Registration Dossier

Administrative data

Description of key information

There are two 90 day studies available, for the read across substance registered under 2,2’-(C16-18 (even numbered, C18 unsaturated) alkyl imino) diethanol CAS No 1218787-32-6. As this contain C18 unsaturated which is more reactive than the C18 saturated in 2,2’-(Octadecylimino)bisethanol) CAS No 10213-78-2 and C16, so it is expected to more toxic and therefore will not underestimate the toxicity of 2,2’-(Octadecylimino)bisethanol.  While these studies are from 1965 and less detailed than current studies, they are considered to be Klimisch 2.  Due to problems with vomiting in the dog study the rat study provides the most reliable repeat dose NOAEL.

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 based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
1965
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: No data on batch no. and composition. Reporting is limited. The study was performed pre-GLP. No guideline was available.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Haematology parameters were limited. No clinical biochemistry was performed. No data reported on frequency and set up of
observations and clinical signs. Ophthalmological examinations and functional observations were not performed.
GLP compliance:
no
Remarks:
pre-GLP
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: own colony maintained at Alderley Park, Cheshire, UK
- Weight at study initiation: 126-310 g (males), 88-232 g (females)
- Age at study initiation: no data
- Fasting period before study: no info
- Housing: 5/cage in Wilmslow-type mobile rat units (wire mesh cages on 3 sides and floor)
- Diet (e.g. ad libitum): ad lib
- Water (e.g. ad libitum): ad lib
- Acclimation period: in house colony


ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data


IN-LIFE DATES: no data
Route of administration:
oral: feed
Vehicle:
maize oil
Details on oral exposure:

DIET PREPARATION:

All diets were pregered in tho central diet room of the Laboratories. The basic ration from which all diets were made was a
standard pulverised rat stock diet. The control diet contained powdered stock diet, 20 parts, malt extract, 18 parts, and maize oil , 2 parts, all by weight. Experimental diets were identical with the control except that the calculated weight of Ethomeen was
incoporated into the diet via the maize oil, in which it was dissolved by gentle heating at 40 degrees C. The ingredients were mixed mechanically and watar added to produce e dough. The mixture was extruded through a sausage-meat machine which moulded thediet into pieces of' 3 - 5 cm long and approximately 1 cm in diameter. These were then dried in a vacuum oven at a temperature of
not more than 40°C.
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
No analyticak verification.
Duration of treatment / exposure:
90 days
Frequency of treatment:
ad libitum in the feed
Remarks:
Doses / Concentrations:
170, 500, 1500, 4500 ppm
Basis:
nominal in diet
No. of animals per sex per dose:
25 sex/group, in the higest dose group 10 sex/group. In addition a further group of 14 rats, 7 males and 7 females, were fed diet
containing 4500 ppm Ethomeen T/12 end killed at intervals up to 6 weeks from the beginning of the experiment. Tissues from these
animals were examined for sudanophilic material.
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: no info
- Rationale for animal assignment (if not random): at random
- Section schedule rationale (if not random): no info
Positive control:
Not used.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes, but frequency of observations not indicated.

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes / No / No data
Initially, the body weight of each rat was determined and recorded. Subsequently all animals were weighed weekly during the course of tho experiment.

FOOD CONSUMPTION:
Measured daily during 1 week in 0, 1500 and 4500 ppm groups.

FOOD EFFICIENCY: No data

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

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY:
Haemoglobin concentrations, packed-cell volumes, white-cell counts and differential white-cell counts were measured pre-
experimentally and imediately prior to killing the animals at the end of the feeding period, These studies were done on individual
samples from 5 male and 5 female rats from each group except-that blood was examined from all animals fed diet containing the
highest level of Ethomeen T/12.

CLINICAL CHEMISTRY: No data

URINALYSIS: No data

NEUROBEHAVIOURAL EXAMINATION: No data

Sacrifice and pathology:
PATHOLOGY:
At the end of the 90-day test period all animals were killed with chloroform and an immediate full post-mortem examination made.
Absolute organ weights were recorded and organ/body weight ratios calculated from a random selection of animals in each group. The following organs wore included: liver, heart, lung, adrenals, kidneys, spleen. Tissues and organs of the remaining animals werefixed in Zenker's fluid except brains, which ware fixed in 10% formal saline, and examined microscopically. The following tissues
and organs were examined: liver, kidney, spleen, heart, lung, adrenals, gonads, thymus, thyroid, pancreas, stomach, duodenum,
jejunum, ileum, caecum, colon, salivary gland, mesenteric lymph nodes, spinal cord and brain (cerebrum, cerebellum and medulla).
Other examinations:
No
Statistics:
Not used.
Clinical signs:
not specified
Mortality:
not specified
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
Mortality and Reactions:
No deaths occurred prematurely and males and females responded similarly. At the highest dose level hairloss was observed and
the animals were generally lethargic throughout the experiment. At the other dose levels no effects were noted.

Body weights:
At 4500 ppm, no body weight gain was observed and at 1500 ppm a decreased body weight gain was observed. At lower levels
no effects were observed.

Food consumption:
The palatability of the diet was affected by the addition of 4500 and 1500 ppm test compound.

Haematology:
No effects were noted.

Pathology:
Changes attributed to treatment were seen only in animals at a dietary level of 4500 ppm. These changes were confined to the
gastrointestinal tract. The stomach and bowel content of all rats at this dietary level was yellow and the mucooa of the small
intestine was thickened and yellow. The mean organ body-weight ratios were not different from controls. Abnormality was seen
only at a dietary level of 4,500 ppm and 1,500 pppm and was confined to the small intestine and regional mesenteric nodes. All
animals fed 4,500 ppm in the diet showed engorgement of the villi and lamina propria of the small intestine with swollen foamy
macrophages. Similar macrophages are occasionally seen to a lesser degree in Peyer's patches and in the regional lymph nodes.
Changes were most proounced In the jejunum and upper ileum but could be detected throughout the small intestine. The
macrophages were sudanophilic and were presumed to contain deposits of Ethomeen. The other components of the intestinal wall
appeared completely normal. A similar change was present to a lesser degree in 31 of 40 rats fed a dietary level of 1,500 ppm. No
changes were present ot a dietary level of 500 ppm or less of Ethomeen T/12.
Dose descriptor:
NOEL
Effect level:
500 ppm
Sex:
male/female
Basis for effect level:
other: At higher levels animals did not gain weight normally and showed pathological lesions confined to the small intestines and its regional lymph nodes. This dietary level of 500 ppm corresponds to ca. 35 mg/kg bw/day (see below)
Critical effects observed:
not specified

A NOEL of 500 ppm in the diet corresponds with:

ca. 33 mg/kg bw in males (taking into account a mean food intake of 20 g/day and a mean BW of 300 g throughout the study period)

ca. 35 mg/kg bw in females (taking into account a mean food intake of 15 g/day and a mean BW of 200 g throughout the study period)

Conclusions:
From this study a NOEL of 500 ppm can be derived. At higher levels animals did not gain weight normally and showed pathological
changes confined to the small intestines and its regional lymph nodes. This dietary level corresponds to ca. 35 mg/kg bw/day.
Executive summary:

The test substance Ethomeen T/12 was administered via the diet daily for 90 days to Wistar rats. One control group and four treated groups were tested, each consisting of 25 males and 25 females (except for the high dose group that had 10 animals per sex). These were exposed to 0, 170, 500, 1500 or 4500 ppm. A further group of 14 rats, 7 males and 7 females, was fed a diet containing 4500 ppm Ethomeen T/12 end killed at intervals up to 6 weeks from the beginning of the experiment. Tissues from these animals were examined for sudanophilic material.

The following parameters were evaluated: clinical signs, body weight and food consumption, haematology, and macroscopy at termination, organ weights and histopathology on a selection of tissues.

No deaths occurred prematurely and males and females responded similarly. At the highest dose level hairloss was observed and the animals were generally lethargic throughout the experiment. At the other dose levels no effects were noted. Body weights: At 4500 ppm, no body weight gain was observed and at 1500 ppm a decreased body weight gain was observed. At lower levels no effects were observed. Food consumption: The palatability of the diet was affected by the addition of 4500 and 1500 ppm test compound. Haematology: No effects were noted. Pathology: Changes attributed to treatment were seen only in animals at a dietary level of 4500 ppm. These changes were confined to the gastrointestinal tract. The stomach and bowel content of all rats at this dietary level was yellow and the mucooa of the small intestine was thickened and yellow. The mean organ body-weight ratios were not different from controls. Abnormality was seen only at a dietary level of 4,500 ppm and 1,500 pppm and was confined to the small intestine and regional mesenteric nodes. All animals fed 4,500 ppm in the diet showed engorgement of the villi and lamina propria of the small intestine with swollen foamy macrophages. Similar macrophages are occasionally seen to a lesser degree in Peyer's patches and in the regional lymph nodes. Changes were most proounced In the jejunum and upper ileum but could be detected throughout the small intestine. The macrophages were sudanophilic and were presumed to contain deposits of Ethomeen T/12. The other components of the intestinal wall appeared completely normal. A similar change was present to a lesser degree in 31 of 40 rats fed a dietary level of 1,500 ppm. No changes were present ot a dietary level of 500 ppm or less of Ethomeen T/12.

From this study a NOEL of 500 ppm can be derived. At higher levels animals did not gain weight normally and showed pathological changes confined to the small intestines and its regional lymph nodes. This dietary level corresponds to ca. 35 mg/kg bw/day.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
35 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
There are two 90 day studies available, for the read across substance registered under 2,2’-(C16-18 (even numbered, C18 unsaturated) alkyl imino) diethanol CAS No 1218787-32-6. While these studies are from 1965 and less detailed than current studies, they are considered to be Klimisch 2. Due to problems with vomiting in the dog study the rat study provides the most reliable repeat dose NOAEL.

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

There are two 90 day studies available, for the read across substance registered under 2,2’-(C16-18 (even numbered, C18 unsaturated) alkyl imino) diethanol CAS No 1218787-32-6. As this contain C18 unsaturated which is more reactive than the C18 saturated in 2,2’-(Octadecylimino)bisethanol CAS No 10213-78-2 and C16, so it is expected to more toxic and therefore will not underestimate the toxicity of 2,2’-(Octadecylimino)bisethanol. The 90 Day dog study, where the dogs were dosed by mixing a solution of the test substance in maize oil into the dry dog diet, suffered from serious problems with acceptance of the diet. This was due to corrosive/irritant nature of the test substance. The highest dose level of 120mg/kg/day vomited 2 to 3 hours after feeding and then began to refuse their food. Despite attempts to acclimatise them to the test substance this group had to be terminated after 5-6 weeks due to their poor condition, with 20% bodyweight loss. There was also sporadic vomiting and reluctance to eat all their food in the 40mg/kg/day group. This left the 13 mg/kg/day group as the NOAEL. In the 90 day rat study which some decrease in food consumption was evident at the higher dose levels the 500ppm in the diet level was a clear NOAEL. This was calculated to be ca. 35 mg/kg/day. Due to these problems with vomiting and reduced food consumption etc. in the dog study, the NOAEL from the rat study has been selected as being more reliable. This is supported by the findings of no clear systemic toxicity in either study, the effects seen in both studies be associated with local irritant effects in the gastrointestinal tract. The 35mg/kg/day NOAEL value will therefore be used for key value for the chemical safety report for read across to, 2,2’-(Octadecylimino)bisethanol CAS No 10213-78-2.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:

There are no repeat dose studies on 2,2’-(Octadecylimino)bisethanol) CAS No 10213-78-2 or the read across substance 2,2’-(C16-18 (evennumbered), alkyl imino) diethanol CAS No 1218787-30-4.  But there are two 90 day oral toxicity studies for the read across substance Ethanol, 2,2’-iminobis-, N-tallow alkyl derives CAS No 61791-44-4 registered under 2,2’-(C16-18 (even numbered, C18 unsaturated) alkyl imino) diethanol CAS No 1218787-32-6.  This is almost the same as 2,2’-(C16-18 (evennumbered), alkyl imino) diethanol CAS No 1218787-30-4, expect of a some of the C18 is unsaturated.  As unsaturation leads to increased reactivity, it is expected that this substance would be more toxic than 2,2’-(Octadecylimino)bisethanol) CAS No 10213-78-2 and therefore it is not expected to underestimate the repeat dose oral toxicity.  There is a 90 day dietary study in rats and a 90 Day study in dogs where the test substance was added to the diet in a solution in maize oil.  The Dog study was dosed at 13, 40, 120 mg/kg/day with a maize oil vehicle control.  The rats were feed diets containing 170, 500, 1’500 and 4’500ppm of the tests substance.  Both studies provide NOAEL values.  These studies were carried out in 1965, but are sufficiently well documented including information on the test substance to be considered suitable for use for REACH. Due to issues with sporadic vomiting and anorexia in the dog study, the data from the rat study will be used as the 90 day NOAEL.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:

We have no repeat dose inhalation study for 2,2’-(Octadecylimino)bisethanol) CAS No 10213-78-2, however it is a waxy solid with a low vapour pressure expected to be less than 0.73 mPa at 20ºC (1.2 mPa at 25ºC), which is based on read across to Bis(2-hydroxyethyl) coco amine CAS No 61791-31-9 which is a worst case due to it being predominantly C12-14, significant exposure to vapours would not be expected at ambient temperatures so it is considered to not be scientifically valid to conduct a repeat dose inhalation study.  Also the irritant nature of the substance would also make this difficult to perform for animal welfare reasons.  The guidance for REACH allows the inhalation long term DNELS to be calculated for the oral repeat dose NOAEL.

Justification for selection of repeated dose toxicity inhalation - local effects endpoint:

We have no study for repeat dose local inhalation effects for 2,2’-(Octadecylimino)bisethanol) CAS No 10213-78-2, however it is a waxy solid with a low vapour pressure expected to be less than 0.73 mPa at 20°C (1.2 mPa at 25°C), ), which is based on read across to Bis(2-hydroxyethyl) coco amine CAS No 61791-31-9 which is a worst case due to it being predominantly C12-14, significant exposure to vapours would not be expected at ambient temperatures so it is considered to not be scientifically valid to conduct a repeat dose inhalation study.  Also the irritant nature of the substance would also make this difficult to perform for animal welfare reasons.  The low possibility of inhalation makes such a test scientifically unjustified.

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:

Due the 2,2’-(Octadecylimino)bisethanol) CAS No 10213-78-2 being considered to be irritating to skin, it is not possible to conduct repeat dose dermal toxicity studies due to animal welfare considerations.  Also it is considered very unlikely that dermal absorption would exceed oral absorption, so it would be expected than the oral NOAEL would be lower than that from a dermal study.  Data from the repeat dose oral studies can be used in the setting of DNELs.

Justification for selection of repeated dose toxicity dermal - local effects endpoint:

2,2’-(Octadecylimino)bisethanol) CAS No 10213-78-2 is classified as a skin irritant, risk management measures such as wearing appropriate gloves and protective clothing will prevent an significant dermal contact.  Local effects on the skin would be expected to be limited to local irritation, which would be dependent on the local concentration rather than the dose or duration of exposure.  This is a medium hazard and does not justify the requirement for an additional dermal animal study to establish a local NOAEL for dermal exposure.

Justification for classification or non-classification

The NOAEL in the 90 day rat study of 35mg/kg/day for 2,2’-(C16-18 (even numbered, C18 unsaturated) alkyl imino) diethanol CAS No 1218787-32-6 the read across substance was for dosing in the diet. Specific Target Organ effects were only seen in the 15000ppm (ca.105mg/kg/day group) and higher groups. The EU CLP (GHS) criteria for classification for Specific Target Organ Toxicity (STOT) are based on data from a 90 day study, for Category 2 the range for such effects is 10-<100 mg/kg/day. As only limited signs of specific target organ toxicity in the gastrointestinal tract were see at higher than 100mg/kg bw/day and there is no evidence of specific target organ toxicity below the 100mg/kg bw/day threshold, therefore there is no requirement for a classification of 2,2’-(Octadecylimino)bisethanol for STOT Category 2 under the EU CLP (GHS) criteria.