to OECD guideline 408 and under GLP conditions (Spurgeon and Hepburn, 1993). Groups of 20 Sprague-Dawley rats per sex and dose were fed the test item at 0.1, 1.0 and 5.0% in diet for 13 weeks. The corresponding dose levels (averaged over the 13-week study period) were ca. 74.1, 740.9, 3591.2 mg/kg bw/day for males and ca. 90.5, 854.9, 4085.5 mg/kg bw/day for females, based on the reported body weight and food intake data.
There were no unscheduled deaths no treatment-related changes in clinical signs, body weight and at ophthalmoscopic examination. Food consumption was lower during the first 4 weeks in males and females of the high-dose group. The lower food intake was likely due to an initial reluctance of the animals to eat the diet. No treatment-related changes were noted in water intake. There were no toxicologically relevant changes in haematological parameters.
Clinical chemistry examination showed an increase in plasma alkaline phosphatase activity (AP) in males and females of the mid- and high-dose groups. Alanine aminotransferase (ALT) was also increased in male and female rats of the high-dose group. An increase in plasma AP activity may reflect induction (increased synthesis) in liver cells rather than increased release from damaged cells.
In males and females of the high-dose group, small reductions in plasma calcium, total serum protein and albumin were observed. It is possible the plasma calcium and serum protein changes may be connected. However, reduction in plasma calcium was not always accompanied by a simultaneous reduction in plasma albumin. Changes in plasma calcium and serum proteins were small, probably representing a physiological rather than a pathological response to treatment.
Total cholesterol and triglycerides in plasma were reduced in male and female of the mid- and high-dose groups. The effect was very small for the mid-dose group. It is possible that the test item blocks the absorption of lipid and other nutrients from the gut. This could also be an explanation for changes in plasma electrolytes and intermediate metabolites.
Decreases in absolute and/or organ weights of spleen (males), kidney (females) and liver (males and females) were observed, mainly in the high- and less in the mid-dos group, but there was no relation to any effect which might have been expected on the basis of the findings at histopathological examination.
Additionally to the increased plasma AP level, treatment-related effects were noted on microscopic examination of the liver. Histopathological examination showed an increase in biliary hyperplasia in the high-dose group. Interference in bile flow could be related to the observed increase in AP, which is a sensitive indicator of cholestasis. Changes in AP develop before any detectable increases in plasma bilirubin levels. It should be noted that while a very small increase in plasma bilirubin was observed in male rats of the mid- and high-dose groups, the bilirubin levels measured were below the sensitivity of the analytical method and must therefore be seen with caution. Although bile duct proliferation and sclerosis noted in the liver may correlate with increased AP, it should be noted that only very minor biliary changes were observed in a few number of females. The reduction in periportal hepatocyte vacuolation found at histopathological examination of the liver could correlate with the reduced plasma lipids, as described above, indicating some alteration in lipid metabolism, another possible explanation for the plasma lipid, serum protein and calcium changes.
Aggregation of macrophages (some of which were pigmented) was seen in the mesenteric lymph nodes of all treated animals. In the spleen, macrophages containing pigment were observed mainly in mid- and high-dose males and females. There was a dose-related increase in the incidence and amount of macrophages. The pigment present in the macrophages in the mesenteric lymph nodes and spleen did not stain with Perls' stain for haemosiderin or aldehyde fuchsin for lipofuscin, but did stain with Schmorl's stain for lipofuscin. Lipofuscin is derived from oxidation of unsaturated lipids or lipoproteins. Given the fact that the test item is composed of a mixture of monomers, dimers and trimers of fatty acids, the pigment likely represented either lipofuscin produced by oxidation of some component of the test substance, or that the test substance itself stained positively.
No evidence of any degenerative effect associated with pigmented macrophages could be found. It is probable that they represented a physiological response to dietary administration of lipid materials such as the test item. Accumulations of macrophages in the mesenteric lymph nodes commonly occur as a result of ageing, or after administration of pigmented or lipid substances in the diet (Gopinath et al., 1987; Mohr et al., 1992). Although the pigment appeared darker in the spleen than in the mesenteric lymph node, this may have reflected a difference in density. The pigment was present alongside normal levels of haemosiderin which appeared tinctorially distinct. The coloured nature of the test substance is also likely to have been responsible for the alteration in the colour of the caecal contents observed at necropsy.
Increased cortical vacuolation in the adrenals was observed in females of the mid and high-dose groups, coupled with a decrease in cytoplasmic rarefaction. This finding probably indicated altered steroidogenesis. However, it was not accompanied by any evidence of degenerative change. The relevance of the reduced extramedullary haemopoiesis seen in the adrenals of the high-dose females is uncertain, but may possibly correlate with the reduction in neutrophil count in females of the mid- and high-dose groups. All of the changes in the adrenal glands were minor in nature and of limited importance.
Increased thyroid follicular epithelial hypertrophy was noted in females of the high-dose group. This finding was unusual and not dose-related. The hypertrophy consisted of a slight increase in height of the follicular epithelium combined with a reduction in the size of the follicular lumen and in the amount of colloid contained therein.
Based on clinical chemistry parameters and histopathological findings considered to be of adaptive nature, the subchronic oral NOAEL for rats was determined to be 1% in diet, corresponding to 741 and 855 mg/kg bw/day for males and females, respectively.
Conclusions for repeated dose toxicity
There are no data available on the repeated dose toxicity of Fatty acids, C18-unsatd., dimers, di-Me esters, hydrogenated. The substance is a UVCB composed of dimerised long-chain fatty alcohols derived from the reduction of the corresponding dimerised fatty acids via an intermediate esterification step with methanol. In general, the initial step in the mammalian metabolism of primary alcohols is the enzymatic oxidation to the corresponding carboxylic acid (see Toxicokinetics). Therefore, hazard assessment is conducted by means of read-across from the source substance Fatty acids, C18-unsatd., dimers.
A subchronic oral toxicity study is available for Fatty acids, C18-unsatd., dimers, in which NOAEL values of 741 and 855 mg/kg bw/day were identified for males and female rats (corresponding to 1% in diet), based on adaptive changes in clinical chemistry parameters and histopathological findings.
Furthermore, in a reproduction/developmental toxicity screening study with Fatty acids, C18-unsatd., dimers, no toxicologically relevant effects were observed in rats treated with the test item up to and including the highest dose level, thus resulting in NOAEL values of at least 1450 (males) and 1692 (females) mg/kg bw/day.
Based on read-across, the substance is considered to be of low toxicity after long-term oral exposure, and no hazard was identified since in a subchronic study toxicologically relevant effects were observed only at dose levels above the currently applicable limit dose level of 1000 mg/kg bw/day.
There are no data available on repeated dose toxicity by the inhalation and dermal routes.
References
Gopinath, C. et al. (1987). In : Atlas of Experimental Toxicological Pathology, pp 112, 113 and 127. MTP Press Ltd., Falcon House, Queen Sq., Lancaster, England.
Mohr, U. et al. (Eds.) (1992). In: Pathobiology of the Ageing Rat, pp 54, 55. ILSI Press, 1126 Sixteenth St., N.W., Washington, D.C.
Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Hazard assessment is conducted by means of read-across from a structural analogue/surrogate. The selected study is the most adequate and reliable study based on the identified similarities in structure and intrinsic properties between source and target substance and overall assessment of quality, duration and dose descriptor level (refer to the endpoint discussion for further details).
Justification for classification or non-classification
Based on read-across from the structurally related substance Fatty acids, C18-unsatd., dimers (CAS No. 61788 -89 -4), the available data on the repeated dose oral toxicity of Fatty acids, C18-unsatd., dimers, di-Me esters, hydrogenated 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.
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